Reduced adenylyl cyclase activation with no decrease in beta-adrenergic receptors in basenji greyhound leukocytes: relevance to beta-adrenergic responses in airway smooth muscle.
Mononuclear leukocytes (MNLs) have been used as a model of beta-adrenergic responsiveness of airway smooth muscle, but the relevance of this model remains controversial. The basenji greyhound (BG) dog model of airway hyperresponsiveness shares some features with human asthma, and airway smooth muscle shows a selective impairment in isoproterenol-stimulated adenylyl cyclase activity. In this study, MNL membranes were obtained from these same dogs, and the beta-adrenergic receptor-adenylyl cyclase cascade function was compared with that in airway smooth muscle. beta-Adrenergic receptor numbers and affinities for iodine 125-cyanopindolol were similar in the two dog groups (receptor numbers [Bmax] = 441 +/- 101 and 447 +/- 61 fmol/mg protein and dissociation constant [Kd] = 269 +/- 44 and 312 +/- 60 pmol/L for mongrel and BG MNLs, respectively). Quantities of the Gs alpha protein were not different in the membranes as determined by immunoblotting. Stimulation of adenylyl cyclase by isoproterenol (100 mumol/L) was impaired in MNL membranes of BG membranes (22% +/- 4% increase over guanosine triphosphate [10 mumol/L]) compared with mongrel membranes (47% +/- 8.6% increase over guanosine triphosphate [10 mumol/L], p < 0.05). Stimulation of adenylyl cyclase by prostaglandin E1 (10 mumol/L), NaF (10 mmol/L), or forskolin (10 mumol/L) did not differ in membranes from the two groups. No difference was found in the lymphocyte subsets in the two groups as determined by flow cytometry. These findings are qualitatively similar to studies of trachealis muscle membranes from these same dogs.(ABSTRACT TRUNCATED AT 250 WORDS)
Emala
C W
CW
Department of Anesthesiology, Johns Hopkins Medical Institutions, Baltimore, MD, USA.
Levine
M A
MA
Aryana
A
A
Margolick
J B
JB
Hirshman
C A
CA
D000262:Adenylyl Cyclases / Q000378:metabolism*; D000319:Adrenergic beta-Antagonists / Q000378:metabolism; D000818:Animals; D004285:Dogs; D004789:Enzyme Activation; D005434:Flow Cytometry; D019204:GTP-Binding Proteins / Q000378:metabolism; D015151:Immunoblotting; D007962:Leukocytes / Q000378:metabolism*; D016131:Lymphocyte Subsets / Q000378:metabolism; D009130:Muscle, Smooth / Q000378:metabolism; D010869:Pindolol / Q000031:analogs & derivatives / Q000378:metabolism; D011869:Radioligand Assay; D011943:Receptors, Adrenergic, beta / Q000378:metabolism*; D014132:Trachea / Q000378:metabolism
D000319:Adrenergic beta-Antagonists; D011943:Receptors, Adrenergic, beta; C047975:cyanopindolol; D010869:Pindolol; D019204:GTP-Binding Proteins; D000262:Adenylyl Cyclases