Effects of surgical sympathetic denervation on G-protein levels, alpha and beta-adrenergic receptors, cAMP production and adenylate cyclase activity in the smooth muscles of rabbit iris.
The relative densities of a number of G protein subunits were quantified in membranes prepared from iris sphincter and dilator muscles of rabbits that have undergone sympathetic denervation and from contralateral innervated controls by immunoblotting with specific polyclonal antibodies against Gs alpha, Gi alpha and Gq alpha protein subunits. In addition, alpha and beta-adrenergic receptor densities, basal and isoproterenol (ISO)-stimulated cAMP production, and basal and ISO+GTP gamma S-stimulated adenylate cyclase (AC) activities were measured in the same tissues. Densitometric analysis of the immunoblot data revealed a 32% reduction in the level of Gi alpha in the denervated sphincter, a 26% increase in the level of Gi alpha in the denervated dilator, and no changes due to denervation were found in the levels of Gs alpha and Gq alpha. Sympathetic denervation had no effect on the densities of alpha- and beta-adrenergic receptors in these tissues, however, it did induce a significant decrease in the KD values of alpha-adrenergic receptors in both dilator and sphincter, and in beta-adrenergic receptors in the sphincter. The basal- and ISO-stimulated cAMP production in the sphincter was 4-fold as high as that of the dilator, and at 0.05 microM ISO the denervation supersensitivity for cAMP production in the sphincter increased by 118% as compared to 36% in the dilator. Sympathetic denervation increased by 19-47% the basal- and ISO-stimulated activity of AC in these tissues, however, it had no effect on the phospholipase C activity.(ABSTRACT TRUNCATED AT 250 WORDS)
Abdel-Latif
A A
AA
Department of Biochemistry and Molecular Biology, Medical College of Georgia, Augusta 30912-2100, USA.
Akhtar
R A
RA
Zhou
C J
CJ
D000262:Adenylyl Cyclases / Q000378:metabolism*; D000818:Animals; D002462:Cell Membrane; D000242:Cyclic AMP / Q000096:biosynthesis*; D003714:Denervation; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D019204:GTP-Binding Proteins / Q000378:metabolism*; D015171:Ganglionectomy; D007498:Iris / Q000294:innervation / Q000378:metabolism*; D008297:Male; D009130:Muscle, Smooth / Q000187:drug effects / Q000378:metabolism*; D011817:Rabbits; D011941:Receptors, Adrenergic / Q000378:metabolism*; D011942:Receptors, Adrenergic, alpha / Q000378:metabolism; D011943:Receptors, Adrenergic, beta / Q000378:metabolism; D017783:Superior Cervical Ganglion / Q000601:surgery; D013566:Sympathomimetics / Q000494:pharmacology
D011941:Receptors, Adrenergic; D011942:Receptors, Adrenergic, alpha; D011943:Receptors, Adrenergic, beta; D013566:Sympathomimetics; D000242:Cyclic AMP; D019204:GTP-Binding Proteins; D000262:Adenylyl Cyclases