AVP及V1a受体对棕色田鼠社会行为及脑区AR表达的影响
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摘要
社会行为是动物行为生态学的基本内容之一，社会行为包括种群内的个体彼此相遇以及相遇以后的一系列相互作用。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。近年来发现，神经肽加压素(AVP)与田鼠的社会组织及社会行为密切相关。AVP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。AVP在影响这些行为的同时，雄激素受体(AR)可能也参与了许多社会行为的调控，本实验将重点研究Vla受体如何影响田鼠的部分社会行为和AR的表达，以探讨Vla受体和AR共同作用下对田鼠社会行为的调控。
由于AVP和雄激素及其受体在雄性动物中的含量较雌性丰富，实验选用雄性棕色田鼠(Microtus mandarinus)。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，通过给雄性棕色田鼠侧脑室中枢连续注射微量的AVP Vla受体mRNA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AVP和雄激素及其受体在雄性棕色田鼠脑内如何共同调节复杂的社会行为。
1．选择成体雄性棕色田鼠侧脑室中枢连续注射微量的AVPVla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低于注射随机序列组，而潜伏期没有显著性的差异。推断AVP VlaR的反义核酸在mRNA水平上阻止其翻译形成VlaR，进而阻碍了AVP调控雄性棕色田鼠的攻击行为。
2．选择成体雄性棕色田鼠侧脑室中枢连续注射微量的AVPVla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：不仅在雄鼠访问时间上有显著差异，还表现在攻击、防御、探究、善待、生殖和非社会行为等方面都发生了显著性的变化。说明给成年雄性棕色田鼠注射AVP VlaR的反义核酸后，降低了雄性棕色田鼠的社会识别能力，以致于减弱了雄性棕色田鼠和配偶鼠之间的配偶联系。
3．选择成体雄性棕色田鼠侧脑室中枢连续注射微量的AVPVla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：不仅表现在雄鼠访问时间上的差异，还表现在防御、探究、善待、和非社会行为等都发生了显著性的变化。说明AVP在田鼠偏好测试中，对同胞鼠和陌生同性鼠的社会识别可能存在着根本不同的神经通路。
4．选择成体雄性棕色田鼠侧脑室中枢连续注射微量的AVPVla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，利用个体社会互作行为学检测平台进行实验，历时30分钟。实验结果显示：在攻击、防御和善待方面发生了显著性变化，而在探究、生殖和非社会行为等方面没有显著性差异。防御行为和善待行为是实验室对刺激鼠的两种相反的表现方式，而在这个行为检测实验中确有相同的变化趋势，提示我们理解AVP VlaR的反义核酸对雄性棕色田鼠作用的时候，要考虑到不同的行为检测模式下，它的作用可能表现不同；同时，也提示我们理解防御行为和善待行为的不同发生机制和外界影响因素。
5．选择成体雄性棕色田鼠侧脑室中枢连续注射微量的AVP Vla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ARmRNA检测。实验结果显示：在棕色田鼠脑区的隔外侧核(LS)、终纹床核(BNST)、下丘脑视前区(AH)、杏仁内侧核(ME)和下丘脑腹内侧核(VMH)ARmRNA的表达有显著性的差异。与ST组相比，AT组雄性棕色田鼠在LS、AH、ME和VMH等四个核团AR的表达均有显著性的差异，明显减少；与CT组相比，AT组雄性棕色田鼠却在这五个核团都有显著性差异，明显减少。说明在这些核团AVP VlaR的反义核酸与AR相互作用影响了雄性田鼠的社会行为，提示我们在理解AVP在介导雄性棕色田鼠的社会识别时，可能是通过雄激素及其受体来发挥作用的。
Neural basis of social behavior has been become one of focus in behavioral neuroscience. Social behaviors conclude a series of social interaction following individual encountering each other in the same population. 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. More recently, neuropeptide AVP have been found to be related closely with social organization in voles and social behaviors. AVP could affect many social behaviors, such as social recognition, aggression, sexual behavior, partner preference, affinity behavior and individual behavior. Besides, AR also participated in modulating animal social behavior. So, the present study will aim at the two problems: 1, how AVP could affect some social behaviors in voles; 2, Is there possibility of effects to AR expression by AVP receptor?
Because the male voles could have abundance of AVP, Androgen and its receptor, we used monogamous male mandarin voles (Microtus mandarinus) as research objects. Combining antisense nucleotides tools with behavioral examination, we examined the correlative behavior after i.c.v, continuous injection of antisense nucleotides of AVP V1a receptor. Meanwhile, Combined antisense nucleotides tools with in situ hybridization, we discussed the problem how AVP, Androgen and its receptor controlled complex social behaviors in male voles.
