Expression and localization of the Na+-H+ exchanger in the guinea pig cochlea

Physiological studies have shown that the Na+-H+ exchanger (NHE) is a major carrier protein regulating the intracellular pH in the cells of the cochlea. The presence of multiple forms of the exchanger has been demonstrated by the recent cloning of four mammalian NHEs, NHE-1 to NHE-4. Despite the structural similarity, these NHE isoforms differ in their tissue distribution, kinetic characteristics, and responses to external stimuli. The present study was undertaken to examine the expression and distribution of four NHE isoforms in the guinea pig cochlea. We used reverse transcription-polymerase chain reaction to assess the expression of NHE-1-4 isoforms and non-radioactive in situ hybridization to examine their localization. Although NHE-2, -3 and -4 isoform mRNAs could be detected in the cochlear tissue, the NHE-1 message was predominant. Cloned guinea pig NHE-1-4 partial cDNA fragments were highly homologous to the corresponding rat NHE isoforms. NHE-1 isoform mRNA was distributed in the hair cells, marginal cells, spiral ligament fibrocytes, spiral prominence cells and spiral ganglion cells. NHE- localized in a variety of cochlear cells would contribute to their differential function.

[1]  J. T. Corwin,et al.  The developing organ of Corti contains retinoic acid and forms supernumerary hair cells in response to exogenous retinoic acid in culture. , 1993, Development.

[2]  J. Pouysségur,et al.  Cloning and expression of a rabbit cDNA encoding a serum-activated ethylisopropylamiloride-resistant epithelial Na+/H+ exchanger isoform (NHE-2). , 1993, The Journal of biological chemistry.

[3]  T. Takasaka,et al.  Expression of voltage-dependent chloride channels in the rat cochlea , 1997, Hearing Research.

[4]  J. Pouysségur,et al.  Functional expression of a human Na+/H+ antiporter gene transfected into antiporter-deficient mouse L cells. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[5]  T. Takasaka,et al.  Involvement of Na(+)-H+ exchange in intracellular pH recovery from acid load in the stria vascularis of the guinea-pig cochlea. , 1994, Acta oto-laryngologica.

[6]  S. Brant,et al.  Cloning and sequencing of a rabbit cDNA encoding an intestinal and kidney-specific Na+/H+ exchanger isoform (NHE-3). , 1992, The Journal of biological chemistry.

[7]  K. Fujimori,et al.  Enhanced expression of a glyceraldehyde-3-phosphate dehydrogenase gene in human lung cancers. , 1987, Cancer research.

[8]  K. Ikeda,et al.  Nonselective cation and Cl channels in luminal membrane of the marginal cell. , 1993, The American journal of physiology.

[9]  T Takasaka,et al.  Intracellular pH regulation in isolated cochlear outer hair cells of the guinea‐pig. , 1992, The Journal of physiology.

[10]  J. T. Corwin,et al.  Regenerative proliferation in inner ear sensory epithelia from adult guinea pigs and humans. , 1993, Science.

[11]  S Grinstein,et al.  Na+/H+ exchange and growth factor-induced cytosolic pH changes. Role in cellular proliferation. , 1989, Biochimica et biophysica acta.

[12]  S. L. Bonting,et al.  Studies on (Na+-K+)-activated ATPase. XXIV. Localization and properties of ATPase in the inner ear of the guinea pig. , 1969, Biochimica et biophysica acta.

[13]  M. Donowitz,et al.  Structure/function studies of mammalian Na‐H exchangers‐‐an update. , 1995, The Journal of physiology.

[14]  D. Copenhagen,et al.  Modulation of neuronal function by intracellular pH , 1996, Neuroscience Research.

[15]  S. Spicer,et al.  The fine structure of spiral ligament cells relates to ion return to the stria and varies with place-frequency , 1996, Hearing Research.

