神经递质对离体培养下丘脑神经元CRF分泌的调控

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	神经递质对离体培养下丘脑神经元CRF分泌的调控
	吴雁
学位类型	硕士
导师	杜继曾
	1992
学位授予单位	中国科学院西北高原生物研究所
学位授予地点	中国科学院西北高原生物研究所
学位专业	动物学
摘要	促肾上腺皮质激素释激素(Corticotropine-Releasing Factor, CRF)的分泌受多种因素的调节。自1981年Wylie Vale等利用高压液相色谱（HPLC）从羊下丘脑提取物中分离纯化并人工合成出具生理活性的CRF 以来，人们发展了CRF的放射免疫测定法（RIA，radioimmunoassay)，在此基础上用细胞免疫化学法明确了CRF神经元在中枢的分布，此后，关于下丘脑CRF分泌的调控以及行为研究便日渐深入发展起来。一系列的研究结果表明，大鼠脑室注射神经递质及神经肽可改变正中隆起处（medin eminence, ME）CRF的含量，同时，垂体门脉血流中ir-CRF的浓度也发生变化。七十年代中期，英国的Jones利用离体培养整个下丘脑的方法以及CRF的生物测定方法，系统地研究了神经质对下丘脑分泌CRF的作用，并提出了一个神经递质对CRF分泌的调控模式。对此模式所描述的神经递质作用和方式由于方法学的差异而有不同的见解。尤其是CRF放射免疫直接测定方法和离体培养下丘脑神经细胞获成功后，这为直接研究下丘脑CRF分泌功能提供了新的手段。本实验旨在利用体外培养新生鼠下丘脑细胞的方法和CRF的放射免疫测定法直接检测神经递质对下丘脑细胞ir－CRF分泌的作用。新生5－9天龄大鼠下丘脑细胞在培养9－16天之后，去甲肾上腺素（norepinephrine, NE）可使ir－CRF的分泌比对照组升高2．68倍，5－羟色氨（5－hydroxytrptamine, t-HT）可使ir－CRF的分泌升高1．95倍，而乙酰胆碱（acetylcholine, Ach）可使ir－CRF的分泌升高近10．48倍NE，5－HT及Ach在1－100 μM 剂量范围内，对下丘脑细胞分泌CRF的刺激作用都随它们的剂量变化而变化。氨基酸类递质γ一氨基丁酸（γ－aminobutyric acid, GABA）；影响CRF的基础分泌，在浓度为100 μM时使CRF的分泌有所下降，但作用不显著。Jones等认为NE对CRF的分泌是抑制的5－HT通过其它神经元对CRF的分泌起作用，本实验的结果与Jones等的观点有所不同。以上结果表明：（1）NE，5－HT，Ach对下丘脑细胞CRF的分泌是兴奋性的，GABA对ir－CRF的分泌的基础影响不显著。（2）NE，5－HT，Ach可直接作用于下丘脑细胞刺激CRF的分泌，其受体在出生后就已出现。并对相应的递质有反应能力。
其他摘要	Secretion of corticotropin-releasing factor (CRF-41) from hypothalamic paraventricular nuclei is modulated by a variety of neurotransmitters. In order to stude the secretory response of hypothalamic cells, a primary cultured cell model has been used in the expriment. The hypothalamus used for culturing were moved from neonatal rat brain of 5-9 days age. All secretory experiments were performed between 9th and 16th days culture. CRF-41 was assayed by RIA. In response to k~+ (56mol/l) depolariztion the 41 amino acid containing peptide, CRF-41 released from cultured cells, the CRF secretion increased up to 7.1 fold comparaing to basal secretion. Several lines of experimental evidence suggest that acetylcholine (Ach), 5-hydroxytryptamine (5-HT) and norepinephrine (NE) are excitatory to the hypothalamic secretion of CRF. As a consequence of some experiment, NE is also inhibitory to CRF secretion from hypothalami. v-aminobutyric acid (GABA), as a amino acic neurotransmitter, dose not affect basal secretion of CRF. In Our present experiment the effects of the putative neurotransmitters Ach, NE, 5-HT and GABA onimmunoreactive CRF-41 release from cultured hypothalamic cells were studied. Our experiments demonstrated that Ach stimulated CRF release in a dose-dependent fashion, at concentrtion ranging from 1 * 10~(-6) to 100 * 10~(-6) M. Ach stimulated CRF secretion incresed up to 10.48 fold as much as control at concentration of 100 μM. NE also stimulated CRF secretion by 3.08 fold in same manner. 5-HT elevated the CRF secretion as did Ach and NE, but it only increase CRF secretion by 1.95 fold at concentration of 100 μM. CABA can not inhibite basal CRF secretion. At concentration of 100 μM the secretion showed slightly decrease in contrast to control, with nonsignificant. The effect of Ach on CRF secretion is the strongest in these neurotransmitters. In summary, all of the three neurotransmitters Ach, NE and 5-HT stimulate directly CRF secretion at cultured cell level. They play a role of excitment on CRF neurones secretion. On the other hand, it is clear that the receptors of Ach, NE and 5-HT which has normal response to their specific neurotansmitters have appeared in neonatal rat hypothalami.
页数	37
语种	中文
文献类型	学位论文
条目标识符	http://210.75.249.4/handle/363003/3316
专题	中国科学院西北高原生物研究所
推荐引用方式 GB/T 7714	吴雁. 神经递质对离体培养下丘脑神经元CRF分泌的调控[D]. 中国科学院西北高原生物研究所. 中国科学院西北高原生物研究所,1992.

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