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棕色田鼠(Microtus mandarinus)和沼泽田鼠(M.fostis)性激素及其受体和Fos在脑内的表达与社会行为的关系

Studies on Relationship between Estrogen, Androgen, and Their Receptor Distribution, FOS Expression and Social Behaviors of the Mandarin Voles (Microtus Mandarinus) and the Reed Voles (M.fostis)

【作者】 何凤琴

【导师】 张育辉; 邰发道;

【作者基本信息】 陕西师范大学 , 动物学, 2004, 硕士

【摘要】 鼠类的社会行为主要受神经内分泌的调节,其中性激素起重要的作用,动物的生存环境、婚配制度和遗传等均为重要的影响因素。因而,研究鼠类的社会行为和激素的关系,不仅能探讨社会行为发生的神经内分泌机制,而且通过研究不同物种不同社会行为发生中激素的调节作用,还能揭示婚配制度及相关社会行为的内在机制、进化机制和适应功能。 田鼠属动物虽然亲缘关系很近,但它们的社会组织和婚配制度却有明显差异。本研究以单配制的棕色田鼠和繁殖特征呈现多配制的沼泽田鼠为研究对象,探讨雄激素、雌激素、雄激素受体、雌激素受体以及Fos蛋白表达和社会行为发生的关系,研究两种田鼠通过两个嗅觉系统发生的社会识别、记忆、交配、攻击行为的神经生物学机制,以探讨两种田鼠的进化机制和适应功能。研究分以下5部分内容: 1.应用行为聚焦取样观察和免疫组织化学相结合的方法,比较了棕色田鼠(Microtus mandarinus)(n=15)和沼泽田鼠(M.fostis)(n=15)在同种雄雄交往中的行为差异,及在雄雄交往前后雌激素β受体(ERβ)和雄激素受体(AR)表达的差异。在2h的雄雄交往中,前1h棕色田鼠对同性入侵者有较多的攻击和防御行为,后1h攻击行为较少,沼泽田鼠前后1h差异不大(p>0.05),整个2h期间,棕色田鼠较沼泽田鼠对同性入侵者有较多的攻击、防御行为,较少的非社会行为。免疫组织化学检测,没有社会交往时棕色田鼠主嗅球系统投射区和犁鼻系统投射区ERβ免疫阳性细胞(ERβ-IRs)明显少于沼泽田鼠,且显色淡,AR免疫阳性细胞(AR-IRs)在两种鼠间差异不大,且都明显少于各自的ERβ-IRs。2h交往后,棕色田鼠主嗅球投射区和犁鼻系统投射区的ERβ-IRs细胞数明显少于交往前,AR-IRs细胞数明显多于交往前;沼泽田鼠交往前与交往后ERβ-IRs和AR-IRs细胞数均无显著差异,且显著多于交往后棕色田鼠ERβ-IRs细胞数,显著少于交往后棕色田鼠AR-IRs细胞数。以上结果表明:两种田鼠在社会交往中社会行为不同;ERβ的减少和AR的增多可能在社会识别及攻击行为中均起一定的作用,可能也是引起两种田鼠社会行为发生差异的原因之一。 2.通过对阉割7、14、30天成年雄性棕色田鼠攻击行为的行为观察,发现在阉割7和14天时,攻击行为没有降低,阉割30时,攻击行为明显降低。阉割7天时,血清中的肇酮(T)和雌二醇(EZ)明显升高,脑区的T免疫阳性细胞 (T-IRs)、AR-IRs、E:免疫阳性细胞(EZ一IRs)、ERp一IRs比假手术组显著升高,血清中高浓度的辜酮和雌二醇以及脑区AR和ERp细胞数目的变化,可能是这一时期攻击行为没有降低的主要原因;阉割14天时,血清中的翠酮明显降低,雌二醇仍高于假手术组,T-IRs、EZ一IRs显著降低,没有检测到AR一IRs、ERp一IRs,ERp没有对攻击行为产生抑制作用,可能是这一时期攻击行为没有降低的主要原因;阉割30天时,血清中没有检测到T,E:也明显降低,T-IRs、AR一IRs、EZ一IRs、ERp一IRs均显著升高,血清中的T和E:明显降低,可能是这一时期攻击行为降低的主要原因。以上结果表明:(l)长期阉割抑制成年棕色田鼠攻击行为;(2)阉割不同时期,攻击行为发生的神经生物学机制可能不同,血清中T、EZ,脑区的T、EZ、AR和ERp可能均起一定的作用。 3.将辜丸下降的成年雄性棕色田鼠分成三组:(l)对照组:嗅闻24h新鲜锯木。(2)暴露组:嗅闻24h动情期雌鼠底物。(3)交配组:与动情期雌鼠交配24ho放射免疫检测血清中的T浓度,交配组高于暴露组,交配组和暴露组都显著高于对照组。用免疫组化检测犁鼻系统部分投射区:终纹床核(B ST)、下丘脑腹内侧核WM均、内侧视前区(Mpo)、隔外侧核(LS)、杏仁内侧核(ME)下IRs、AR一IRs、Fos免疫阳性细胞(Fos一IRs),T-IRS、AR一IRs、Fos一IRS在这5个脑区内,交配组显著高于对照组,交配组在内侧一视前区和杏仁内侧核显著高于暴露组,两组在终纹床核、下丘脑腹内侧核、隔外侧核没有显著差异;暴露组在终纹床核、下丘脑腹内侧核、隔外侧核显著高于对照组,内侧视前区和杏仁内侧核两组没有显著差异。表明:(l)内源性雄激素对其受体有上调作用。(2)脑内的雄激素通过其受体的调节作用和血清中较高浓度雄激素可能是雄性棕色田鼠完成交配活动的两个必要条件。(3)不同的脑区在交配活动中可能起不同的作用。 4.放射免疫检测血清中的EZ浓度,交配组比暴露组、对照组显著增高,暴露组和对照组没有显著差异。通过免疫组化检测与性行为有关的脑区:弓状核(ARc)、终纹床核、下丘脑腹内侧核、内侧视前区、隔外侧核、杏仁内侧核EZ、ERp免疫阳性细胞,EZ一IRs在交配组比对照组和暴露组各区域都显著增多,暴露组比对照组在隔外侧核显著增多外,其它区域没有显著差异。ERp一IRs在这三组之间均没有显著差异,而且ERp免疫阳性细胞颜色浅淡。表明:雌激素对雄性棕色田鼠的交配活动起一定的作用,但可能通过其他受体,ERp在雄性棕色田鼠的交配活动中可能没有起重要作用。 5.用免疫组织化学检测T、AR、Fos在对照组、暴露组、交配组嗅球(MOB)乡和副嗅球(AOB)僧帽细胞和颗粒细胞免疫反应,在MOB三组之间没有显著差异,交配组在AOB僧帽细胞和颗?

