突触传递：一个信号怎么跨过 20 纳米
这不是朱涵老师的论文。她本人的文章发在中文期刊，国际索引里查不到；这里只是一篇开放获取综述，用来给这一节配英文原话。下划线 = 中文容易"变味"或丢信息的词句：红线带编号 = 12 条注释卡（悬停看、点击跳转）。这一页的中文主干是教科书的突触生理学，英文原话来自一篇前额叶（PFC）综述——它讲的是这套机器怎么被调坏。两者我会分开标。英文原文附于文末
这一页是什么，不是什么
一、不是她的论文。朱涵老师是教学型老师，20 年上课，参编过 3 部生理学中英文教材，但国际索引里查不到她的第一作者论文。这一页的中文主干，是《生理学》里"突触传递"那一章她讲的内容；英文对照取自 Yan 与 Rein 2022 年发在 Molecular Psychiatry 的一篇开放获取综述，讲的是前额叶皮层突触传递的调控与失调。
二、为什么拿一篇"疾病综述"配"生理学基础"？因为突触生理学最枯燥的那部分（SNARE、量子、总和）在教科书里是名词表；而这个领域真正的前沿问题是：这些参数在活体里被谁调、调坏了会怎样。这篇综述每一节都在回答后者。所以写法是——教材 给你骨架，原文 告诉你这套骨架在 2022 年的研究里被问到哪一步。
三、先给你一个不方便的事实。这篇原文里没有出现 SNARE、量子释放（quantal release）、总和（summation）、重摄取（reuptake）、EPSP/IPSP 这些教科书词——它是给同行看的，默认你懂。这些词我会写，但会标 教材，而且不会伪造英文原句来"配"它们。你能看到的每一个英文引号，都能在这篇 md 里逐字搜到。
原文怎么说 · 这篇综述的立论
“Dysregulation of PFC synaptic transmission is strongly linked to the social deficits, affective disturbance, and memory loss in brain disorders”
译：前额叶突触传递的失调，与脑疾病中的社交缺陷、情感障碍和记忆丧失密切相关。
看它的用词：不是"突触没了"，是 dysregulation（失调）。失调意味着这套机器还在转，只是参数不对——这跟 synaptic transmission 这个词的"效能可调"性质是同一件事。这一页后面所有内容，都是在给"参数"列清单。
这一页想让你记住的一句话：突触不是一根导线，是一个可调的增益旋钮。（这句是我说的，不是原文。你要能在考试里把它展开成五个可以调的地方：释放概率、受体数目、受体亚型、清运速度、以及突触长在哪儿。）
一、20 纳米：化学突触长什么样
教材 化学突触三件套：
- 突触前终末：内含突触小泡（直径约 40 nm），其中一部分"停靠"在活性带（active zone）旁边随时可释放——这个子集叫可立即释放池（readily releasable pool, RRP）。一个典型 bouton 有几百个小泡，RRP 只占很小一部分，其余是"储备池"。
- 突触间隙：20–25 nm。这个宽度是化学传递的关键——够窄，递质扩散到对岸只要微秒级；又够宽，两侧的膜不接触，信号必须先"翻译"成化学物质才能过去。题目里说的"跨过 20 纳米"，就是这一段。
- 突触后膜：受体加支架蛋白构成的突触后致密物（postsynaptic density, PSD，厚几十纳米）。PSD 里最著名的一类支架蛋白是 Shank 家族——这篇综述开篇就讲它。
形态上还分兴奋性与抑制性：兴奋性突触（Gray I 型）不对称，PSD 厚、小泡圆而清亮；抑制性突触（Gray II 型）对称，两侧致密物差不多厚、小泡多为椭圆。你在电镜图里就是这么认的。
至于电突触（缝隙连接，gap junction）：间隙只有约 1.5 nm，两侧 connexin 直接对接成孔，电流双向、几乎无延迟、不能放大也不能反转符号。它快，但它没有"增益旋钮"——这就是为什么神经系统的主力是化学突触。
下面这句原文，是把"解剖位置"直接翻译成"功能分工"的最好例子：
原文怎么说 · 长在哪儿，就管什么
“PFC fast spiking (FS) interneurons (presumably expressing parvalbumin, PV+) form contacts on the perisomatic domains of pyramidal neurons, while non-FS interneurons (presumably expressing somatostatin, SST+) contact peridendritic domains”
“enabling PV+ and SST+ interneurons to specialize in the control of the output and input of principal neurons, respectively.”
译：前额叶的快速放电（FS）中间神经元（推测表达小清蛋白 PV）与锥体神经元的胞体周围区域形成接触；而非 FS 中间神经元（推测表达生长抑素 SST）接触树突周围区域——这使得 PV+ 与 SST+ 中间神经元分别专司对主神经元"输出"与"输入"的控制。
这一句值一整节课。同一个递质（GABA）、同一类受体，突触长在胞体上还是树突上，功能完全不同：胞体周围抑制直接管"发不发动作电位"（输出），树突抑制管"能收到多少输入"（输入）。第四节的总和问题，根子在这里——总和不是纯数学，是几何。
另一个"解剖即功能"的入口，是 PSD 的分子组成：
原文怎么说 · PSD 里的一块砖
“Haploinsufficiency of the Shank3 gene, which encodes a scaffolding protein at postsynaptic density of glutamatergic synapses, has been causally linked to autism”
译：Shank3 基因编码谷氨酸能突触突触后致密物中的一种支架蛋白，该基因的单倍剂量不足（haploinsufficiency）与孤独症存在因果关联。
"支架蛋白"（scaffolding protein）听着无聊，但它是把受体"钉"在正确位置的那层结构。Shank3 少一半 → PSD 结构松 → NMDA 受体在突触上的定位变差 → 电生理上看到 NMDA 电流变小。你以后做 WB 测 PSD95、Shank3 这类蛋白，测的就是这层"钉子"。
二、释放：钙是扳机，SNARE 是绞盘
教材 一次释放的完整链条，五步：
- 动作电位到达突触前终末 → 去极化 → 电压门控 Ca²⁺ 通道开放（中枢突触以 P/Q 型为主，N 型也参与）。
- Ca²⁺ 在活性带的微域内瞬间升高到几十微摩尔。注意：不是胞浆整体升高，是"局部喷泉"——这也是为什么突触前终末的钙通道必须紧贴囊泡。
- 钙感受器 synaptotagmin-1 结合 Ca²⁺，解除对融合机器的抑制。
- SNARE 复合物（突触前膜上的 syntaxin-1 与 SNAP-25，囊泡上的 synaptobrevin-2/VAMP2）像拉链一样从 N 端向 C 端"拉合"，把囊泡膜拽向突触前膜。
- 形成融合孔 → 递质外排（"全融合"与"kiss-and-run"两种模式并存）。
SNARE 模型最强的一类证据来自毒素：破伤风毒素和肉毒毒素都是锌依赖蛋白酶，专切 VAMP 或 SNAP-25，切完释放就没了。你以后看到"肉毒杆菌中毒表现为弛缓性麻痹"，那就是 SNARE 被切开、乙酰胆碱放不出来的临床版本。
三个参数决定"这一次能放多少"：
- 释放概率 pr：动作电位到来时，一个停靠好的囊泡真的融合的概率。不同突触差别很大，从 0.05 到 0.9 都有。
- RRP 大小：有多少囊泡在"待命"。
- 残余钙：前一个动作电位留下的 Ca²⁺ 没清干净，下一个动作电位的释放就被易化（facilitation）。这是成对脉冲易化（PPF）的主要机制之一。
原文在讲急性应激时，正好用了 RRP 这个词：
原文怎么说 · 应激把"待命的囊泡"变多
“One mechanism is related to the enhancement of readily releasable pool of glutamate vesicles via a non-genomic mechanism mediated by membrane receptors”
译：其中一个机制，与谷氨酸囊泡的可立即释放池被增大有关，这通过膜受体介导的非基因组机制实现。
"非基因组机制"（non-genomic）是关键词：糖皮质激素通过膜受体在几分钟内起效，不经过核内转录。这告诉你突触强度的快速变化可以完全不碰基因——你若只测蛋白总量，这类变化是测不到的。
还有一个必须知道的点：突触前标记物是可以测量的，而且在人的尸检脑里测过：
原文怎么说 · 一个 SNARE 成员在人脑里的变化
“Reduced expression of the presynaptic vesicle protein VAMP has also been reported in SZ PFC”
“while other presynaptic markers appear unchanged”
译：在精神分裂症（SZ）患者的前额叶中，还报告了突触前囊泡蛋白 VAMP 的表达降低，而其他突触前标记物看起来没有变化。
VAMP（= synaptobrevin）就是 SNARE 三根绳子里囊泡那一根。"VAMP 降、其他突触前标记没变"是一个很讲究的表述：它指向"某个特定分子变了"，而不是"突触前终末整体少了"。做 WB 的人要能读出这层区别（见注 5）。
三、从一个囊泡到一个电位
教材 递质结合受体 → 配体门控离子通道打开 → 膜电导改变 → 膜电位朝该通道的反转电位移动。符号由反转电位决定，不由"哪种递质"决定。这是整章最容易被背反的一句话。
兴奋性（EPSP）：谷氨酸作用于 AMPA 受体（Na⁺/K⁺ 通道，反转电位约 0 mV）→ 去极化。NMDA 受体是另一种东西：它通透 Ca²⁺，但在静息电位被 Mg²⁺ 堵住，必须"先去极化"才能开，而且还需要甘氨酸或 D-丝氨酸做共激动剂。所以 NMDA 受体是一个符合检测器（coincidence detector）：突触前释放 + 突触后已经去极化，两个条件同时满足才放 Ca²⁺ 进来。第五节的 LTP 全靠这个性质。
抑制性（IPSP）：GABA 作用于 GABA_A 受体（Cl⁻ 通道）→ 膜电位朝 Cl⁻ 反转电位移动。这里有个教科书图常画错的地方：GABA_A 的反转电位不是固定的 −70 mV，它取决于胞内 Cl⁻ 浓度。成熟神经元靠 KCC2 把 Cl⁻ 泵出，反转电位低于静息电位，GABA 才产生超极化；发育早期 NKCC1 占优，胞内 Cl⁻ 高，GABA 反而去极化——这不是"抑制性递质失灵"，是同一个递质在不同 Cl⁻ 梯度下符号相反。GABA_B 受体则通过 G 蛋白打开 K⁺ 通道、抑制 Ca²⁺ 通道，慢（几百毫秒）、持久。
自发的小事件叫 mEPSP / mIPSP（俗称 mini），它对应单个囊泡的释放。这个对应关系是量子假说的核心：mini 的幅度约等于一个量子的大小（由受体数目和密度决定），mini 的频率约等于释放概率 × 突触数目。第七节会用到这个区分——它是你判读电生理数据的第一把钥匙。
原文怎么说 · 抑制有两种，快慢强弱不同
“PV+ interneurons, which could fire in millisecond synchrony, exert fast and powerful inhibition on principal neuron firing, whereas SST+ interneurons exert weak and more variable inhibitory effects on firing output in behaving mice”
译：PV+ 中间神经元能够以毫秒级同步放电，对主神经元的放电施加快速而强大的抑制；而 SST+ 中间神经元的抑制作用较弱、在行为中的小鼠身上变化更大。
