ACTIVATION OF INOSITOL PHOSPHOLIPID TURNOVER AND CALCIUM SIGNALING IN RAT SERTOLI CELLS BY P2-PURINERGIC RECEPTORS - MODULATION OF FOLLICLE-STIMULATING-HORMONE RESPONSES
ACTIVATION OF INOSITOL PHOSPHOLIPID TURNOVER AND CALCIUM SIGNALING IN RAT SERTOLI CELLS BY P2-PURINERGIC RECEPTORS - MODULATION OF FOLLICLE-STIMULATING-HORMONE RESPONSES
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DOI：
10.1210/en.134.3.1537
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发表时间：
1994-03-01
期刊：
影响因子：
4.8
通讯作者：
ZIPARO, E
中科院分区：
文献类型：
--
作者：
FILIPPINI, A;RICCIOLI, A;ZIPARO, E
为了研究细胞外核苷酸在支持细胞调控中的作用，我们测试了ATP及其类似物对Ca2+-磷脂和camp依赖通路的影响。培养的未成熟动物支持细胞与ATP或结构相关化合物孵育，测定磷酸肌苷（PI）的周转或cAMP的积累。在测试的几种核苷酸磷酸类似物中，腺苷5'- o -（3-硫代三磷酸）是刺激肌醇磷酸积累最有效的激动剂。嘌呤核苷酸对PI周转率的影响具有时间和浓度依赖性。由于不可水解的ATP类似物也刺激PI转换，ATP代谢物或代谢产物不负责观察到的刺激。不同ATP类似物[腺苷5′- o-（3-硫代三磷酸）> ATP与UTP > β， γ -亚甲基腺苷5′-三磷酸，2-甲基硫代ATP >腺苷]的效价顺序与支持细胞表面P-2U受体（核苷酸受体）的存在一致。在携带细胞内Ca2+指示剂fura-2的单个Sertoli细胞中，PI周转增加伴随着Ca2+浓度的短暂增加。单独使用时，ATP及其类似物对支持细胞中的cAMP水平没有直接影响。然而，ATP或其类似物抑制fsh依赖性cAMP积累超过70%。嘌呤核苷酸也有效地阻断了FSH远端对cAMP积累的影响，因为细胞外ATP完全逆转了FSH诱导的支持细胞形状的变化。根据使用的嘌呤或嘧啶核苷酸不同，百日咳毒素对cAMP积累的核苷酸依赖性抑制在不同程度上被阻断。这表明不止一种机制有助于嘌呤核苷酸依赖性cAMP积累的抑制。这些数据证明，嘌呤核苷酸受体与多种途径偶联存在于培养的Sertoli细胞中，并且细胞外ATP对Sertoli细胞的FSH反应性具有深远的生物学作用。
To study the role of extracellular nucleotides in the regulation of Sertoli cells, the effects of ATP and its analogs on the Ca2+-phospholipid- and cAMP-dependent pathways were tested. Cultured Sertoli cells from immature animals were incubated with ATP or structurally related compounds, and phosphoinositide (PI) turnover or cAMP accumulation was measured. Among the several nucleotide phosphate analogs tested, adenosine 5'-O-(3-thiotriphosphate) was the agonist most potent in stimulating inositol phosphate accumulation. The effects of purine nucleotides on PI turnover were time and concentration dependent. Because nonhydrolizable ATP analogs also stimulated PI turnover, ATP metabolites or metabolic products are not responsible for the observed stimulation. The order of potency of the different ATP analogs [adenosine 5'-O-(3-thiotriphosphate) > ATP congruent to UTP > beta,gamma-methyleneadenosine 5'-triphosphate, 2-methylthio-ATP > adenosine] was consistent with the presence of P-2U receptors (nucleotide receptors) on the surface of the Sertoli cell. Augmented PI turnover was accompanied by a transient increase in Ca2+ concentration, measured in single Sertoli cells loaded with the intracellular Ca2+ indicator fura-2. When used alone, ATP and its analogs did not have a direct effect on cAMP levels in the Sertoli cell. However, ATP or its analogs inhibited FSH-dependent cAMP accumulation by more than 70%. Purine nucleotides also efficiently blocked the effects of FSH distal to cAMP accumulation, because extracellular ATP completely reversed the changes in Sertoli cell shape induced by FSH. The nucleotide-dependent inhibition of cAMP accumulation was blocked by pertussis toxin to a different degree depending on the purine or pirimidine nucleotide used. This indicated that more than one mechanism contributes to the purine nucleotide-dependent inhibition of cAMP accumulation. These data provide evidence that purine nucleotide receptors coupled to multiple pathways are present on the Sertoli cell in culture, and that extracellular ATP has profound biological effects on the FSH responsiveness of the Sertoli cell.