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Nucleus incertus promotes cortical desynchronization and behavioral arousal

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Abstract

Arousal and vigilance are essential for survival and relevant regulatory neural circuits lie within the brainstem, hypothalamus and forebrain. The nucleus incertus (NI) is a distinct site within the pontine periventricular gray, containing a substantial population of GABAergic neurons with long-range, ascending projections. Existing neuroanatomical data and functional studies in anesthetized rats, suggest the NI is a central component of a midline behavioral control network well positioned to modulate arousal, vigilance and exploratory navigation, yet none of these roles have been established experimentally. We used a chemogenetic approach—clozapine-N-oxide (CNO) activation of virally delivered excitatory hM3Dq-DREADDs—to activate the NI in rats and examined the behavioral and physiological effects, relative to effects in naïve rats and appropriate viral-treated controls. hM3Dq activation by CNO resulted in long-lasting depolarization of NI neurons with action potentials, in vitro. Peripheral injection of CNO significantly increased c-Fos immunoreactivity in the NI and promoted cortical electroencephalograph (EEG) desynchronization. These brain changes were associated with heightened arousal, and increased locomotor activity in the homecage and in a novel environment. Furthermore, NI activation altered responses in a fear conditioning paradigm, reflected by increased head-scanning, vigilant behaviors during conditioned fear recall. These findings provide direct evidence that the NI promotes general arousal via a broad behavioral activation circuit and support early hypotheses, based on its connectivity, that the NI is a modulator of cognition and attention, and emotional and motivated behaviors.

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References

  • Anaclet C, Pedersen NP, Ferrari LL, Venner A, Bass CE, Arrigoni E, Fuller PM (2015) Basal forebrain control of wakefulness and cortical rhythms. Nat Commun 6:8744. doi:10.1038/ncomms9744

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Alexander GM, Rogan SC, Abbas AI, Armbruster BN, Pei Y, Allen JA, Nonneman RJ, Hartmann J, Moy SS, Nicolelis MA, McNamara JO, Roth BL (2009) Remote control of neuronal activity in transgenic mice expressing evolved G protein-coupled receptors. Neuron 63:27–39. doi:10.1016/j.neuron.2009.06.014

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bathgate RAD, Samuel CS, Burazin TCD, Layfield S, Claasz AA, Reytomas IG, Dawson NF, Zhao C, Bond C, Summers RJ, Parry LJ, Wade JD, Tregear GW (2002) Human relaxin gene 3 (H3) and the equivalent mouse relaxin (M3) gene. Novel members of the relaxin peptide family. J Biol Chem 277:1148–1157. doi:10.1074/jbc.M107882200

    Article  CAS  PubMed  Google Scholar 

  • Bittencourt JC, Sawchenko PE (2000) Do centrally administered neuropeptides access cognate receptors? An analysis in the central corticotropin-releasing factor system. J Neurosci 20:1142–1156

    CAS  PubMed  Google Scholar 

  • Blasiak A, Siwiec M, Grabowiecka A, Blasiak T, Czerw A, Blasiak E, Kania A, Rajfur Z, Lewandowski MH, Gundlach AL (2015) Excitatory orexinergic innervation of rat nucleus incertus—Implications for ascending arousal, motivation and feeding control. Neuropharmacology 99:432–447. doi:10.1016/j.neuropharm.2015.08.014

    Article  CAS  PubMed  Google Scholar 

  • Brown RE, McKenna JT (2015) Turning a negative into a positive: ascending GABAergic control of cortical activation and arousal. Front Neurol 6:135. doi:10.3389/fneur.2015.00135

    PubMed  PubMed Central  Google Scholar 

  • Brown M, Renehan WE, Schweitzer L (2000) Changes in GABA-immunoreactivity during development of the rostral subdivision of the nucleus of the solitary tract. Neuroscience 100:849–859