"fast and powerful" 对应一个具体现象：胞体周围抑制能在 1–2 ms 内把兴奋性输入"短路"掉（分流抑制），不需要等膜电位被超极化。这就是为什么 PV 细胞常被形容成"给锥体细胞装了个闸门"。
原文怎么说 · 一句话概括这篇的模型
“Collectively, the existing evidence suggests a model whereby the disruption of NMDA and GABA transmission in PFC, which is partially driven by aberrant dopamine and serotonin neuromodulation, produces excitation/inhibition imbalance and network dysconnectivity in SZ”
译：综合来看，现有证据支持这样一个模型：前额叶中 NMDA 与 GABA 传递的破坏（部分由异常的多巴胺与 5-羟色胺神经调制驱动）造成兴奋/抑制失衡，以及精神分裂症中的网络连接失谐。
注意"失衡"在这里是有具体归属的：E（NMDA 介导的兴奋）与 I（GABA 介导的抑制）两头都被动了，而不仅仅是某一头。你以后在文献里看到 E/I imbalance，先问它测的是哪一头、用什么指标测的（见注 6）。
四、总和：时间、空间，还有"长在哪儿"
教材 单个 EPSP 通常只有零点几到几毫伏，离阈值（轴突始段，约 −55 到 −50 mV）差得远。所以必须总和：
- 时间总和：同一个突触连续来几个动作电位，间隔小于膜时间常数（约 10–20 ms），EPSP 叠起来。
- 空间总和：不同突触在同一时间窗内一起开，电流在胞体相加。
- 衰减：树突上的 EPSP 传到胞体要按长度常数 λ 衰减，远端突触的"话语权"天然更小。树突棘的细颈还有额外电阻，进一步隔离。
还有一层容易被忽略：抑制不一定表现为超极化。打开 Cl⁻ 通道可以只是把膜电导提高，让同时到来的兴奋性电流"漏掉"——这叫分流抑制（shunting inhibition）。所以你在电压轨迹上看不到明显的下凹，EPSP 却已经被压小了。这也是为什么"抑制性突触数目"和"抑制强度"不能简单换算。
而总和的结果，强烈依赖突触长在哪——这正是第一节那句原文的深意。胞体周围的抑制只要能压住轴突始段附近的一小段，就能决定这个神经元发不发；树突上的抑制则是在"输入端"做加权，改变的是这个神经元对哪一路输入更敏感。
再往上一层，是"抑制的抑制"：
原文怎么说 · 去抑制：负负得正
“PFC in awake mice also has a basic disinhibitory circuit module in which activation of VIP+ interneurons transiently suppresses primarily SST+ and a fraction of PV+ interneurons”
译：清醒小鼠的前额叶还存在一个基本的去抑制环路模块：VIP+ 中间神经元的激活会短暂抑制 SST+ 中间神经元以及一部分 PV+ 中间神经元。
"去抑制"（disinhibition）不是"没有抑制"，是三级串联：VIP 抑制 SST，SST 本来抑制锥体细胞，于是 VIP 兴奋 = 锥体细胞被放开。你要习惯这种"负负得正"的电路语法，皮层里到处都是。
五、可塑性入门：LTP 与 LTD
教材 突触强度不是固定的，它会按使用历史被长期改写。两个方向：
LTP（长时程增强）：高频刺激（如 100 Hz × 1 s，或 theta-burst）→ 突触后大幅去极化解除 NMDA 的 Mg²⁺ 阻滞，同时突触前释放谷氨酸 → NMDA 开放 → 突触后 Ca²⁺ 大幅升高 → CaMKII 自磷酸化 → AMPA 受体磷酸化并插入膜 → 突触后反应变大。早期 LTP（1–3 小时内）不依赖蛋白合成；晚期 LTP 需要转录翻译和结构改变（树突棘变大、出现穿孔突触）。
LTD（长时程抑制）：低频刺激（如 1 Hz × 15 min）→ Ca²⁺ 升高幅度较小但持久 → 磷酸酶（calcineurin / PP1）占优 → AMPA 受体去磷酸化、内吞 → 突触后反应变小。
所以"Ca²⁺ 进来"不决定可塑性的方向，Ca²⁺ 的幅度与动力学才决定：大而快 → LTP；小而持久 → LTD。这条"钙阈值"规则是整章的核心，比背"高频 LTP、低频 LTD"重要得多。
再加两条性质：输入特异性（只有被刺激的那条输入变强，邻居不变）与协同性（弱输入单独不行，和强输入同时来就能被带上）。以及 STDP：突触前放电略早于突触后（10–20 ms）→ 增强；反序 → 减弱。时间顺序本身成了信息。
原文怎么说 · 同一个受体，两个方向
“D1R activation in PFC facilitates the maintenance of long-term potentiation (LTP) of glutamatergic transmission, as well as the induction of long-term depression (LTD) of glutamatergic transmission, indicating its bidirectional modulation of synaptic plasticity in mPFC”
译：前额叶 D1 受体的激活既促进谷氨酸能传递长时程增强（LTP）的维持，也促进其长时程抑制（LTD）的诱导，说明 D1 受体对内侧前额叶的突触可塑性具有双向调制作用。
注意 "bidirectional"（双向）：同一个 G 蛋白偶联受体，按剂量和状态不同，可以推 LTP 也可以推 LTD。这跟下一段的倒 U 曲线是同一件事的两面。教科书常把递质写成"兴奋性/抑制性"，但调制系统（多巴胺、去甲肾上腺素、5-HT）没有固定符号。
原文怎么说 · 倒 U 形
“Stimulation of D1R produces an inverted-U dose response on PFC neuronal firing and cognitive performance during working memory tasks”
译：D1 受体的刺激对前额叶神经元放电以及工作记忆任务中的认知表现，产生倒 U 形的剂量反应。
倒 U 的意思是：中间最好，两头都差。多巴胺太少（前额叶功能低下）和太多（应激、高 D1 刺激）都会损害工作记忆。这张曲线是所有"给某递质增强剂就能提高认知"的论文最容易被忽略的反例。
原文怎么说 · 慢性应激关掉 LTP
“Chronic stress impairs synaptic plasticity by reducing LTP induction in the hippocampus-PFC connection”
译：慢性应激通过降低海马—前额叶通路中 LTP 的诱导，损害突触可塑性。
这句是"生理学 → 病理学"的接口：可塑性不是抽象概念，它在整体动物身上可以被一个环境因素压下去。你若做行为学加电生理，这就是最常见的表型。
六、突触不是一个人在战斗
三套调制系统，一层套一层。
一、受体与胞内信号。教材 代谢型受体（GPCR）不直接开门，它改的是"这台机器的工作参数"：磷酸化受体、改通道开放概率、把受体调上膜或调下膜。原文给了一个漂亮的例子——α1 和 α2 肾上腺素受体都能抑制前额叶锥体细胞的 NMDA 电流，但机制完全不同（α1 走 PLC/IP3/Ca²⁺，α2 走 PKA 加微管运输）：
原文怎么说 · 同一个表型，两条通路
“Activation of α1-ARs or α2-ARs suppresses NMDAR currents in PFC pyramidal neurons by distinct mechanisms”
译：α1 或 α2 肾上腺素受体的激活都会抑制前额叶锥体神经元的 NMDA 受体电流，但机制不同。
这一句是你做 WB 时的护身符：同样的"NMDA 电流下降"，可以是通道被磷酸化，也可以是受体被运走了。前者测总蛋白不变，后者测膜蛋白会降。不分开测，你永远说不清机制。
二、转运体（重摄取）。教材 递质释放后，靠转运体从间隙收回：谷氨酸主要靠星形胶质细胞的 GLT-1 / GLAST（神经元侧有 EAAT3），GABA 靠 GAT-1 / GAT-3，多巴胺、去甲肾上腺素、5-HT 分别靠 DAT、NET、SERT；囊泡膜上的 VMAT 负责把递质装进囊泡。转运体决定三件事：信号持续多久、能扩散多远、下次释放的囊泡里装多少。哌甲酯、安非他明、SSRI、可卡因的作用点全在这一层。
这里我要老实说一句：这篇原文没有出现"转运体"或"重摄取"（我搜过）。它讲多巴胺时只说"提高前额叶内的儿茶酚胺传递"，没交代经过哪个转运体。这一段是我按教科书补的——考试用没问题，写论文引用时要另找文献。
三、胶质细胞与细胞外基质。教材 星形胶质细胞的突起包裹突触，形成"三部分突触"（tripartite synapse）：它清谷氨酸、释放 D-丝氨酸（NMDA 的共激动剂）、通过 Ca²⁺ 波释放胶质递质。小胶质细胞则做突触修剪：用补体（C1q / C3）标记"该删的"突触，再吞噬掉。还有一种被低估的结构：神经元周围网（perineuronal net, PNN），一层细胞外基质，主要包在 PV 中间神经元外面，起"稳定突触、限制可塑性"的作用。
原文怎么说 · 修剪
“It has been suggested that SZ is driven by disrupted cortical synaptic pruning”
译：有观点认为，精神分裂症由皮层突触修剪的异常所驱动。
注意 "It has been suggested"：这是假说，不是结论。但它的逻辑值得学——把"发病年龄"当成一条机制线索：青春期正是修剪最猛的时候，症状也正好在那时出现。原文下一句就把这两件事连起来了。
原文怎么说 · 一层"网"
“lower density of perineuronal nets (PNN) - extracellular structures that stabilize synapses, also suggesting synaptic loss or destabilization”
译：神经元周围网（PNN）的密度降低——这是一类稳定突触的细胞外结构——同样提示突触丢失或失稳。
原文用破折号顺手给 PNN 下定义，说明作者知道读者不熟。你要记住它的功能标签是"稳定、限制可塑性"：PNN 被降解（例如用 chondroitinase ABC 处理）之后，那一段的"可塑性窗口"会重新打开，这是发育与学习研究里的常用手法。
七、实验室里怎么"看见"突触
三个层次，从粗到细。各自能看到什么、看不到什么，我列成一张表——这张表比任何一段文字都实用：
| 方法 | 记什么 | 能说明 | 不能说明 |
|---|---|---|---|
| 场电位 fEPSP | 一群突触同步产生的细胞外场电位，量 slope（上升斜率） | 整群突触的效能；LTP / LTD 的常规判据 | 单个突触、单个受体亚型 |
| 成对脉冲比 PPR | 两个间隔约 50 ms 的刺激，第二个 fEPSP 除以第一个 | 释放概率 pr（PPR 小 → pr 高） | 突触后受体数目 |
| 全细胞 mEPSC | 加 TTX 后自发的小事件：频率与幅度 | 频率 ≈ pr × 突触数；幅度 ≈ 量子大小（受体数 / 密度） | 具体是哪条输入（除非用光遗传或电刺激分离） |
| 诱发 EPSC 的 NMDAR / AMPAR 比 | +40 mV 记 NMDA、−70 mV 记 AMPA，取比值 | 受体组成（如 GluN2B 比例） | 受体在突触内还是突触外 |
| WB / 免疫组化 | 蛋白总量或定位 | 分子表达量、分布 | 功能强弱——总量与功能不同向是常态 |
几个操作要点，都是新手最容易翻车的地方：
