Thyroid hormone up-regulates Na+/K+ pump alpha2 mRNA but not alpha2 protein isoform in cultured skeletal muscle.
Thyroid hormone (T(3)) is known to up-regulate the physiological expression of the Na(+)/K(+) pump in cultured skeletal muscle. We recently reported that primary cultured rat skeletal muscle expresses only the alpha(1), beta(1) and beta(2) protein isoforms of Na(+)/K(+) pump. Interestingly, alpha(2) mRNA is detectable while the alpha(2) protein isoform is not. We therefore examined whether T(3) might up-regulate the expression of Na(+)/K(+) pump alpha(2) isoform at the protein and mRNA level. We also examined the regulation by this hormone of the other isoforms of the pump. Primary cultures were treated with T3 for 48 h from day 4 to day 6 of differentiation. Protein and mRNA isoforms of Na(+)/K(+) pump were identified by Western blotting and Northern blotting, respectively. T(3) induced a marked increase in the beta(1) protein and a slight increase in the alpha(1) protein. T(3) did not affect expression of the beta(2) protein. The alpha(2) protein was not detected in either untreated or T(3)-treated cells. In contrast, alpha(2) mRNA was highly up-regulated by T(3) treatment compared to the other isoforms. The lack of expression of the alpha(2) protein isoform following T(3) treatment suggests that posttranscriptional events related to this isoform may be dependent on other growth factors or hormones.
Sharabani-Yosef
Orna
O
Faculty of Life Sciences, Gonda-Goldschmeid Center, Bar-Ilan University, Ramat-Gan 52900, Israel.
Nir
Uri
U
Sampson
Sanford R
SR
D000818:Animals; D046508:Culture Techniques; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D007527:Isoenzymes / Q000378:metabolism; D018482:Muscle, Skeletal / Q000187:drug effects* / Q000201:enzymology; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D000254:Sodium-Potassium-Exchanging ATPase / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D014284:Triiodothyronine / Q000494:pharmacology*
D007527:Isoenzymes; D012333:RNA, Messenger; D014284:Triiodothyronine; C498532:Atp1a2 protein, rat; D000254:Sodium-Potassium-Exchanging ATPase