Abstract
The gaseous messenger nitric oxide (NO) has been implicated in a wide range of behaviors, including aggression, anxiety, depression, and cognitive functioning. To further elucidate the physiological role of NO and its down-stream mechanisms, we conducted behavioral and expressional phenotyping of mice lacking the neuronal isoform of nitric oxide synthase (NOS-I), the major source of NO in the central nervous system. No differences were observed in activity-related parameters; in contrast to the a priori hypothesis, derived from pharmacological treatments, depression-related tests (Forced Swim Test, Learned Helplessness) also yielded no significantly different results. A subtle anxiolytic phenotype however was present, with knockdown mice displaying a higher open arm time as compared to their respective wildtypes, yet all other investigated anxiety-related parameters were unchanged. The most prominent feature however was gender-independent cognitive impairment in spatial learning and memory, as assessed by the Water Maze test and an automatized holeboard paradigm. No significant dysregulation of monoamine transporters was evidenced by qRT PCR. To further examine the underlying molecular mechanisms, the transcriptome of knockdown animals was thus examined in the hippocampus, striatum and cerebellum by microarray analysis. A set of > 120 differentially expressed genes was identified, whereat the hippocampus and the striatum showed similar expressional profiles as compared to the cerebellum in hierarchical clustering.
Access provided by Autonomous University of Puebla. Download to read the full chapter text
Chapter PDF
Similar content being viewed by others
References
Anneser JM, Cookson MR, Ince PG, Shaw PJ, Borasio GD (2001) Glial cells of the spinal cord and subcortical white matter up-regulate neuronal nitric oxide synthase in sporadic amyotrophic lateral sclerosis. Exp Neurol 171: 418–421
Beijamini V, Guimaraes FS (2006) Activation of neurons containing the enzyme nitric oxide synthase following exposure to an elevated plus maze. Brain Res Bull 69: 347–355
Ben-Arie N, Bellen HJ, Armstrong DL, McCall AE, Gordadze PR, Guo Q, Matzuk MM, Zoghbi HY (1997) Mathl is essential for genesis of cerebellar granule neurons. Nature 390: 169–172
Bernstein HG, Bogerts B, Keilhoff G (2005) The many faces of nitric oxide in schizophrenia. A review. Schizophr Res 78: 69–86
Bilbo SD, Hotchkiss AK, Chiavegatto S, Nelson RJ (2003) Blunted stress responses in delayed type hypersensitivity in mice lacking the neuronal isoform of nitric oxide synthase. J Neuroimmunol 140: 41–48
Bird DC, Bujas-Bobanovic M, Robertson HA, Dursun SM (2001) Lack of phencyclidine-induced effects in mice with reduced neuronal nitric oxide synthase. Psychopharmacology (Berl) 155: 299–309
Chiang MC, Chen HM, Lee YH, Chang HH, Wu YC, Soong BW, Chen CM, Wu YR, Liu CS, Niu DM, Wu JY, Chen YT, Chern Y (2007) Dysregulation of C/EBPalpha by mutant Huntingtin causes the urea cycle deficiency in Huntington’s disease. Hum Mol Genet (in press)
Chiavegatto S, Nelson RJ (2003) Interaction of nitric oxide and serotonin in aggressive behavior. Horm Behav 44: 233–241
Chiavegatto S, Dawson VL, Mamounas LA, Koliatsos VE, Dawson TM, Nelson RJ (2001) Brain serotonin dysfunction accounts for aggression in male mice lacking neuronal nitric oxide synthase. Proc Natl Acad Sci USA 98: 1277–1281
Chourbaji S, Zacher C, Sanchis-Segura C, Dormann C, Vollmayr B, Gass P (2005) Learned helplessness: validity and reliability of depressive-like states in mice. Brain Res Brain Res Protoc 16: 70–78