1. Selected male adult mandarin voles were given i.c.v, continuous injections of antisense nucleotides of AVP V1a receptor respectively 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, anrisense group showed significant reduction in frequency and total duration in aggression behavior. We suggested that antisense nucleotides of AVP V1a receptor inhibit the translation of V1aR in mRNA levels. Furthermore, it blocked AVP to modulate aggression behavior in male voles.
2. Selected male adult mandarin voles were given i.c.v, continuous injections of antisense nucleotides of AVP V1a receptor in 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 not only in visiting duration but also in aggression, defensive behavior, investigating behavior, amicable behavior, reproductive behavior and nonsocial behavior. It is suggested that continuous injection of antisense nucleotides of AVP V1a receptor can suppress social recognition after i.c.v, continuous injection of antisense nucleotides of AVP V1a receptor in male adult voles and reduced the pair bond formation of male and female voles.
3. Selected male adult mandarin voles were given i.c.v, continuous injections of antisense nucleotides of AVP V1a receptor 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 differences not only in visiting duration, but also in defensive behavior, investigating behavior, amicable behavior, and nonsocial behavior. It suggested that there were different nervous circuit, which male voles recognized socially its sibling and stranger voles in the preference behavior examination.
4. Selected male adult mandarin voles were given i.c.v, continuous injections of antisense nucleotides of AVP V1a receptor in first day, third day and fifth day. After six hours, we examined individual behavior during 30 minutes. The results were following: There were significant differences in aggression, defensive behavior and amicable behavior but not in investigating behavior, reproductive behavior and nonsocial behavior. There were the two opposite manners about defensive behavior and amicable behavior, but there were similar changing tendency in the behavior examination. It might show us that there were different roles in consideration of different behavior examinations while antisense nucleotides of AVP V1a receptor affected behaviors of male mandarin voles. What's more, it also suggested us that there were different mechanisms and environment factors on defensive behavior and amicable behavior.
5. Selected male adult mandarin voles were given i.c.v, continuous injections of antisense nucleotides of AVP V1a receptor in first day, third day and fifth day. After six hours, AR mRNA was examnined using in situ hybridization method in male voles. The results were following: There were significant differences in the LS, BNST, AH, ME and VMH. Compared with injection of scramble nucleotides, there were significant reduction in the number of AR mRNA in LS, AH, ME and VMH. Compared with injection of saline, there were significant reductions in the number of AR in these nucleuses. These data showed that the roles between antisense nucleotides of AVP V1a receptor and AR affected social behavior in the male voles. It explained that AVP V1a receptor modulated social behavior in male voles possibly through Androgen and its receptor.
由于AVP和雄激素及其受体在雄性动物中的含量较雌性丰富，实验选用雄性棕色田鼠(Microtus mandarinus)。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，通过给雄性棕色田鼠侧脑室中枢连续注射微量的AVP Vla受体mRNA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AVP和雄激素及其受体在雄性棕色田鼠脑内如何共同调节复杂的社会行为。
1．选择成体雄性棕色田鼠侧脑室中枢连续注射微量的AVPVla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低于注射随机序列组，而潜伏期没有显著性的差异。推断AVP VlaR的反义核酸在mRNA水平上阻止其翻译形成VlaR，进而阻碍了AVP调控雄性棕色田鼠的攻击行为。
2．选择成体雄性棕色田鼠侧脑室中枢连续注射微量的AVPVla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：不仅在雄鼠访问时间上有显著差异，还表现在攻击、防御、探究、善待、生殖和非社会行为等方面都发生了显著性的变化。说明给成年雄性棕色田鼠注射AVP VlaR的反义核酸后，降低了雄性棕色田鼠的社会识别能力，以致于减弱了雄性棕色田鼠和配偶鼠之间的配偶联系。
3．选择成体雄性棕色田鼠侧脑室中枢连续注射微量的AVPVla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：不仅表现在雄鼠访问时间上的差异，还表现在防御、探究、善待、和非社会行为等都发生了显著性的变化。说明AVP在田鼠偏好测试中，对同胞鼠和陌生同性鼠的社会识别可能存在着根本不同的神经通路。
4．选择成体雄性棕色田鼠侧脑室中枢连续注射微量的AVPVla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，利用个体社会互作行为学检测平台进行实验，历时30分钟。实验结果显示：在攻击、防御和善待方面发生了显著性变化，而在探究、生殖和非社会行为等方面没有显著性差异。防御行为和善待行为是实验室对刺激鼠的两种相反的表现方式，而在这个行为检测实验中确有相同的变化趋势，提示我们理解AVP VlaR的反义核酸对雄性棕色田鼠作用的时候，要考虑到不同的行为检测模式下，它的作用可能表现不同；同时，也提示我们理解防御行为和善待行为的不同发生机制和外界影响因素。
5．选择成体雄性棕色田鼠侧脑室中枢连续注射微量的AVP Vla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ARmRNA检测。实验结果显示：在棕色田鼠脑区的隔外侧核(LS)、终纹床核(BNST)、下丘脑视前区(AH)、杏仁内侧核(ME)和下丘脑腹内侧核(VMH)ARmRNA的表达有显著性的差异。与ST组相比，AT组雄性棕色田鼠在LS、AH、ME和VMH等四个核团AR的表达均有显著性的差异，明显减少；与CT组相比，AT组雄性棕色田鼠却在这五个核团都有显著性差异，明显减少。说明在这些核团AVP VlaR的反义核酸与AR相互作用影响了雄性田鼠的社会行为，提示我们在理解AVP在介导雄性棕色田鼠的社会识别时，可能是通过雄激素及其受体来发挥作用的。
