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The FASEB Journal, Vol 1, 380-387, Copyright © 1987 by The Federation of American Societies for Experimental Biology

RESEARCH COMMUNICATIONS

Expression of rat mRNA coding for hormone-stimulated adenylate cyclase in Xenopus oocytes

AA Smith, T Brooker and G Brooker
Fidia-Georgetown Institute for the Neurosciences, Georgetown University Medical Center, Washington, DC 20007.

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.


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Copyright © 1987 by The Federation of American Societies for Experimental Biology.