GRK5过表达对P301S Tau转基因小鼠抑郁样行为的影响及其机制

doi:10.13481/j.1671-587X.20230304

摘要/Abstract

摘要：

目的探讨G蛋白偶联受体激酶5（GRK5）过表达对P301S Tau转基因小鼠抑郁样行为的影响，并阐明其可能的作用机制。方法 17只雄性P301S Tau小鼠随机分为空白对照组（n=6，不做任何处理）、阴性对照组（n=5，双侧海马区给予空载对照病毒）和过表达组（n=6，双侧海马区给予GRK5过表达病毒）。糖水偏好实验检测各组小鼠糖水偏好率，悬尾实验（TST）和强迫游泳实验（FST）检测各组小鼠不动时间百分率，Western blotting法检测各组小鼠海马组织中神经功能相关蛋白表达水平，免疫荧光染色检测各组小鼠海马组织中小胶质细胞标记物抗体（IBA1）、神经元特异性核蛋白（NeuN）和胶质纤维酸蛋白（GFAP）表达情况。结果糖水偏好实验检测，与空白对照组和阴性对照组比较，过表达组小鼠糖水偏好率升高（P<0.05）。TST和FST检测，与空白对照组和阴性对照组比较，过表达组小鼠不动时间百分率升高（P<0.05）。Western blotting法检测，与空白对照组和阴性对照组比较，过表达组小鼠海马组织中GRK5、Tau T205和IBA1蛋白表达水平升高（P<0.05），突触小泡蛋白（SYN）蛋白表达水平降低（P<0.05）。免疫荧光染色检测，与空白对照组比较，阴性对照组和过表达组小鼠海马组织中均有增强绿色荧光蛋白（EGFP）表达，EGFP主要与NeuN发生共定位，小胶质细胞和星形胶质细胞中几乎未观察到EGFP；与空白对照组和阴性对照组比较，过表达组小鼠海马组织中小胶质细胞数增加（P<0.05）。结论海马组织中GRK5过表达诱导P301S Tau小鼠抑郁样行为，其机制可能与GRK5在神经元核中表达水平升高有关。

关键词: 小鼠，P301S, G蛋白偶联受体激酶5, 抑郁样行为, Tau蛋白

Abstract:

Objective To discuss the effect of G protein-coupled receptor kinase5（GRK5） over-expression on the depression-like behaviors of the P301S Tau transgenic mice，and to clarify its possible mechanism. Methods Seventeen male P301S Tau mice were randomly divided into blank control group （given no treatment） and negative control group （given negative control virus in bilateral hippocampus regions），over-expression group （given GRK5 over-expression virus in bilateral hippocampus regions）. The sugar-water preference test was used to detect the sugar-water preference rates of the mice in various groups， the percentages of immobility time of the mice in various groups were detected by tail suspension test（TST） and forced swimming test（FST）； the expression levels of neurological function-related proteins in hippocampus tissue of the mice in various groups were dectated by Western blotting method； the expressions of microglia marker protein （IBA1）， neuronal marker protein （NeuN）， and astrocyte marker protein （GFAP） in hippocampus tissue of the mice in various groups were detected by immunofluorescence staining. Result The sugar-water preference assay results showed that compared with blank control and negative control groups， the percentage of sugar-water preference of the mice in over-expression group was decreased（P<0.05）. The TST and FST results showed that the percentages of immobility time of the mice in over-expression group were increased compared with blank control and negative control groups （P<0.05）. The Western blotting results showed that the expression levels of GRK5， Tau T205， and IBA1 proteins in hippocampus tissue of the mice in over-expression group were increased （P<0.05） and the expression level of synatotagmin（SYN） protein was decreased （P<0.05）.The immunofluorescence staining results showed that compared with blank control group， the fluorescence expression of enhanced green fluoorescence protein（EGFP） in hippocampus tissue of the mice in negative control and over-expression groups could be seen and the EGFP was mainly co-localized with the NeuN； EGFP was hardly found in the microglia and astrocytes；compared with blank control and negative control groups， the number of microglia in hippocampus tissue of the mice in over-expression group was increased（P<0.05）. Conclusion Over-expression of GRK5 in hippocampus tissue of the P301S Tau mice induces the depressive-like behaviors，and its mechanism may be related to the increasing of the expression of GRK5 in nucleus of the neurons.

Key words: Mice， P301S, G protein-coupled receptor kinase 5, Depression-like behavior, Tau protein

中图分类号:

R749.4

章天珍,沈洪涛,龚正,赵斌,王岩. GRK5过表达对P301S Tau转基因小鼠抑郁样行为的影响及其机制[J]. 吉林大学学报(医学版), 2023, 49(3): 573-579.

Tianzhen ZHANG,Hongtao SHEN,Zheng GONG,Bin ZHAO,Yan WANG. Effect of GRK5 over-expression on depression- like behavior in P301S Tau transgenic mice and its mechanism[J]. Journal of Jilin University(Medicine Edition), 2023, 49(3): 573-579.

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Group	n	Sugar-water perference rate	Percentage of immobility time in FST	Percentage of immobility time in TST
Blank control	6	43.40±19.88	13.65±11.77	14.70±13.46
Negative control	5	38.56±13.21	15.46±3.85	12.64±6.07
Over-expression	5	78.65±5.47^*Δ	46.46±16.83^*Δ	44.03±16.17^*Δ

Group	GRK5	Tau T205	IBA1	GFAP	NeuN	PSD95	SYN
Blank control	0.98±0.02	1.08±0.10	1.07±0.13	1.05±0.05	0.85±0.22	1.02±0.03	1.05±0.07
Negative control	1.25±0.30	1.24±0.04	1.29±0.12	1.14±0.08	0.69±0.09	1.12±0.09	1.05±0.08
Over-expression	2.56±0.14^*Δ	1.54±0.11^*Δ	2.03±0.27^*Δ	1.05±0.10	0.57±0.13	1.06±0.10	0.74±0.10^*Δ