Cryan JF, Kaupmann K (2005) Don’t worry ‘B’ happy!: a role for GABA(B) receptors in anxiety and depression. Trends Pharmacol Sci 26: 36–43
Cryan JF, Page ME, Lucki I (2002) Noradrenergic lesions differentially alter the antidepressant-like effects of reboxetine in a modified forced swim test. Eur J Pharmacol 436: 197–205
Czech DA, Jacobson EB, LeSueur-Reed KT, Kazel MR (2003) Putative anxiety-linked effects of the nitric oxide synthase inhibitor L-NAME in three murine exploratory behavior models. Pharmacol Biochem Behav 75: 741–748
da Silva GD, Matteussi AS, dos Santos AR, Calixto JB, Rodrigues AL (2000) Evidence for dual effects of nitric oxide in the forced swimming test and in the tail suspension test in mice. Neuroreport 11: 3699–3702
De Oliveira CL, Del Bel EA, Guimaraes FS (1997) Effects of L-NOARG on plus-maze performance in rats. Pharmacol Biochem Behav 56: 55–59
Del Bel EA, Guimaraes FS, Bermudez-Echeverry M, Gomes MZ, Schiaveto-de-souza A, Padovan-Neto FE, Tumas V, Barion-Cavalcanti AP, Lazzarini M, Nucci-da-Silva LP, de Paula-Souza D (2005) Role of nitric oxide on motor behavior. Cell Mol Neurobiol 25: 371–392
Demas GE, Kriegsfeld LJ, Blackshaw S, Huang P, Gammie SC, Nelson RJ, Snyder SH (1999) Elimination of aggressive behavior in male mice lacking endothelial nitric oxide synthase. J Neurosci 19: RC30
Dere E, Frisch C, De Souza Silva MA, Godecke A, Schrader J, Huston JP (2001) Unaltered radial maze performance and brain acetylcholine of the endothelial nitric oxide synthase knockout mouse. Neuroscience 107: 561–570
Dere E, De Souza Silva MA, Topic B, Fiorillo C, Li JS, Sadile AG, Frisch C, Huston JP (2002) Aged endothelial nitric oxide synthase knockout mice exhibit higher mortality concomitant with impaired open-field habituation and alterations in forebrain neurotransmitter levels. Genes Brain Behav 1: 204–213
Dunn RW, Reed TA, Copeland PD, Frye CA (1998) The nitric oxide synthase inhibitor 7-nitroindazole displays enhanced anxiolytic efficacy without tolerance in rats following subchronic administration. Neuropharmacology 37: 899–904
Elfline GS, Branda EM, Babich M, Quock RM (2004) Antagonism by NOS inhibition of the behavioral effects of benzodiazepine and GABAA receptor agonists in the mouse elevated plus-maze. Neuropsychopharmacology 29: 1419–1425
Ergun Y, Ergun UG (2007) Prevention of pro-depressant effect of Larginine in the forced swim test by NG-nitro-L-arginine and [1H-[l,2,4]Oxadiazole[4,3-a]quinoxalin-l-one]. Eur J Pharmacol 554: 150–154
Faria MS, Muscara MN, Moreno Junior H, Teixeira SA, Dias HB, De Oliveira B, Graeff FG, De Nucci G (1997) Acute inhibition of nitric oxide synthesis induces anxiolysis in the plus maze test. Eur J Pharmacol 323: 37–43
Fedele E, Conti A, Raiteri M (1997a) The glutamate receptor/NO/cyclic GMP pathway in the hippocampus of freely moving rats: modulation by cyclothiazide, interaction with GABA and the behavioural consequences. Neuropharmacology 36: 1393–1403
Fedele E, Varnier G, Raiteri M (1997b) In vivo microdialysis study of GABA(A) and GABA(B) receptors modulating the glutamate receptor/NO/cyclic GMP pathway in the rat hippocampus. Neuropharmacology 36: 1405–1415
Focking M, Holker I, Trapp T (2003) Chronic glucocorticoid receptor activation impairs CREB transcriptional activity in clonal neurons. Biochem Biophys Res Commun 304: 720–723
Forestiero D, Manfrim CM, Guimaraes FS, de Oliveira RM (2006) Anxio-lytic-like effects induced by nitric oxide synthase inhibitors microinjected into the medial amygdala of rats. Psychopharmacology (Berl) 184: 166–172
