Thyroid hormone modulates ClC-2 chloride channel gene expression in rat renal proximal tubules.
Thyroid hormones has its main role in controlling metabolism, but it can also modulate extracellular fluid Volume (ECFV) through its action on the expression and activity of Na(+) transporters. Otherwise, chloride is the main anion in the ECFV and the influence of thyroid hormones in the regulation of chloride transporters is not yet understood. In this work, we studied the effect of thyroid hormones in the expression of ClC-2, a cell Volume-, pH- and voltage-sensitive Cl(-) channel, in rat kidney. To analyze the modulation of ClC-2 gene expression by thyroid hormones, we used hypothyroid (Hypo) rats with or without thyroxine (T(4)) replacement and hyperthyroid (Hyper) rats as our experimental models. Total RNA was isolated and the expression of ClC-2 mRNA was evaluated by a ribonuclease protection assay, and/or semi-quantitative RT-PCR. Renal ClC-2 expression decreased in Hypo rats and increased in Hyper rats. In addition, semi-quantitative RT-PCR of different nephron segments showed that these changes were due exclusively to the modulation of ClC-2 mRNA expression by thyroid hormone in convoluted and straight proximal tubules. To investigate whether thyroid hormones action was direct or indirect, renal proximal tubule primary culture cells were prepared and subjected to different T(4) concentrations. ClC-2 mRNA expression was increased by T(4) in a dose-dependent fashion, as analyzed by RT-PCR. Western blotting demonstrated that ClC-2 protein expression followed the same profile of mRNA expression.
Santos Ornellas
D
D
Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, RJ, Brazil, CEP 21949-900.
Grozovsky
R
R
Goldenberg
R C
RC
Carvalho
D P
DP
Fong
P
P
Guggino
W B
WB
Morales
M
M
D000818:Animals; D015153:Blotting, Western / Q000379:methods; D000090007:CLC-2 Chloride Channels; D002478:Cells, Cultured; D018118:Chloride Channels / Q000032:analysis / Q000235:genetics*; D005786:Gene Expression Regulation / Q000187:drug effects; D006980:Hyperthyroidism / Q000378:metabolism*; D007037:Hypothyroidism / Q000378:metabolism*; D007687:Kidney Tubules, Proximal / Q000378:metabolism*; D008297:Male; D023421:Models, Animal; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017208:Rats, Wistar; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013963:Thyroid Hormones / Q000502:physiology*; D013974:Thyroxine / Q000494:pharmacology
D000090007:CLC-2 Chloride Channels; D018118:Chloride Channels; D012333:RNA, Messenger; D013963:Thyroid Hormones; D013974:Thyroxine