[16]  M. Leppert,et al.  Molecular cloning and physical and genetic mapping of a novel human Na+/H+ exchanger (NHE5/SLC9A5) to chromosome 16q22.1. , 1995, Genomics.

[17]  P. Aronson Kinetic properties of the plasma membrane Na+-H+ exchanger. , 1985, Annual review of physiology.

[18]  C. Sardet,et al.  Molecular cloning, primary structure, and expression of the human growth factor-activatable Na+ H+ , 1989, Cell.

[19]  G. Shull,et al.  Molecular cloning of putative members of the Na/H exchanger gene family. cDNA cloning, deduced amino acid sequence, and mRNA tissue expression of the rat Na/H exchanger NHE-1 and two structurally related proteins. , 1992, The Journal of biological chemistry.

[20]  P. Wangemann,et al.  Vestibular dark cells contain the Na + H + exchanger NHE-1 in , 1996, Hearing Research.

[21]  T. Takasaka,et al.  The Na-K-Cl cotransporters in the rat cochlea: RT-PCR and partial sequence analysis. , 1996, Biochemical and biophysical research communications.

[22]  J. Pouysségur,et al.  Growth factor activation and "H(+)-sensing" of the Na+/H+ exchanger isoform 1 (NHE1). Evidence for an additional mechanism not requiring direct phosphorylation. , 1994, The Journal of biological chemistry.

[23]  S. Brant,et al.  Mammalian Na+/H+ exchanger gene family: structure and function studies. , 1995, The American journal of physiology.

[24]  C. Rose,et al.  pH regulation and proton signalling by glial cells , 1996, Progress in Neurobiology.

[25]  C. Sardet,et al.  Growth factors induce phosphorylation of the Na+/H+ antiporter, glycoprotein of 110 kD. , 1990, Science.

[26]  T. Takasaka,et al.  Alternatively spliced isoforms of the Na+/Ca2+ exchanger in the guinea pig cochlea. , 1997, Biochemical and biophysical research communications.

[27]  E. Ma,et al.  Expression and localization of Na+/H+ exchangers in rat central nervous system , 1997, Neuroscience.

[28]  G. Shull,et al.  Primary structure and functional expression of a novel gastrointestinal isoform of the rat Na/H exchanger. , 1993, The Journal of biological chemistry.

[29]  S. Grinstein,et al.  Activation of the Na+/H+ antiporter during cell volume regulation. Evidence for a phosphorylation-independent mechanism. , 1992, The Journal of biological chemistry.

[30]  J. Collins,et al.  Molecular cloning, sequencing, tissue distribution, and functional expression of a Na+/H+ exchanger (NHE-2). , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[31]  P. Bernd,et al.  Nerve growth factor and serum differentially regulate development of the embryonic otic vesicle and cochleovestibular ganglion in vitro. , 1989, Developmental biology.

[32]  R. Macleod,et al.  Activation of Na+/H+ Exchange Is Required for Regulatory Volume Decrease after Modest “Physiological” Volume Increases in Jejunal Villus Epithelial Cells* , 1996, The Journal of Biological Chemistry.

[33]  P. Wilson,et al.  NHE2 and NHE3 are human and rabbit intestinal brush-border proteins. , 1996, The American journal of physiology.

[34]  W. Frankel,et al.  Sodium/Hydrogen Exchanger Gene Defect in Slow-Wave Epilepsy Mutant Mice , 1997, Cell.

[35]  S. Brant,et al.  Cloning, tissue distribution, and functional analysis of the human Na+/N+ exchanger isoform, NHE3. , 1995, The American journal of physiology.

[36]  P. Wangemann,et al.  The Na+/H+ exchanger in transitional cells of the inner ear , 1993, Hearing Research.

[37]  G. Moonen,et al.  Retinoic acid stimulates regeneration of mammalian auditory hair cells. , 1993, Science.

[38]  Jochen Schacht,et al.  Homeostatic Mechanisms in the Cochlea , 1996 .