- fEPSP 量 slope，不量 peak。因为 peak 常常混着 population spike（一群细胞同步放电的尖峰），而 slope 只反映突触电流。这一条你第一次做 LTP 就会被人纠正。
- 切片要"活着"。人工脑脊液（ACSF）要持续通 95% O₂ / 5% CO₂；32–34 ℃ 与室温下的动力学不一样；灌注速度、切片厚度、从断头到入液的时间，都影响结果。
- 看兴奋性突触要阻断抑制。不加 picrotoxin 或 bicuculline，你记到的 EPSP 里混着 GABA_A 的分流效应，幅度偏小而且不稳定。
- 对照要配对。同窝野生型、同一天、同一批切片、同一支电极；能配对的就配对，不能配对的要在统计里说清楚。
原文里有一条数据，正好是"频率还是幅度"这个区分的好例子：
原文怎么说 · 频率升高意味着什么
“Electrophysiological recordings similarly indicate that ketamine elevates the frequency of excitatory postsynaptic currents (EPSC) in mouse PFC pyramidal neurons”
译：电生理记录同样显示，氯胺酮提高了小鼠前额叶锥体神经元兴奋性突触后电流（EPSC）的频率。
"频率升高"在量子分析里的标准解释是：释放概率升高，或者功能性突触数目增多——不是"单个突触变强"（那是幅度升高）。这一个词的差别，决定你后面补什么实验：补 PPR？补 mEPSC 频率？还是补膜蛋白？
最后一条，是给以 WB 为主的本科生的桥：
原文怎么说 · 突触蛋白可以当"指纹"
“Studies of postmortem human brains have found the significant association between cognitive decline and the loss of synaptic proteins in PFC, such as SNAP25, SNAP47, SV2C, SYT2 and GRIA3”
译：对死后人脑的研究发现，认知衰退与前额叶突触蛋白的丢失显著相关，例如 SNAP25、SNAP47、SV2C、SYT2 与 GRIA3。
这五个名字你迟早会跑：SNAP25 与 SYT2 属于释放机器（SNARE 与钙感受器），SV2C 是囊泡蛋白，GRIA3 是 AMPA 的一个亚基。它们分属三类不同的东西，但在"突触丢了"这件事上会一起降。你在 WB 上看到它们同向变化，说明的是"突触数量 / 完整性"，不是"某个机制"。
给刚进组的你
这一页对你最直接的三件事：
- 先问通路，再问指标。"突触传递增强"这句话不成立，除非你说清：哪条通路（海马→前额叶？Schaffer 侧支→CA1？）、哪个指标（fEPSP slope / mEPSC 频率 / mEPSC 幅度 / PPR）、哪种刺激（高频？配对？）。指标不同，结论的机制层级就不同——这是这一页最想让你带走的方法学。
- 你 WB 测的每一个蛋白，都要能说出它站在突触哪一侧。GluA1 / GluA2 = 突触后 AMPA 亚基（注意膜表面与总蛋白要分开提，否则"上调"可能只是转位）；NR1 / NR2A / NR2B = NMDA 亚基；PSD95 = 突触后支架（常被当"突触量"指标，但它不是万能内参）；SNAP25、synaptophysin、SV2C = 突触前；GAD67 = GABA 合成。测之前先问：这个蛋白变了，我预期的电生理表型是什么？反过来问更狠：如果电生理没变，我还能怎么解释？
- 读文献先看 Methods 的哪一段。看切片厚度与 ACSF 配方、看刺激参数（频率、串长、间隔）、看有没有加 GABA 阻断剂、看动物年龄。这四项里任何一项不同，两篇论文的"LTP"就不是同一个量。
组会上可以问的三个问题：(1) 我们记的是 fEPSP 还是全细胞？(2) LTP 的判据是诱导后多久、增幅多少？(3) 有没有做 PPR 来区分突触前还是突触后？
最后一句：教科书把突触画成一个开关，实验室里它更像一个音量旋钮——而你要做的，是证明你拧的是哪个旋钮，不是你感觉声音变大了。
▲ 本篇注释（挂在英文术语上，对照学习用）
附录 · 英文原文
Mechanisms of Synaptic Transmission Dysregulation in the Prefrontal Cortex: Pathophysiological Implications
Abstract
Prefrontal cortex (PFC) serves as the chief executive officer of the brain, controlling the highest level cognitive and emotional processes. Its local circuits among glutamatergic principal neurons and GABAergic interneurons, as well as its long-range connections with other brain regions, have been functionally linked to specific behaviors, ranging from working memory to reward seeking. The efficacy of synaptic signaling in PFC network is profundedly influenced by monoaminergic inputs via the activation of dopamine, adrenergic or serotonin receptors. Stress hormones and neuropeptides also exert complex effects on the synaptic structure and function of PFC neurons. Dysregulation of PFC synaptic transmission is strongly linked to the social deficits, affective disturbance, and memory loss in brain disorders including autism, schizophrenia, depression and Alzheimer’s disease. Critical neural circuits, biological pathways, and molecular players that go awry in these mental illnesses have been revealed by integrated electrophysiological, optogenetic, biochemical, and transcriptomic studies of PFC. Novel epigenetic mechanism-based strategies are proposed as potential avenues of therapeutic intervention for PFC-involved diseases. This review provides an overview on PFC network organization and synaptic modulation, as well as the mechanisms linking PFC dysfunction to the pathophysiology of neurodevelopmental, neuropsychiatric and neurodegenerative diseases. Insights from the preclinical studies offer the potential for discovering new medical treatment of human patients with these brain disorders.
Prefrontal cortex (PFC) is the central hub for high-level executive functions critically involved in mental health and diseases. In this review, we will provide an overview on how synaptic transmission in PFC is regulated in normal and pathological conditions. We will first summarize PFC functional organization by describing the local and long-range neuronal circuits and the behavioral correlates to these top-down and bottom-up synaptic connections. Next, we will summarize PFC modulation by describing the influence of monoaminergic systems, stress hormones and neuropeptides on the synaptic structure and function of PFC neurons. Finally, we will summarize the PFC synaptic dysregulation in a variety of brain disorders and potential treatment strategies. Because of the broad scope of this review, only selective neuronal circuits, signaling molecules and disease mechanisms are included. Interested readers are encouraged to find more comprehensive details on specific aspects of the topic in the orignial papers and review articles cited here.
PFC Organization and Function
PFC network organization and functional implications
PFC is the cortical region located at the anterior part of the frontal lobe. PFC of humans is delineated into two functionally, morphologically, and evolutionarily different regions: ventromedial PFC and lateral PFC. PFC is the last portion of the brain to fully develop. In monkey PFC neurons, the rapid phase of synaptogenesis reaches the plateau at 2 months old, and maintains the consistently high synaptic density from early adolescence through puberty (2 months to 3 years), the formative time when learning experiences are most intense [1].
