伏隔核中型多棘神經元興奮性及海馬長時程增強在抑郁癥發(fā)病中的作用
發(fā)布時間：2018-09-13 11:37
【摘要】：抑郁癥是現今世界上最流行的精神疾病，在中國有超過2600萬人患有抑郁癥，具有高發(fā)病率及低診斷率的特點，對當今社會帶來了嚴重的影響。同時，抑郁癥的產生原因復雜，遺傳和環(huán)境因素兼而有之，而且在病理上不易顯現出可檢測的實質性病變特征，因此對人們研究抑郁癥的發(fā)病機制造成了極大的困擾，直接影響到抗抑郁藥物的開發(fā)。因此闡明抑郁癥的發(fā)病機制，為抑郁癥的臨床治療提供有效的思路與方法是當今中國乃至世界精神疾病研究領域亟待解決的科學問題之一。近年來負責獎賞及成癮機制的重要腦區(qū)——伏隔核（Nuclear accumbens, NAc）在抑郁發(fā)病和抗抑郁藥物治療中的作用受到越來越多的關注。NAc是基底核區(qū)域的一個信息整合核團，主要接受來自中腦腹側背蓋區(qū)（ventraltegmental area, VTA）的多巴胺能神經投射，參與與情緒調節(jié)相關的神經環(huán)路。同時，抑郁癥發(fā)病假說之一的單胺類假說認為多巴胺在抑郁癥發(fā)病及抗抑郁治療中有著不可忽視的作用，但是多巴胺對腦內相關腦區(qū)神經活動的影響仍不明了，因此如果能夠從電生理學的角度，對NAc的神經元放電特點進行研究，并結合行為學和生化學的研究，則能夠幫助我們更加全面深入的了解腦內多巴胺系統中的NAc神經元活性在抑郁癥的發(fā)病過程中的作用及變化情況。另外，我們也對來自于郭熙志教授實驗室的一種干細胞因子（stem cell factor, SCF）及其酪氨酸激酶受體c-Kit通路功能受損的基因突變小鼠進行了電生理學的研究。這種基因突變小鼠具有顯著的抑郁樣行為表現，并且伴隨著學習與記憶功能的缺陷和海馬區(qū)域的神經元再生障礙。因此通過對其海馬區(qū)突觸可塑性的研究，能夠幫助我們了解這種基因突變對海馬的突觸傳遞及其可塑性的影響，從電生理學的角度解釋這種抑郁小鼠在學習與記憶方面出現缺陷的原因。 在本研究中，我們選取了兩種抑郁癥模型鼠Wistar Kyoto (WKY)大鼠和c-KitV922G/+雜合突變小鼠，在進行行為學檢測驗證抑郁樣行為的基礎上，1）研究幼年期WKY大鼠在抑郁發(fā)生及藥物治療過程中，多巴胺NAc的中型多棘神經元（Medium spiny neurons, MSNs）興奮性的變化；2）研究成年c-KitV922G/+雜合突變小鼠海馬（hippocampus, HP）的CA1區(qū)及CA3區(qū)突觸傳遞及長時程增強（long-term potentiation, LTP）形成的變化。通過以上檢測，我們探討了NAc的MSNs放電特點及海馬神經環(huán)路的信號傳導變化在抑郁癥發(fā)病中的作用。 主要的方法及結果： 1檢測幼年期WKY大鼠在抑郁發(fā)生及藥物治療過程中，多巴胺D2類受體（dopamine D2-like receptor）介導的NAc MSNs膜興奮性的變化。 （1）制作4~5周健康雄性WKY大鼠和其正常對照組Wistar大鼠的大腦冠狀切片（含有NAc），切片厚度300μm。 （2）通過可視化全細胞膜片鉗的方法，記錄并分析在給予多巴胺D2類受體激動劑喹吡羅（quinpirole）前后，NAc中心核（core）區(qū)域的MSNs動作電位的發(fā)放情況。在記錄過程中，始終在灌流液中加入伽馬氨基丁酸A（GABAA）受體阻斷劑木防己苦毒素（picrotoxin, PTX）和興奮性谷氨酸受體阻斷劑犬尿喹啉酸（kynurenic acid）。在電流鉗模式下給予細胞膜間隔20sec、強度呈步進遞增形式的電流刺激，電流強度范圍為0-500pA，步幅為50pA，記錄其誘發(fā)的動作電位數量及產生閾值。結果發(fā)現在幼年期WKY大鼠中，多巴胺D2類受體介導的信號通路對NAc core區(qū)域的MSNs膜興奮性的抑制作用缺失； （3）給予幼年期WKY大鼠持續(xù)12天的諾米芬新（nomifensine，一種多巴胺和去甲腎上腺素重攝取抑制劑）治療，通過一系列的行為學檢測發(fā)現治療前的WKY大鼠在糖水偏好、開場實驗和強迫游泳的行為學實驗中均表現出顯著的抑郁樣行為，而nomifensine治療后其在糖水偏好和強迫游泳實驗中的抑郁樣行為顯著減少。 （4）對治療后的WKY大鼠NAc core區(qū)域的MSNs進行電生理學檢測，并與未治療組的WKY大鼠以及正常對照組Wistar大鼠比較，結果發(fā)現在接受nomifensine治療后WKY大鼠NAc core區(qū)域的MSNs膜興奮性減弱。 2檢測成年c-KitV922G/+雜合突變小鼠在抑郁發(fā)生過程中，，Mossyfiber-CA3(MF-CA3)和Schaffer collateral-commissural fiber-CA1(Sch-CA1)通路上高頻刺激誘導的LTP形成情況。 （1）制作2~3月齡的健康雄性c-KitV922G/+雜合突變小鼠和其野生型對照小鼠的急性海馬切片，切片厚度400μm。 （2）在場電位模式下給予海馬切片MF-CA3通路100Hz/1sec兩次，以及Sch-CA1通路100Hz/1sec1次的高頻刺激，觀察LTP的形成情況。實驗結束后灌流含有0.1μM DCG IV的人工腦脊液（artificialcerebrospinal fluid, ACSF)，觀察突觸反應的被抑制情況，以驗證所記錄的通路確實為MF-CA3。結果發(fā)現c-KitV922G/+雜合突變小鼠的MF-CA3通路的LTP形成受損，而Sch-CA1通路的LTP形成未受影響。主要結論： （1）Nomifensine能夠改善幼年期WKY大鼠的抑郁樣行為； （2）多巴胺D2類受體介導的對MSNs興奮性的抑制的缺失可能參與幼年期WKY大鼠抑郁樣行為的發(fā)生。 （3）Nomifensine可能通過影響幼年期WKY大鼠海馬和紋狀體區(qū)域的DRD2發(fā)揮抗抑郁作用。 （4）成年小鼠海馬的SCF/c-Kit通路異常對MF-CA3通路LTP形成的抑制，可能參與其學習與記憶的形成障礙。