Frisch C, Dere E, Silva MA, Godecke A, Schrader J, Huston JP (2000) Superior water maze performance and increase in fear-related behavior in the endothelial nitric oxide synthase-deficient mouse together with monoamine changes in cerebellum and ventral striatum. J Neurosci 20: 6694–6700
Gabbott PL, Bacon SJ (1995) Co-localisation of NADPH diaphorase activity and GABA immunoreactivity in local circuit neurones in the medial prefrontal cortex (mPFC) of the rat. Brain Res 699: 321–328
Galimberti D, Scarpini E, Venturelli E, Strobel A, Herterich S, Fenoglio C, Guidi I, Scalabrini D, Cortini F, Bresolin N, Lesch K-P, Reif A (2007) As Association of a NOS1 promoter repeat with Alzheimer’s disease. Neurobiol Ageing (in press)
Gammie SC, Nelson RJ (1999) Maternal aggression is reduced in neuronal nitric oxide synthase-deficient mice. J Neurosci 19: 8027–8035
Gammie SC, Huang PL, Nelson RJ (2000) Maternal aggression in endothelial nitric oxide synthase-deficient mice. Horm Behav 38: 13–20
Gerlach M, Blum-Degen D, Ransmayr G, Leblhuber F, Pedersen V, Riederer P (2001) Expression, but not activity of neuronal NOS is regionally increased in the alcoholic brain. Alcohol Alcoholism 36: 65–69
Golfier G, Dang MT, Dauphinot L, Graison E, Rossier J, Potier MC (2004) VARAN: a web server for variability analysis of DNA microarray experiments. Bioinformatics 20: 1641–1643
Hammond DL, Ruda MA (1989) Developmental alterations in thermal nociceptive threshold and the distribution of immunoreactive calcitonin gene-related peptide and substance P after neonatal administration of capsaicin in the rat. Neurosci Lett 97: 57–62
Harkin AJ, Bruce KH, Craft B, Paul IA (1999) Nitric oxide synthase inhibitors have antidepressant-like properties in mice. 1. Acute treatments are active in the forced swim test. Eur J Pharmacol 372: 207–213
Harkin A, Connor TJ, Walsh M, St John N, Kelly JP (2003) Serotonergic mediation of the antidepressant-like effects of nitric oxide synthase inhibitors. Neuropharmacology 44: 616–623
Harkin A, Connor TJ, Burns MP, Kelly JP (2004) Nitric oxide synthase inhibitors augment the effects of serotonin re-uptake inhibitors in the forced swimming test. Eur Neuropsychopharmacol 14: 274–281
Hattori F, Murayama N, Noshita T, Oikawa S (2003) Mitochondrial peroxiredoxin-3 protects hippocampal neurons from excitotoxic injury in vivo. J Neurochem 86: 860–868
Hawkins RD, Son H, Arancio O (1998) Nitric oxide as a retrograde messenger during long-term potentiation in hippocampus. Prog Brain Res 118: 155–172
Holmes A, Murphy DL, Crawley JN (2002) Reduced aggression in mice lacking the serotonin transporter. Psychopharmacology (Berl) 161: 160–167
Huang PL, Huang Z, Mashimo H, Bloch KD, Moskowitz MA, Bevan JA, Fishman MC (1995) Hypertension in mice lacking the gene for endothelial nitric oxide synthase. Nature 377: 239–242
Hunot S, Boissiere F, Faucheux B, Brugg B, Mouatt-Prigent A, Agid Y, Hirsch EC (1996) Nitric oxide synthase and neuronal vulnerability in Parkinson’s disease. Neuroscience: 72: 355–363
Inan SY, Yalcin I, Aksu F (2004) Dual effects of nitric oxide in the mouse forced swimming test: possible contribution of nitric oxide-mediated serotonin release and potassium channel modulation. Pharmacol Biochem Behav 77: 457–464
Javitt DC, Zukin SR (1991) Recent advances in the phencyclidine model of schizophrenia. Am J Psychiatry 148: 1301–1308
Joca SR, Guimaraes FS (2006) Inhibition of neuronal nitric oxide synthase in the rat hippocampus induces antidepressant-like effects. Psychopharmacology (Berl) 185: 298–305
