γ-氨基丁酸能神经元条件性敲除PGC-1α基因对小鼠焦虑抑郁样行为的影响

发布时间：2017-12-27 05:10

本文关键词：γ-氨基丁酸能神经元条件性敲除PGC-1α基因对小鼠焦虑抑郁样行为的影响　出处：《江苏大学》2017年硕士论文　论文类型：学位论文

【摘要】：【目的】构建条件性敲除γ-氨基丁酸能神经元中过氧化物酶体增殖物激活受体γ辅激活因子-1α(peroxisom proliferator-activated receptor-γcoactivator-1 alpha,PGC-1α)基因的小鼠,并探索条件性敲除γ-氨基丁酸能神经元中过氧化物酶体增殖物激活受体γ辅激活因子-1α对小鼠焦虑抑郁样行为的影响。【方法】从美国Jackson Lab引进SPF级PGC-1αflox/+杂合子小鼠,通过饲养、繁育获得PGC-1αflox/flox纯合子小鼠。将其与特异性在γ-氨基丁酸能神经元表达Cre重组酶的Dlx5/6-Cre-IRES-EGFP小鼠合笼,产生的子代进行基因型鉴定,即可获得特异性在γ-氨基丁酸能神经元中敲除PGC-1α的纯合子小鼠,杂合子小鼠和野生型小鼠。纯合子9只,杂合子16只,野生型10只。分别做高架十字实验,垂直型开场实验,悬尾实验,强迫游泳实验。用Etho Vision软件采集小鼠行为数据,用Stat View软件进行统计分析。【结果】(1)成功构建敲除PGC-1α的纯合子小鼠(同时表达Loxp/loxp Cre),敲除PGC-1α的杂合子小鼠(同时表达Loxp/-Cre)及野生型小鼠(只表达Loxp或者Cre)。(2)青年期(4周)行为学实验结果:在垂直型开场实验中,纯合子小鼠运动总距离与运动速度比杂合子(P0.05)及野生型(P0.01)显著增加,差异有统计学意义;而杂合子与野生型之间无统计学意义。在高架十字迷宫中纯合子小鼠进入开放臂频率百分比和运动总距离比杂合子(P0.05)与野生型(P0.05)都要高,差异有统计学意义;杂合子和野生型之间无统计学意义。悬尾实验中野生型挣扎时间比纯合子(P0.05)和杂合子(P0.05)小鼠要短,差异有统计学意义。而纯合子和杂合子小鼠之间无统计学意义;野生型静止时间比纯合子和杂合子长(P0.05),差异有统计学意义。纯合子和杂合子之间无统计学意义。强迫游泳实验中我们得到了与悬尾实验相同的结果。成年期(8周)行为学实验结果:在垂直型开场实验中,纯合子小鼠运动总距离比野生型(P0.01)显著增加,差异有统计学意义;纯合子与杂合子小鼠之间差异无统计学意义;杂合子与野生型之间无统计学意义。高架十字迷实验中纯合子小鼠进入开放臂频率百分比和运动总距离比杂合子(P0.05)与野生型(P0.05)都要高,差异有统计学意义;杂合子和野生型之间差异无统计学意义。悬尾实验中野生型挣扎时间比纯合子(P0.05)和杂合子(P0.05)小鼠要短,差异有统计学意义。而纯合子和杂合子小鼠之间无统计学意义;野生型静止时间比纯合子和杂合子长(P0.05),差异有统计学意义;纯合子和杂合子之间无统计学意义。强迫游泳实验中我们得到了与悬尾实验相同的结果。4周与8周实验对比结果:垂直型开场实验3组小鼠运动总距离和运动速度对比都无统计学意义。高架十字迷宫实验野生型和杂合子小鼠4周进入开放臂频率百分比比8周减少,差异有统计学意义(P0.01);纯合子小鼠对比无统计学意义。野生型和杂合子小鼠4周运动总距离比8周长有显著统计学意义(P0.01),纯合子小鼠4周运动总距离比8周长有显著统计学意义(P0.05)。悬尾实验和强迫游泳实验结果一样,野生型小鼠4周挣扎时间比8周短,差异有统计学意义(P0.01);野生型小鼠4周静止时间比8周长,差异有统计学意义(P0.01)。而杂合子和纯合子小鼠之间差异无统计学意义。【结论】以上结果说明,条件性敲除γ-氨基丁酸能神经元中PGC-1α基因后小鼠表现出抗焦虑抑郁样行为。野生型小鼠的抗焦虑抑郁能力随年龄增长而变强,但是杂合子与纯合子抗焦虑抑郁样行为无改变。
[Abstract]:[Objective] to construct conditional knockout of GABAergic neurons in peroxisome proliferator activated receptor gamma coactivator -1 alpha (peroxisom proliferator-activated receptor- coactivator-1 alpha PGC-1 gamma, alpha) gene knockout mice, and explore the GABAergic neurons of peroxisome proliferator activated receptor gamma coactivator -1 alpha effect the anxiety and depression like behaviors in mice. [Methods] SPF PGC-1 alpha flox/+ heterozygote mice were introduced from Jackson Lab in the United States, and PGC-1 alpha flox/flox homozygote mice were obtained by feeding and breeding. The specificity of GABAergic Cre recombinase expression neurons of the Dlx5/6-Cre-IRES-EGFP mice in cages, produce offspring for genotype identification, can be obtained in specific GABAergic neurons in the knockout mice homozygous except PGC-1 alpha, heterozygous mice and wild type mice. 9 homozygotes, 16 heterozygotes and 10 wild type. The cross test of viaduct, vertical opening experiment, tail suspension experiment, and forced swimming test were done respectively. The behavior data of mice were collected by Etho Vision software, and the statistical analysis was carried out by Stat View software. [results] (1) we successfully constructed homozygous mice with knockout PGC-1 alpha (simultaneously expressing Loxp/loxp Cre), knockout PGC-1 alpha heterozygous mice (simultaneously expressing Loxp/-Cre) and wild type mice (expressing Loxp or Cre only). (2) the behavioral experiment results in the youth stage (4 weeks): in the vertical opening experiment, the total distance and speed of homozygous mice were significantly higher than those of heterozygote (P0.05) and wild type (P0.01), but the difference was statistically significant, but there was no significant difference between heterozygote and wild type. In the elevated cross