黑质致密部-纹状体多巴胺能通路的上游脑区追踪  

A TRACING STUDY OF UPSTREAM BRAIN REGIONS OF DOPAMINERGIC PATHWAY PROJECTING FROM SUBSTANTIA NIGRA PARS COMPACTA TO STRIATUM

作　　者：商晓钰 张腾元 谢俊霞 徐华敏 SHANG Xiaoyu;ZHANG Tengyuan;XIE Junxia;XU Huamin(Department of Physiology and Pathophysiology,School of Basic Medicine,Medical College of Qingdao University,Qingdao 266071,China)

机构地区：[1]青岛大学医学部基础医学院生理学与病理生理学系,山东青岛266071

出　　处：《青岛大学学报（医学版）》2023年第3期349-352,共4页Journal of Qingdao University(Medical Sciences)

基　　金：国家自然科学基金项目(31871202)。

摘　　要：目的探究投射到纹状体(Str)的黑质致密部(SNpc)多巴胺(DA)能神经元的上游脑区。方法采用逆向病毒示踪技术逆向追踪SNpc-Str环路的上游脑区。应用立体定位技术在8周龄C57BL/6雄性小鼠Str区注射逆行追踪病毒rAAV-hsyn-cre-wpre-pA(retro);同时在SNpc注射病毒AAV-DIO-EF1α-GFP-TVA和AAV-DIO-EF1α-RVG(1∶2混合);3周后在SNpc注射逆行跨突触狂犬病毒RV-Enva-ΔG-dsRed。1周后取脑进行冷冻切片,应用Olympus VS120荧光显微镜观察表达dsRed阳性细胞的脑区。结果应用荧光显微镜在SNpc可以观察到GFP(绿色)和dsRed(红色)阳性细胞,证明依赖Cre重组酶表达的AAV-DIO-EF1α-GFP-TVA病毒以及依赖TVA元件表达的狂犬病毒均在SNpc正常表达。同时在外侧下丘脑(LH)、前连合后肢间质核(IPAC)脑区观察到dsRed(红色)阳性细胞。结论LH和IPAC脑区可能是SNpc-Str环路DA能神经元的上游脑区。Objective To investigate the upstream brain regions of dopaminergic neurons projecting from the substantia nigra pars compacta(SNpc)to the striatum(Str).Methods The retrograde virus tracing technique was used to identify the upstream brain regions of the SNpc-Str circuit.Eight-week-old male C57BL/6 mice received stereotactic injection of the retrograde virus rAAV-hsyn-cre-wpre-pA(retro)into the Str and also a 1∶2 mixture of AAV-DIO-EF1α-GFP-TVA and AAV-DIO-EF1α-RVG into the SNpc.After three weeks,the retrograde transsynaptic rabies virus RV-Enva-ΔG-dsRed was injected into the SNpc.After another one week,the brain was taken to prepare slices to observe dsRed-positive brain regions using an Olympus VS120 fluorescence microscope.Results Cells positive for GFP(green)and dsRed(red)were observed in the SNpc under the fluorescence microscope,demonstrating that both Cre recombinase-dependent AAV-DIO-EF1α-GFP-TVA virus and TVA-dependent ra-bies virus were normally expressed in the SNpc.At the same time,dsRed-positive cells were observed in the lateral hypothalamus(LH)and the interstitial nucleus of the posterior limb of the anterior commissure(IPAC).Conclusion LH and IPAC might be the upstream brain regions of dopaminergic neurons of the SNpc-Str circuit.

关 键 词：多巴胺能神经元 密部 纹状体 下丘脑区 侧 前连合 大脑 神经解剖束追踪技术 

分 类 号：R338.2[医药卫生—人体生理学]