Characterization of a Specific Antibody to the Rat Angiotensin II AT₁ Receptor

Abstract

We raised a polyclonal antibody against a decapeptide corresponding to the carboxyl terminus of the rat angiotensin II AT1 receptor. This antibody was demonstrated to be specific for the rat receptor according to a number of approaches. These included (a) the ultrastructural localization of immunogold-labeled receptor on the surfaces of zona glomerulosa cells in the adrenal cortex, (b) the specific labeling of Chinese hamster ovarian (CHO) cells transfected with AT1 receptors, (c) the identification of a specific band on Western blots, (d) the immunocytochemical co-localization of angiotensin receptors on neurons in the lamina terminalis of the brain shown to be responsive to circulating angiotensin II, as shown by the expression of c-fos, and (e) the correlation between the expression of the mRNA of the AT1 receptor and AT1 receptor immunoreactivity.(J Histochem Cytochem 47:507-515, 1999)

Angiotensin-II Is a Putative Neurotransmitter in Lactate-Induced Panic-Like Responses in Rats with Disruption of GABAergic Inhibition in the Dorsomedial Hypothalamus

“…Overall, These data support the notion that the primary antibody used was immunostaining the AT 1 R antigen. The results of the immunostaining are represented in Figure 7D-G and clearly demonstrate the presence of AT 1 Rs on cell bodies and fibers in the DMH, LH, and median eminence, which is consistent with previous data describing AT 1 R mRNA (Lenkei et al, 1995(Lenkei et al, , 1998 and protein in the adult male rat brain (Giles et al, 1999). In addition, we also detected AT 1 Rs on cell bodies in CVOs such as the SFO and in other regions of the hypothalamus [i.e., the paraventricular hypothalamic nucleus and median preoptic area (MPOA); data not shown], which is also consistent with previous findings (Lenkei et al, 1995(Lenkei et al, , 1998Giles et al, 1999).…”

Age-related renal disease in female Dahl salt-sensitive rats is attenuated with 17²-estradiol supplementation by modulating nitric oxide synthase expression

“…Phospho-AKt has been reported to increase eNOS activity by enhancing its phosphorylation 23 . We found no significant difference in the expression of pAKt protein with aging, ovariectomy or E 2 supplementation.…”

Angiotensin II Binding to Angiotensin I–Converting Enzyme Triggers Calcium Signaling

“…Figure S1A) and confirmed by earlier data. 11,12 After the stable transfection of these cells with an empty plasmid (CHO-mock) and a plasmid containing a DNA cassette for somatic human ACE expression (CHO-ACE), specific ACE activity could be measured in CHO-ACE cells but not in the CHO-mock cells ( Figure S1B). To exclude any possible involvement of the AT 1 receptor in this effect, we confirmed its absence in CHO-ACE cells by flow cytometry using specific AT 1 antibodies ( Figure S1C).…”