Karolewicz B, Paul IA, Antkiewicz-Michaluk L (2001) Effect of NOS inhibitor on forced swim test and neurotransmitters turnover in the mouse brain. Pol J Pharmacol 53: 587–596
Khan SA, Lee K, Minhas KM, Gonzalez DR, Raju SV, Tejani AD, Li D, Berkowitz DE, Hare JM (2004) Neuronal nitric oxide synthase negatively regulates xanthine oxidoreductase inhibition of cardiac excitation-contraction coupling. Proc Natl Acad Sci USA 101: 15944–15948
Kilic F, Murphy DL, Rudnick G (2003) A human serotonin transporter mutation causes constitutive activation of transport activity. Mol Pharmacol 64: 440–446
Kirchner L, Weitzdoerfer R, Hoeger H, Url A, Schmidt P, Engelmann M, Villar SR, Fountoulakis M, Lubec G, Lubec B (2004) Impaired cognitive performance in neuronal nitric oxide synthase knockout mice is associated with hippocampal protein derangements. Nitric Oxide 11: 316–330
Kiss JP, Vizi ES (2001) Nitric oxide: a novel link between synaptic and nonsynaptic transmission. Trends Neurosci 24: 211–215
Koylu EO, Kanit L, Taskiran D, Dagci T, Balkan B, Pogun S (2005) Effects of nitric oxide synthase inhibition on spatial discrimination learning and central DA2 and mACh receptors. Pharmacol Biochem Behav 81: 32–40
Kriegsfeld LJ, Dawson TM, Dawson VL, Nelson RJ, Snyder SH (1997) Aggressive behavior in male mice lacking the gene for neuronal nitric oxide synthase requires testosterone. Brain Res 769: 66–70
Kriegsfeld LJ, Demas GE, Lee SE Jr, Dawson TM, Dawson VL, Nelson RJ (1999) Circadian locomotor analysis of male mice lacking the gene for neuronal nitric oxide synthase (nNOS-/-). J Biol Rhythms 14: 20–27
Kubota Y, Mikawa S, Kawaguchi Y (1993) Neostriatal GABAergic interneurones contain NOS, calretinin or parvalbumin. Neuroreport 5: 205–208
La Y, Wan C, Zhu H, Yang Y, Chen Y, Pan Y, Ji B, Feng G, He L (2006) Hippocampus protein profiling reveals aberration of malate dehydrogenase in chlorpromazine/clozapine treated rats. Neurosci Lett 408: 29–34
Law A, Gauthier S, Quirion R (2001) Say NO to Alzheimer’s disease: the putative links between nitric oxide and dementia of the Alzheimer’s type. Brain Res Brain Res Rev 35: 73–96
Le Roy I, Pothion S, Mortaud S, Chabert C, Nicolas L, Cherfouh A, Roubertoux PL (2000) Loss of aggression, after transfer onto a C57BL/6J background, in mice carrying a targeted disruption of the neuronal nitric oxide synthase gene. Behav Genet 30: 367–373
Majlessi N, Kadkhodaee M, Parviz M, Naghdi N (2003) Serotonin depletion in rat hippocampus attenuates L-NAME-induced spatial learning deficits. Brain Res 963: 244–251
Monzon ME, Varas MM, De Barioglio SR (2001) Anxiogenesis induced by nitric oxide synthase inhibition and anxiolytic effect of melaninconcentrating hormone (MCH) in rat brain. Peptides 22: 1043–1047
Nelson RJ, Demas GE, Huang PL, Fishman MC, Dawson VL, Dawson TM, Snyder SH (1995) Behavioural abnormalities in male mice lacking neuronal nitric oxide synthase. Nature 378: 383–386
Nelson RJ, Trainor BC, Chiavegatto S, Demas GE (2006) Pleiotropic contributions of nitric oxide to aggressive behavior. Neurosci Biobehav Rev 30: 346–355
Nonn L, Berggren M, Powis G (2003) Increased expression of mitochondrial peroxiredoxin-3 (thioredoxin peroxidase-2) protects cancer cells against hypoxia and drug-induced hydrogen peroxide-dependent apoptosis. Mol Cancer Res 1: 682–689
Packer MA, Stasiv Y, Benraiss A, Chmielnicki E, Grinberg A, Westphal H, Goldman SA, Enikolopov G (2003) Nitric oxide negatively regulates mammalian adult neurogenesis. Proc Natl Acad Sci USA 100: 9566–9571
