{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2023,10,28]],"date-time":"2023-10-28T19:24:16Z","timestamp":1698521056639},"reference-count":67,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2005,2,4]],"date-time":"2005-02-04T00:00:00Z","timestamp":1107475200000},"content-version":"vor","delay-in-days":4297,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Journal Cellular Physiology"],"published-print":{"date-parts":[[1993,5]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>In the present investigation, a hCG sensitive glycosyl\u2010phosphatidylinositol (GPI) was isolated from cultured rat granulosa cells obtained from the ovaries of diethylstilbestrol (DES) implanted immature rats. The inositol\u2010phosphoglycan (IPG) moiety of the GPI\u2010lipid contains galactose, glucosamine, and myoinositol as demonstrated by metabolic labelling of granulosa cells for different time periods (5\u201396 h) with [<jats:sup>3<\/jats:sup>H]galactose, [<jats:sup>3<\/jats:sup>H]glucosamine, or [<jats:sup>3<\/jats:sup>H]myoinositol and treatment of the purified [<jats:sup>3<\/jats:sup>H]GPI with phosphatidylinositol\u2010specific phospholipase C. Labelling equilibrium of the GPI\u2010lipid was achieved after 24 h ([<jats:sup>3<\/jats:sup>H]galactose and [<jats:sup>3<\/jats:sup>H]myoinositol) or 72 h ([<jats:sup>3<\/jats:sup>H]glucosamine) incubation, whereas incorporation of other labelled carbohydrates tested ([<jats:sup>3<\/jats:sup>H]galactosamine, [<jats:sup>3<\/jats:sup>H]mannose, and [<jats:sup>3<\/jats:sup>H]sorbitol) was negligible throughout the time period studied. The glucosamine C\u20101 appears to be linked through a glycosidic bond to the myoinositol molecule of the IPG moiety as revealed by the generation of phosphatidylinositol (PtdIns) after nitrous acid deamination of dual labelled ([<jats:sup>3<\/jats:sup>H]glucosamine\/[<jats:sup>14<\/jats:sup>C]palmitate or [<jats:sup>3<\/jats:sup>H]glucosamine\/[<jats:sup>14<\/jats:sup>C]myristate) glycosyl\u2010phosphatidylinositol. To investigate the fatty acid composition of the diacylglycerol (DAG) backbone of the GPI, granulosa cells were also labelled (5\u201372 hr) with [<jats:sup>14<\/jats:sup>C]linoleate, [<jats:sup>3<\/jats:sup>H]myristate, [<jats:sup>3<\/jats:sup>H]\u2010oleate, [<jats:sup>3<\/jats:sup>H]palmitate, or [<jats:sup>3<\/jats:sup>H]stearate and the radioactivity associated with the purified glycosyl\u2010phosphatidylinositol determined. Incorporation of [<jats:sup>3<\/jats:sup>H]palmitate and [<jats:sup>3<\/jats:sup>H]myristate into the GPI\u2010lipid peaked after 8 h and 24 h of labelling, respectively, and both fatty acids were partially released after PLA<jats:sub>2<\/jats:sub> treatment of the dual labelled ([<jats:sup>3<\/jats:sup>H]glucosamine\/[<jats:sup>14<\/jats:sup>C]palmitate or [<jats:sup>3<\/jats:sup>H]glucosamine\/[<jats:sup>14<\/jats:sup>C]myristate) GPI. In parallel experiments no significant incorporation of labelled stearate, oleate, or linoleic acid into the DAG backbone of the glycosylphosphatidylinositol could be detected. Granulosa cells were also labelled with [<jats:sup>3<\/jats:sup>H]glucosamine in the presence of FSH (30 ng\/ml), cholera toxin (1 \u03bcg\/ml), or the membrane permeable cAMP analog (but)<jats:sub>2<\/jats:sub> cAMP (1 mM). Time related increases in GPI\u2010labelling were apparent after 48 h and reached a maximum level (3\u2010, 5\u2010, and 7\u2010fold for FSH, CT, and (but)<jats:sub>2<\/jats:sub> cAMP, respectively) after 72 h in culture. In another set of experiments, granulosa cells were labelled for 72 h with [<jats:sup>3<\/jats:sup>H]glucosamine in the presence of (but)<jats:sub>2<\/jats:sub>cAMP (1 mM), TPA (10<jats:sup>\u22127<\/jats:sup> M), or combination thereof. The effect of treatment with the membrane permeable cAMP analog on GPI labelling was prevented in the presence of TPA, whereas no differences in [<jats:sup>3<\/jats:sup>H]GPI content could be observed in untreated granulosa cells or cells cultured in the presence of the protein kinase C\u2010activating phorbol ester alone. In cells differentiated with FSH (30 ng\/ml for 3 days) to induce LH receptors, treatment with hCG (100 ng\/ml) induced a rapid (60 sec) and transient (5 min) decrease in the GPI content, whereas no efect of the hormone on undifferentiated granulosa cells could be observed. The rapid effect elicited by hCG on GPI content and turnover may be an early transduction mechanism involved in the biological effects of LH\/hCG in differentiated granulosa cells. \u00a9 1993 Wiley\u2010Liss, Inc.<\/jats:p>","DOI":"10.1002\/jcp.1041550208","type":"journal-article","created":{"date-parts":[[2005,2,26]],"date-time":"2005-02-26T05:47:20Z","timestamp":1109396840000},"page":"273-281","source":"Crossref","is-referenced-by-count":8,"title":["Follicle\u2010stimulating hormone and human chorionic gonadotropin induced changes in granulosa cell glycosyl\u2010phosphatidylinositol concentration"],"prefix":"10.1002","volume":"155","author":[{"given":"Luisa F.","family":"Fanjul","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Isabel","family":"Marrero","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"F.","family":"Estevez","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"J.","family":"Gonzalez","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"J.","family":"Quintana","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pino","family":"Santana","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"C. 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