Early studies of macaque monkeys have revealed PFC as a crucial hub necessary for successful maintenance of working memory (WM), a cognitive process involving information holding and manipulation, which serves as the fundamental backbone for executive functions [2],[3]. The key findings is that in a delayed reaching task, some primate dorsolateral PFC neurons exhibit the increased firing during the delay period when response-related information needs to be held in mind to guide the subsequent execution of correct responses [4],[5]. The delayed and persistent spiking activity that encodes spatial “working memory” signals arises from recurrent excitation among PFC pyramidal neurons [5],[6]. Cortical network models predict that NMDA receptors subserve the recurrent synaptic excitation and persist firing of PFC pyramidal neurons during working memory [7],[8]. In parallel with these neurophysiological studies, neuroimaging studies also implicate the human PFC in higher-order cognitive processing including working memory [9] and sustained attention [10].
Based on anatomical, functional and connectivity homologies, it is thought that rodent PFC is composed of three cytoarchitecturally defined parts: the prelimbic (PL), infralimbic (IL), and anterior cingulate cortex (ACC) [11],[12]. Electro-physiological and pharmacological evidence further supports the existence of a frontal cortical area in the rat that exhibits increased firing during the delay period in the absence of sensory stimulus that instruct the orienting response, as in primate PFC [13]. Recordings of neuronal ensembles in rats during new task learning further find that the dynamic changes in PFC neuronal activity parallel with behavioral decoding, suggesting that working memory can be mediated by dynamic activation of different neural populations in PFC [14].
PFC projects to distributed subcortical autonomic, motor, and limbic regions (). Tracing studies have found that deep layer PFC pyramidal neurons project to mediodorsal thalamus, lateral hypothalamus, ventral and dorsal striatum, and basolateral amygdala (BLA) [15]. Prefrontal circuitry can dynamically control reward-seeking behaviour through projection-specific cue encoding [16]: stimulation of PFC to nucleus accumbens (NAc) neurons promotes conditioned reward-seeking behaviour, while activation of PFC to thalamic neurons suppresses both the acquisition and expression of conditioned reward seeking. A specific subset of mPFC projections to NAc neurons encodes the decision to initiate or suppress reward seeking when faced with risk of punishment [17].
Dorsomedial PFC exerts the top-down control of motor cortex ensembles to inhibit inappropriate responses during a delayed-response task [18]. The neural projection from medial PFC (mPFC) to a brainstem area has been found to control behavioral adaptation to social defeat [19]: social defeat weakens functional connectivity of these two areas, while selective inhibition of this pathway recaptures behavioral effects of social defeat. A spatiotemporal dynamic network, which originates in PFC and ventral striatum, relays through the amygdala and ventral tegmental area (VTA), and converges in ventral hippocampus (vHIP), is found to encode depression vulnerability in mice subjected to social defeat stress [20].
PFC also receives inputs from diverse regions, including hippocampus, thalamus, and BLA (). The direct hippocampal to prefrontal afferents in mice are found to be critical for encoding of spatial cues during working memory, and successful encoding of task-related information is mediated by gamma-frequency synchrony between hippocampus and PFC [21]. Proper vHIP-mPFC signaling is necessary to recall social memories [22]. Reciprocal interactions between PFC and thalamus play a critical role in high-level cognitive function [23]. Thalamic input amplifies PFC connectivity, sustaining attentional control [24] and enabling sensory selection in divided attention [25]. The reciprocal connection between PFC and BLA is implicated in reward-seeking and fear-related responses, and optogenetic studies have revealed that BLA to PL circuit is critical in governing the selection of behavioral responses amid conflicting cues of reward and punishment [26].
PFC cellular components and synaptic circuits
PFC is composed of pyramidal neurons with wide arbor axons, wide spikes, low discharge rates, and interneurons, many of which having horizontal axonal arbors, narrow action potentials (APs) and high evoked firing rates [27]. PFC fast spiking (FS) interneurons (presumably expressing parvalbumin, PV+) form contacts on the perisomatic domains of pyramidal neurons, while non-FS interneurons (presumably expressing somatostatin, SST+) contact peridendritic domains [28], enabling PV+ and SST+ interneurons to specialize in the control of the output and input of principal neurons, respectively. PV+ interneurons, which could fire in millisecond synchrony, exert fast and powerful inhibition on principal neuron firing, whereas SST+ interneurons exert weak and more variable inhibitory effects on firing output in behaving mice [29]. PFC in awake mice also has a basic disinhibitory circuit module in which activation of VIP+ interneurons transiently suppresses primarily SST+ and a fraction of PV+ interneurons [30]. Such disinhibitory control may serve to amplify local processing and modulate information gating. A subclass of VIP neurons expressing ChAT, ChAT-VIP, directly excites neurons throughout cortical layers via fast synaptic transmission of acetylcholine in mPFC, providing a local source to control attention [31].
Opotogenetics, in vivo electrophysiological recording and single-cell microendoscopic Ca2+ imaging of PFC neurons in behaving animals have revealed the cell-type functional response diversity that contributes to diverse behavioral correlates (). PL pyramidal neurons are found to contain anatomically and molecularly distinct subpopulations that target three downstream regions implicated in social behavior: NAc, BLA, and VTA. Activation of NAc-projecting PL neurons, but not the other subpopulations, decreases the preference for a social target [32]. PV+ interneurons in mPFC also play a key role in social behaviors: activation of mPFC PV+ neurons rescues social memory impairment caused by vHIP inhibition [33]. Dampening thalamic activity causes significant reductions of GABA signaling in the mPFC and concomitant abnormalities in cognition and social interaction, which is ameliorated by selectively activating mPFC PV+ interneurons [34]. BLA inputs also make strong connections onto PFC PV+ and SST+ interneurons, driving feedforward inhibition of PFC pyramidal neurons [35],[36].
In mice performing a reward foraging task, SST+ interneurons in ACC are found to selectively respond at reward approach, whereas PV+ interneurons respond at reward leaving [29]. In mice performing a PFC-dependent sensory discrimination task, PV+ interneurons are responsive to sensory cues, motor action, and trial outcomes, while SST+ and VIP+ interneurons respond more selectively to motor action or trial outcomes, respectively, and pyramidal neurons show much greater functional heterogeneity with varied responses across cortical layers [37]. This cell type-specific encoding of task-related signals may be crucial for local computation within the PFC microcircuit to guide goal-directed behavior. Furthermore, SST+ interneurons in mPFC of mice are found to play a primary role in orchestrating the ability to discriminate positive and negative affective states [38]: inhibition of mPFC SST+ interneurons abolishes affective state discrimination, while an increased synchronous activity of PFC SST+ interneurons, which guides the inhibition of PFC pyramidal neurons, is associated with this social cognitive function.
More recent studies have further revealed the functional role of PFC interneurons in working memory. During a spatial WM task, optogenetically inhibiting PFC SST+, but not PV+, interneurons decreases the long-range synchrony between hippocampus and PFC and impairs working memory accuracy [39], suggesting that SST+ interneurons are uniquely involved in facilitating hippocampal-prefrontal synchrony and prefrontal spatial encoding. Inhibiting PFC pyramidal neurons by optogenetically activating SST+ or PV+ interneurons during the delay impairs working memory performance, while activating VIP+ interneurons in PFC improves memory retention [40], which has provided a circuit mechanism underlying memory-guided behavior. PV+ and SST+ interneurons in mPFC are also found to have distinct roles in WM: PV interneurons show weak target-dependent delay-period activity and are strongly inhibited by reward; while SST+ interneurons show strong target-dependent delay-period activity, and only a subtype of them is inhibited by reward [41].
Moreover, in a social fear conditioning paradigm, SST+ interneurons suppress PV+ interneurons and disinhibit pyramidal cells, which consequently enhances PFC output to mediate social fear responses [42]. This disinhibitory microcircuit in PFC through interactions between interneuron subtypes provides an important circuit mechanism in gating social fear behavior. The relapse of extinguished fear is found to be mediated by a feedforward inhibitory pathway from ventral hippocampus to IL cortex, which involves the recruitment of PV+ interneurons in IL [43]. In the avoidance behavior of elevated plus maze, PFC VIP+ interneurons enable cortical circuits to integrate hippocampal inputs, therefore effectively gating the transmission of signals related to open arm avoidance across hippocampal-prefrontal network [44] ().
Regulation of PFC Synaptic Function
Monoaminergic regulation of PFC synaptic transmission
PFC is very sensitive to neurochemical environment (), and monoaminergic modulation of the efficacy of synaptic connections in PFC circuits has profound effects on PFC-mediated executive function, including working memory, regulation of attention, inhibition of inappropriate behaviors, and impulsivity control [45]. Reduced catecholamine transmission has been linked to ADHD, while overamplified noradrenergic transmission is associated with PTSD [46]. The therapeutic actions of psychostimulant methylphenidate for ADHD involve the preferential increase of catecholamine neurotransmission within PFC [47] and dose-dependent, bi-directional regulation of PFC glutamate receptors [48].
Dopamine
Serial section electron microscopy studies suggest that the majority of dopamine synapses in all PFC layers are on pyramidal neurons, but a significant fraction are on GABAergic interneurons [49]. Electrophysiological studies revealed that dopamine selectively modulates PFC excitatory and inhibitory microcircuits. DA depressed inhibitory transmission between FS interneurons and pyramidal neurons, but enhanced inhibition between non-FS interneurons and pyramidal cells [28]. Dopamine also reduces recurrent excitation in layer V pyramidal neurons [50], which may involve target-specific expression of presynaptic and postsynaptic D1Rs [50],[51]. Dopamine has no effect on excitatory transmission from pyramidal neurons to FS interneurons, but increases the excitability of FS interneurons [52]. In an experimental condition mimicking NMDA receptor hypofunction and GABAA receptor deficiency in schizophrenia, dopamine, by acting on D2 receptors, promoted burst firing in a subset of PFC pyramidal neurons [53].
Stimulation of D1R produces an inverted-U dose response on PFC neuronal firing and cognitive performance during working memory tasks [46]. High levels of D1R activation of cAMP signaling reduces PFC neuronal firing and impairs working memory via opening of HCN channels at excitatory synapses [54]. It suggests that optimal levels of dopamine are essential for PFC function, while excessive dopamine stimulation could be detrimental.
