Skip to main content
SpringerLink
Log in

Hypothermia translocates nitric oxide synthase from cytosol to membrane in snail neurons

  • Regular Article
  • Published:
Cell and Tissue Research Aims and scope Submit manuscript

Abstract

Neuronal nitric oxide (NO) levels are modulated through the control of catalytic activity of NO synthase (NOS). Although signals limiting excess NO synthesis are being extensively studied in the vertebrate nervous system, our knowledge is rather limited on the control of NOS in neurons of invertebrates. We have previously reported a transient inactivation of NOS in hibernating snails. In the present study, we aimed to understand the mechanism leading to blocked NO production during hypothermic periods of Helix pomatia. We have found that hypothermic challenge translocated NOS from the cytosol to the perinuclear endoplasmic reticulum, and that this cytosol to membrane trafficking was essential for inhibition of NO synthesis. Cold stress also downregulated NOS mRNA levels in snail neurons, although the amount of NOS protein remained unaffected in response to hypothermia. Our studies with cultured neurons and glia cells revealed that glia–neuron signaling may inhibit membrane binding and inactivation of NOS. We provide evidence that hypothermia keeps NO synthesis “hibernated” through subcellular redistribution of NOS.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Abbreviations

BH4:

Tetrahydrobiopterin

CNS:

Central nervous system (cerebral and subesophageal ganglia)

CYTRI :

Relative intensity of cytoplasmic NADPHd staining

DIC:

Differential interference contrast

DXM:

Dexamethasone

FMN:

Flavin mononucleotide

FrK:

Forskolin

FAD:

Flavin adenin dinucleotide

HelixNOS:

Helix pomatia neuronal NOS

NMDA:

N-methyl-D-aspartate

NADPH:

Reduced nicotinamide-dinucleotide-phosphate

NADPHd:

NADPH diaphorase

NOS:

Nitric oxide synthase

PNRRI :

Relative intensity of perinuclear NADPHd staining

QRC:

Quercetin

References

  • Aras-López R, Xavier FE, Ferrer M, Balfagón G (2009) Dexamethasone decreases neuronal nitric oxide release in mesenteric arteries from hypertensive rats through decreased protein kinase C activation. Clin Sci (Lond) 117:305–312

    Article  Google Scholar 

  • Biber K, Neumann H, Inoue K, Boddeke HW (2007) Neuronal ‘On’ and ‘Off’ signals control microglia. Trends Neurosci 30:596–602

    Article  CAS  PubMed  Google Scholar 

  • Bicker G (2001) Sources and targets of nitric oxide signalling in insect nervous systems. Cell Tissue Res 303:137–146

    Article  CAS  PubMed  Google Scholar 

  • Bishop CD, Pires A, Norby SW, Boudko D, Moroz LL, Hadfield MG (2008) Analysis of nitric oxide-cyclic guanosine monophosphate signaling during metamorphosis of the nudibranch Phestilla sibogae Bergh (Gastropoda: Opisthobranchia). Evol Dev 10:288–299

    Article  CAS  PubMed  Google Scholar 

  • Bodnárová M, Martásek P, Moroz LL (2005) Calcium/calmodulin-dependent nitric oxide synthase activity in the CNS of Aplysia californica: biochemical characterization and link to cGMP pathways. J Inorg Biochem 99:922–928

    Article  PubMed  Google Scholar 

  • Boehning D, Snyder SH (2003) Novel neural modulators. Annu Rev Neurosci 26:105–131

    Article  CAS  PubMed  Google Scholar 

  • Bolshakov VYu, Gapon SA, Magazanik LG (1992) Transduction mechanism for glutamate-induced potassium current in neurones of the mollusc Planorbarius corneus. J Physiol 455:33–50

    CAS  PubMed  Google Scholar 

  • Bougie JK, Lim T, Farah CA, Manjunath V, Nagakura I, Ferraro GB, Sossin WS (2009) The atypical protein kinase C in Aplysia can form a protein kinase M by cleavage. J Neurochem 109:1129–1143

