Skip to main content
Top
Published in: European Archives of Oto-Rhino-Laryngology 3/2004

01-03-2004 | Otology

Evidence for a possible NOS back-up system in the organ of Corti of the guinea pig

Authors: Ulf-Rüdiger Heinrich, Jan Maurer, Wolf Mann

Published in: European Archives of Oto-Rhino-Laryngology | Issue 3/2004

Login to get access

Abstract

Recently, the two Ca2+/calmodulin-regulated nitric oxide synthase isoforms, nNOS and eNOS, and NO itself have been identified in the cochlea of vertebrates using specific antibodies and a new fluorescence indicator. In order to acquire more information about the quantitative and spatial distribution of these two constitutively expressed NOS isoforms (cNOS) in the organ of Corti at the cellular and subcelluar levels, ultrathin sections of London resin (LR) White-embedded cochleae of the guinea pig were incubated with various concentrations of commercially available antibodies to nNOS and eNOS. The immunoreactivity was visualized by a gold-labeled secondary antibody and the amount of the immunoreactions/μm2 was quantified for different cell types and subcellular regions. Both NOS isoforms were identified to varying degrees in the same cell types and subcellular regions. A prominent eNOS immunoreactivity was identified in nearly every cell type. In all analyzed animals the highest number of gold-coupled anti-eNOS antibodies was always seen in the cells of the reticular lamina, especially in the cuticular structures of outer and inner hair cells, pillar cells and apical Deiters' cells. Also the microtubuli-containing cytoplasmic regions of Deiters' cells were scattered with gold-coupled anti-eNOS antibodies. A clear eNOS immunoreaction was also found in the remaining cytoplasm of inner and outer hair cells and in the apical Deiters' cells. Numerous anti-nNOS antibodies were located in the outer hair cells and in the cuticular structures of the apical Deiters' cells. The amount of the gold-labeled anti-nNOS antibodies in the cuticular plates of the pillar cells and outer hair cells and in the cytoplasm of inner hair cells and apical Deiters' cells were clearly less but still above unspecific background labeling. The spatial co-localization of the two NOS isotypes in the same cell regions was proven in double-labeling experiments. The spatial distribution of the two cNOS isoforms confirmed recent findings of other authors who localized NO distribution and production sites. The cNOS co-expression with similar function in the same cell type and subcellular regions may represent a functional "back-up system" in which one NOS isoform can replace the other in case of pathophysiological malfunction.
Literature
1.
go back to reference Boissel JP, Schwarz PM, Förstermann U (1998) Neuronal-type NO synthase: transcript diversity and expressional regulation. Nitric Oxide 2:337–349CrossRefPubMed Boissel JP, Schwarz PM, Förstermann U (1998) Neuronal-type NO synthase: transcript diversity and expressional regulation. Nitric Oxide 2:337–349CrossRefPubMed
2.
go back to reference Brenman JE, Chao DS, Gee SH, McGee AW, Craven SE, Santillano DR, Wu Z, Huang F, Xia H, Peters MF, Froehner SC, Bredt DS (1996) Interaction of nitric oxide synthase with the postsynaptic density protein PSD-95 and alpha-syntrophin mediated by PDZ domains. Cell 84:757–767PubMed Brenman JE, Chao DS, Gee SH, McGee AW, Craven SE, Santillano DR, Wu Z, Huang F, Xia H, Peters MF, Froehner SC, Bredt DS (1996) Interaction of nitric oxide synthase with the postsynaptic density protein PSD-95 and alpha-syntrophin mediated by PDZ domains. Cell 84:757–767PubMed
3.
go back to reference Dallos P, He DZZ, Sziklai I, Metha S, Evans BN (1997) Acetylcholine, outer hair cell electromotility, and the cochlear amplifier. J Neurosci 17:2212–2226PubMed Dallos P, He DZZ, Sziklai I, Metha S, Evans BN (1997) Acetylcholine, outer hair cell electromotility, and the cochlear amplifier. J Neurosci 17:2212–2226PubMed
4.