Packer MA, Hemish J, Mignone JL, John S, Pugach I, Enikolopov G (2005) Transgenic mice overexpressing nNOS in the adult nervous system. Cell Mol Biol (Noisy-le-grand) 51: 269–277
Pitsikas N, Rigamonti AE, Cella SG, Muller EE (2003) The GABAB receptor and recognition memory: possible modulation of its behavioral effects by the nitrergic system. Neuroscience 118: 1121–1127
Pokk P, Vali M (2002a) Effects of nitric oxide synthase inhibitors 7-NI, LNAME, and L-NOARG in staircase test. Arch Med Res 33: 265–268
Pokk P, Vali M (2002b) The effects of the nitric oxide synthase inhibitors on the behaviour of small-platform-stressed mice in the plus-maze test. Prog Neuropsychopharmacol Biol Psychiatry 26: 241–247
Prendergast MA, Buccafusco JJ, Terry AV Jr (1997) Nitric oxide synthase inhibition impairs spatial navigation learning and induces conditioned taste aversion. Pharmacol Biochem Behav 57: 347–352
Reddy DS, Kulkarni SK (1998) Inhibition of neuronal nitric oxide synthase (n-cNOS) reverses the corticotrophin-induced behavioral effects in rats. Mol Cell Biochem 183: 25–38
Reif A, Schmitt A, Fritzen S, Chourbaji S, Bartsch C, Urani A, Wycislo M, Mossner R, Sommer C, Gass P, Lesch KP (2004) Differential effect of endothelial nitric oxide synthase (NOS-III) on the regulation of adult neurogenesis and behaviour. Eur J Neurosci 20: 885–895
Reif A, Herterich S, Strobel A, Ehlis AC, Saur D, Jacob CP, Wienker T, Topner T, Fritzen S, Walter U, Schmitt A, Fallgatter AJ, Lesch KP (2006a) A neuronal nitric oxide synthase (NOS-I) haplotype associated with schizophrenia modifies prefrontal cortex function. Mol Psychiatry: 286–300
Reif A, Strobel A, Jacob CP, Herterich S, Freitag CM, Topner T, Mossner R, Fritzen S, Schmitt A, Lesch KP (2006b) A NOS-III haplotype that includes functional polymorphisms is associated with bipolar disorder. Int J Neuropsychopharmacol 9: 13–20
Ridder S, Chourbaji S, Hellweg R, Urani A, Zacher C, Schmid W, Zink M, Hortnagl H, Flor H, Henn FA, Schutz G, Gass P (2005) Mice with genetically altered glucocorticoid receptor expression show altered sensitivity for stress-induced depressive reactions. J Neurosci 25: 6243–6250
Salchner P, Lubec G, Engelmann M, Orlando GF, Wolf G, Sartori SB, Hoeger H, Singewald N (2004) Genetic functional inactivation of neuronal nitric oxide synthase affects stress-related Fos expression in specific brain regions. Cell Mol Life Sci 61: 1498–1506
Schafe GE, Bauer EP, Rosis S, Farb CR, Rodrigues SM, LeDoux JE (2005) Memory consolidation of Pavlovian fear conditioning requires nitric oxide signaling in the lateral amygdala. Eur J Neurosci 22: 201–211
Schilling K, Schmidt HH, Baader SL (1994) Nitric oxide synthase expression reveals compartments of cerebellar granule cells and suggests a role for mossy fibers in their development. Neuroscience 59: 893–903
Schulte-Herbraggen O, Chourbaji S, Ridder S, Brandwein C, Gass P, Hortnagl H, Hellweg R (2006) Stress-resistant mice overexpressing glucocorticoid receptors display enhanced BDNF in the amygdala and hippocampus with unchanged NGF and serotonergic function. Psychoneuroendocrinology 31: 1266–1277
Semba J, Sakai M, Miyoshi R, Kito S (1995) NG-monomethyl-L-arginine, an inhibitor of nitric oxide synthase, increases extracellular GABA in the striatum of the freely moving rat. Neuroreport 6: 1426–1428
Silva AJ, Kogan JH, Frankland PW, Kida S (1998) CREB and memory. Annu Rev Neurosci 21: 127–148
Snyder SH, Ferris CD (2000) Novel neurotransmitters and their neuropsychiatric relevance. Am J Psychiatry 157: 1738–1751
