雌激素受体β（ERβ）对雌性棕色田鼠社会行为及其脑区OTR和AVPV1aR表达的影响

【作者】 刘利敏；

【导师】 邰发道；

【作者基本信息】 陕西师范大学 ， 神经生物学， 2008， 硕士

【摘要】 动物的社会行为与神经系统、内分泌系统等密切相关。神经肽、性激素及很多内分泌因子,与动物的生存环境共同影响着动物的社会行为。大量实验证据表明,雌激素(E)、中枢加压素(AVP)和催产素(OT)是调控社会识别的关键物质。它们可以影响动物的社会识别、繁殖行为、性行为、攻击行为、配偶选择行为、亲密行为和个体社会互作行为等等,并且,ERβ可以调控OTmRNA和AVPmRNA的转录。ERβ在社会行为中是否可以调节OTRmRNA和AVPV1aRmRNA还未见报道。雌激素可以调控下丘脑中的OT和AVP基因的表达,这种调控作用在雌激素受体β基因敲除的小鼠中完全不存在,即在此调控中雌激素受体β(ERβ)是否被激活是个关键因素。而AVP在神经系统中发挥作用主要是依赖于V1a受体的存在,OT的作用是通过与其受体结合,经下丘脑—垂体—肾上腺(HPA)轴或自主神经等系统来改变动物的生理和行为。但是关于ERβ在OTR和AVPV1aR调控棕色田鼠(Microtus mandarinus)社会行为中的作用还不清楚。因此,本实验将重点研究ERβ如何影响田鼠的部分社会行为和OTR和AVPVlaR的表达,以探讨ERβ与OTR和AVPV1aR共同作用下对田鼠社会行为的调控。本实验利用反义核酸技术和行为学检测平台相结合进行研究,通过给雌性棕色田鼠侧脑室中枢连续注射微量的雌激素受体βmRNA反义核酸,然后对实验鼠进行相关的行为检测;同时利用反义核酸技术和原位杂交组织化学技术相结合,探讨雌激素及其受体和催产素、加压素及其受体在雌性棕色田鼠脑内如何共同调节复杂的社会行为。1.选择成体雌性棕色田鼠侧脑室中枢连续注射微量的ERβ受体mRNA反义核酸,分别在第一日、第三日及第五日同一时间麻醉注射。六小时后,利用攻击行为学检测平台进行实验,历时10分钟。实验结果显示:注射反义核酸组攻击行为的频次和持续时间明显高于注射随机序列组,而潜伏期没有显著性的差异。推断ERβmRNA的反义核酸在mRNA水平上阻止其翻译形成ERβ,进而阻碍了E及受体调控雌性棕色田鼠的攻击行为。2.选择成体雌性棕色田鼠侧脑室中枢连续注射微量的ERβ受体mRNA反义核酸,分别在第一日、第三日及第五日同一时间麻醉注射。六小时后,利用配偶选择行为学检测平台进行实验,历时60分钟。实验结果显示:雌鼠在对陌生异性鼠箱、配偶鼠箱和中立箱中访问时间上有显著差异。说明给成年雌性棕色田鼠注射ERβmRNA的反义核酸后,使BST、CE、LSD、PVN和ME核团AVPV1aRmRNA和OTRmRNA表达增强,导致与其形成的神经投射功能发挥,从而表现出与之相应的行为活性增强,以致于增强了雌性棕色田鼠和配偶鼠之间的配偶联系。3.选择成体雌性棕色田鼠侧脑室中枢连续注射微量的ERβ受体mRNA反义核酸,分别在第一日、第三日及第五日同一时间麻醉注射。六小时后,利用亲密行为学检测平台进行实验,历时60分钟。实验结果显示:雌鼠对配偶鼠的善待行为的增加,防御行为减弱。对陌生异性鼠的探究行为减弱,防御行为的增强。这些变化说明ERβ在田鼠偏好测试中,对配偶鼠和陌生异性鼠的社会识别可能存在着根本不同的神经通路,推测ERβ反义核酸可加强配偶关系。4.选择成体雌性棕色田鼠侧脑室中枢连续注射微量的ERβmRNA反义核酸,分别在第一日、第三日及第五日同一时间麻醉注射。六小时后,利用原位杂交组织化学技术进行OTmRNA检测。实验结果显示:与对照组相比在棕色田鼠脑区核团中终纹床核(BST)和杏仁中央核(CE)的OTR的表达有显著性增强,在杏仁内侧核(ME)、下丘脑视前区(AH)和下丘脑室旁核(PVN)的OTR的表达有显著性减弱。说明在这些核团ERβ的反义核酸与OTR相互作用影响了雌性田鼠的社会行为,提示我们在理解E在介导雌性棕色田鼠的社会识别时,可能是通过催产素及其受体来发挥作用的。5.选择成体雌性棕色田鼠侧脑室中枢连续注射微量的ERβ受体mRNA反义核酸,分别在第一日、第三日及第五日同一时间麻醉注射。六小时后,利用原位杂交组织化学技术进行AVPV1RmRNA检测。实验结果显示:与对照组相比在棕色田鼠脑区核团中终纹床核(BST)、下丘脑室旁核(PVN)和隔外侧核背部(LSD)核团中AVPV1aR的表达有显著性增加,在杏仁中央核(CE)的AVPV1aR表达显著性减弱,由此推测在这些核团ERβ的反义核酸与AVPR相互作用影响了雌性田鼠的社会行为。由以上结论可得出:ERβ、OTR和AVPV1aR与雌性棕色田鼠的社会行为密切相关。ERβ可在一定程度上降低其攻击行为。ERβ对雌性棕色田鼠的配偶关系的调节及社会识别活动的调控是与OTR和AVPV1aR相关联的,ERβ反义核酸可提高雌性棕色田鼠的配偶识别能力,加强配偶之间的联系,即任何一种行为都受到复杂的神经内分泌机制的调控。更多还原