    Article  CAS  PubMed  Google Scholar 

  • Burazin TCD, Bathgate RAD, Macris M, Layfield S, Gundlach AL, Tregear GW (2002) Restricted, but abundant, expression of the novel rat gene-3 (R3) relaxin in the dorsal tegmental region of brain. J Neurochem 82:1553–1557

    Article  CAS  PubMed  Google Scholar 

  • Butler WN, Taube JS (2015) The nucleus prepositus hypoglossi contributes to head direction cell stability in rats. J Neurosci 35:2547–2558. doi:10.1523/JNEUROSCI.3254-14.2015

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Callander GE, Ma S, Ganella DE, Wimmer VC, Gundlach AL, Thomas WG, Bathgate RAD (2012) Silencing relaxin-3 in nucleus incertus of adult rodents: a viral vector-based approach to investigate neuropeptide function. PLoS One 7:e42300. doi:10.1371/journal.pone.0042300

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Calvez J, de Avila C, Matte LO, Guevremont G, Gundlach AL, Timofeeva E (2016) Role of relaxin-3/RXFP3 system in stress-induced binge-like eating in female rats. Neuropharmacology 102:207–215. doi:10.1016/j.neuropharm.2015.11.014

    Article  CAS  PubMed  Google Scholar 

  • Cervera-Ferri A, Guerrero-Martinez J, Bataller-Mompean M, Taberner-Cortes A, Martinez-Ricos J, Ruiz-Torner A, Teruel-Marti V (2011) Theta synchronization between the hippocampus and the nucleus incertus in urethane-anesthetized rats. Exp Brain Res 211:177–192. doi:10.1007/s00221-011-2666-3

    Article  CAS  PubMed  Google Scholar 

  • Cervera-Ferri A, Rahmani Y, Martinez-Bellver S, Teruel-Marti V, Martinez-Ricos J (2012) Glutamatergic projection from the nucleus incertus to the septohippocampal system. Neurosci Lett 517:71–76. doi:10.1016/j.neulet.2012.04.014

    Article  CAS  PubMed  Google Scholar 

  • Chang YC, Gottlieb DI (1988) Characterization of the proteins purified with monoclonal antibodies to glutamic acid decarboxylase. J Neurosci 8:2123–2130

    CAS  PubMed  Google Scholar 

  • Cheron G, Saussez S, Gerrits N, Godaux E (1995) Existence in the nucleus incertus of the cat of horizontal-eye-movement-related neurons projecting to the cerebellar flocculus. J Neurophysiol 74:1367–1372

    CAS  PubMed  Google Scholar 

  • Farooq U, Rajkumar R, Sukumaran S, Wu Y, Tan WH, Dawe GS (2013) Corticotropin-releasing factor infusion into nucleus incertus suppresses medial prefrontal cortical activity and hippocampo-medial prefrontal cortical long-term potentiation. Eur J Neurosci 38:2516–2525. doi:10.1111/ejn.12242

    Article  PubMed  Google Scholar 

  • French JD, Magoun HW (1952) Effects of chronic lesions in central cephalic brain stem of monkeys. AMA Arch Neurol Psychiatry 68:591–604

    Article  CAS  PubMed  Google Scholar 

  • Ford B, Holmes CJ, Mainville L, Jones BE (1995) GABAergic neurons in the rat pontomesencephalic tegmentum: codistribution with cholinergic and other tegmental neurons projecting to the posterior lateral hypothalamus. J Comp Neurol 363:177–196. doi:10.1002/cne.903630203

    Article  CAS  PubMed  Google Scholar 

  • Fuhrmann F, Justus D, Sosulina L, Kaneko H, Beutel T, Friedrichs D, Schoch S, Schwarz MK, Fuhrmann M, Remy S (2015) Locomotion, theta oscillations, and the speed-correlated firing of hippocampal neurons are controlled by a medial septal glutamatergic circuit. Neuron 86:1253–1264. doi:10.1016/j.neuron.2015.05.001

