Molecular and functional evidence for Na+-K+-2Cl- cotransporter expression in rat skeletal muscle

Jennifer A. Wong, Liusong Fu, Edward G. Schneider, Donald Thomason

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36 Citations (Scopus)

Abstract

Doubt has been raised about the expression of a functional Na+-K+2Cl- cotransporter in rat skeletal muscle. In this study we present molecular and functional evidence for expression of a protein having the characteristics of a cotransporter. RT-PCR of RNA isolated from rat soleus muscle with primers to a conserved putative membrane-spanning domain resulted in a single product of predicted size. Sequencing of the product showed that it bears >90% homology with known rodent NKCC1 (BSC2) cotransporters. RNase protection assay of RNA isolated from the rat soleus muscle also identified this sequence. Immunologic detection of the cotransporter with two different antibodies indicated the presence of cotransporter protein, perhaps more than one, in blots of total muscle protein. Immunohistochemical detection by confocal microscopy localized the majority of expression of the protein to the muscle fibers. Functional studies of cotransport activity also indicate the appropriate sensitivity to inhibitors and ion dependence. Taken together, these data support the presence and function of Na+-K+-2Cl- cotransporter activity in the soleus muscle of the rat.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Regulatory Integrative and Comparative Physiology
Volume277
Issue number1 46-1
StatePublished - Jul 1 1999

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Skeletal Muscle
Muscle Proteins
Member 2 Solute Carrier Family 12
RNA
Ribonucleases
Confocal Microscopy
Rodentia
Proteins
Ions
Polymerase Chain Reaction
Membranes
Antibodies

All Science Journal Classification (ASJC) codes

  • Physiology
  • Physiology (medical)

Cite this

Molecular and functional evidence for Na+-K+-2Cl- cotransporter expression in rat skeletal muscle. / Wong, Jennifer A.; Fu, Liusong; Schneider, Edward G.; Thomason, Donald.

In: American Journal of Physiology - Regulatory Integrative and Comparative Physiology, Vol. 277, No. 1 46-1, 01.07.1999.

Research output: Contribution to journalArticle

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