[Abstract]:Depression is the most prevalent psychiatric disorder in the world today. In China, more than 26 million people suffer from depression, which has the characteristics of high incidence and low diagnostic rate. It has brought serious impact on today's society. Qualitative pathological changes cause great trouble to the study of the pathogenesis of depression and directly affect the development of antidepressants. Therefore, to clarify the pathogenesis of depression and provide effective ideas and methods for the clinical treatment of depression is an urgent scientific question to be solved in the field of psychiatric diseases in China and even in the world. In recent years, more and more attention has been paid to the role of the nucleus accumbens (NAc), an important brain region responsible for reward and addiction mechanisms, in the pathogenesis of depression and antidepressant drugs. NAc is an information-integrating nucleus in the basal nucleus region, which mainly receives information from the ventral tegmental area (VTA). The monoamine hypothesis, one of the hypotheses of depression, holds that dopamine plays an important role in the pathogenesis and antidepressant therapy of depression, but the effect of dopamine on neurological activity in the brain-related areas is still unknown, so if it can be induced from electricity From the point of view of physiology, studying the characteristics of NAc neuronal firing and combining behavioral and biochemical studies can help us understand the role and changes of NAc neuronal activity in the brain dopamine system in the pathogenesis of depression. The mutant mice with a stem cell factor (SCF) and its tyrosine kinase receptor c-Kit pathway dysfunction were electrophysiologically studied. The study of synaptic plasticity in the hippocampus can help us to understand the effect of this gene mutation on synaptic transmission and plasticity in the hippocampus, and explain the reason for the deficiency of learning and memory in the depressive mice from the perspective of electrophysiology.