Vale AL, Green S, Montgomery AM, Shaft S (1998) The nitric oxide synthesis inhibitor L-NAME produces anxiogenic-like effects in the rat elevated plus-maze test, but not in the social interaction test. J Psychopharmacol 12: 268–272
Valtschanoff JG, Weinberg RJ, Kharazia VN, Nakane M, Schmidt HH (1993a) Neurons in rat hippocampus that synthesize nitric oxide. J Comp Neural 331: 111–121
Valtschanoff JG, Weinberg RJ, Kharazia VN, Schmidt HH, Nakane M, Rustioni A (1993b) Neurons in rat cerebral cortex that synthesize nitric oxide: NADPH diaphorase histochemistry, NOS immunocytochemistry, and colocalization with GABA. Neurosci Lett 157: 157–161
van Amsterdam JG, Opperhuizen A (1999) Nitric oxide and biopterin in depression and stress. Psychiatry Res 85: 33–38
Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A, Speleman F (2002) Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol 3: RESEARCH0034
Volke V, Koks S, Vasar E, Bourin M, Bradwejn J, Mannisto PT (1995) Inhibition of nitric oxide synthase causes anxiolytic-like behaviour in an elevated plus-maze. Neuroreport 6: 1413–1416
Volke V, Soosaar A, Koks S, Bourin M, Mannisto PT, Vasar E (1997) 7-Nitroindazole, a nitric oxide synthase inhibitor, has anxiolytic-like properties in exploratory models of anxiety. Psychopharmacology (Berl) 131: 399–405
Volke V, Wegener G, Bourin M, Vasar E (2003) Antidepressant-and anxiolytic-like effects of selective neuronal NOS inhibitor l-(2-trifluoromethylphenyl)-imidazole in mice. Behav Brain Res 140: 141–147
Wass C, Archer T, Palsson E, Fejgin K, Alexandersson A, Klamer D, Engel JA, Svensson L (2006a) Phencyclidine affects memory in a nitric oxide-dependent manner: working and reference memory. Behav Brain Res 174: 49–55
Wass C, Archer T, Palsson E, Fejgin K, Klamer D, Engel JA, Svensson L (2006b) Effects of phencyclidine on spatial learning and memory: nitric oxide-dependent mechanisms. Behav Brain Res 171: 147–153
Wegener G, Volke V, Rosenberg R (2000) Endogenous nitric oxide decreases hippocampal levels of serotonin and dopamine in vivo. Br J Pharmacol 130: 575–580
Wegener G, Volke V, Harvey BH, Rosenberg R (2003) Local, but not systemic, administration of serotonergic antidepressants decreases hippocampal nitric oxide synthase activity. Brain Res 959: 128–134
Weitzdoerfer R, Hoeger H, Engidawork E, Engelmann M, Singewald N, Lubec G, Lubec B (2004) Neuronal nitric oxide synthase knockout mice show impaired cognitive performance. Nitric Oxide 10: 130–140
Wood-Allum CA, Barber SC, Kirby J, Heath P, Holden H, Mead R, Higginbottom A, Allen S, Beaujeux T, Alexson SE, Ince PG, Shaw PJ (2006) Impairment of mitochondrial anti-oxidant defence in SOD1-related motor neuron injury and amelioration by ebselen. Brain 129: 1693–1709
Yildiz F, Ulak G, Erden BF, Gacar N (2000) Anxiolytic-like effects of 7-nitroindazole in the rat plus-maze test. Pharmacol Biochem Behav 65: 199–202
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Additional information
Dedicated to Prof. Dr. Peter Riederer on occasion of his 65th birthday.
Rights and permissions
Copyright information
© 2007 Springer-Verlag
About this paper
Cite this paper
Wultsch, T. et al. (2007). Behavioural and expressional phenotyping of nitric oxide synthase-I knockdown animals. In: Gerlach, M., Deckert, J., Double, K., Koutsilieri, E. (eds) Neuropsychiatric Disorders An Integrative Approach. Journal of Neural Transmission. Supplementa, vol 72. Springer, Vienna. https://doi.org/10.1007/978-3-211-73574-9_10
Download citation
DOI: https://doi.org/10.1007/978-3-211-73574-9_10
Publisher Name: Springer, Vienna
Print ISBN: 978-3-211-73573-2
Online ISBN: 978-3-211-73574-9
eBook Packages: MedicineMedicine (R0)