    Article  CAS  PubMed  Google Scholar 

  • Bredt DS, Snyder SH (1992) Nitric oxide, a novel neuronal messenger. Neuron 8:3–11

    Article  CAS  PubMed  Google Scholar 

  • Broillet MC (1999) S-nitrosylation of proteins. Cell Mol Life Sci 55:1036–1042

    Article  CAS  PubMed  Google Scholar 

  • Cohan CS, Karnes JL, Zhou FQ (2003) Culturing neurons from the snail Helisoma. Methods Cell Biol 71:157–170

    Article  PubMed  Google Scholar 

  • Cooke IRC, Edwards SL, Anderson CR (1994) The distribution of NADPH diaphorase activity and immunoreactivity to nitric oxide synthase in the nervous system of the pulmonate mollusc Helix aspersa. Cell Tissue Res 277:565–572

    Article  CAS  Google Scholar 

  • Cristino LV, Guglielmotti V, Cotugno A, Musio C, Santillo S (2008) Nitric oxide signaling pathways at neural level in invertebrates: functional implications in cnidarians. Brain Res 1225:17–25

    Article  CAS  PubMed  Google Scholar 

  • Davies SA (2006) Signalling via cGMP: lessons from Drosophila. Cell Signal 18:409–421

    Article  CAS  PubMed  Google Scholar 

  • Elphick M, Williams L, Shea M (1996) New features of the locust optic lobe: evidence of a role for nitric oxide in insect vision. J Exp Biol Pt 11:2395–23407

    Google Scholar 

  • Gelperin A (1994) Nitric oxide mediates network oscillations of olfactory interneurons in a terrestrial mollusc. Nature 369:61–63

    Article  CAS  PubMed  Google Scholar 

  • Gervasi N, Tchénio P, Preat T (2010) PKA dynamics in a Drosophila learning center: coincidence detection by rutabaga adenylyl cyclase and spatial regulation by dunce phosphodiesterase. Neuron 65:516–529

    Article  CAS  PubMed  Google Scholar 

  • Giraldi-Guimarães A, Batista CM, Carneiro K, Tenório F, Cavalcante LA, Mendez-Otero R (2007) A critical survey on nitric oxide synthase expression and nitric oxide function in the retinotectal system. Brain Res Rev 56:403–426

    Article  PubMed  Google Scholar 

  • Gschwendt M, Horn F, Kittstein W, Marks F (1983) Inhibition of the calcium- and phospholipid-dependent protein kinase activity from mouse brain cytosol by quercetin. Biochem Biophys Res Commun 117:444–447

    Article  CAS  PubMed  Google Scholar 

  • Guevara I, Iwanejko J, Dembinska-Kiec A, Pankiewicz J, Wanat A, Anna P, Golabek I, Bartus S, Malczewska-Malec M, Szczudlik A (1998) Determination of nitrite/nitrate in human biological material by the simple Griess reaction. Clin Chim Acta 274:177–188

    Article  CAS  PubMed  Google Scholar 

  • Halm MP, Chichery MP, Chichery R (2003) Effect of nitric oxide synthase inhibition on the manipulative behaviour of Sepia officinalis. Comp Biochem Physiol C Toxicol Pharmacol 134:139–146

    Article  CAS  PubMed  Google Scholar 

  • Hatcher NG, Sudlow LC, Moroz LL, Gillette R (2006) Nitric oxide potentiates cAMP-gated cation current in feeding neurons of Pleurobranchaea californica independent of cAMP and cGMP signaling pathways. J Neurophysiol 95:3219–3227

    CAS  PubMed  Google Scholar 

  • Hegde AN (2010) The ubiquitin-proteasome pathway and synaptic plasticity. Learn Mem 17:314–327

    Article  PubMed  Google Scholar 

  • Hirata K, Kuroda R, Sakoda T, Katayama M, Inoue N, Suematsu M, Kawashima S, Yokoyama M (1995) Inhibition of endothelial nitric oxide synthase activity by protein kinase C. Hypertension 25:180–185

