Adenylyl cyclase type 5 protein expression during cardiac development and stress.
Adenylyl cyclase (AC) types 5 and 6 (AC5 and AC6) are the two major AC isoforms expressed in the mammalian heart that mediate signals from beta-adrenergic receptor stimulation. Because of the unavailability of isoform-specific antibodies, it is difficult to ascertain the expression levels of AC5 protein in the heart. Here we demonstrated the successful generation of an AC5 isoform-specific mouse monoclonal antibody and studied the expression of AC5 protein during cardiac development in different mammalian species. The specificity of the antibody was confirmed using heart and brain tissues from AC5 knockout mice and from transgenic mice overexpressing AC5. In mice, the AC5 protein was highest in the brain but was also detectable in all organs studied, including the heart, brain, lung, liver, stomach, kidney, skeletal muscle, and vascular tissues. Western blot analysis showed that AC5 was most abundant in the neonatal heart and declined to basal levels in the adult heart. AC5 protein increased in the heart with pressure-overload left ventricular hypertrophy. Thus this new AC5 antibody demonstrated that this AC isoform behaves similarly to fetal type genes, such as atrial natriuretic peptide; i.e., it declines with development and increases with pressure-overload hypertrophy.
Hu
Che-Lin
CL
Department of Cell Biology and Molecular Medicine and the Cardiovascular Research Institute at the University of Medicine and Dentistry of New Jersey, New Jersey Medical School, Newark, NJ, USA.
Chandra
Rachna
R
Ge
Hui
H
Pain
Jayashree
J
Yan
Lin
L
Babu
Gopal
G
Depre
Christophe
C
Iwatsubo
Kousaku
K
Ishikawa
Yoshihiro
Y
Sadoshima
Junichi
J
Vatner
Stephen F
SF
Vatner
Dorothy E
DE
D000262:Adenylyl Cyclases / Q000172:deficiency / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D000367:Age Factors; D000375:Aging / Q000378:metabolism; D000818:Animals; D000831:Animals, Newborn; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D019556:COS Cells; D002522:Chlorocebus aethiops; D004195:Disease Models, Animal; D004285:Dogs; D018507:Gene Expression Regulation, Developmental; D006321:Heart / Q000254:growth & development* / Q000503:physiopathology; D017379:Hypertrophy, Left Ventricular / Q000201:enzymology* / Q000503:physiopathology; D007527:Isoenzymes / Q000172:deficiency / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D009206:Myocardium / Q000201:enzymology*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D013552:Swine; D014162:Transfection
D000911:Antibodies, Monoclonal; D007527:Isoenzymes; D012333:RNA, Messenger; D000262:Adenylyl Cyclases; C121100:adenylyl cyclase 6; C419510:adenylyl cyclase type V