【Abstract】 Social behaviors of rodent are not only related with existence environment, mating system, heredity et al., but also are regulated by neuroendocrine, particularly by androgen and estrogen. Therefore, studies the relation between hormone and social behaviors of different species with different mating system, can not only explore the neuroendocrinical mechanism of social behaviors, but also can reveal the intrinsic mechanism, evolution mechanism and adaptation function.The microtine rodents show profound differences in reproductive biology and social organization in spite of their taxonomic relationship. In order to explore the relation between androgen, estrogen, androgen receptor, estrogen receptor, expression of Fos protein and social behaviors, using monogamous mandarin voles (Microtus mandarinus) and reed voles (Microtus fortis) with promiscuous reproductive characteristics as two study objects, the neurobiological basis of social organization, memory, mating, aggressive behavior of two species are studied through two olfactory systems, and explore evolution mechanism and adaptation function of two species voles. The study is divided in five parts.1. In order to investigate the neurobiological mechanism of the social behavior difference in the adult male mandarin voles (Microtus mandarinus) and reed voles(M.fostis), the behavior focal sampling method and immunohistochemistry method were used to investigate these differences in the behaviors of intraspecies male-male interactions and the expressions of estrogen receptor P (ER0) and androgen receptor (AR) between two species, respectively (n=15). During the two hour interaction of male-male individuals, the mandarin voles spent significantly more time on the aggressive and defensive behavior to homosexual invaders in the first one hour than in the last hour, while the reed voles didn’t have significantly behavioral difference between the first and the last one hour (p>0.05); During the whole two-hour interaction of male-male individuals, the mandarin voles spent significantly more time on theaggressive and defensive behaviors to homosexual invaders and less time on the nonsocial behaviors than the reed voles. Before the two-hour interaction of male-male individuals , we found there were obviously fewer and lighter stained ERp immunoreactive neurons (ER|3-IRs) in both main olfactory bulb system project regions and vomeronasal system project regions in the mandarin voles than in the reed voles, but no significant difference in numbers of AR immunoreactive neurons (AR-IRs) between the two species through the examining of immunoreactive neurons, moreover, there were significantly fewer AR-IRs than ERp-IRs in two species of vole before the two-hour interactions. There were significantly fewer ER-IRs in both the main olfactory bulb project regions and the vomeronasal system project regions in the mandarin voles after interactions than before interactions, but there were significantly more AR-IRs after interactions than before interactions in the mandarin voles; There were not significant difference in both numbers of ER|3-IRs and AR-IRs between before and after interactions in the reed voles, and there were greatly significantly more ERp-IRs neurons in the reed voles than in the mandarin voles which had gone through interactions, there were significantly fewer AR-IRs in the reed voles than in the mandarin voles which had gone through interactions. The results indicate that mandarin voles and reed voles had different social behaviors in social interactions. The result suggest that the decrease of ERp-IRs and increase of AR-IRs might affect social recognition and aggressive behavior, and this might also be one of reasons of social behaviors difference in two vole species.2. Aggressive behavior of the adult male mandarin was conducted on 7,14 and 30 days after castration, respectively, the aggressive behavior was not decreased on 7 and 14 days after castration, however, the aggressive behavior was significantly decreased on 30 days after castration. 7 days after cast

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