We evaluated the effects of culturing mouse MTAL cells under conditions that suppressed steady-state cytosolic Cl− on chloride channels fused into bilayers from basolateral vesicles of cultured MTAL cells. We used two agents to suppress Cl− entry: 10−6 M PGE2 and 10−4 M bumetanide. Basolateral Cl− channels from control cultured MTAL cells exhibited the signature characteristics of mmClC-Ka channels: increased open-time probability (P o) either by raising cytosolic-face [Cl−] or, at 2 mM cytosolic Cl−, by adding (ATP + PKA), and first-order conductance kinetics. Either 10−6 M PGE2 or 10−4 M bumetanide in culture media reduced steady-state MTAL cytosolic Cl−. Chloride channels from these cells exhibited characteristics unique to CTAL mcClC-Ka channels, namely: no augmentation of P o either by raising cytosolic Cl− or with cytosolic (ATP + PKA), and multi-ion occupancy. Semi-quantitative RT-PCR and real-time quantitative PCR showed that culturing MTAL cells with 10−6 M PGE2 or 10−4 M bumetanide reduced mRNA levels encoding mmClC-Ka but not mRNA levels encoding mcClC-Ka. However, when MTAL cells were cultured under control conditions, and then pre-incubated for 60 minutes with 10−4 M bumetanide, cytosolic Cl− fell acutely but Cl− channels exhibited characteristics of mmClC-Ka channels. Thus PGE2 and bumetanide, both of which lower steady-state MTAL cytosolic Cl− concentrations, inhibit either the transcriptional and/or the translational processes for mmClC-Ka synthesis.
The Journal of Membrane Biology – Springer Journals
Published: Oct 9, 2003
It’s your single place to instantly
discover and read the research
that matters to you.
Enjoy affordable access to
over 18 million articles from more than
15,000 peer-reviewed journals.
All for just $49/month
Query the DeepDyve database, plus search all of PubMed and Google Scholar seamlessly
Save any article or search result from DeepDyve, PubMed, and Google Scholar... all in one place.
Get unlimited, online access to over 18 million full-text articles from more than 15,000 scientific journals.
Read from thousands of the leading scholarly journals from SpringerNature, Elsevier, Wiley-Blackwell, Oxford University Press and more.
All the latest content is available, no embargo periods.
“Hi guys, I cannot tell you how much I love this resource. Incredible. I really believe you've hit the nail on the head with this site in regards to solving the research-purchase issue.”Daniel C.
“Whoa! It’s like Spotify but for academic articles.”@Phil_Robichaud
“I must say, @deepdyve is a fabulous solution to the independent researcher's problem of #access to #information.”@deepthiw
“My last article couldn't be possible without the platform @deepdyve that makes journal papers cheaper.”@JoseServera