In this study, we selected Wistar Kyoto (WKY) rats and c-Kit V922G /+ heterozygous mice from two depression models. On the basis of behavioral tests to verify depression-like behavior, we studied the Medium Spiny Neurons (MSN) of dopamine NAc during depression onset and drug treatment in young WKY rats. 2) To study the changes of synaptic transmission and long-term potentiation (LTP) in CA1 and CA3 regions of adult c-KitV922G/+ heterozygous mice hippocampus (HP). The role of.
Main methods and results:
1 To detect the changes of dopamine D2-like receptor-mediated NAc-MSNs membrane excitability during depression and drug therapy in juvenile WKY rats.
(1) Coronary sections (containing NAc) of the brain of healthy male WKY rats and Wistar rats were made at 4-5 weeks and the thickness of the sections was 300 microns.
(2) Visual whole-cell patch clamp technique was used to record and analyze the release of action potential of MSNs in the central nucleus of NAc before and after administration of quinpirol, a dopamine D2 receptor agonist. Xin, PTX and excitatory glutamate receptor blocker kynurenic acid were administered at a 20 sec interval with a step-by-step incremental current stimulus. The current intensity ranged from 0 to 500 pA with a 50 pA stride. The number and threshold of evoked action potentials were recorded in young WKY rats. Dopamine D2 receptor-mediated signaling pathway has no inhibitory effect on the excitability of MSNs membrane in NAc core region.
(3) Treated with nomifensine (a dopamine and norepinephrine reuptake inhibitor) for 12 days in juvenile WKY rats, a series of behavioral tests showed significant depression-like behaviors in glycemic preference, open-field experiment and forced swimming behavior tests in WKY rats before treatment. After treatment with sine, depressive behaviors in sugar water preference and forced swimming test were significantly reduced.
(4) After treatment, MSNs in the NAc core region of WKY rats were detected electrophysiologically, and compared with WKY rats without treatment group and Wistar rats with normal control group. The results showed that the excitability of MSNs membrane in the NAc core region of WKY rats decreased after treatment with nomifensine.
2. To detect the formation of LTP induced by high frequency stimulation in Mossyfiber-CA3 (MF-CA3) and Schaffer collateral-commissural fiber-CA1 (Sch-CA1) pathways during depression in adult c-KitV922G/+ heterozygous mice.
(1) Acute hippocampal slices of healthy male c-KitV922G/+ heterozygous mutant mice aged 2-3 months and wild type control mice were prepared. The slices were 400 microns thick.
(2) The formation of LTP was observed by stimulating the hippocampal MF-CA3 pathway 100 Hz/1 sec twice and the Sch-CA1 pathway 100 Hz/1 sec twice under the field potential mode. It was found that LTP formation in the MF-CA3 pathway was impaired in c-KitV922G/+ heterozygous mice, but not in the Sch-CA1 pathway.
(1) Nomifensine can improve depressive behavior in juvenile WKY rats.
(2) The absence of dopamine D2 receptor-mediated inhibition of MSNs excitability may be involved in the development of depression-like behavior in young WKY rats.
(3) Nomifensine may play an antidepressant role by influencing DRD2 in hippocampus and striatum of juvenile WKY rats.
(4) The abnormal SCF/c-Kit pathway in the hippocampus of adult mice inhibits the formation of LTP in MF-CA3 pathway, which may be involved in the formation of learning and memory disorders.
【學位授予單位】：上海交通大學
【學位級別】：博士
【學位授予年份】：2013
【分類號】：R749.41
[Abstract]:Depression is the most prevalent psychiatric disorder in the world today. In China, more than 26 million people suffer from depression, which has the characteristics of high incidence and low diagnostic rate. It has brought serious impact on today's society. Qualitative pathological changes cause great trouble to the study of the pathogenesis of depression and directly affect the development of antidepressants. Therefore, to clarify the pathogenesis of depression and provide effective ideas and methods for the clinical treatment of depression is an urgent scientific question to be solved in the field of psychiatric diseases in China and even in the world. In recent years, more and more attention has been paid to the role of the nucleus accumbens (NAc), an important brain region responsible for reward and addiction mechanisms, in the pathogenesis of depression and antidepressant drugs. NAc is an information-integrating nucleus in the basal nucleus region, which mainly receives information from the ventral tegmental area (VTA). The monoamine hypothesis, one of the hypotheses of depression, holds that dopamine plays an important role in the pathogenesis and antidepressant therapy of depression, but the effect of dopamine on neurological activity in the brain-related areas is still unknown, so if it can be induced from electricity From the point of view of physiology, studying the characteristics of NAc neuronal firing and combining behavioral and biochemical studies can help us understand the role and changes of NAc neuronal activity in the brain dopamine system in the pathogenesis of depression. The mutant mice with a stem cell factor (SCF) and its tyrosine kinase receptor c-Kit pathway dysfunction were electrophysiologically studied. The study of synaptic plasticity in the hippocampus can help us to understand the effect of this gene mutation on synaptic transmission and plasticity in the hippocampus, and explain the reason for the deficiency of learning and memory in the depressive mice from the perspective of electrophysiology.
