【摘要】：研究背景及目的：新鑒定的神經(jīng)肽S(neuropeptide S, NPS)前體mRNA主要表達(dá)于藍(lán)斑核與Barrington核之間的無名核、三叉神經(jīng)感覺主核及臂旁外側(cè)核,而NPSR mRNA廣范分布于多個(gè)腦區(qū),如：大腦皮質(zhì)、杏仁核復(fù)合體、海馬下托、丘腦、下丘腦。NPS與NPS受體(neuropeptide S receptor, NPSR)高選擇性結(jié)合引起細(xì)胞Ca2+內(nèi)流及胞內(nèi)cAMP水平增高。近期研究證實(shí)NPS參與睡眠與覺醒時(shí)相變化、運(yùn)動(dòng)、焦慮、學(xué)習(xí)記憶、攝食與能量代謝的調(diào)節(jié),還與成癮、鎮(zhèn)痛、神經(jīng)內(nèi)分泌等多種生理與病理過程有關(guān)。上述研究提示NPS可能經(jīng)其受體介導(dǎo)參與生理調(diào)節(jié)和病理失調(diào)過程。本研究運(yùn)用神經(jīng)系統(tǒng)功能活動(dòng)形態(tài)定位法確定NPS在大鼠腦的作用靶點(diǎn),利用即刻早期基因(immediate-early gene, IEGS) c-fos作為神經(jīng)功能活動(dòng)的標(biāo)識(shí)物,揭示NPS中樞注射后活動(dòng)神經(jīng)元的分布,為闡明NPS參與生理與病理調(diào)節(jié)過程提供形態(tài)定位。 方法：健康成年SD雄性大鼠(250-320g,n=12)隨機(jī)分為兩組：生理鹽水組(n=6),NPS組(n=6)。經(jīng)注射導(dǎo)管(直徑0.6mm,長(zhǎng)度28mm；AP：-0.9mm；ML+1.5mm；DV-3.3mm)10:00h分別注射生理鹽水2.5μ1或等容量NPS1nmol,1.5h后,水合氯醛(420mg/kg)麻醉下經(jīng)升主動(dòng)脈灌注0.9%NaCl200m1,4%多聚甲醛磷酸鹽緩沖液300m1(0.1mol,pH7.4,4℃),腦分離后35%蔗糖磷酸鹽緩沖液4-C48h。恒冷箱-20-C行全腦冠狀切35μm。行浮片c-fos免疫組織化學(xué),光鏡下觀察Fos免疫反應(yīng)(immunoreactivity-IR)(?)中經(jīng)元在全腦的分布,計(jì)數(shù)并統(tǒng)計(jì)分析兩組間Fos-IR神經(jīng)元數(shù)量在各核的變化。 結(jié)果： 1.端腦Fos-IR神經(jīng)元分布 與生理鹽水組比較,NPS促Fos-IR神經(jīng)元數(shù)量增加分別為端腦的梨形皮質(zhì)399%(P0.0001)；皮層第一感覺區(qū)200%、第二感覺區(qū)198%、第一運(yùn)動(dòng)區(qū)197%、第二運(yùn)動(dòng)區(qū)198%　(P0.0001)；杏仁核皮質(zhì)部296%、基底部214%、外側(cè)區(qū)305%、中央?yún)^(qū)413%　(P0.0001)、終紋床核110%(P0.05)；海馬CA1區(qū)203%、CA2區(qū)203%、CA3562%區(qū)、海馬下托268%(P0.0001)。 2.間腦Fos-IR神經(jīng)元分布 與生理鹽水組比較,NPS促Fos-IR神經(jīng)元數(shù)量增加分別為間腦的下丘腦視交叉上核322%、室旁核108%、背內(nèi)側(cè)核274%、腹內(nèi)側(cè)核126%、弓狀核267%、穹窿周核520%、結(jié)節(jié)乳頭體腹側(cè)核641%和背側(cè)核586%、外側(cè)區(qū)378%(P0.0001)。 3.腦干Fos-IR神經(jīng)元分布 與生理鹽水組比較,NPS促Fos-IR神經(jīng)元數(shù)量增加分別為腦干的上丘灰質(zhì)層203%、下丘灰質(zhì)層479%、導(dǎo)水管周圍灰質(zhì)263%、藍(lán)斑202%、中縫背核210%、嘴側(cè)中縫線核157%(P0.0001)。 結(jié)論： 1、NPS激活下丘腦組胺能神經(jīng)元結(jié)節(jié)乳頭體核、orexin能神經(jīng)元穹窿周核和下丘腦外側(cè)區(qū)、視交叉上核、腦干去甲腎上腺素能神經(jīng)元藍(lán)斑核、5-羥色胺能神經(jīng)元中縫背核和嘴側(cè)中縫線核參與睡眠覺醒周期的調(diào)節(jié)。 2、NPS激活杏仁核、下丘腦室旁核、腦干藍(lán)斑核、中縫背核、嘴側(cè)中縫線核神經(jīng)元參與情緒的調(diào)節(jié)。 3、NPS激活梨形皮質(zhì)、杏仁體神經(jīng)元參與嗅覺調(diào)節(jié)。 4、NPS激活海馬、杏仁體神經(jīng)元參與學(xué)習(xí)記憶的調(diào)節(jié)。 5、NPS激活下丘腦弓狀核、背內(nèi)側(cè)核、腹內(nèi)側(cè)核、室旁核和下丘腦外側(cè)區(qū)神經(jīng)元參與攝食調(diào)節(jié)。 6、NPS激活下丘腦orexin能神經(jīng)元、室旁核神經(jīng)元參與藥物成癮及依賴調(diào)節(jié)。 7、NPS激活上丘、下丘神經(jīng)元參與聽覺、視覺調(diào)節(jié)。 8、NPS激活導(dǎo)水管周圍灰質(zhì)、弓狀核參與痛覺調(diào)節(jié)。 9、NPS激活弓狀核參與神經(jīng)內(nèi)分泌及植物神經(jīng)調(diào)節(jié)。
[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.
【學(xué)位授予單位】：蘭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R392.1

【共引文獻(xiàn)】

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本文編號(hào)：2221192