【Abstract】 Animal social behavior,was controlled and regulated by centre nervous system and neuroendocrine system.Neuropeptide,sex hormone,a number of endocrine factors and environments where animal lives cooperate to affect social behavior of animals.Many experimental evidences indicate that E,OT and AVP acting in the brain appear to be critical for social discrimination.They could affect many social behaviors,such as social recognition,aggression, sexual behavior,partner preference,affinity behavior and individual behavior.Whether ERβcan regulate the OTRmRNA and AVPV1aRmRNA has not been reported.We preit that estrogen may show much regulation on the levels of OT and AVP messenger RNA.So,the present study will aim at the two problems:1,how E could affect some social behaviors in voles;2,Is there effects to OTR and AVPV1a expression by E receptor?we used monogamous female mandarin voles(Microtus mandarinus) as research objects. Combining antisense nucleotides technic with behavioral observation,we examined the correlative behavior after i.c.v,continuous injection of antisense nucleotides of estrogen receptor-β.Meanwhile, Combined antisense nucleotides technic with in situ hybridization,we investigated how ERβ, vasopressin,oxytocin and its receptors controlled complex social behaviors in female voles.1.Selected female adult mandarin voles were given i.c.v,continuous injections of antisense nucleotides of estrogen receptor-βrespectively in first day,third day and fifth day.After six hours, we examined aggression behavior during 10 minutes.The results were following:Compared with group of injection of scramble nucleotides,anisense group showed significant increase in frequency and total duration in aggression behavior.We suggested that antisense nucleotides of ERβinhibit the translation of ERβin mRNA levels.Furthermore,it blocked E to modulate aggression behavior in male voles.2.Selected female adult mandarin voles were given i.c.v,continuous injections of antisense nucleotides of estrogen receptor-βin first day,third day and fifth day.After six hours,we examined partner preference behavior during 60 minutes.The results were following:There were significant differences in visiting duration in mate’s compartment,strange heterosexual vole’s compartment and neutral compartment,and significant increase in the number of AVPV1RmRNA and OTRmRNA in BST、CE、LSD、PVN and ME,It is suggested that continuous injection of antisense nucleotides of ERβcan enhance social recognition,and can enhance the pair bond formation of male and female voles. 3.Selected female adult mandarin voles were given i.c.v,continuous injections of antisense nucleotides of estrogen receptor-βrespectively in first day,third day and fifth day.After six hours, we examined affinity behavior during 60 minutes.The results were following:There were significant enhance in amicable behavior and decrease in defensive behavior with its mate.At the same time,there were significant enhance in defensive behavior and decrease in investigating behavior with the strange heterosexual voles.It suggested that there were different nervous circuit, which female voles recognized socially its mate and stranger voles in the preference behavior examination.4.Selected female adult mandarin voles were given i.c.v,continuous injections of antisense nucleotides of estrogen receptor-βin first day,third day and fifth day.After six hours,OTR mRNA was examnined using in situ hybridization method in female voles.The results were following: There were significant differences in the BST,CE,ME,PVN and AH.Compared with injection of scramble nucleotides and injection of isotonic saline treatment,there were significant reduction in the number of OTR mRNA in AH,PVN and ME,and significant increase in the number of OTR mRNA in BST and CE Compared with injection of saline.These data suggested that the roles between antisense nucleotides of ERβand OTR affected social behavior in the female voles.It explained that ERβmodulated social behavior in female voles possibly through oxytocin and its receptor.5.Selected female adult mandarin voles were given i.c.v,continuous injections of antisense nucleotides of estrogen receptor-βin first day,third day and fifth day.After six hours, AVPV1RmRNA was examnined using in situ hybridization method in female voles.The results were following:There were significant differences in the BST,CE,PVN and LSD.Compared with injection of scramble nucleotides and injection of isotonic saline treatment,there were significant increase in the number of AVPV1RmRNA in BST、PVN and LSD,and significant reduction in the number of AVPV1R mRNA in CE.These data suggested that the roles between antisense nucleotides of ERβand AVPV1R affected social behavior in the female voles.Results from those conclusion that ERβ、OTR and AVPV1aR have nearly relation with female adult mandarin voles.ERβcan reduce the aggression behavior at a certain extent.The antisense nucleotides of estrogen receptorβcan enhance the preference of mandarin voles for mate or strange heterosexual vole,and strengthen the relation with mate in female adult mandarin voles.So,all behaviors were regulated by the nerve internal secretion mechanism.更多还原