    CAS  PubMed  Google Scholar 

  • Huang S, Kerschbaum NH, Engel E, Hermann A (1997) Biochemicl characteriztion and histochemicl localization of nitric oxide synthase in the nervous system of the snail Helix pomatia. J Neurochem 69:2516–2528

    Article  CAS  PubMed  Google Scholar 

  • Jacklet JW (1997) Nitric oxide signaling in invertebrates. Invertebr Neurosci 3:1–14

    Article  CAS  Google Scholar 

  • Jiang J, Cyr D, Babbitt RW, Sessa WC, Patterson C (2003) Chaperone-dependent regulation of endothelial nitric-oxide synthase intracellular trafficking by the co-chaperone/ubiquitin ligase CHIP. J Biol Chem 278:49332–49341

    Article  CAS  PubMed  Google Scholar 

  • Kim S, Gailite I, Moussian B, Luschnig S, Goette M, Fricke K, Honemann-Capito M, Grubmüller H, Wodarz A (2009) Kinase-activity-independent functions of atypical protein kinase C in Drosophila. J Cell Sci 122:3759–3771

    Article  CAS  PubMed  Google Scholar 

  • Kojima S, Ogawa H, Kouuchi T, Nidaira T, Hosono T, Ito E (2000) Neuron-independent Ca(2+) signaling in glial cells of snail’s brain. Neuroscience 100:893–900

    Article  CAS  PubMed  Google Scholar 

  • Korneev SA, Piper MR, Picot J, Phillips R, Korneeva EI, O’Shea M (1998) Molecular characterization of NOS in a mollusc: expression in a giant modulatory neuron. J Neurobiol 35:65–76

    Article  CAS  PubMed  Google Scholar 

  • Kuang Z, Lewis RS, Curtis JM, Zhan Y, Saunders BM, Babon JJ, Kolesnik TB, Low A, Masters SL, Willson TA, Kedzierski L, Yao S, Handman E, Norton RS, Nicholson SE (2010) The SPRY domain-containing SOCS box protein SPSB2 targets iNOS for proteasomal degradation. J Cell Biol (in press)

  • Lieberman DN, Mody I (1999) Casein kinase-II regulates NMDA channel function in hippocampal neurons. Nat Neurosci 2:125–132

    Article  CAS  PubMed  Google Scholar 

  • Lin LH (2009) Glutamatergic neurons say NO in the nucleus tractus solitarii. J Chem Neuroanat 38:154–165

    Article  CAS  PubMed  Google Scholar 

  • Lipton SA (1999) Neuronal protection and destruction by NO. Cell Death Differ 6:943–951

    Article  CAS  PubMed  Google Scholar 

  • Magee AI (1990) Lipid modification of proteins and its relevance to protein targeting. J Cell Sci 97:581–584

    CAS  PubMed  Google Scholar 

  • Manseau F, Fan X, Hueftlein T, Sossin W, Castellucci VF (2001) Ca2+-independent protein kinase C Apl II mediates the serotonin-induced facilitation at depressed aplysia sensorimotor synapses. J Neurosci 21:1247–1256

    CAS  PubMed  Google Scholar 

  • Marchetti B, Serra PA, Tirolo C, L’episcopo F, Caniglia S, Gennuso F, Testa N, Miele E, Desole S, Barden N, Morale MC (2005) Glucocorticoid receptor-nitric oxide crosstalk and vulnerability to experimental parkinsonism: pivotal role for glia-neuron interactions. Brain Res Brain Res Rev 48:302–321

    Article  CAS  PubMed  Google Scholar 

  • Martínez A (1995) Nitric oxide synthase in invertebrates. Histochem J 27:770–776

