Involvement of GABAergic projection from central amygdaloid nucleus to paraventricular nucleus of hypothalamus in electroacupuncture induced regulation of gastric function in GAD2-Cre mice

WANG Hao; SHEN Guo-ming; WANG Xi-yang; ZHANG Meng-ting

doi:10.13702/j.1000-0607.190898

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Involvement of GABAergic projection from central amygdaloid nucleus to paraventricular nucleus of hypothalamus in electroacupuncture induced regulation of gastric function in GAD2-Cre mice

更新时间：2023-08-11

- Involvement of GABAergic projection from central amygdaloid nucleus to paraventricular nucleus of hypothalamus in electroacupuncture induced regulation of gastric function in GAD2-Cre mice
- Acupuncture Research Vol. 45, Issue 5, Pages: 351-356(2020)
- 作者机构：
  
  1. 安徽中医药大学中西医结合学院(生命科学学院)
  2. 安徽中医药大学中西医结合研究所
- 作者简介：
- 基金信息：
- DOI：10.13702/j.1000-0607.190898
  CLC：
- Published：2020
- 稿件说明：
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WANG Hao, SHEN Guo-ming, WANG Xi-yang, et al. Involvement of GABAergic projection from central amygdaloid nucleus to paraventricular nucleus of hypothalamus in electroacupuncture induced regulation of gastric function in GAD2-Cre mice[J]. Acupuncture research, 2020, 45(5): 351-356.
DOI：

WANG Hao, SHEN Guo-ming, WANG Xi-yang, et al. Involvement of GABAergic projection from central amygdaloid nucleus to paraventricular nucleus of hypothalamus in electroacupuncture induced regulation of gastric function in GAD2-Cre mice[J]. Acupuncture research, 2020, 45(5): 351-356. DOI： 10.13702/j.1000-0607.190898.

摘要

目的:观察杏仁中央核(CeA)γ-氨基丁酸(GABA)能神经元及下丘脑室旁核(PVN)在胃俞募配穴针刺调节胃功能中的作用

基于CeA

(GABA)

-PVN环路探讨胃俞募配穴针刺调节胃功能的中枢机制。方法:实验分3部分进行:第1部分选取C57BL/6小鼠

随机分为正常组和电针组(电针"中脘"和右侧"胃俞"20 min)

每组6只

用免疫荧光染色法检测CeA、PVN区c-fos表达;第2部分用GAD2-Cre小鼠4只

在CeA区注射顺行示踪病毒(rAAV-EF1α-DIO-mcherry-WPRE-pA)

观察CeA中GABA能神经元是否投射到PVN区;第3部分选取GAD2-Cre小鼠

随机分为空载病毒组(CeA注射rAAV-EF1α-DIO-mcherry-WPRE-pA)、化学遗传激活病毒组[CeA注射rAAV-EF1α-DIO-hM3D(Gq)-mcherry-WPRE-pA]、化学遗传激活病毒+电针组[CeA注射rAAV-EF1α-DIO-hM3D(Gq)-mcherry-WPRE-pA+电针"中脘"和右侧"胃俞"20 min]

每组6只

记录各组小鼠的进食量

检测胃排空率

同时用高效液相色谱法测定PVN区GABA含量。结果:1)与正常组比较

电针组CeA、PVN区c-fos表达均明显增加(P

0.01)。2)小鼠脑区切片可见病毒在CeA区GABA能神经元胞体大量表达

在PVN区可见大量有病毒表达的投射纤维

表明CeA中GABA能神经元可投射到PVN区

即CeA到PVN有直接的纤维联系。3)与空载病毒组比较

化学遗传激活病毒组小鼠进食量和胃排空率明显下降(P

0.01)

PVN区GABA含量明显升高(P

0.01);与化学遗传激活病毒组比较

化学遗传激活病毒+电针组进食量和胃排空率明显增加(P

0.01)

PVN区GABA含量明显下降(P

0.01)。结论:电针胃俞募配穴对胃功能的调控可通过CeA(GABA)-PVN环路探讨胃俞募配穴针刺调节胃功能的中枢机制。方法:实验分3部分进行:第1部分选取C57BL/6小鼠

随机分为正常组和电针组(电针"中脘"和右侧"胃俞"20 min)

每组6只

用免疫荧光染色法检测CeA、PVN区c-fos表达;第2部分用GAD2-Cre小鼠4只

在CeA区注射顺行示踪病毒(rAAV-EF1α-DIO-mcherry-WPRE-pA)