Optogenetic stimulation of VTA neurons to trigger the synaptically released dopamine has revealed a marked and prolonged enhancement of the excitability of PFC PV+ interneurons and a modest and short-lived enhancement of the excitability of PFC principal neurons [55]. Blocking dopamine receptor activation in PFC shortens the VTA excitation of PFC PV+ interneurons and prolongs the VTA excitation of PFC principal neurons, and pharmacological evidence suggests that the dopaminergic effect on PFC principal neurons is through influencing the inhibitory transmission system [55]. These results have unmasked a role of dopamine in regulating the temporal dynamics of excitation/inhibition balance in VTA-PFC circuit.
Dopamine D4 receptor, which is largely restricted to PFC neurons, is highly implicated in ADHD and schizophrenia [56],[57]. It has been revealed that D4 receptors serve as a homeostatic synaptic factor to stabilize cortical excitability by dynamically regulating CaMKII [58],[59] and the downstream NMDA and AMPA receptor channels [60]–[62]. In contrast to the activity-dependent, bi-directional effect of D4R stimulation on AMPARs in PFC pyramidal neurons [62],[63], D4 receptors suppress AMPARs in PFC GABAergic interneurons via the unique calcineurin/Slingshot/cofilin signaling mechanism [64]. Moreover, D4R stimulation elicits distinct effects on synaptic-driven action potential firing in PFC projection neurons versus fast-spiking interneurons, which are differentially altered in neuropsychiatric disorder-related conditions [65]. Stress exposure in D4R knockout mice induces schizophrenia-like behaviors via disruption of GABAergic transmission in PFC [66]. These results have provided a framework for understanding how D4R signaling is involved in the regulation of PFC functions.
D4R is one of the most variable proteins (and genes) in humans. A unique primate-specific feature of D4R is the additional 2 to 11 proline-rich repeats (most commonly 4) located in the third intracellular loop, which allows more complex simultaneous interactions with other proteins containing the SH3 domain. Interestingly, genetic studies have linked novelty-seeking behavior and ADHD to long-repeat D4R allele (hD4.7) [57],[67]. Using D4R knockout mice with PFC expression of human D4R variants containing different repeats (hD4.4 and hD4.7), researchers have revealed the impact of hD4 variants on synaptic transmission and network activity in PFC neurons, the proteins interacting with hD4 variants and the signaling pathways activated by hD4 variants, as well as the behavioral changes caused by hD4 variants [68],[69]. These studies provide novel insights into the functional roles of the remarkable polymorphism of human D4 receptors, and shed light on the development of new therapeutic agents for diseases associated with dopaminergic dysfunction ().
Noradrenaline
The noradrenergic system in PFC is involved in many physiological and psychological processes, including working memory and mood control [46]. Endogenous norepinephrine (NE, also called noradrenaline), by acting on α1-ARs, exerts complex effects in PFC: NE increases the firing of some PFC pyramidal neurons via presynaptic excitation of glutamate release under basal conditions, but when high levels of α1-AR stimulation occurs, such as with stress exposure, NE suppresses PFC neuronal firing through opening of K+ channels on spines [70]. Stimulation of α2A-ARs enhances the spatially-tuned, delay-related firing of PFC neurons and strengthens working memory networks through inhibition of cAMP and closing HCN (Hyperpolarization-activated Cyclic Nucleotide-gated) channels [71]. The interaction of HCN channels and cAMP regulating proteins in dendritic spines of PFC is considered to be potential substrate of working memory [72].
Activation of α1-ARs or α2-ARs suppresses NMDAR currents in PFC pyramidal neurons by distinct mechanisms: the α1-AR effect depends on the PLC/IP3/Ca2+ pathway, whereas the α2-AR effect depends on PKA and the microtubule-based transport of NMDARs that is regulated by ERK signaling. Moreover, the effects of α1-ARs and α2-ARs are differentially modified by RGS4 [73], a member of the RGS family protein that plays an important role in modulating GPCR signaling [74] and an identified schizophrenia susceptibility gene [75],[76] ().
Serotonin
PFC receives a major serotonergic projection, which is dysfunctional in individuals who show impulsive aggression and violence [77]. Aberrant serotonergic neurotransmission has long been implicated in the pathogenesis of neuropsychiatric disorders associated with PFC dysfunction, including schizophrenia, depression and anxiety [78]. Most clinically effective antipsychotic drugs, antidepressants and anxiolytics all exert potent effects on the serotonin system. The pleiotropic functions of serotonin are afforded by the concerted actions of multiple serotonin receptor subtypes [79].
A series of studies have revealed the synaptic functions of serotonin receptors in PFC and how dysfunction of the serotonin system contributes to mental illnesses [80]–[89]. One of the key targets of serotonin receptor signaling is the glutamatergic system. It has been found that 5-HT1A receptors in PFC pyramidal neurons suppress NMDAR function by disrupting the microtubule/kinesin-based dendritic transport of NMDA receptors [83], while this effect of 5-HT1A is opposed by 5-HT2A/C signaling [82]. In the human PFC, serotonin suppresses monosynaptic excitatory connections from pyramidal cells to interneurons [90]. At thalamocortical synapses, 5-HT2A activation enhances NMDA transmission, gates the induction of NMDAR-mediated temporal plasticity, and facilitates associated cognitive functions [91].
Another main target of serotonin receptor signaling is the GABAergic system. 5-HT2 receptors inhibit GABAAR-mediated currents in PFC pyramidal neurons through phosphorylation of GABAA receptors by the activation of anchored PKC [87], while 5-HT4 receptors modulate GABAergic signaling bi-directionally, depending on the basal PKA activation levels that are determined by neuronal activity [80]. Serotonergic regulation of GABA transmission in PFC is subject to the alteration by corticotropin-releasing factor (CRF) in response to stressful stimuli [89] or chronic antidepressant treatment [86]. In addition to pyramidal neurons, serotonin also affects GABAergic fast-spiking interneurons in PFC by enhancing their excitability and gamma frequency temporal integration via 5-HT2A suppression of an inward-rectifying potassium conductance [92]. These studies have provided unique and complex mechanisms for serotonin receptors to dynamically regulate synaptic transmission and neuronal excitability in the PFC network in physiological and pathophysiological conditions ().
Hormonal regulation of PFC synaptic transmission
Stress hormones
Stress hormones exert complex and profound effects on the synaptic structure and function in PFC neurons (). Glucocorticoid action in the PFC shows a marked functional heterogeneity. Glucocorticoid receptor (GR) knockdown in IL induces compromised stress adaptation and depression-like behavior, while GR knockdown in PL increases HPA axis responses to acute stress but does not affect stress sensitization or helplessness behavior [93], highlighting the region-specific impact of PFC on stress coping and emotional control.
Exposing animals to an acute stressor, such as forced swim, foot shock or restraint, produces significantly enhanced glutamatergic transmission in PFC circuitry [94]–[97]. One mechanism is related to the enhancement of readily releasable pool of glutamate vesicles via a non-genomic mechanism mediated by membrane receptors [95]. Acute stress, via acting on GR, also induces a delayed potentiation of NMDAR-and AMPAR-mediated synaptic currents, which is correlated with the increased level of synaptic NMDAR and AMPAR subunits [96],[98]. Serum-and glucocorticoid-inducible kinases (SGKs), an immediate early gene activated by stress hormone, is shown to be involved by enhancing Rab4-mediated trafficking of glutamate receptors from early endosomes to plasma membrane in PFC neurons [98],[99]. The GR-induced upregulation of SGK and glutamatergic signaling in acutely stressed animals is blocked by inhibition of HDAC6, a unique member of the HDAC family that directly regulates the GR chaperone protein, heat shock protein 90 (HSP90) [97]. Interestingly, reduced SGK expression is found in postmortem brains of PTSD patients, and inhibition of SGK in rat PFC produces helplessness- and anhedonia-like phenotypes [100].
Adhesion molecules that anchor glutamate receptors at the synaptic surface are also implicated in the acute stress-induced enhancement of GluR2 membrane trafficking and memory facilitation [101]. In addition, acute footshock stress increases spine density and induces dendritic remodeling in medial PFC, which can be partially blocked by chronic treatment with the antidepressant desipramine [102],[103]. This effect of stress and antidepressant is linked to the alteration of key genes involved in synaptic plasticity and spine structure [104].
In contrast to the positive effects of acute stress, chronic stress (e.g. 21-day restraint stress) produces impaired dendritic branching, atrophy and spine loss in PFC pyramidal neurons [102],[105], and such structural reorganization is reversed after 3-week cessation of stress [106]. Chronic stress impairs synaptic plasticity by reducing LTP induction in the hippocampus-PFC connection [107], and repeated social stress during mid-adolescence significantly decreases synaptic activity and intrinsic excitability of mPFC neurons [108]. Chronic exposure to corticosterone also causes the reduction of NR2B and GluR2/3 subunit expression in ventromedial PFC [109].
A prominent loss of GluR1 and NR1 subunit expression has been found in PFC pyramidal neurons from repeatedly stressed animals, which leads to the depression of AMPAR- and NMDAR-mediated synaptic currents [110]. This stress-induced loss of glutamate receptor expression is attributable to the increased ubiquitin/proteasome-mediated degradation of GluR1 and NR1 that is controlled by E3 ubiquitin ligases Nedd4 and Fbx2, and inhibition of proteasomes or knockdown of these E3 ligases in PFC prevents the loss of glutamatergic responses and recognition memory in stressed animals [110]. The transcription of Nedd4 is upregulated by repeated stress via an epigenetic mechanism involving the elevated histone deacetylase 2 (HDAC2) and the ensuing suppression of histone methyltransferase Ehmt2 (also known as G9a) that catalyzes the repressive mark H3K9me2 [111]. The impairment of glutamate receptors and excitatory transmission in PFC of chronically stressed animals is blocked by HDAC2 inhibitors [111].