    PubMed  Google Scholar 

  • Marzinzig M, Nussler AK, Stadler J, Marzinzig E, Barthlen W, Nussler NC, Beger HG, SMJr M, Bruckner UB (1997) Improved methods to measure end products of nitric oxide in biological fluids: nitrite, nitrate, S-nitrosothiols. Nitric Oxide 1:177–189

    Article  CAS  PubMed  Google Scholar 

  • Matsuo R, Ito E (2009) A novel nitric oxide synthase expressed specifically in the olfactory center. Biochem Biophys Res Commun 386:724–728

    Article  CAS  PubMed  Google Scholar 

  • Mattiello T, Fiore G, Brown ER, d’Ischia M, Palumbo A (2010) Nitric oxide mediates the glutamate-dependent pathway for neurotransmission in Sepia officinalis chromatophore organs. J Biol Chem (in press)

  • Mayer B (1994) Nitric oxide/cyclic GMP-mediated signal transduction. Ann NY Acad Sci 733:357–364

    Article  CAS  PubMed  Google Scholar 

  • Michel T, Li GK, Busconi L (1993) Phosphorylation and subcellular translocation of endothelial nitric oxide synthase. Proc Natl Acad Sci USA 90:6252–6256

    Article  CAS  PubMed  Google Scholar 

  • Moroz LL (2006) Localization of putative nitrergic neurons in peripheral chemosensory areas and the central nervous system of Aplysia californica. J Comp Neurol 495:10–20

    Article  CAS  PubMed  Google Scholar 

  • Moroz LL, Gillette R (1995) From Polyplacophora to Cephalopoda: comparative analysis of nitric oxide signalling in mollusca. Acta Biol Hung 46:169–182

    CAS  PubMed  Google Scholar 

  • Moroz LL, Chen D, Gillette MU, Gillette R (1996) Nitric oxide synthase activity in the molluscan CNS. J Neurochem 66:873–876

    Article  CAS  PubMed  Google Scholar 

  • Müller U, Bicker G (1994) Calcium-activated release of nitric oxide and cellular distribution of nitric oxide-synthesizing neurons in the nervous system of the locust. J Neurosci 14:7521–7528

    PubMed  Google Scholar 

  • Nakamura T, Lipton SA (2010) Preventing Ca2+-mediated nitrosative stress in neurodegenerative diseases: possible pharmacological strategies. Cell Calcium 47:190–197

    Article  CAS  PubMed  Google Scholar 

  • Newland PL, Yates P (2008) Nitric oxide modulates salt and sugar responses via different signaling pathways. Chem Senses 33:347–356

    Article  CAS  PubMed  Google Scholar 

  • Niederwieser A, Curtius HC (1970) Amino acid analysis in clinical chemistry. Z Klin Chem Klin Biochem 7:404–426

    CAS  PubMed  Google Scholar 

  • Nowakowska A, Caputa M, Rogalska J (2006) Seasonal changes in cryoprotectants concentrations in Helix pomatia snails. J Physiol Pharmacol 57:93–105

    PubMed  Google Scholar 

  • Olgart C, Gustaffson LE, Wiklund NP (2000) Evidence for nonvesicular nitric oxide release evoked by nerve activation. Eur J Neurosci 12:1303–1309

    Article  CAS  PubMed  Google Scholar 

  • Palumbo A (2005) Nitric oxide in marine invertebrates: a comparative perspective. Comp Biochem Physiol A Mol Integr Physiol 142:241–248

    Article  PubMed  Google Scholar 

  • Palumbo A, Di Cosmo A, Poli A, Di Cristo C, d’Ischia M (1999) A calcium/calmodulin-dependent nitric oxide synthase, NMDAR2/3 receptor subunits, and glutamate in the CNS of the cuttlefish Sepia officinalis: localization in specific neural pathways controlling the inking system. J Neurochem 73:1254–1263

    Article  CAS  PubMed  Google Scholar 

  • Peng HM, Morishima Y, Jenkins GJ, Dunbar AY, Lau M, Patterson C, Pratt WB, Osawa Y (2004) Ubiquitylation of neuronal nitric-oxide synthase by CHIP, a chaperone-dependent E3 ligase. J Biol Chem 279:52970–52977