In this study, we selected Wistar Kyoto (WKY) rats and c-Kit V922G /+ heterozygous mice from two depression models. On the basis of behavioral tests to verify depression-like behavior, we studied the Medium Spiny Neurons (MSN) of dopamine NAc during depression onset and drug treatment in young WKY rats. 2) To study the changes of synaptic transmission and long-term potentiation (LTP) in CA1 and CA3 regions of adult c-KitV922G/+ heterozygous mice hippocampus (HP). The role of.
Main methods and results:
1 To detect the changes of dopamine D2-like receptor-mediated NAc-MSNs membrane excitability during depression and drug therapy in juvenile WKY rats.
(1) Coronary sections (containing NAc) of the brain of healthy male WKY rats and Wistar rats were made at 4-5 weeks and the thickness of the sections was 300 microns.
(2) Visual whole-cell patch clamp technique was used to record and analyze the release of action potential of MSNs in the central nucleus of NAc before and after administration of quinpirol, a dopamine D2 receptor agonist. Xin, PTX and excitatory glutamate receptor blocker kynurenic acid were administered at a 20 sec interval with a step-by-step incremental current stimulus. The current intensity ranged from 0 to 500 pA with a 50 pA stride. The number and threshold of evoked action potentials were recorded in young WKY rats. Dopamine D2 receptor-mediated signaling pathway has no inhibitory effect on the excitability of MSNs membrane in NAc core region.
(3) Treated with nomifensine (a dopamine and norepinephrine reuptake inhibitor) for 12 days in juvenile WKY rats, a series of behavioral tests showed significant depression-like behaviors in glycemic preference, open-field experiment and forced swimming behavior tests in WKY rats before treatment. After treatment with sine, depressive behaviors in sugar water preference and forced swimming test were significantly reduced.
(4) After treatment, MSNs in the NAc core region of WKY rats were detected electrophysiologically, and compared with WKY rats without treatment group and Wistar rats with normal control group. The results showed that the excitability of MSNs membrane in the NAc core region of WKY rats decreased after treatment with nomifensine.
2. To detect the formation of LTP induced by high frequency stimulation in Mossyfiber-CA3 (MF-CA3) and Schaffer collateral-commissural fiber-CA1 (Sch-CA1) pathways during depression in adult c-KitV922G/+ heterozygous mice.
(1) Acute hippocampal slices of healthy male c-KitV922G/+ heterozygous mutant mice aged 2-3 months and wild type control mice were prepared. The slices were 400 microns thick.
(2) The formation of LTP was observed by stimulating the hippocampal MF-CA3 pathway 100 Hz/1 sec twice and the Sch-CA1 pathway 100 Hz/1 sec twice under the field potential mode. It was found that LTP formation in the MF-CA3 pathway was impaired in c-KitV922G/+ heterozygous mice, but not in the Sch-CA1 pathway.
(1) Nomifensine can improve depressive behavior in juvenile WKY rats.
(2) The absence of dopamine D2 receptor-mediated inhibition of MSNs excitability may be involved in the development of depression-like behavior in young WKY rats.
(3) Nomifensine may play an antidepressant role by influencing DRD2 in hippocampus and striatum of juvenile WKY rats.
(4) The abnormal SCF/c-Kit pathway in the hippocampus of adult mice inhibits the formation of LTP in MF-CA3 pathway, which may be involved in the formation of learning and memory disorders.
【學位授予單位】：上海交通大學
【學位級別】：博士
【學位授予年份】：2013
【分類號】：R749.41
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