中枢注射神经肽S引起的活动神经元分布-c-fos免疫组织化学证据
发布时间：2018-09-04 05:47
【摘要】：研究背景及目的：新鉴定的神经肽S(neuropeptide S, NPS)前体mRNA主要表达于蓝斑核与Barrington核之间的无名核、三叉神经感觉主核及臂旁外侧核,而NPSR mRNA广范分布于多个脑区,如：大脑皮质、杏仁核复合体、海马下托、丘脑、下丘脑。NPS与NPS受体(neuropeptide S receptor, NPSR)高选择性结合引起细胞Ca2+内流及胞内cAMP水平增高。近期研究证实NPS参与睡眠与觉醒时相变化、运动、焦虑、学习记忆、摄食与能量代谢的调节,还与成瘾、镇痛、神经内分泌等多种生理与病理过程有关。上述研究提示NPS可能经其受体介导参与生理调节和病理失调过程。本研究运用神经系统功能活动形态定位法确定NPS在大鼠脑的作用靶点,利用即刻早期基因(immediate-early gene, IEGS) c-fos作为神经功能活动的标识物,揭示NPS中枢注射后活动神经元的分布,为阐明NPS参与生理与病理调节过程提供形态定位。 方法：健康成年SD雄性大鼠(250-320g,n=12)随机分为两组：生理盐水组(n=6),NPS组(n=6)。经注射导管(直径0.6mm,长度28mm；AP：-0.9mm；ML+1.5mm；DV-3.3mm)10:00h分别注射生理盐水2.5μ1或等容量NPS1nmol,1.5h后,水合氯醛(420mg/kg)麻醉下经升主动脉灌注0.9%NaCl200m1,4%多聚甲醛磷酸盐缓冲液300m1(0.1mol,pH7.4,4℃),脑分离后35%蔗糖磷酸盐缓冲液4-C48h。恒冷箱-20-C行全脑冠状切35μm。行浮片c-fos免疫组织化学,光镜下观察Fos免疫反应(immunoreactivity-IR)(?)中经元在全脑的分布,计数并统计分析两组间Fos-IR神经元数量在各核的变化。 结果： 1.端脑Fos-IR神经元分布 与生理盐水组比较,NPS促Fos-IR神经元数量增加分别为端脑的梨形皮质399%(P0.0001)；皮层第一感觉区200%、第二感觉区198%、第一运动区197%、第二运动区198% (P0.0001)；杏仁核皮质部296%、基底部214%、外侧区305%、中央区413% (P0.0001)、终纹床核110%(P0.05)；海马CA1区203%、CA2区203%、CA3562%区、海马下托268%(P0.0001)。 2.间脑Fos-IR神经元分布 与生理盐水组比较,NPS促Fos-IR神经元数量增加分别为间脑的下丘脑视交叉上核322%、室旁核108%、背内侧核274%、腹内侧核126%、弓状核267%、穹窿周核520%、结节乳头体腹侧核641%和背侧核586%、外侧区378%(P0.0001)。 3.脑干Fos-IR神经元分布 与生理盐水组比较,NPS促Fos-IR神经元数量增加分别为脑干的上丘灰质层203%、下丘灰质层479%、导水管周围灰质263%、蓝斑202%、中缝背核210%、嘴侧中缝线核157%(P0.0001)。 结论： 1、NPS激活下丘脑组胺能神经元结节乳头体核、orexin能神经元穹窿周核和下丘脑外侧区、视交叉上核、脑干去甲肾上腺素能神经元蓝斑核、5-羟色胺能神经元中缝背核和嘴侧中缝线核参与睡眠觉醒周期的调节。 2、NPS激活杏仁核、下丘脑室旁核、脑干蓝斑核、中缝背核、嘴侧中缝线核神经元参与情绪的调节。 3、NPS激活梨形皮质、杏仁体神经元参与嗅觉调节。 4、NPS激活海马、杏仁体神经元参与学习记忆的调节。 5、NPS激活下丘脑弓状核、背内侧核、腹内侧核、室旁核和下丘脑外侧区神经元参与摄食调节。 6、NPS激活下丘脑orexin能神经元、室旁核神经元参与药物成瘾及依赖调节。 7、NPS激活上丘、下丘神经元参与听觉、视觉调节。 8、NPS激活导水管周围灰质、弓状核参与痛觉调节。 9、NPS激活弓状核参与神经内分泌及植物神经调节。
[Abstract]:BACKGROUND AND OBJECTIVE: The newly identified precursor mRNA of neuropeptide S (NPS) is mainly expressed in the innominate nucleus between locus coeruleus and Barrington nucleus, the main trigeminal sensory nucleus and the lateral parabrachial nucleus, while NPSR mRNA is widely distributed in many brain regions, such as: cerebral cortex, amygdala complex, hippocampal hypothalamus, thalamus, hypothalamus. High selective binding of neuropeptide S receptor (NPSR) results in increased intracellular Ca2+ influx and intracellular cAMP levels. Recent studies have confirmed that NPS is involved in the regulation of sleep and wakefulness phase change, movement, anxiety, learning and memory, food intake and energy metabolism, and is also related to addiction, analgesia, neuroendocrine and other physiological and pathological processes. It is suggested that NPS may be involved in physiological regulation and pathological disorders mediated by its receptors. In this study, the target of action of NPS in rat brain was determined by means of functional morphological localization of nervous system. Immediate-early gene (IEGS) c-fos was used as a marker of neurological activity to reveal the active neurons after central injection of NPS. The distribution provides a morphological location for elucidating the involvement of NPS in physiological and pathological regulation.