观察CeA中GABA能神经元是否投射到PVN区;第3部分选取GAD2-Cre小鼠

每组6只

记录各组小鼠的进食量

检测胃排空率

同时用高效液相色谱法测定PVN区GABA含量。结果:1)与正常组比较

电针组CeA、PVN区c-fos表达均明显增加(P

0.01)。2)小鼠脑区切片可见病毒在CeA区GABA能神经元胞体大量表达

在PVN区可见大量有病毒表达的投射纤维

表明CeA中GABA能神经元可投射到PVN区

即CeA到PVN有直接的纤维联系。3)与空载病毒组比较

化学遗传激活病毒组小鼠进食量和胃排空率明显下降(P

0.01)

PVN区GABA含量明显升高(P

0.01);与化学遗传激活病毒组比较

化学遗传激活病毒+电针组进食量和胃排空率明显增加(P

0.01)

PVN区GABA含量明显下降(P

0.01)。结论:电针胃俞募配穴对胃功能的调控可通过CeA

(GABA)

-PVN环路实现。Objective To explore the effect of γ-aminobutyric acid(GABA)ergic neuronal circuit of the central amygdaloid nucleus(CeA) and the paraventricular nucleus of hypothalamus(PVN) on electroacupuncture(EA)-induced regulation of gastric function by way of CeA-PVN projection. Methods The present study included 3 parts: 1) C57 BL/6 mice were randomly divided into control and EA groups(n=6 in each group). EA was applied to right "Weishu"(BL21

Back-shu point) and "Zhongwan"(CV12

Front-mu point) for 20 min

followed by detecting the expression of c-fos in the CeA and PVN by using immunofluorescence staining; 2) Microinjection of anterograde tracer(rAAV-EF1α-DIO-mcherry-WPRE-pA) into the CeA was conducted in GAD2-Cre mice for confirming the projection of GABAergic neurons from CeA to PVN; 3) GAD2-Cre mice were randomly divided into rAAV-DIO-mcherry(intra-CeA injection of rAAV-EF1α-DIO-mcherry-WPRE-pA)

rAAV-DIO-hM3 D(Gq)-mcherry(intra-CeA injection of rAAV-EF1α-DIO-hM3 D(Gq)-mcherry-WPRE-pA) and rAAV-DIO-hM3 D(Gq)-mcherry+EA groups(n=6 in each group). The food intake and gastric empty were detected

and the concentration of GABA in the PVN was assayed by using high performance liquid chromatography on the 28(GABA)-PVN环路实现。

Abstract

Objective To explore the effect of γ-aminobutyric acid(GABA)ergic neuronal circuit of the central amygdaloid nucleus(CeA) and the paraventricular nucleus of hypothalamus(PVN) on electroacupuncture(EA)-induced regulation of gastric function by way of CeA-PVN projection. Methods The present study included 3 parts: 1) C57 BL/6 mice were randomly divided into control and EA groups(n=6 in each group). EA was applied to right "Weishu"(BL21

Back-shu point) and "Zhongwan"(CV12

Front-mu point) for 20 min

and the concentration of GABA in the PVN was assayed by using high performance liquid chromatography on the 28

(th)

day after intra-CeA injection. Results 1) The expression of c-fos in the CeA and PVN was significantly increased in the EA group relevant to the control group(P

0.01)

suggesting an activation of neurons in both CeA and PVN after EA. 2) Following CeA injection of rAAV-EF1α-DIO-mcherry-WPRE-pA

the densely expressed virus GABAergic neurons were found in CeA and large number of projection fibers found in the PVN

suggesting a direct connection between CeA and PVN. 3) After activating the GABAergic neurons of CeA

the concentration of GABA in the PVN was obviously increased(P

0.01)

the food intake and the gastric empty were considerably decreased relevant to the rAAV-DIO-mcherry group(P

0.01). Following EA intervention

the concentration of GABA in the PVN was obviously decreased(P

0.01)

the food intake and the gastric empty were significantly increased relevant to the rAAV-DIO-hM3 D(Gq)-mcherry group(P

0.01). Conclusion EA of BL21 and CV12(Back-shu and Front-mu acupoints) can increase food intake and gastric empty in GAD2-Cre mice

which may be achieved by suppressing the release of GABA in PVN through CeA-PVN GABAergic neural circuit.

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Related Institution

Department of Thoracic Surgery, The First Affiliated Hospital of Anhui Medical University

College of Acupuncture-Moxibustion and Tuina, Chengdu University of Traditional Chinese Medicine

College of Basic Medicine,Chengdu University of Traditional Chinese Medicine

Department of Anatomy,Wannan Medical College

College of Acupuncture-Moxibustion and Tuina， Chengdu University of Traditional Chinese Medicine

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