maze, the percentage and total distance of homozygous mice entering the open arms were higher than those of heterozygote (P0.05) and wild type (P0.05), the difference was statistically significant, and there was no significant difference between heterozygote and wild type. The struggle time of the wild type in the tail suspension experiment was shorter than that of the homozygote (P0.05) and the heterozygote (P0.05) mice, and the difference was statistically significant. There was no statistically significant difference between homozygote and heterozygote mice, and there was a significant difference between the homozygote and heterozygote length (P0.05) of the wild type. There was no statistical significance between the homozygote and the heterozygote. In the forced swimming experiment, we got the same result as the tail suspension experiment. Adult (8 weeks) the results of behavioral experiments: in the vertical opening experiment, mice homozygous for the motor total distance than the wild type (P0.01) increased significantly, the difference was statistically significant; there was no significant difference between homozygous and heterozygous mice; no statistical significance between heterozygotes and wild type. The percentage and total distance of homozygous mice entering the open arms were higher than those of heterozygote (P0.05) and wild type (P0.05). The struggle time of the wild type in the tail suspension experiment was shorter than that of the homozygote (P0.05) and the heterozygote (P0.05) mice, and the difference was statistically significant. There was no statistical significance between homozygote and heterozygous mice. The resting time of wild type was longer than that of homozygote and heterozygote (P0.05), and the difference was statistically significant. Homozygote and heterozygote were not statistically significant. In the forced swimming experiment, we got the same result as the tail suspension experiment. The results of comparison between 4 weeks and 8 weeks were compared: there was no significant difference between the total distance and the speed of movement of the 3 groups of mice in the vertical open field experiment. In the elevated maze test, the percentage of open arms in wild type and heterozygous mice decreased 4 weeks to 8 weeks, and the difference was statistically significant (P0.01). Homozygous mice had no significant difference. The total exercise distance of wild type and heterozygous mice in 4 weeks was significantly higher than that of 8 Zhou Changyou (P0.01), and the total exercise distance of homozygous mice in 4 weeks was significantly higher than that of 8 Zhou Changyou (P0.05). The results of tail suspension test and forced swimming test were the same. Wild type mice had shorter 4 weeks' struggle time than 8 weeks, and the difference was statistically significant (P0.01). The resting time of 4 weeks in wild type mice was 8 weeks, and the difference was statistically significant (P0.01). There was no significant difference between the heterozygote and the homozygote mice. [Conclusion] the above results suggest that the conditioned knockout of the PGC-1 - alpha gene in the neurons of gamma aminobutyric acid shows anti - anxiety and depressive behavior. The anti anxiety and depression ability of wild type mice increased with age, but the antidepressant behavior of heterozygotes and homozygotes did not change.
【学位授予单位】：江苏大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R749.4

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