METHODS: Healthy adult male SD rats (250-320 g, n=12) were randomly divided into two groups: normal saline group (n=6) and NPS group (n=6). The ascending aorta was perfused with chloral hydrate (420 mg/kg) anesthesia at 10:00 h after injection of normal saline (2.5 u 1) or constant volume NPS 1 nmol (1.5 h). 1% NaCl200m1,4% paraformaldehyde phosphate buffer 300m1 (0.1 mol, pH 7.4,4 C), 35% sucrose phosphate buffer 4-C48h after brain isolation. The whole brain was coronal sectioned at 35 micron by constant cooling chamber-20-C. Immunohistochemistry of c-fos was performed. The distribution of the meridians in the whole brain of Fos immunoreactivity-IR was observed under light microscope, and the Fos immunoreactivity-IR was counted and statistically analyzed between the two groups. The number of s-IR neurons in different nuclei.
Result:
Distribution of Fos-IR neurons in 1. terminals
Compared with the normal saline group, the number of Fos-IR neurons increased by 399% in the pyriform cortex (P 0.0001), 200% in the first sensory cortex, 198% in the second sensory cortex, 197% in the first motor cortex, 198% in the second motor cortex, 296% in the amygdala cortex, 214% in the basal cortex, 305% in the lateral cortex, 413% in the central cortex (P 0.0001), 110% in the bed nucleus terminalis (P 0.05); hippocampal CA1 area 203%, CA2 area 203%, CA3562% area, lower hippocampal 268% (P0.0001).
2. the distribution of Fos-IR neurons in the diencephalon
Compared with normal saline group, the number of Fos-IR neurons in mesencephalic hypothalamic suprachiasmatic nucleus, paraventricular nucleus, dorsomedial nucleus, ventromedial nucleus, arcuate nucleus, arcuate nucleus, perifornical nucleus, ventral papillary nucleus, dorsal nucleus and lateral area increased by 32.2%, 108%, 274%, 126%, 267%, 520%, 641% and 586%, respectively (P 0.0001).
Distribution of Fos-IR neurons in the 3. brainstem
Compared with the normal saline group, the number of Fos-IR neurons stimulated by NPS increased by 203% in the superior colliculus, 479% in the inferior colliculus, 263% in the periaqueductal gray, 202% in the locus coeruleus, 210% in the dorsal raphe nucleus and 157% in the rostral raphe nucleus respectively (P 0.0001).
Conclusion:
1. NPS activates the nucleus papillaris, perifornical nucleus of orexinergic neurons, lateral hypothalamic area, suprachiasmatic nucleus, locus coeruleus of noradrenergic neurons in the brain stem, dorsal raphe nucleus of 5-HT neurons and raphe nucleus in the mouth to regulate the sleep wake cycle.
2. NPS activates amygdala, paraventricular nucleus of hypothalamus, locus coeruleus, dorsal raphe nucleus and raphe nucleus of the mouth to regulate emotion.
3, NPS activated piriform cortex and amygdala neurons involved in olfactory regulation.
4, NPS activates the hippocampus and amygdala neurons in the regulation of learning and memory.
5. NPS activates neurons in hypothalamic arcuate nucleus, dorsomedial nucleus, ventromedial nucleus, paraventricular nucleus and lateral hypothalamic area.
6, NPS activates hypothalamic orexin neurons and paraventricular neurons participate in drug addiction and dependence regulation.
7, NPS activates the superior colliculus, and the inferior colliculus neurons participate in auditory and visual regulation.
8, NPS activates periaqueductal gray and arcuate nucleus participates in pain regulation.