go back to reference Fessenden JD, Schacht J (1997) Localization of soluble guanylate cyclase activity in the guinea pig cochlea suggests involvement in regulation of blood flow and supporting cell physiology. J Histochem Cytochem 45:1401–1408PubMed Fessenden JD, Schacht J (1997) Localization of soluble guanylate cyclase activity in the guinea pig cochlea suggests involvement in regulation of blood flow and supporting cell physiology. J Histochem Cytochem 45:1401–1408PubMed
5.
go back to reference Fessenden JD, Schacht J (1998) The nitric oxide/cyclic GMP pathway: a potential major regulator of cochlear physiology. Hear Res 118:168–176CrossRefPubMed Fessenden JD, Schacht J (1998) The nitric oxide/cyclic GMP pathway: a potential major regulator of cochlear physiology. Hear Res 118:168–176CrossRefPubMed
6.
go back to reference Fessenden JD, Atschuler RA, Seasholtz AF, Schacht J (1999) Nitric oxide/cyclic guanosine monophosphate pathway in the peripheral and central auditory system of the rat. J Comp Neurology 404:52–63CrossRef Fessenden JD, Atschuler RA, Seasholtz AF, Schacht J (1999) Nitric oxide/cyclic guanosine monophosphate pathway in the peripheral and central auditory system of the rat. J Comp Neurology 404:52–63CrossRef
7.
go back to reference Flock Å, Block B, Fridberger A, Scarfone E, Ulfendahl M (1999) Supporting cells contribute to control of hearing sensitivity. J Neurosci 19:4498–4507PubMed Flock Å, Block B, Fridberger A, Scarfone E, Ulfendahl M (1999) Supporting cells contribute to control of hearing sensitivity. J Neurosci 19:4498–4507PubMed
8.
go back to reference Förstermann U, Kleinert H (1995) Nitric oxide synthase: expression and expressional control of the three isoforms. Naunyn-Schmiedebergs Arch Pharmacol 352:351–364 Förstermann U, Kleinert H (1995) Nitric oxide synthase: expression and expressional control of the three isoforms. Naunyn-Schmiedebergs Arch Pharmacol 352:351–364
9.
go back to reference Förstermann U, Boissel J-P, Kleinert H (1998) Expressional control of the constitutive isoforms of nitric oxide synthase (nNOS and eNOS). FASEB J 12:773–779PubMed Förstermann U, Boissel J-P, Kleinert H (1998) Expressional control of the constitutive isoforms of nitric oxide synthase (nNOS and eNOS). FASEB J 12:773–779PubMed
10.
go back to reference Franz P, Hauser-Kronberger C, Böck P, Quint C, Baumgartner WD (1996) Localization of nitric oxide synthase I and III in the cochlea. Acta Otolaryngol (Stockh) 116:726–731 Franz P, Hauser-Kronberger C, Böck P, Quint C, Baumgartner WD (1996) Localization of nitric oxide synthase I and III in the cochlea. Acta Otolaryngol (Stockh) 116:726–731
11.
go back to reference Frolenkov GI, Atzori M, Kalinec F, Mammano F (1998) The membrane-based mechanism of cell motility in cochlear outer hair cells. Mol Biol Cell 9:1961–1968PubMed Frolenkov GI, Atzori M, Kalinec F, Mammano F (1998) The membrane-based mechanism of cell motility in cochlear outer hair cells. Mol Biol Cell 9:1961–1968PubMed
12.
go back to reference Fulton D, Gratton J-P, Sessa WC (2001) Post-translational control of endothelial nitric oxide synthase: Why isn't calcium/calmodulin enough? J Pharmacol Exp Therapeut 299:818–824 Fulton D, Gratton J-P, Sessa WC (2001) Post-translational control of endothelial nitric oxide synthase: Why isn't calcium/calmodulin enough? J Pharmacol Exp Therapeut 299:818–824
13.
go back to reference Gosepath K, Gath I, Maurer J, Pollock JS, Amedee R, Förstermann U, Mann W (1997) Characterization of NO synthase isoforms expressed in different structures of the guinea pig cochlea. Brain Res 747:26–33CrossRefPubMed Gosepath K, Gath I, Maurer J, Pollock JS, Amedee R, Förstermann U, Mann W (1997) Characterization of NO synthase isoforms expressed in different structures of the guinea pig cochlea. Brain Res 747:26–33CrossRefPubMed
14.
go back to reference Heinrich U-R, Maurer J, Gosepath K, Mann W (1997) Electron microscopic localization of nitric oxide I synthase in the organ of Corti of the guinea pig. Eur Arch Otorhinolaryngol 254:396–400PubMed Heinrich U-R, Maurer J, Gosepath K, Mann W (1997) Electron microscopic localization of nitric oxide I synthase in the organ of Corti of the guinea pig. Eur Arch Otorhinolaryngol 254:396–400PubMed
15.