    Article  CAS  PubMed  Google Scholar 

  • Geiling B, Vandal G, Posner AR, de Bruyns A, Dutchak KL, Garnett S, Dankort D (2013) A modular lentiviral and retroviral construction system to rapidly generate vectors for gene expression and gene knockdown in vitro and in vivo. PLoS One 8:e76279. doi:10.1371/journal.pone.0076279

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Goto M, Swanson LW, Canteras NS (2001) Connections of the nucleus incertus. J Comp Neurol 438:86–122

    Article  CAS  PubMed  Google Scholar 

  • Hangya B, Borhegyi Z, Szilagyi N, Freund TF, Varga V (2009) GABAergic neurons of the medial septum lead the hippocampal network during theta activity. J Neurosci 29:8094–8102. doi:10.1523/JNEUROSCI.5665-08.2009

    Article  CAS  PubMed  Google Scholar 

  • Harris KD, Thiele A (2011) Cortical state and attention. Nat Rev Neurosci 12:509–523. doi:10.1038/nrn3084

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Heinke B, Ruscheweyh R, Forsthuber L, Wunderbaldinger G, Sandkuhler J (2004) Physiological, neurochemical and morphological properties of a subgroup of GABAergic spinal lamina II neurones identified by expression of green fluorescent protein in mice. J Physiol 560:249–266. doi:10.1113/jphysiol.2004.070540

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hikosaka O (2010) The habenula: from stress evasion to value-based decision-making. Nat Rev Neurosci 11:503–513. doi:10.1038/nrn2866

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ikeda H, Kiritoshi T, Murase K (2008) Effect of excitatory and inhibitory agents and a glial inhibitor on optically-recorded primary-afferent excitation. Mol Pain 4:39. doi:10.1186/1744-8069-4-39

    Article  PubMed  PubMed Central  Google Scholar 

  • Kaufman DL, Erlander MG, Clare-Salzler M, Atkinson MA, Maclaren NK, Tobin AJ (1992) Autoimmunity to two forms of glutamate decarboxylase in insulin-dependent diabetes mellitus. J Clin Invest 89:283–292. doi:10.1172/JCI115573

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kizawa H, Nishi K, Ishibashi Y, Harada M, Asano T, Ito Y, Suzuki N, Hinuma S, Fujisawa Y, Onda H, Nishimura O, Fujino M (2003) Production of recombinant human relaxin 3 in AtT20 cells. Regul Pept 113:79–84

    Article  CAS  PubMed  Google Scholar 

  • Krashes MJ, Koda S, Ye C, Rogan SC, Adams AC, Cusher DS, Maratos-Flier E, Roth BL, Lowell BB (2011) Rapid, reversible activation of AgRP neurons drives feeding behavior in mice. J Clin Invest 121:1424–1428. doi:10.1172/JCI46229

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kubota Y, Inagaki S, Shiosaka S, Cho HJ, Tateishi K, Hashimura E, Hamaoka T, Tohyama M (1983) The distribution of cholecystokinin octapeptide-like structures in the lower brain stem of the rat: an immunohistochemical analysis. Neuroscience 9:587–604

    Article  CAS  PubMed  Google Scholar 

  • Lawther AJ, Clissold ML, Ma S, Kent S, Lowry CA, Gundlach AL, Hale MW (2015) Anxiogenic drug administration and elevated plus-maze exposure in rats activate populations of relaxin-3 neurons in the nucleus incertus and serotonergic neurons in the dorsal raphe nucleus. Neuroscience 303:270–284. doi:10.1016/j.neuroscience.2015.06.052

    Article  CAS  PubMed  Google Scholar 

  • Lee SH, Dan Y (2012) Neuromodulation of brain states. Neuron 76:209–222. doi:10.1016/j.neuron.2012.09.012

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lee LC, Rajkumar R, Dawe GS (2014) Selective lesioning of nucleus incertus with corticotropin releasing factor-saporin conjugate. Brain Res 1543:179–190. doi:10.1016/j.brainres.2013.11.021