Neural basis of social behavior has been become one of focus in behavioral neuroscience. Social behaviors conclude a series of social interaction following individual encountering each other in the same population. 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. More recently, neuropeptide AVP have been found to be related closely with social organization in voles and social behaviors. AVP could affect many social behaviors, such as social recognition, aggression, sexual behavior, partner preference, affinity behavior and individual behavior. Besides, AR also participated in modulating animal social behavior. So, the present study will aim at the two problems: 1, how AVP could affect some social behaviors in voles; 2, Is there possibility of effects to AR expression by AVP receptor?
Because the male voles could have abundance of AVP, Androgen and its receptor, we used monogamous male mandarin voles (Microtus mandarinus) as research objects. Combining antisense nucleotides tools with behavioral examination, we examined the correlative behavior after i.c.v, continuous injection of antisense nucleotides of AVP V1a receptor. Meanwhile, Combined antisense nucleotides tools with in situ hybridization, we discussed the problem how AVP, Androgen and its receptor controlled complex social behaviors in male voles.
1. Selected male adult mandarin voles were given i.c.v, continuous injections of antisense nucleotides of AVP V1a receptor respectively 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, anrisense group showed significant reduction in frequency and total duration in aggression behavior. We suggested that antisense nucleotides of AVP V1a receptor inhibit the translation of V1aR in mRNA levels. Furthermore, it blocked AVP to modulate aggression behavior in male voles.
2. Selected male adult mandarin voles were given i.c.v, continuous injections of antisense nucleotides of AVP V1a receptor in 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 not only in visiting duration but also in aggression, defensive behavior, investigating behavior, amicable behavior, reproductive behavior and nonsocial behavior. It is suggested that continuous injection of antisense nucleotides of AVP V1a receptor can suppress social recognition after i.c.v, continuous injection of antisense nucleotides of AVP V1a receptor in male adult voles and reduced the pair bond formation of male and female voles.
3. Selected male adult mandarin voles were given i.c.v, continuous injections of antisense nucleotides of AVP V1a receptor 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 differences not only in visiting duration, but also in defensive behavior, investigating behavior, amicable behavior, and nonsocial behavior. It suggested that there were different nervous circuit, which male voles recognized socially its sibling and stranger voles in the preference behavior examination.
4. Selected male adult mandarin voles were given i.c.v, continuous injections of antisense nucleotides of AVP V1a receptor in first day, third day and fifth day. After six hours, we examined individual behavior during 30 minutes. The results were following: There were significant differences in aggression, defensive behavior and amicable behavior but not in investigating behavior, reproductive behavior and nonsocial behavior. There were the two opposite manners about defensive behavior and amicable behavior, but there were similar changing tendency in the behavior examination. It might show us that there were different roles in consideration of different behavior examinations while antisense nucleotides of AVP V1a receptor affected behaviors of male mandarin voles. What's more, it also suggested us that there were different mechanisms and environment factors on defensive behavior and amicable behavior.
5. Selected male adult mandarin voles were given i.c.v, continuous injections of antisense nucleotides of AVP V1a receptor in first day, third day and fifth day. After six hours, AR mRNA was examnined using in situ hybridization method in male voles. The results were following: There were significant differences in the LS, BNST, AH, ME and VMH. Compared with injection of scramble nucleotides, there were significant reduction in the number of AR mRNA in LS, AH, ME and VMH. Compared with injection of saline, there were significant reductions in the number of AR in these nucleuses. These data showed that the roles between antisense nucleotides of AVP V1a receptor and AR affected social behavior in the male voles. It explained that AVP V1a receptor modulated social behavior in male voles possibly through Androgen and its receptor.
引文
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