There are many additional molecules that have been implicated in governing the effects of chronic stress in PFC. One key player is the mTORC (mammalian target of rapamycin complex) signaling pathway. Decreased levels of mTORC are found in PFC of humans with major depressive disorder [112], and the fast-acting antidepressant ketamine increases mTORC signaling in rat PFC [113]. REDD1, an endogenous inhibitor of mTOR, shows the elevated expression in PFC of animals exposed to chronic unpredictable stress and PFC of post-partum depressed humans [114]. REDD1 knockdown renders animals to have greater resilience to the chronic stress-induced spine shrinkage and AMPAR current reduction [114].
Neuropeptides
The neuropeptide precursor VGF plays a critical role in depression and antidepressant efficacy [115]. Reduced levels of VGF have been found in PFC of MDD patients and PFC of chronically stressed mice [116]. VGF in PFC regulates susceptibility to stress and the antidepressant response to ketamine, and infusion of VGF C-terminal peptide TLQP-62 to PFC produces sustained antidepressant responses [116].
Neuropeptide Y (NPY), a stress modulatory transmitter, has been associated with PTSD. NPY in IL cortex, acting on NPY Y1 receptors, increases inhibitory synaptic transmission onto IL projection neurons and dampens their excitability, resulting in the significant impairment of fear extinction memory [117].
The “prosocial” hormone oxytocin (OT) plays a key role in regulating social and emotional behaviors through a population of SST+ interneurons that express the oxytocin receptor (OxtrIN) in mPFC. Silencing OxtrIN of female mice or deleting Oxtr gene in mPFC results in sociosexual deficits [118], while OxtrIN in male mice regulate anxiety-related behaviors by interacting with the co-expressed corticotropin-releasing-hormone-binding protein (CRHBP), an antagonist of the stress hormone CRH [119] ().
PFC Dysregulation and Diseases
Transcriptomic meta-analyses of human cerebral cortex across major neuropsychiatric disorders, including autism (ASD), schizophrenia (SZ), bipolar disorder (BD) and depression (MDD), have found the shared transcriptional dysregulation underlying convergent molecular neuropathology, such as a gradient of synaptic gene down-regulation and astroglial gene up-regulation in ASD, SZ and BD (ASD > SZ = BD), and dysregulation of HPA-axis and hormonal signaling in MDD [120]. Synaptic plasticity deficits in PFC of animal models have been linked to clinical findings from human patients with frontal cortical dysfunction, including the decreased functional connectivity in prefrontal-limbic circuits, executive function and memory deficits, inflexible and maladaptive behaviors [121]. In what follows, we will summarize mechanisms of synaptic transmission dysregulation in the PFC and pathophysiological implications in a few neurodevelopmental, neuropsychiatric and neurodegenerative diseases linked to the impairment of social, emotional, and cognitive processes.
Autism
Autism is a prevalent neurodevelopmental disorder characterized by core symptoms like social deficits and stereotypic behaviors. The importance of PFC in autism is demonstrated by structural, functional, and genomic studies. Postmortem studies have found focal patches of abnormal laminar cytoarchitecture and cortical disorganization of neurons, but not glia, in prefrontal and temporal cortical tissue from a majority of young children with autism, with the clearest signs of abnormal expression of excitatory neuronal markers in deep cortical layers 4 and 5 [122]. Many of the high-level executive functions controlled by PFC are impaired in autism. Transcriptomic analysis of autistic brains has found 444 genes showing significant expression changes in PFC, but only 2 in cerebellum [123].
Haploinsufficiency of the Shank3 gene, which encodes a scaffolding protein at postsynaptic density of glutamatergic synapses, has been causally linked to autism [124],[125]. High-resolution MRI has found that loss of Shank3 in mice results in the disrupted local and long-range prefrontal and frontostriatal functional connectivity, which is predictive of social communication deficits [126]. In PFC pyramidal neurons of Shank3-deficient mice, NMDAR function and synaptic distribution are significantly diminished, which is caused by a marked loss of actin filaments at glutamatergic synapses due to the altered Rac1/PAK/cofilin signaling [127]. Inhibiting cofilin or activating Rac1 rescues the social deficits and NMDAR hypofunction in Shank3-deficient mice [127] (), suggesting that targeting actin regulators to restore actin-based NMDAR synaptic delivery could be a therapeutic option for autism [128]. Chemogenetic activation of PFC pyramidal neurons in Shank3-deficient mice also restores social preference behaviors and elevates glutamatergic synaptic function [129]. Moreover, the reduced synaptic plasticity in hippocampus-mPFC pathway and impaired social recognition memory and attention in Shank3-deficient rats are attenuated by oxytocin treatment [130].
Another prominent genetic link to autism spectrum disorders and intellectual disability is the microdeletion or microduplication of the human 16p11.2 gene locus [131],[132]. A 16p11.2 deletion mouse model, 16p11+/−, exhibits NMDAR hypofunction in PFC pyramidal neurons [133]. Chemogenetic activation of PFC pyramidal neurons leads to the amelioration of cognitive and social impairments and the restoration of NMDAR function in 16p11+/− mice [133]. On the other hand, a 16p11.2 duplication mouse model, 16p11dp/+, exhibits deficient GABAergic synaptic transmission and elevated excitability in PFC pyramidal neurons [134]. Restoring the expression of Npas4, a key regulator of GABA synapses [135],[136], ameliorates the social and cognitive deficits, as well as GABAergic synaptic impairment and neuronal hyperexcitability in 16p11dp/+ mice [134]. Synaptic dysfunction and excitation/inhibition imbalance is identified as a common pathophysiological feature of multiple ASD models [137]–[139].
Large scale genetic screenings have revealed that many of the identified top-ranking autism risk factors are genes implicated in synaptic homeostasis, transcriptional regulation and chromatin remodeling pathways [140]–[142]. Epigenomic studies show that PFC neurons from subjects with autism exhibit the altered H3K4me3 peaks at numerous genes regulating neuronal connectivity, social behaviors, and cognition, often in conjunction with altered expression of the corresponding transcripts [143]. A recent series of studies have shown that targeting epigenetic enzymes to adjust gene expression and ameliorate synaptic defects in PFC is a potential strategy to normalize behavioral phenotypes associated with autism [144]–[147].
In PFC of Shank3-deficient mice, the loss of histone acetylation and upregulation of HDAC2 have been identified [145]. Systemic administration of the class I HDAC inhibitor romidepsin or MS-275 leads to the robust and persistent alleviation of social deficits, as well as the restoration of actin regulators and NMDAR function in PFC [144],[145]. A histone acetylome-wide association study (H3K27ac ChIP-seq) of human ASD postmortem samples also reveals common “epimutations” on genes involved in synaptic transmission, ion transport, and histone deacetylation [148]. Moreover, the repressive histone mark H3K9me2 and its catalyzing enzymes EHMT1/2 are selectively increased in the PFC of Shank3-deficient mice and autistic human postmortem brains [146]. Inhibition of EHMT1/2 rescues social deficits and restores PFC NMDAR function in Shank3-deficient mice via restoring the expression of plasticity gene Arc, which encodes the activity-regulated cytoskeleton-associated protein Arc [146]. The nucleocytoplasmic shuttle protein involved in both cell adhesion and transcriptional regulation, β-catenin, is found to link the loss of Shank3 in the synapse to the upregulation of HDAC2 and EHMT1/2 transcription in the nucleus [145],[146] ().
Another identified high-risk factor for autism is the Cullin 3 (Cul3) gene [141], which encodes a core component of the E3 ubiquitin ligase complex that mediates proteasomal degradation [149]. Loss of Cul3 in forebrain or PFC leads to social deficits and NMDAR hypofunction, while loss of Cul3 in striatum leads to stereotypic behaviors and cell type-specific alteration of neuronal excitability in striatal circuits [147]. One of the misregulated proteins resulting from forebrain Cul3-deficiency is Smyd3, a histone methyltransferase involved in gene transcription. Inhibition or knockdown of Smyd3 ameliorates social deficits and restores PFC NMDAR function in forebrain Cul3-deficient mice [147]. These studies have demonstrated how aberrations in molecular and epigenetic pathways implicated in autism are interconnected.
Schizophrenia
Schizophrenia (SZ) is a chronic psychiatric disorder that is characterized by delusions, hallucinations, disorganized speech and behavior, and negative symptoms [150]. SZ has been linked to both aberrant PFC development during embryonic stages and disrupted PFC maturation in later adolescence driven by environmental stressors, making SZ a unique disorder with two critical susceptibility periods [151]. SZ patients exhibit reduced gray matter in frontal cortex, evidenced by MRI studies [152],[153]. It has been suggested that SZ is driven by disrupted cortical synaptic pruning [154] because its symptoms tend to appear late in adolescence corresponding with the developmental period in which PFC connectivity undergoes maturation [155]. One hypothesis is that psychosocial stress in late development drives microglial overactivation, leading to the increased synaptic pruning in PFC [156]. In accordance with these theories, SZ patients display significant reductions in dendritic spine density in PFC [157], and lower density of perineuronal nets (PNN) - extracellular structures that stabilize synapses, also suggesting synaptic loss or destabilization [158]. SZ patients show reduced blood flow in PFC during task performance [159] and reduced functional connectivity between PFC and other brain regions when performing a cognitive task [160].
SZ etiology has a strong genetic component, and many high-risk SZ genes identified by GWAS studies are highly enriched in mid-frontal lobe [161], form protein interactions in fetal dorsolateral (DL) and ventrolateral PFC [162], and/or are functionally implicated in synaptic transmission [162],[163]. Proteomic analysis of SZ DLPFC samples identified 15 differentially expressed proteins, 7 of which were synaptic [164]. These studies suggest synaptic dysfunction in PFC as a core mechanism in SZ.