    Article  CAS  PubMed  Google Scholar 

  • Percival JM, Anderson KN, Huang P, Adams ME, Froehner SC (2010) Golgi and sarcolemmal neuronal NOS differentially regulate contraction-induced fatigue and vasoconstriction in exercising mouse skeletal muscle. J Clin Invest 120:816–826

    Article  CAS  PubMed  Google Scholar 

  • Peruzzi E, Fontana G, Sonetti D (2004) Presence and role of nitric oxide in the central nervous system of the freshwater snail Planorbarius corneus: possible implication in neuron-microglia communication. Brain Res 1005:9–20

    Article  CAS  PubMed  Google Scholar 

  • Pfaffl MW (2006) Relative quantification. In: Dorak T (ed) Real-time PCR. International University Line, La Jolla, CA, pp 63–82

    Google Scholar 

  • Pisu MB, Conforti E, Fenoglio C, Necchi D, Scherini E, Bernocchi G (1999) Nitric-oxide containing neurons in the nervous ganglia of Helix aspersa during rest and activity: immunocytochemical and enzyme histochemical detection. J Comp Neurol 409:274–284

    Article  CAS  PubMed  Google Scholar 

  • Reihill JA, Ewart MA, Hardie DG, Salt IP (2007) AMP-activated protein kinase mediates VEGF-stimulated endothelial NO production. Biochem Biophys Res Commun 354:1084–1088

    Article  CAS  PubMed  Google Scholar 

  • Rőszer T, Zs C, Szentmiklósi AJ, Bánfalvi G (2004) Nitric oxide synthesis is blocked in the enteral nervous system during dormant periods of the snail, Helix lucorum. Cell Tissue Res 316:255–262

    Article  PubMed  Google Scholar 

  • Rőszer T, Kiss-Tóth É, Szentmiklósi AJ, Bánfalvi G (2005) Seasonal periodicity of enteric nitric oxide synthesis and its regulation in Helix lucorum. Invertebr Biol 124:18–24

    Article  Google Scholar 

  • Rőszer T, Józsa T, Szentmiklósi AJ, Bánfalvi G (2009) Acetylcholine inhibits nitric oxide (NO) synthesis in the gastropod nervous system. Cell Tissue Res 336:325–335

    Article  PubMed  Google Scholar 

  • Rzymowska J, Gawron A, Pawlikowska-Pawlega B, Jakubowicz-Gil J, Wojcierowski J (1999) The effect of quercetin on induction of apoptosis. Folia Histochem Cytobiol 37:125–126

    CAS  PubMed  Google Scholar 

  • Salzman AL (1995) Nitric oxide in the gut. New Horiz 3:352–364

    CAS  PubMed  Google Scholar 

  • Scheinker V, Fiore G, Di Cristo C, Di Cosmo A, d’Ischia M, Enikolopov G, Palumbo A (2005) Nitric oxide synthase in the nervous system and ink gland of the cuttlefish Sepia officinalis: molecular cloning and expression. Biochem Biophys Res Commun 338:1204–1215

    Article  CAS  PubMed  Google Scholar 

  • Schmidt HH, Walter U (1994) NO at work. Cell 78:919–925

    Article  CAS  PubMed  Google Scholar 

  • Serfőző Z, Szentmiklósi AJ, Elekes K (2008) Characterization of nitric oxidergic neurons in the alimentary tract of the snail Helix pomatia L.: histochemical and physiological study. J Comp Neurol 506:801–821

    Article  PubMed  Google Scholar 

  • Sifuentes-Romero I, Merchant-Larios H, García-Gasca A (2010) Hox gene expression in the embryonic genital system of the sea turtle Lepidochelys olivacea (Eschscholt, 1829), a species with temperature-dependent sex determination. Gene Expr Patterns (in press)

