Molecular and functional characterization of sodium-hydrogen exchanger in skin as well as cultured keratinocytes and melanocytes

Rangaprasad Sarangarajan*, Holli Shumaker, Manoocher Soleimani, Caroline Le Poole, Raymond E. Boissy

*Corresponding author for this work

Research output: Contribution to journalArticle

29 Scopus citations

Abstract

The sodium-hydrogen (Na+/H+) exchanger is one of the few transporter proteins involved in the regulation and maintenance of intracellular pH and cell volume in most eukaryotic cell types. The current study investigates the expression of isoforms of the Na+/H+ exchanger (NHE) in human skin and in cultured keratinocytes, melanocytes, and melanoma cells by reverse transcription-polymerase chain reaction (RT-PCR), immunohistochemical analysis and functional studies. Neonatal foreskins were used to isolate RNA from epidermis and dermis, and to initiate cultures of keratinocytes and melanocytes. RT-PCR on RNA isolated from epidermis, dermis, keratinocytes, melanocytes and melanoma cells using PCR primers specific for NHE-1 yielded a 463 bp PCR product. RT-PCR performed using primers specific for NHE isoforms 2, 3, 4 and 5 did not yield any products. Western blotting analysis (of keratinocyte and melanocyte cell cultures) and indirect immunohistochemistry on neonatal foreskin, keratinocytes, melanocytes and melanoma cells using a NHE-1-specific polyclonal antibody demonstrated NHE-1 expression at the protein level. Physiological regulation of intracellular pH using a pH-sensitive dye, BCECF, detected an amiloride-sensitive NHE activity in human keratinocyte, melanocyte and melanoma cell cultures. These results indicate that cultures of human keratinocytes and melanocytes established from human skin and melanoma cells express the NHE-1 isoform of the sodium-hydrogen exchanger.

Original languageEnglish (US)
Pages (from-to)181-192
Number of pages12
JournalBiochimica et Biophysica Acta - Biomembranes
Volume1511
Issue number1
DOIs
StatePublished - Mar 9 2001

Keywords

  • (Skin)
  • Keratinocyte
  • Melanocyte
  • Melanoma
  • Na/H exchanger

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Cell Biology

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