The synaptic pathology of SZ in the PFC is complex, involving disrupted glutamatergic, GABAergic, dopaminergic, and serotonergic transmission (). There is strong support for NMDA hypofunction in SZ, as treatment with NMDAR antagonists, such as MK-801 or phencyclidine, can induce SZ-like symptoms in both human subjects [165] and in animals [166]. NMDAR-mediated connectivity between PFC and hippocampus is impaired in SZ, which is suggested as a mechanism underlying working memory deficits [167]. Blocking NMDARs with MK-801 injection leads to the impairment of mPFC-dependent cognitive flexibility and hippocampus-mPFC pathway-dependent spatial working memory [168].
Paradoxically, MK-801 drives the increased release of glutamate in the mPFC, though the antipsychotic medications clozapine and haloperidol are both able to block MK-801-induced glutamate release in PFC of rats [169]. NMDAR antagonists may elicit this paradoxical response by primarily blocking NMDA receptors on GABAergic interneurons, thereby promoting disinhibition of pyramidal neurons [170]. It has thus been postulated that the NMDA hypofunction in SZ mainly occurs in cortical GABAergic interneurons [165]. Postmortem schizophrenia studies have revealed supporting evidence [171], including the reduced density of NR2A-expressing PV interneurons in cortical layers 3 and 4 [172] and the reduced density of GAD67- and NR2A-coexpressing neurons in layers 2 and 5 of anterior cingulate cortex [173].
In rats treated with NMDAR antagonists, PFC neurons display the elevated and irregular spiking activity and the reduced coordinated burst firing, suggesting that NMDA hypofunction in PFC may drive disorganized network activity [174]. In agreement with this, monkeys performing a SZ-relevant cognitive task exhibit the reduced synchronous firing and disrupted connectivity in PFC neurons after the administration of an NMDAR antagonist [175]. PFC-specific inhibition of either NMDA or GABA transmission produces working memory deficits in rats, suggesting that either system could contribute to working memory deficits in SZ [176]. Several biochemical indicators of NMDA dysfunction are reported in SZ PFC, including the reduced expression of several synaptic plasticity-related genes [177], and the significantly lower postsynaptic expression of the NMDAR subunit NR1 and PSD95 [178]. Reduced expression of the presynaptic vesicle protein VAMP has also been reported in SZ PFC [179], while other presynaptic markers appear unchanged [177],[179].
Several well-replicated cellular indications of disrupted GABAergic synaptic transmission have been reported in PFC of SZ patients, including the decreased GAD67 expression and reduced gamma oscillations, suggesting deficient GABA synthesis and GABAergic synaptic transmission, respectively [180]. While no change in the number of PV+ cells [181] or the density of GABAergic synaptic inputs on DLPFC neurons [182] is observed in SZ patients, the reduced PV expression is reported both at the individual cellular level [181] and specifically within presynaptic boutons of PV+ basket cells [182]. Furthermore, an expression analysis of DLPFC from SZ patients indicated the reduced expression of several GABA-related genes, including neuropeptides and both presynaptic and postsynaptic markers [183]. Inhibiting GABAergic transmission in PFC of rats produces several behavioral phenotypes mirroring SZ symptomology [184]. The evidence implicating GABAergic deficits in PFC as a core mechanism in SZ-related cognitive dysfunction has been reviewed previously [185].
Dopamine modulation of PFC, which also plays a major role in working memory [186], is thought to contribute to PFC dysfunction in SZ. In a mouse model carrying a genetic risk factor for neuropsychiatric disorders, adolescent isolation stress induces DNA hypermethylation of the tyrosine hydroxylase (TH) gene in mesocortical dopaminergic neurons, causing the reduced TH expression and cortical dopamine release, leading to SZ-associated behavioral abnormalities [187]. D2R activation leads to long-lasting NMDAR hypofunction, resulting in the profound disruption of functional connectivity between hippocampus and PFC [167]. D2R overexpression in mouse striatum causes the reduced inhibitory synaptic input to PFC pyramidal neurons, suggesting that the deficit in PFC GABAergic function in schizophrenia could be secondary to alterations in the striatal dopamine system [188]. On the other hand, D1R activation in PFC facilitates the maintenance of long-term potentiation (LTP) of glutamatergic transmission, as well as the induction of long-term depression (LTD) of glutamatergic transmission, indicating its bidirectional modulation of synaptic plasticity in mPFC [189]. Computational models predict that an imbalance of D1R/D2R activation in the PFC may underlie the associated cognitive, positive and negative symptoms of SZ [190]. Positron emission tomography studies indicate that radioligand binding to D1R is reduced in PFC of SZ patients, and the extent of the reduction correlates directly with the severity of SZ-related negative symptoms [191]. Further postmortem studies have confirmed the reduced D1R expression in PFC of SZ patients [192].
In addition to dopamine, serotonin dysfunction in PFC is also implicated in SZ, as 5-HT receptors are the major targets of many atypical antipsychotics used to treat SZ, such as clozapine and olanzapine. A systematic review of literatures reported the increased expression of 5-HT1A and the reduced expression of 5-HT2A receptors in PFC of SZ patients [193]. PET studies in SZ patients have similarly shown the reduced 5-HT2A availability in frontal cortex [194]. The novel antipsychotic drug lurasidone restores the diminished NMDAR function in PFC pyramidal neurons from the phencyclidine (PCP) model of schizophrenia via antagonizing 5-HT7 receptors [195], which may underlie its effect on reversing cognitive impairment in SZ. The exact mechanisms by which serotonin dysfunction in PFC contributes to SZ pathophysiology remains to be further elucidated. Collectively, the existing evidence suggests a model whereby the disruption of NMDA and GABA transmission in PFC, which is partially driven by aberrant dopamine and serotonin neuromodulation, produces excitation/inhibition imbalance and network dysconnectivity in SZ ().
Emerging studies have revealed the epigenetic alterations in SZ [196]. ATAC-seq analysis of SZ DLPFC shows that genetic regions with high chromatin accessibility are highly enriched in heritable genetic risk factors for SZ [197]. In agreement with this, two histone markers associated with transcriptional activation (H3K4me3 and H3K27ac) are enriched at SZ genetic risk variants [198]. Broadly disrupted DNA methylation profiles are also reported in SZ PFC [199]. Epigenetic dysregulation of 5-HT2A receptor has been demonstrated in human frontal cortex [200]. In a rat model of SZ, abnormal epigenetic repression of the NMDAR subunit Grin2b in PFC drives glutamatergic synaptic deficits [201]. Upregulation of HDAC1 is found in PFC of human SZ patients and Hdac1 overexpression in PFC is associated with SZ-like phenotypes in mice [202]. These findings suggest that epigenetic dysregulation of selective genes may play an important role in SZ.
Depression
Substantial evidence indicates that mPFC dysfunction plays a critical role in the pathology of major depressive disorder (MDD) [203]. MDD patients display the reduced glucose metabolism in DLPFC [204], and the significantly reduced mPFC volume spanning across dorsomedial PFC, ventromedial (VM) PFC and ACC [205]. While both DLPFC and VMPFC are involved in MDD, they play different roles: damage to VMPFC is associated with low levels of depression, and damage to DLPFC is associated with high levels of depression [206]. The PFC has also emerged as a hub for MDD treatment, as repeated transcranial magnetic stimulation (rTMS) therapy targeting the left PFC appears effective in mitigating depression symptoms [207].
MDD is associated with aberrant synaptic transmission in PFC, affecting both GABA and glutamate systems (). A meta-analysis of GWAS studies identified 102 genetic risk variants in MDD patients, many of which display high expression in frontal cortex and are enriched in biological pathways related to synaptic transmission [208]. Diminished expression of several synaptic genes, including CALM2, SYN1, RAB3A, RAB4B and TUBB4, is observed in PFC of MDD patients, along with reduced spine density in layers II/III of DLPFC [209]. Protein expression levels of the NMDA subunits NR2A/NR2B and PSD95 are also reduced in PFC of MDD patients [210], and RNA-sequencing analyses of postmortem DLPFC from MDD patients similarly revealed the downregulation of genes related to glutamatergic transmission [211]. Moreover, an optogenetic study reveals that activating D1R-, but not D2R-, expressing PFC pyramidal neurons produces rapid and long-lasting antidepressant and anxiolytic responses, and disruption of D1R activity blocks the rapid antidepressant effects of ketamine [212].
In addition to the disrupted glutamatergic transmission, there is also evidence for GABAergic impairment in MDD. Microarray analyses of PFC tissue from MDD patients indicate the downregulation of genes related to GABAergic signaling, including multiple GABAA receptor subunits [213]. Additionally, protein levels of GAD67, the primary enzyme responsible for GABA synthesis, are reduced in MDD PFC, and PET imaging studies reveal lower GABA levels in PFC of MDD patients [214]. RNA-seq of postmortem DLPFC tissue from individuals who died by suicide also revealed the downregulation of GABAAR γ2 subunit [215]. Interestingly, mice carrying heterozygous deficiency of the GABAAR γ2 subunit in forebrain display depression-like behavioral phenotypes [216] and the reduced surface expression of NMDA and AMPA receptor subunits in mPFC that could be restored via ketamine treatment [217], suggesting that GABAergic impairment could affect glutamatergic signaling in MDD PFC. Studies in mice indicate that scopolamine, an effective antidepressant medication acting on muscarinic acetylcholine receptors, enacts its antidepressant effects via targeting somatostatin-expressing GABAergic interneurons in mPFC [218], further implicating the prefrontal cortical GABA system in MDD. The density of PNNs is also reduced in PFC of stressed mice [219] and rats [220], suggesting that PNNs may be disrupted in MDD, similar to SZ.
