Cyclic AMP binding to the amphibian oocyte plasma membrane: possible interrelationship between meiotic arrest and membrane fluidity.
Cyclic AMP, which maintains the vertebrate oocyte in prophase arrest under physiological conditions, exhibits specific and saturable binding to the cytoplasmic face of the prophase-arrested Rana pipiens oocyte plasma membrane. Scatchard type analyses of [3H]cAMP binding to isolated plasma membranes indicate a single class of binding sites with a Kd = 19.3 +/- 7.0 nM at cAMP concentrations below 10(-6) M and additional low affinity site(s) and/or non-specific binding at concentrations above 10(-6) M. Photoaffinity labeling of prophase oocyte plasma membranes with [32P]-8-N3cAMP demonstrates cAMP/cGMP-displacable binding of 8-N3[32P]cAMP to a 100-110 kDa peptide doublet. Plasma membrane fluidity was monitored by electron spin resonance in isolated plasma-vitelline membranes using a 5-doxyl stearic acid probe. Exogenous dibutyryl cAMP (dbcAMP) produces an increase in membrane fluidity within minutes and blocks and/or reverses the progesterone-induced decrease in plasma membrane fluidity. The dbcAMP concentration that produced half-maximal fluidity increase (10 microM) corresponds to the half-maximal inhibiting dose of dbcAMP for progesterone induction of meiosis. Cholera toxin, which elevates intracellular cAMP and blocks meiosis, also increases membrane fluidity and inhibits progesterone-induced decrease in membrane fluidity. Elevated levels of intracellular cAMP thus appear to maintain meiotic arrest by binding to specific plasma membrane site(s) and maintaining the plasma membrane in a relatively fluid state. The progesterone-induced fall in intracellular cAMP first reported in Rana thus appears to be responsible for the progesterone-induced increase in membrane fluidity and further suggests that the change in membrane order is essential for the resumption of the meiotic divisions.
Morrill
G A
GA
Department of Physiology and Biophysics, Albert Einstein College of Medicine, Bronx, NY 10461.
Doi
K
K
Erlichman
J
J
Kostellow
A B
AB
D000345:Affinity Labels; D000818:Animals; D003994:Bucladesine / Q000494:pharmacology; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism*; D000242:Cyclic AMP / Q000378:metabolism* / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004578:Electron Spin Resonance Spectroscopy; D005260:Female; D007328:Insulin / Q000494:pharmacology; D008560:Membrane Fluidity / Q000187:drug effects; D009865:Oocytes / Q000378:metabolism*; D011374:Progesterone / Q000494:pharmacology; D011418:Prophase; D011894:Rana pipiens; D013997:Time Factors; D014316:Tritium
D000345:Affinity Labels; D007328:Insulin; D014316:Tritium; D011374:Progesterone; D003994:Bucladesine; D000242:Cyclic AMP