Expression of transforming growth factor-alpha mRNA and peptide in rodent kidneys.
The expression and function of transforming growth factor alpha (TGF-(alpha) in the kidney are not fully characterised. There exists controversy concerning the detection of renal TGF-alpha mRNA and the localisation of its immunoreactivity. In attempts to clarify the detection and localisation issue, the present study aimed to detect TGF-alpha mRNA in neonate and adult rat kidneys, to examine the specificity of two commonly used anti-TGF-alpha antibodies and finally to localise renal TGF-alpha immunoreactivity using a specific antibody. TGF-alpha mRNA of around 4.8 kb was readily detected with a sensitive nonradioactive northern analysis, with a similar abundance in neonatal and adult rat kidneys. Renal TGF-alpha peptide of the 6-kDa mature form was identified by western blotting. By using various controls, including specimens from TGF-alpha knock out mice in comparison with wild-type mice, the present study has confirmed the specificity of a polyclonal anti-human recombinant TGF-alpha antibody. With this antibody, TGF-alpha immunoreactivity was localised to the proximal tubules in renal cortex. In addition, the present study has also demonstrated a non-specificity in localising TGF-alpha in rodent kidneys by the most commonly used monoclonal anti-human TGF-alpha C-terminal peptide antibody, which stained collecting ducts in renal cortex and medulla.
Xian
C J
CJ
Child Health Research Institute, North Adelaide, South Australia, Australia.
D000818:Animals; D000911:Antibodies, Monoclonal; D019084:Fluorescent Antibody Technique, Indirect; D015870:Gene Expression; D006801:Humans; D007668:Kidney / Q000378:metabolism* / Q000473:pathology; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D010455:Peptides / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger; D051381:Rats; D017207:Rats, Sprague-Dawley; D016211:Transforming Growth Factor alpha / Q000235:genetics / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D010455:Peptides; D012333:RNA, Messenger; D016211:Transforming Growth Factor alpha