The NMDAR antagonist ketamine produces rapid antidepressant effects in those with treatment-resistant depression [221], and its therapeutic efficacy appears to be dependent upon its effects on PFC, along with its connecting regions. In rodents, low doses of ketamine have been shown to drive the increased glutamate release [222] and glutamate cycling [223] in PFC. Electrophysiological recordings similarly indicate that ketamine elevates the frequency of excitatory postsynaptic currents (EPSC) in mouse PFC pyramidal neurons [224]. In human MDD patients, ketamine administration similarly increases glutamate/glutamine cycling in PFC [225]. The paradoxical increase in glutamate activity driven by ketamine initially appeared to be mediated by the loss of NMDA-mediated excitation of GABA interneurons, resulting in disinhibition of pyramidal neurons [170] (). However, deletion of the NMDAR subunit GluN2B from mPFC pyramidal neurons in vivo produces antidepressant-like effects, while also preventing ketamine’s effects on behavior, EPSC and the induction of mTOR activation in mouse mPFC [224], suggesting that ketamine enacts direct effects on pyramidal neurons via GluN2B antagonism.
The ketamine-induced elevation in PFC glutamate release promotes AMPA receptor activation, which is necessary for ketamine’s antidepressant effects; pre-administration of the AMPAR receptor antagonist NBQX prevents ketamine’s antidepressant-like behavioral effects in mice [226]. NBQX treatment also prevents ketamine’s induction of mTOR signaling, suggesting that mTOR activation is mediated by AMPA receptor activation [227]. mTOR inhibition is sufficient to prevent the ketamine-dependent induction of synaptogenesis in PFC and antidepressant-like behavioral effects in rats [113]. The mTOR activation of BDNF and synaptogenesis in PFC appear to be involved in Ketamine’s therapeutic function, because transgenic mice carrying a mutant form of BDNF that cannot translocate to dendrites do not display ketamine-dependent synaptogenesis nor antidepressant-like behavioral responses [228]. Notably, the significantly reduced protein expression level of mTOR and several downstream signaling molecules is reported in postmortem PFC tissue from depressed patients [112]. Mice expressing a non-phosphorylatable mutant copy of the signaling molecule eIF4E (eukaryotic initiation factor 4E), which is activated downstream of mTORC1, display depression-like behaviors [229], further implicating the mTOR signaling pathway in MDD. The ketamine metabolite hydroxynorketamine similarly produces antidepressant effects in mice when administered exclusively in the PFC, and this effect is dependent upon the activation of BDNF, and downstream TrkB and mTORC1 signaling [230]. Furthermore, direct activation of the mTORC1 pathway via NV-5138 produces antidepressant effects in rats, suggesting that mTORC1 activation alone may be sufficient to alleviate depressive symptoms [231].
Collectively, these studies suggest that in MDD patients, the PFC is characterized by diminished expression of glutamatergic and GABAergic receptors, accompanied by a reduction in dendritic spine density. Ketamine treatment thus enacts its therapeutic effects by transiently promoting glutamate release in PFC to activate AMPA receptors, which in turn activates the downstream mTOR signaling pathway and promotes BDNF expression, leading to synaptogenesis and elevated glutamatergic transmission ().
In addition to the involvement of PFC synaptic dysregulation in depression, epigenetic changes have been described in PFC of individuals with MDD. Twenty genes are found to be differentially methylated in PFC of MDD patients from a genome-wide methylation analysis, including hypermethylation of Grin2a [232]. HDAC inhibitor produces antidepressant-like effects in mice [233]. DNA methyltransferases have also been implicated in depression [234]. Elevated DNA methylation is observed in PFC of rats subjected to learned helplessness stress, which is reversed by antidepressant imipramine treatment [235]. Moreover, systemic administration of DNA methyltransferase inhibitors induces antidepressant-like effects in rats [236]. These findings suggest that pharmacological targeting of epigenetic enzymes may be a novel treatment strategy for depressive symptoms.
Alzheimer’s Disease
PFC is one of the first regions to be affected in clinical AD, which may explain the early occurrence of the impairment of PFC-mediated cognitive processes, such as working memory and attention. Aging is the leading risk factor for developing AD, so many studies have focused on revealing neurobiological changes that affect synaptic integrity with aging and why aged brain is vulnerable to AD. In aged PFC, the cAMP/PKA pathway is disinhibited, and PKA inhibition provides an effective approach for treating age-related cognitive decline [237]. The marked loss of PFC persistent firing in aged monkeys is also partially restored by inhibiting cAMP signaling or blocking HCN or KCNQ channels [238]. Age-related increase in PKA phosphorylation of tau at serine 214 in monkey PFC is thought to confer risk for degeneration of glutamatergic synapses [239]. From aging monkey studies, it is found that PFC is particularly vulnerable to calcium dysregulation and tau phosphorylation [240]. The loss of thin spines in monkey PFC, which are particularly plastic and linked to learning, is selectively correlated with age-related cognitive impairment [241]. Aβ infused into the monkey brain induces accelerated cortical aging by triggering the specific loss of dendritic thin spines in the PFC [242].
Cholinergic system is crucial for cognitive processes, and deficient acetylcholine (ACh) function has been implicated in AD. A major AD therapy is acetylcholinesterase (AChE) inhibitors, which act to enhance cholinergic function by prolonging the action of endogenously released ACh. AChE inhibitors produce a strong and persistent reduction of synaptic NMDAR responses in PFC pyramidal neurons via nicotinic ACh receptors [243]. Activation of muscarinic or nicotinic ACh receptors in PFC pyramidal neurons also enhances inhibitory synaptic transmission through PKC-dependent mechanisms [244],[245]. The cholinergic regulation of NMDA and GABA function is impaired in transgenic mice overexpressing mutant β-amyloid precursor protein (APP) [243],[245],[246]. Aβ-induced loss of RACK1 (an anchoring protein for activated PKC) distribution in the membrane fraction of cortical neurons is found to underlie the impairment of muscarinic regulation of PKC [247]. In aged monkeys with naturally occurring cholinergic depletion, low-dose muscarinic M1R stimulation enhances delay firing in dorsolateral PFC neurons and improves working memory via a mechanism involving KCNQ channels [248]. These studies suggest that restoring cholinergic regulation of PFC synaptic activity and neuronal excitability might help to treat AD-associated cognitive disorders.
The brain noradrenergic system is also critical for cognition and noradrengergic dysfunction is a critical early step in AD progression [249]. A recent study has found that β-amyloid oligomers bind to an allosteric site on α2-ARs to redirect norepinephrine signaling to activate the pathogenic GSK3β/tau cascade [250]. Guanfacine, a specific α2-AR antagonist, has been used to treat cognitive disorders linked to PFC dysfunction [251].
Systematic search for global gene expression changes in PFC during the course of AD has found a temporally orchestrated increase of genes involved in synaptic activity during the very early pre-symptomatic stage, probably representing a coping mechanism against increased β-amyloid levels, and the decreased expression of these synaptic genes in later Braak stages, suggesting a reduction in synaptic activity that coincides with AD neuropathology and cognitive impairment [252]. Studies of postmortem human brains have found the significant association between cognitive decline and the loss of synaptic proteins in PFC, such as SNAP25, SNAP47, SV2C, SYT2 and GRIA3 [253],[254], suggesting that these PFC synaptic markers are predictive molecular fingerprint for neurodegenerative diseases, and understanding the basis of synaptic impairment is the most effective route for early intervention and prevention of cognitive decline.
Both AD patients and symptomatic AD mouse models show impaired synaptic plasticity in PFC, which is related to the impaired NMDAR function [255]. The diminished transcription of NMDA and AMPA receptors in late-stage AD mouse models has been linked to the EHMT1/2-mediated repressive histone methylation at their promoters [256]. Treatment of AD models with specific EHMT1/2 inhibitors reverses the increased H3K9me2 enrichment at genes involved in neuronal signaling (e.g. Grin2b encoding NR2B subunit and Gria2 encoding GluR2 subunit), restores glutamate receptor expression and excitatory synaptic function in PFC, and rescues the impaired recognition memory, working memory, and spatial memory [256] ().
In addition to the downregulation of genes involved in synaptic transmission and synapse organization, there is also prominent upregulation of genes involved in cell stress, immune response activation and cell death in PFC of AD patients [257]–[259]. A recent studie has found that H3K4 trimethylation (H3K4me3), a histone mark for gene activation, is significantly elevated in PFC of human AD postmortem tissues and AD mouse models, and inhibiting H3K4-specific methyltransferases, including MLL1-4 (also named as KMT2A-D) and SETD1a/b, leads to the substantial recovery of glutamatergic synaptic function in PFC pyramidal neurons, and the significant improvement of memory-related behaviors in mutant Tau transgenic mice [260]. One of the top-ranking genes elevated by the abnormally high H3K4me3 in PFC of AD patients and mouse models is SGK1, which encodes the stress-responsive kinase SGK1. A short treatment with the specific SGK1 inhibitor results in the significant reduction of hyper-phosphorylated tau (a pathological hallmark of AD) and the recovery of synaptic receptor expression and glutamatergic transmission in PFC, as well as the restoration of recognition and spatial memories, in mutant Tau transgenic mice [260] (). These results suggest that targeting histone methylation enzymes may represent a novel therapeutic avenue for normalizing the aberrant epigenetic regulation of gene transcription in PFC that underlies cognitive deficits in AD.
Concluding Remarks
Overwhelming evidence has indicated that PFC is a uniquely important brain region for the highest level of excecutive function, which drives self-regulatory and goal-directed behaviors [261]–[263]. Dysregulation of PFC synaptic transmission directly contributes to the impairment of social, affective, motivational, and cognitive function associated with brain disorders including autism, schizophrenia, depression and Alzheimer’s disease. Studies in animal models have revealed critical neural circuits, biological pathways, and molecular players that converge to control the excitatory and inhibitory signals in PFC networks. Novel epigenetic mechanism-based strategies have been proposed as potential avenues of therapeutic intervention for mental diseases linked to PFC dysfunction.
There are many open questions that await to be further explored in future studies, and a few examples are listed as follows:
Acknowledgements
Acklowdgements:We are grateful to former and current members in Dr. Yan’s laboratory for their contributions to the original findings and NIH grants (MH108842, MH112237, DA037618, AG056060, AG064656 to Z.Y.) for their support of some of the work reviewed here.