INVITRO EVIDENCE FOR MODULATION OF MORPHOLOGICAL AND FUNCTIONAL-DEVELOPMENT OF HYPOTHALAMIC IMMUNOREACTIVE ATRIAL-NATRIURETIC-PEPTIDE NEURONS BY CYCLIC 3',5'-ADENOSINE-MONOPHOSPHATE
INVITRO EVIDENCE FOR MODULATION OF MORPHOLOGICAL AND FUNCTIONAL-DEVELOPMENT OF HYPOTHALAMIC IMMUNOREACTIVE ATRIAL-NATRIURETIC-PEPTIDE NEURONS BY CYCLIC 3',5'-ADENOSINE-MONOPHOSPHATE
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DOI：
10.1210/en.131.2.911
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发表时间：
1992-08-01
期刊：
影响因子：
4.8
通讯作者：
LIM, AT
中科院分区：
文献类型：
--
作者：
LEE, D;HUANG, WQ;LIM, AT
在大鼠的下丘脑中，心房钠尿肽（ANP）的前体被产生并加工成其较小的3 K mol wt种类的同类物，其由具有室周区域和室旁核中的细胞体的神经元分泌。采用长期单层培养的新生大鼠下丘脑细胞，我们已经确定了一个小的人口的细胞染色阳性的免疫反应（IR）心钠素。72 +/- 7%（平均值+/- SF; n = 4/1000个细胞）的irANP阳性细胞与神经元特异性烯醇化酶染色共定位;一些细胞具有多个神经突，并在胞体、沿着神经元突起的静脉曲张和长神经突的末端显示irANP染色。在10（-6）-10（-4）M范围内，forskolin、3-异丁基-1-甲基黄嘌呤或8-溴-cAMP以剂量相关和时间依赖的方式显著增加irANP阳性细胞和具有神经突的细胞的总数。在10（-4）M时，forskolin处理4天使irANP阳性神经元的数量增加了4倍（P < 0.01），而长突起细胞的数量增加了3倍（P < 0.01）。此外，通过使用地高辛标记的30个碱基对反义寡核苷酸探针进行比色原位杂交确定，它使显示pro-ANP mRNA阳性信号的细胞数量增加了约三倍（P < 0.01）。与上述观察结果一致，毛喉素、3-异丁基-1-甲基黄嘌呤或8-溴-cAMP处理显著增加培养物中存在的irANP总量，毛喉素的ED 50约为5 × 10 - 5 M。虽然用10（-7）M佛波醇12-肉豆蔻酸酯13-乙酸酯处理使培养物中irANP的产生大约加倍（P < 0.05），但佛波醇12-肉豆蔻酸酯13-乙酸酯对调节irANP神经元的数量或轴突生长几乎没有影响。因此，我们目前的研究结果表明，蛋白激酶-A途径比蛋白激酶-C途径更重要的是在调节ANP产生的神经元的功能和形态发育过程中的个体发育的大鼠下丘脑。
In the hypothalamus of the rat, the precursor of atrial natriuretic peptide (ANP) is produced and processed into its smaller congeners of 3K mol wt species, which are secreted from neurons with cell bodies in the periventricular areas and the paraventricular nuclei of the tissue. Employing long term monolayer cultures of neonatal rat hypothalamic cells, we have identified a small population of cells that stained positive for immunoreactive (ir) ANP. Seventy-two +/- 7% (mean +/- SF; n = 4 per 1000 cells) of the irANP positive cells were colocalized with the staining of neuron-specific enolase; some of the cells possessed multiple neurites and showed irANP staining in the perikarya, in the varicosities along neuronal processes, and at the terminals of long neurites. Over the range of 10(-6)-10(-4) M, forskolin, 3-isobutyl-1-methylxanthine, or 8-bromo-cAMP significantly augmented the total number of irANP-positive cells and those possessing neurites in a dose-related and time-dependent manner. At 10(-4) M, 4 days of forskolin treatment increased the number of irANP-positive neurons 4-fold (P < 0.01) while tripling that of the cells with long neurites (P < 0.01). Furthermore, it approximately tripled the number of cells (P < 0.01) showing positive signals for pro-ANP mRNA, as ascertained by colorimetric in situ hybridization using a 30-basepair antisense oligonucleotide probe labeled with digoxigenin. Consistent with the above observation, forskolin, 3-isobutyl-1-methylxanthine, or 8-bromo-cAMP treatment significantly augmented the total amount of irANP present in the cultures, with an ED50 of forskolin approximating 5 X 10(-5) M. Although treatment with 10(-7) M phorbol 12-myristate 13-acetate approximately doubled the production of irANP in the cultures (P < 0.05), phorbol 12-myristate 13-acetate had little effect on modulating the number or neurite out growth of irANP neurons. Thus, our present findings suggest that protein kinase-A pathways are of greater importance than protein kinase-C pathways in regulating both the functional and morphological development of ANP-producing neurons during the ontogenesis of the rat hypothalamus.