Epidermal growth factor elevates intracellular pH in chicken granulosa cells by activating protein kinase C.


:Previous studies from our laboratory have demonstrated that epidermal growth factor (EGF), induces intracellular alkalinization in chicken granulosa cells by activating a sodium-dependent and amiloride-sensitive Na+/H+ antiporter. In the present investigation we have examined the possible involvement of protein kinase C (PKC) in the regulation of intracellular pH (pHi) by EGF in chicken granulosa cells. Intracellular pH in granulosa cells obtained from the two largest preovulatory follicles was determined spectrofluorometrically using the dye 2',7'-bis-(carboxyethyl)-5(6)-carboxyfluorescein. The resting pHi was 6.81 +/- 0.01 (n = 30) when the extracellular pH and sodium concentration were 7.3 and 144 mM, respectively. 12-O-Tetradecanoyl-phorbol-13-acetate (TPA; 50-400 ng/ml) and 1-oleoyl-2-acetylglycerol (OAG; 1-75 micrograms/ml) mimicked the actions of EGF by inducing a concentration-dependent increase in pHi which reached a maximum of 0.25-0.30 pH units. 4 alpha-Phorbol 12,13-didecanoate, a phorbol ester with no tumor promoting activity had no effect on pHi. Cytosolic alkalinization was observed within 10 min of the addition of each agent and increased over the 60-min observation period. Like EGF-induced cytosolic alkalinization, the increases in pHi in response to TPA or OAG were dependent on the presence of sodium concentration and were inhibited by amiloride, an inhibitor of the Na+/H+ antiporter. The effects of EGF, TPA, and OAG were attenuated by the PKC inhibitors 5-isoquinolinylsulfonyl-2-methyl piperazine and trifluoperazine. Down-regulation of granulosa cell PKC by pretreatment with TPA (200 ng/ml) for 2.5 h inhibited EGF-, TPA-, and OAG-induced cytosolic alkalinization. The effects of maximally stimulatory concentrations of EGF and TPA on cytosolic alkalinization were not additive. The increases in pHi induced by TPA and OAG, but not by EGF, were dependent on the presence of extracellular Ca++. These studies suggest that the EGF-induced intracellular alkalinization in chicken granulosa cells involves a PKC-mediated activation of the Na+/H+ antiporter.






Li M,Morley P,Tsang BK




Has Abstract


1991-12-01 00:00:00












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    pub_type: 杂志文章


    authors: Botero LF,Roberts CT Jr,LeRoith D,Adashi EY,Hernandez ER

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  • Active intestinal calcium transport in the absence of transient receptor potential vanilloid type 6 and calbindin-D9k.

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    authors: Benn BS,Ajibade D,Porta A,Dhawan P,Hediger M,Peng JB,Jiang Y,Oh GT,Jeung EB,Lieben L,Bouillon R,Carmeliet G,Christakos S

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    authors: Gangrade BK,Davis JS,May JV

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  • TGF-β effects on prostate cancer cell migration and invasion are mediated by PGE2 through activation of PI3K/AKT/mTOR pathway.

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    authors: Wilks JW,Hui OJ

    更新日期:1987-03-01 00:00:00

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    authors: Wuttke M,Papewalis C,Meyer Y,Kessler C,Jacobs B,Willenberg HS,Schinner S,Kouatchoua C,Baehring T,Scherbaum WA,Schott M

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    authors: Siragy H,Howell N,Rose CE Jr,Khosla MC,Peach MJ,Carey RM

    更新日期:1988-01-01 00:00:00

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    authors: Bambino TH,Hsueh AJ

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    authors: Tanji N,Yokoyama M,Takeuchi M,Terada N,Dahiya R,Cunha GR

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    authors: Barseghian G,Levine R,Epps P

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    authors: Georgiou M,Payne AH

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  • Tissue distribution, turnover, and glycosylation of the long and short growth hormone receptor isoforms in rat tissues.

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    pub_type: 杂志文章


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    pub_type: 杂志文章


    authors: Resko JA,Pleom JG,Stadelman HL

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    authors: Dahl GE,Evans NP,Moenter SM,Karsch FJ

    更新日期:1994-07-01 00:00:00

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    pub_type: 杂志文章


    authors: Pauliina Markkula S,Lyons D,Yueh CY,Riches C,Hurst P,Fielding B,Heisler LK,Evans ML

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    pub_type: 杂志文章


    authors: Calduch-Giner J,Duval H,Chesnel F,Boeuf G,Pérez-Sánchez J,Boujard D

    更新日期:2001-07-01 00:00:00

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    pub_type: 杂志文章


    authors: Orwig KE,Dai G,Rasmussen CA,Soares MJ

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    pub_type: 杂志文章


    authors: Wada T,Naito M,Kenmochi H,Tsuneki H,Sasaoka T

    更新日期:2007-02-01 00:00:00