9, NPS activates arcuate nucleus to participate in neuroendocrine and autonomic regulation.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R392.1
本文编号：2221192
[Abstract]:BACKGROUND AND OBJECTIVE: The newly identified precursor mRNA of neuropeptide S (NPS) is mainly expressed in the innominate nucleus between locus coeruleus and Barrington nucleus, the main trigeminal sensory nucleus and the lateral parabrachial nucleus, while NPSR mRNA is widely distributed in many brain regions, such as: cerebral cortex, amygdala complex, hippocampal hypothalamus, thalamus, hypothalamus. High selective binding of neuropeptide S receptor (NPSR) results in increased intracellular Ca2+ influx and intracellular cAMP levels. Recent studies have confirmed that NPS is involved in the regulation of sleep and wakefulness phase change, movement, anxiety, learning and memory, food intake and energy metabolism, and is also related to addiction, analgesia, neuroendocrine and other physiological and pathological processes. It is suggested that NPS may be involved in physiological regulation and pathological disorders mediated by its receptors. In this study, the target of action of NPS in rat brain was determined by means of functional morphological localization of nervous system. Immediate-early gene (IEGS) c-fos was used as a marker of neurological activity to reveal the active neurons after central injection of NPS. The distribution provides a morphological location for elucidating the involvement of NPS in physiological and pathological regulation.
METHODS: Healthy adult male SD rats (250-320 g, n=12) were randomly divided into two groups: normal saline group (n=6) and NPS group (n=6). The ascending aorta was perfused with chloral hydrate (420 mg/kg) anesthesia at 10:00 h after injection of normal saline (2.5 u 1) or constant volume NPS 1 nmol (1.5 h). 1% NaCl200m1,4% paraformaldehyde phosphate buffer 300m1 (0.1 mol, pH 7.4,4 C), 35% sucrose phosphate buffer 4-C48h after brain isolation. The whole brain was coronal sectioned at 35 micron by constant cooling chamber-20-C. Immunohistochemistry of c-fos was performed. The distribution of the meridians in the whole brain of Fos immunoreactivity-IR was observed under light microscope, and the Fos immunoreactivity-IR was counted and statistically analyzed between the two groups. The number of s-IR neurons in different nuclei.
Result:
Distribution of Fos-IR neurons in 1. terminals
Compared with the normal saline group, the number of Fos-IR neurons increased by 399% in the pyriform cortex (P 0.0001), 200% in the first sensory cortex, 198% in the second sensory cortex, 197% in the first motor cortex, 198% in the second motor cortex, 296% in the amygdala cortex, 214% in the basal cortex, 305% in the lateral cortex, 413% in the central cortex (P 0.0001), 110% in the bed nucleus terminalis (P 0.05); hippocampal CA1 area 203%, CA2 area 203%, CA3562% area, lower hippocampal 268% (P0.0001).
2. the distribution of Fos-IR neurons in the diencephalon
Compared with normal saline group, the number of Fos-IR neurons in mesencephalic hypothalamic suprachiasmatic nucleus, paraventricular nucleus, dorsomedial nucleus, ventromedial nucleus, arcuate nucleus, arcuate nucleus, perifornical nucleus, ventral papillary nucleus, dorsal nucleus and lateral area increased by 32.2%, 108%, 274%, 126%, 267%, 520%, 641% and 586%, respectively (P 0.0001).
Distribution of Fos-IR neurons in the 3. brainstem
Compared with the normal saline group, the number of Fos-IR neurons stimulated by NPS increased by 203% in the superior colliculus, 479% in the inferior colliculus, 263% in the periaqueductal gray, 202% in the locus coeruleus, 210% in the dorsal raphe nucleus and 157% in the rostral raphe nucleus respectively (P 0.0001).
Conclusion:
1. NPS activates the nucleus papillaris, perifornical nucleus of orexinergic neurons, lateral hypothalamic area, suprachiasmatic nucleus, locus coeruleus of noradrenergic neurons in the brain stem, dorsal raphe nucleus of 5-HT neurons and raphe nucleus in the mouth to regulate the sleep wake cycle.
2. NPS activates amygdala, paraventricular nucleus of hypothalamus, locus coeruleus, dorsal raphe nucleus and raphe nucleus of the mouth to regulate emotion.
3, NPS activated piriform cortex and amygdala neurons involved in olfactory regulation.
4, NPS activates the hippocampus and amygdala neurons in the regulation of learning and memory.
5. NPS activates neurons in hypothalamic arcuate nucleus, dorsomedial nucleus, ventromedial nucleus, paraventricular nucleus and lateral hypothalamic area.
6, NPS activates hypothalamic orexin neurons and paraventricular neurons participate in drug addiction and dependence regulation.
7, NPS activates the superior colliculus, and the inferior colliculus neurons participate in auditory and visual regulation.
8, NPS activates periaqueductal gray and arcuate nucleus participates in pain regulation.
9, NPS activates arcuate nucleus to participate in neuroendocrine and autonomic regulation.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R392.1
【共引文献】
相关期刊论文 前2条
1 李昀;肖云虹;张磊;;促肾上腺皮质激素释放因子及其受体的研究[J];山东医药;2009年48期
2 罗涛;郝伟;;反奖赏系统与成瘾行为[J];中国药物依赖性杂志;2010年04期
相关博士学位论文 前1条
1 韩仁文;神经肽S对记忆、结肠运动和摄食的调节作用[D];兰州大学;2009年
，本文编号：2221192
本文链接：https://www.wllwen.com/xiyixuelunwen/2221192.html
最近更新
教材专著