go back to reference Heinrich U-R, Maurer J, Gosepath K, Mann W (1998a) Immunoelectron microscopic localization of nitric oxide synthase III in the guinea pig organ of Corti. Eur Arch Otorhinolaryngol 255:483–490CrossRefPubMed Heinrich U-R, Maurer J, Gosepath K, Mann W (1998a) Immunoelectron microscopic localization of nitric oxide synthase III in the guinea pig organ of Corti. Eur Arch Otorhinolaryngol 255:483–490CrossRefPubMed
16.
go back to reference Heinrich U-R, Maurer J, Mann W (1998b) Possible Ca2+ -dependent mechanism of apical outer hair cell modulation within the cochlea of the guinea pig. Cell Tiss Res 292:57–65CrossRef Heinrich U-R, Maurer J, Mann W (1998b) Possible Ca2+ -dependent mechanism of apical outer hair cell modulation within the cochlea of the guinea pig. Cell Tiss Res 292:57–65CrossRef
17.
go back to reference Heinrich U-R, Maurer J, Koesling D, MannW, Förstermann U (2000) Immuno-electron microscopic localization of the α1 and β1 subunits of soluble guanylyl cyclase in the guinea pig organ of Corti. Brain Res 885:6–13CrossRefPubMed Heinrich U-R, Maurer J, Koesling D, MannW, Förstermann U (2000) Immuno-electron microscopic localization of the α1 and β1 subunits of soluble guanylyl cyclase in the guinea pig organ of Corti. Brain Res 885:6–13CrossRefPubMed
18.
go back to reference Hess A, Bloch W, Arnhold S, Andressen C, Stennert E, Addicks K, Michel O (1998) Nitric oxide synthase in the vestibulocochlear system of mice. Brain Res 813:97–102CrossRefPubMed Hess A, Bloch W, Arnhold S, Andressen C, Stennert E, Addicks K, Michel O (1998) Nitric oxide synthase in the vestibulocochlear system of mice. Brain Res 813:97–102CrossRefPubMed
19.
go back to reference Kachar B, Brownell WE, Altschuler R, Fex J (1986) Elektrokinetic shape changes of cochlear outer hair cells. Nature 322:365–368PubMed Kachar B, Brownell WE, Altschuler R, Fex J (1986) Elektrokinetic shape changes of cochlear outer hair cells. Nature 322:365–368PubMed
20.
go back to reference Kojima H, Nakatsubo N, Kikuchi K, Urano Y, Higuchi T, Tanaka J, Kudo Y, Nagano T (1998) Direct evidence of NO production in rat hippocampus and cortex using a new fluorescent indicator: DAF-2 DA. Neuro Report 9:3345–3348 Kojima H, Nakatsubo N, Kikuchi K, Urano Y, Higuchi T, Tanaka J, Kudo Y, Nagano T (1998) Direct evidence of NO production in rat hippocampus and cortex using a new fluorescent indicator: DAF-2 DA. Neuro Report 9:3345–3348
21.
go back to reference Lagostena L, Ashmore JF, Kachar B, Mammano F (2001) Purinergic control of intercellular communication bentween Hensen's cells of the guinea-pig cochlea. J Physiol 531:693–706PubMed Lagostena L, Ashmore JF, Kachar B, Mammano F (2001) Purinergic control of intercellular communication bentween Hensen's cells of the guinea-pig cochlea. J Physiol 531:693–706PubMed
22.
go back to reference Lenzi D, Roberts WM (1994) Calcium signalling in hair cells: multiple roles in a compact cell. Curr Opin Neurobiol 4:496–502PubMed Lenzi D, Roberts WM (1994) Calcium signalling in hair cells: multiple roles in a compact cell. Curr Opin Neurobiol 4:496–502PubMed
23.
go back to reference Li H, Wallerath T, Förstermann U (2002) Physiological mechanisms regulating the expression of endothelial-type NO synthase. Nitric Oxide Biol Chem 7:1432–1476 Li H, Wallerath T, Förstermann U (2002) Physiological mechanisms regulating the expression of endothelial-type NO synthase. Nitric Oxide Biol Chem 7:1432–1476
24.
go back to reference Matsunobu T, Schacht J (2000) Nitric oxide/cyclic GMP pathway attenuates ATP-evoked intracellular calcium increase in supporting cells of the guinea pig cochlea. J Comp Neurol 423:452–461CrossRefPubMed Matsunobu T, Schacht J (2000) Nitric oxide/cyclic GMP pathway attenuates ATP-evoked intracellular calcium increase in supporting cells of the guinea pig cochlea. J Comp Neurol 423:452–461CrossRefPubMed
25.
go back to reference Michel O, Hess A, Bloch W, Stennert E, Su J, Addicks K (1999) Localization of the NO/cGMP-pathway in the cochlea of guinea pigs. Hear Res 133:1–9CrossRefPubMed Michel O, Hess A, Bloch W, Stennert E, Su J, Addicks K (1999) Localization of the NO/cGMP-pathway in the cochlea of guinea pigs. Hear Res 133:1–9CrossRefPubMed
26.
