Expression of rat mRNA coding for hormone-stimulated adenylate cyclase in Xenopus oocytes.
Beta-Adrenergic agonist-stimulated hyperpolarization, whole-cell cAMP accumulation, and activity of isoproterenol-stimulated membrane-bound adenylate cyclase (EC 4.6.1.1) in Xenopus laevis ovarian oocytes are entirely dependent on the presence of nascent follicle cells. A method was developed to remove rapidly and completely all extra-oocyte cell types to yield defolliculated oocytes that exhibited normal viability and resting membrane potentials yet lacked beta-adrenergic receptor (beta AR)-stimulated responses. Purified follicle membranes contained beta AR-stimulated adenylate cyclase activity, whereas oocyte cell membranes did not. Purified oocyte membrane preparations from X. laevis oocytes previously microinjected with C6-2B rat astrocytoma mRNA, and subsequently defolliculated, exhibited novel beta AR and forskolin-stimulated adenylate cyclase activity. These experiments demonstrate that oocytes expressed rat C6-2B mRNA coding for the beta-adrenergic receptor and the components necessary for forskolin-stimulated adenylate cyclase activity.
Smith
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Fidia-Georgetown Institute for the Neurosciences, Georgetown University Medical Center, Washington, DC 20007.
Brooker
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Brooker
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D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology; D000262:Adenylyl Cyclases / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D001254:Astrocytoma; D002462:Cell Membrane / Q000201:enzymology; D005576:Colforsin / Q000494:pharmacology; D005260:Female; D007545:Isoproterenol / Q000494:pharmacology; D008564:Membrane Potentials / Q000187:drug effects; D008845:Microinjections; D009865:Oocytes / Q000201:enzymology*; D006080:Ovarian Follicle / Q000502:physiology; D012333:RNA, Messenger / Q000235:genetics*; D051381:Rats; D011943:Receptors, Adrenergic, beta / Q000502:physiology*; D014170:Transformation, Genetic; D014407:Tumor Cells, Cultured; D014982:Xenopus laevis
D012333:RNA, Messenger; D011943:Receptors, Adrenergic, beta; D005576:Colforsin; D015124:8-Bromo Cyclic Adenosine Monophosphate; D000262:Adenylyl Cyclases; D007545:Isoproterenol