Beta 2-adrenergic receptor-stimulated increase in cAMP in rat heart cells is not coupled to changes in Ca2+ dynamics, contractility, or phospholamban phosphorylation.
Previous studies have shown that both beta 1- and beta 2-adrenergic receptors (AR) are present in rat ventricular myocytes, but stimulation of these receptor subtypes elicits qualitatively different cellular responses (Xiao, R.-P., and Lakatta, E. G. (1993) Circ. Res. 73, 286-300). In the present study, the biochemical mechanism underlying the distinct beta AR subtype actions have been investigated. Although both beta 1AR and beta 2AR stimulation increased total cellular cAMP in suspensions of rat ventricular myocytes to a similar extent, the maximum elevation of the membrane bound cAMP by beta 2AR stimulation was only half of that induced by beta 1AR stimulation, suggesting that stimulation the beta AR subtypes leads to different compartmentation of cAMP. The effects of beta 1AR stimulation on Ca2+ transient (indexed by the transient increase in indo-1 fluorescence ration after excitation) and contraction amplitude (measured via photodiode array) and their kinetics closely paralleled the increase in cAMP. In contrast, the increase in both membrane bound and total cAMP content after beta 2AR stimulation were completely dissociated from the effects of beta 2AR stimulation to increase the amplitudes of cytosolic Ca2+ transient and contraction. Furthermore, beta 2AR stimulation did not phosphorylate phospholamban to the same extent as did beta 1AR stimulation. This finding provides a mechanism for the failure of beta 2AR stimulation to accelerate the kinetics of the Ca2+i (cytosolic Ca2+) transient and contraction. These results indicate that the effects of beta 2AR stimulation on Ca2+i transient and contraction are uncoupled from the cAMP production and cAMP-dependent protein phosphorylation and indicate that, in addition to coupling to adenylate cyclase, beta 2AR stimulation also activates other signal transduction pathway(s) to produce changes in cytosolic Ca2+ and contraction.
Gerontology Research Center, National Institute on Aging, National Institutes of Health, Baltimore, Maryland 21224.
R P
RP
Xiao
C
C
Hohl
R
R
Altschuld
L
L
Jones
B
B
Livingston
B
B
Ziman
B
B
Tantini
E G
EG
Lakatta
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000378:metabolism*; D066298:In Vitro Techniques; D008297:Male; D009200:Myocardial Contraction; D009206:Myocardium / Q000378:metabolism*; D010766:Phosphorylation; D051381:Rats; D017208:Rats, Wistar; D011943:Receptors, Adrenergic, beta / Q000145:classification / Q000502:physiology*; D000099211:Phospholamban
D002118:Calcium; D002135:Calcium-Binding Proteins; D000242:Cyclic AMP; D011943:Receptors, Adrenergic, beta; D000099211:Phospholamban