  • Sonetti D, Peruzzi E (2004) Neuron-microglia communication in the CNS of the freshwater snail Planorbarius corneus. Acta Biol Hung 55:273–285

    Article  CAS  PubMed  Google Scholar 

  • Sossin WS, Abrams TW (2009) Evolutionary conservation of the signaling proteins upstream of cyclic AMP-dependent kinase and protein kinase C in gastropod mollusks. Brain Behav Evol 74:191–205

    Article  PubMed  Google Scholar 

  • Sugita S, Baxter DA, Byrne JH (1994) cAMP-independent effects of 8-(4-parachlorophenylthio)-cyclic AMP on spike duration and membrane currents in pleural sensory neurons of Aplysia. J Neurophysiol 72:1250–1259

    CAS  PubMed  Google Scholar 

  • Venardos K, Enriquez C, Marshall T, Chin-Dusting JP, Ahlers B, Kaye DM (2009) Protein kinase C mediated inhibition of endothelial L-arginine transport is mediated by MARCKS protein. J Mol Cell Cardiol 46:86–92

    Article  CAS  PubMed  Google Scholar 

  • Vignola C, Fenoglio C, Scherini E, Bernocchi G (1995) The cerebral neurons of Helix aspersa during hibernation. Changes in the cytochemical detection of calmodulin, cytoskeletal components and phosphatases. Tissue Cell 27:185–196

    Article  CAS  PubMed  Google Scholar 

  • Villanueva C, Giulivi C (2010) Subcellular and cellular locations of nitric oxide synthase isoforms as determinants of health and disease. Free Radic Biol Med (in press)

  • Vincent SR (2010) Nitric oxide neurons and neurotransmission. Prog Neurobiol 90:246–255

    Article  CAS  PubMed  Google Scholar 

  • Winfield MO, Lu C, Wilson ID, Coghill JA, Edwards KJ (2010) Plant responses to cold: transcriptome analysis of wheat. Plant Biotechnol J (in press)

  • Zhou L, Zhu DY (2009) Neuronal nitric oxide synthase: structure, subcellular localization, regulation, and clinical implications. Nitric Oxide 20:223–230

    Article  CAS  PubMed  Google Scholar 

  • Zhuo M, Hawkins RD (1995) Long-term depression: a learning-related type of synaptic plasticity in the mammalian central nervous system. Rev Neurosci 6:259–277

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We are grateful to Dr. Lucía Fuentes and Dr. Christian Hellriegel for valuable discussion.

Author information

Author notes

  1. Tamás Rőszer

    Present address: Department of Regenerative Cardiology, Spanish National Cardiovascular Research Center, 28029, Madrid, Melchor Fernandez Almagro 3, Spain

Authors and Affiliations

  1. Department of Microbial Biotechnology and Cell Biology (formerly Animal Anatomy and Physiology), Faculty of Science and Technology, University of Debrecen, Debrecen, Hungary

    Tamás Rőszer, Éva Kiss-Tóth, Dávid Rózsa & Gáspár Bánfalvi

  2. Department of Pediatrics, Medical and Health Science Center, University of Debrecen, Debrecen, Hungary

    Tamás Józsa

  3. Department of Pharmacology and Pharmacotherapy, Medical and Health Science Center, University of Debrecen, Debrecen, Hungary

    A. József Szentmiklósi

Authors
  1. Tamás Rőszer
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Éva Kiss-Tóth
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Dávid Rózsa
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Tamás Józsa
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. A. József Szentmiklósi
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Gáspár Bánfalvi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Tamás Rőszer.

Electronic supplementary material

Rights and permissions

Reprints and permissions

About this article

Cite this article

Rőszer, T., Kiss-Tóth, É., Rózsa, D. et al. Hypothermia translocates nitric oxide synthase from cytosol to membrane in snail neurons. Cell Tissue Res 342, 191–203 (2010). https://doi.org/10.1007/s00441-010-1063-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00441-010-1063-8

Keywords

Search

Navigation