The expression of gastric H+-K+-ATPase mRNA and protein in developing rat fundic gland.
The proton pump H+-K+-ATPase is the final common pathway mediating the production and secretion of hydrochloric acid by gastric parietal cells. The present studies were undertaken to examine whether the expression of gastric H+-K+-ATPase mRNA and protein changes are associated with the development of H+-K+-ATPase activity in the rat fundic gland. H+-K+-ATPase activity was examined in rat fundic gland at different stages from gestational day 18.5 to postnatal 8 weeks. The expression of H+-K+-ATPase mRNA was detected by in situ hybridization using a digoxigenin-labelled RNA probe with a tyramide signal amplification system. The expression of H+-K+-ATPase protein was evaluated by immunoblotting and immunohistochemistry using antibodies against H+-K+-ATPase alpha- and beta-subunits. We found that H+-K+-ATPase enzyme activity was detectable from the onset of gland formation (day 19.5 of gestation) and increased with age in the developing rat fundic gland. Expression of mRNA and protein was also discernible at the same time, and a progressive increase in expressions was observed as rats developed. Our results suggested that in developing rat fundic gland, the expression of both mRNA and protein of H+-K+-ATPase increased with age in a manner that parallels the development of H+-K+-ATPase enzyme activity.
Yang
D H
DH
Department of Anatomy, Kagoshima University Faculty of Medicine, Japan.
Tsuyama
S
S
Murata
F
F
D009597:4-Nitrophenylphosphatase / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D005088:Exocrine Glands / Q000201:enzymology* / Q000254:growth & development; D005260:Female; D005748:Gastric Fundus / Q000201:enzymology / Q000254:growth & development; D005753:Gastric Mucosa / Q000201:enzymology* / Q000254:growth & development; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D017506:H(+)-K(+)-Exchanging ATPase / Q000096:biosynthesis*; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D011247:Pregnancy; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000096:biosynthesis*; D051381:Rats; D017208:Rats, Wistar; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects
D012333:RNA, Messenger; D009597:4-Nitrophenylphosphatase; D017506:H(+)-K(+)-Exchanging ATPase