    Article  CAS  PubMed  Google Scholar 

  • Lein ES, Hawrylycz MJ, Ao N et al (2007) Genome-wide atlas of gene expression in the adult mouse brain. Nature 445:168–176. doi:10.1038/nature05453

    Article  CAS  PubMed  Google Scholar 

  • Lin SC, Brown RE, Hussain Shuler MG, Petersen CC, Kepecs A (2015) Optogenetic dissection of the basal forebrain neuromodulatory control of cortical activation, plasticity, and cognition. J Neurosci 35:13896–13903. doi:10.1523/JNEUROSCI.2590-15.2015

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lizee G, Aerts JL, Gonzales MI, Chinnasamy N, Morgan RA, Topalian SL (2003) Real-time quantitative reverse transcriptase-polymerase chain reaction as a method for determining lentiviral vector titers and measuring transgene expression. Hum Gene Ther 14:497–507. doi:10.1089/104303403764539387

    Article  CAS  PubMed  Google Scholar 

  • Ma S, Gundlach AL (2015) Ascending control of arousal and motivation: role of nucleus incertus and its peptide neuromodulators in behavioural responses to stress. J Neuroendocrinol 27:457–467. doi:10.1111/jne.12259

    Article  CAS  PubMed  Google Scholar 

  • Ma S, Bonaventure P, Ferraro T, Shen PJ, Burazin TCD, Bathgate RAD, Liu C, Tregear GW, Sutton SW, Gundlach AL (2007) Relaxin-3 in GABA projection neurons of nucleus incertus suggests widespread influence on forebrain circuits via G-protein-coupled receptor-135 in the rat. Neuroscience 144:165–190. doi:10.1016/j.neuroscience.2006.08.072

    Article  CAS  PubMed  Google Scholar 

  • Ma S, Olucha-Bordonau FE, Hossain MA, Lin F, Kuei C, Liu C, Wade JD, Sutton SW, Nunez A, Gundlach AL (2009a) Modulation of hippocampal theta oscillations and spatial memory by relaxin-3 neurons of the nucleus incertus. Learn Mem 16:730–742. doi:10.1101/lm.1438109

    Article  CAS  PubMed  Google Scholar 

  • Ma S, Sang Q, Lanciego JL, Gundlach AL (2009b) Localization of relaxin-3 in brain of Macaca fascicularis: identification of a nucleus incertus in primate. J Comp Neurol 517:856–872. doi:10.1002/cne.22197

    Article  CAS  PubMed  Google Scholar 

  • Ma S, Blasiak A, Olucha-Bordonau FE, Verberne AJ, Gundlach AL (2013) Heterogeneous responses of nucleus incertus neurons to corticotrophin-releasing factor and coherent activity with hippocampal theta rhythm in the rat. J Physiol 591:3981–4001. doi:10.1113/jphysiol.2013.254300

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Martinez-Bellver S, Cervera-Ferri A, Martinez-Ricos J, Ruiz-Torner A, Luque-Garcia A, Blasco-Serra A, Guerrero-Martinez J, Bataller-Mompean M, Teruel-Marti V (2015) Regular theta-firing neurons in the nucleus incertus during sustained hippocampal activation. Eur J Neurosci 41:1049–1067. doi:10.1111/ejn.12884

    Article  PubMed  Google Scholar 

  • Morozov A, Kellendonk C, Simpson E, Tronche F (2003) Using conditional mutagenesis to study the brain. Biol Psychiatry 54:1125–1133

    Article  CAS  PubMed  Google Scholar 

  • Niell CM, Stryker MP (2010) Modulation of visual responses by behavioral state in mouse visual cortex. Neuron 65:472–479. doi:10.1016/j.neuron.2010.01.033

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Nunez A, Cervera-Ferri A, Olucha-Bordonau F, Ruiz-Torner A, Teruel V (2006) Nucleus incertus contribution to hippocampal theta rhythm generation. Eur J Neurosci 23:2731–2738. doi:10.1111/j.1460-9568.2006.04797.x