go back to reference Nobili R, Mammano F, Ashmore JF (1998) How well do we understand the cochlea? Trends Neurosci 21:159–167 Nobili R, Mammano F, Ashmore JF (1998) How well do we understand the cochlea? Trends Neurosci 21:159–167
27.
go back to reference Popa R, Anniko M, Takumida M, Arnold W (2001) Localization of nitric oxide synthase isoforms in the human cochlea. Acta Otolaryngol 121:454–459CrossRefPubMed Popa R, Anniko M, Takumida M, Arnold W (2001) Localization of nitric oxide synthase isoforms in the human cochlea. Acta Otolaryngol 121:454–459CrossRefPubMed
28.
go back to reference Ricci AJ, Fettiplace R (1998)Calcium permeation of the turtle hair cell mechanotransducer channel and its relation to the composition of endolymph. J Physiol 506:159–173PubMed Ricci AJ, Fettiplace R (1998)Calcium permeation of the turtle hair cell mechanotransducer channel and its relation to the composition of endolymph. J Physiol 506:159–173PubMed
29.
go back to reference Ruan R-S (2002) Possible roles of nitric oxide in the physiology and pathophysiology of the mammalian cochlea. Ann NY Acad Sci 962:260–274PubMed Ruan R-S (2002) Possible roles of nitric oxide in the physiology and pathophysiology of the mammalian cochlea. Ann NY Acad Sci 962:260–274PubMed
30.
go back to reference Shi X, Ren T, Nuttall AL (2001) Nitric oxide distribution and production in the guinea pig cochlea. Hear Res 153:23–31CrossRefPubMed Shi X, Ren T, Nuttall AL (2001) Nitric oxide distribution and production in the guinea pig cochlea. Hear Res 153:23–31CrossRefPubMed
31.
go back to reference Shi X, Ren T, Nuttall AL (2002) The electrochemical and fluorescence detection of nitric oxide in the cochlea and its increase following loud sound. Hear Res 164:49–58PubMed Shi X, Ren T, Nuttall AL (2002) The electrochemical and fluorescence detection of nitric oxide in the cochlea and its increase following loud sound. Hear Res 164:49–58PubMed
32.
go back to reference Sönyi M, He DZZ, Ribári O, Sziklai I, Dallos P (1999) Cyclic GMP and outer hair cell electromotility. Hear Res 137:29–42CrossRefPubMed Sönyi M, He DZZ, Ribári O, Sziklai I, Dallos P (1999) Cyclic GMP and outer hair cell electromotility. Hear Res 137:29–42CrossRefPubMed
33.
go back to reference Son H, Hawkins RD, Martin K, Kiebler M, Huang PL, Fishman MC, Kandel ER (1996) Long-term potentiation is reduced in mice that are doubly mutant in endothelial and neuronal nitric oxide synthase. Cell 87:1015–1023PubMed Son H, Hawkins RD, Martin K, Kiebler M, Huang PL, Fishman MC, Kandel ER (1996) Long-term potentiation is reduced in mice that are doubly mutant in endothelial and neuronal nitric oxide synthase. Cell 87:1015–1023PubMed
34.
go back to reference Takumida A, Anniko M (2001a) Direct evidence of nitric oxide production in the guinea pig organ of Corti. Acta Otolaryngol 121:342–345CrossRefPubMed Takumida A, Anniko M (2001a) Direct evidence of nitric oxide production in the guinea pig organ of Corti. Acta Otolaryngol 121:342–345CrossRefPubMed
35.
go back to reference Takumida A, Anniko M (2001b) Detection of nitric oxide in the guinea pig inner ear, using a combination of aldehyde fixative and 4,5 diaminofluorescein diaceate. Acta Otolaryngol 121:460–464CrossRefPubMed Takumida A, Anniko M (2001b) Detection of nitric oxide in the guinea pig inner ear, using a combination of aldehyde fixative and 4,5 diaminofluorescein diaceate. Acta Otolaryngol 121:460–464CrossRefPubMed
36.
go back to reference Tian F, Fessenden JD, Schacht J (1999) Cyclic GMP-dependent protein kinase-I in the guinea pig cochlea. Hear Res 131:63–70CrossRefPubMed Tian F, Fessenden JD, Schacht J (1999) Cyclic GMP-dependent protein kinase-I in the guinea pig cochlea. Hear Res 131:63–70CrossRefPubMed
Metadata
Title
Evidence for a possible NOS back-up system in the organ of Corti of the guinea pig
Authors
Ulf-Rüdiger Heinrich
Jan Maurer
Wolf Mann
Publication date
01-03-2004
Publisher
Springer-Verlag
Published in
European Archives of Oto-Rhino-Laryngology / Issue 3/2004
Print ISSN: 0937-4477
Electronic ISSN: 1434-4726
DOI
https://doi.org/10.1007/s00405-003-0648-1

Other articles of this Issue 3/2004

European Archives of Oto-Rhino-Laryngology 3/2004 Go to the issue