    Article  CAS  PubMed  Google Scholar 

  • Olbrich S, Olbrich H, Jahn I, Sander C, Adamaszek M, Hegerl U, Reque F, Stengler K (2013) EEG-vigilance regulation during the resting state in obsessive-compulsive disorder. Clin Neurophysiol 124:497–502. doi:10.1016/j.clinph.2012.08.018

    Article  PubMed  Google Scholar 

  • Olucha-Bordonau FE, Teruel V, Barcia-Gonzalez J, Ruiz-Torner A, Valverde-Navarro AA, Martinez-Soriano F (2003) Cytoarchitecture and efferent projections of the nucleus incertus of the rat. J Comp Neurol 464:62–97. doi:10.1002/cne.10774

    Article  PubMed  Google Scholar 

  • Pang KC, Jiao X, Sinha S, Beck KD, Servatius RJ (2011) Damage of GABAergic neurons in the medial septum impairs spatial working memory and extinction of active avoidance: effects on proactive interference. Hippocampus 21:835–846. doi:10.1002/hipo.20799

    CAS  PubMed  Google Scholar 

  • Paulson JC, McClure WO (1975) Inhibition of axoplasmic transport by colchicine, podophyllotoxin, and vinblastine: an effect on microtubules. Ann N Y Acad Sci 253:517–527

    Article  CAS  PubMed  Google Scholar 

  • Paxinos G, Watson C (2007) The rat brain in stereotaxic coordinates, 4th edn. Academic Press, San Diego

    Google Scholar 

  • Paxinos G, Carrive P, Wang H, Wang PY (1999) Chemoarchitectonic atlas of the rat brainstem. Academic Press, San Diego

    Google Scholar 

  • Pereira CW, Santos FN, Sanchez-Perez AM, Otero-Garcia M, Marchioro M, Ma S, Gundlach AL, Olucha-Bordonau FE (2013) Electrolytic lesion of the nucleus incertus retards extinction of auditory conditioned fear. Behav Brain Res 247:201–210. doi:10.1016/j.bbr.2013.03.025

    Article  CAS  PubMed  Google Scholar 

  • Pfaff DW (2006) Brain arousal and information theory: neural and genetic mechanisms. Harvard University Press, Cambridge

    Book  Google Scholar 

  • Potter E, Sutton S, Donaldson C, Chen R, Perrin M, Lewis K, Sawchenko PE, Vale W (1994) Distribution of corticotropin-releasing factor receptor mRNA expression in the rat brain and pituitary. Proc Natl Acad Sci USA 91:8777–8781

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Quinkert AW, Vimal V, Weil ZM, Reeke GN, Schiff ND, Banavar JR, Pfaff DW (2011) Quantitative descriptions of generalized arousal, an elementary function of the vertebrate brain. Proc Natl Acad Sci USA 108(Suppl 3):15617–15623. doi:10.1073/pnas.1101894108

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Rajkumar R, Wu Y, Farooq U, Tan WH, Dawe GS (2016) Stress activates the nucleus incertus and modulates plasticity in the hippocampo-medial prefrontal cortical pathway. Brain Res Bull 120:83–89. doi:10.1016/j.brainresbull.2015.10.010

    Article  CAS  PubMed  Google Scholar 

  • Richichi C, Lin EJ, Stefanin D, Colella D, Ravizza T, Grignaschi G, Veglianese P, Sperk G, During MJ, Vezzani A (2004) Anticonvulsant and antiepileptogenic effects mediated by adeno-associated virus vector neuropeptide Y expression in the rat hippocampus. J Neurosci 24:3051–3059. doi:10.1523/JNEUROSCI.4056-03.2004

    Article  CAS  PubMed  Google Scholar 

  • Robinson J, Manseau F, Ducharme G, Amilhon B, Vigneault E, El Mestikawy S, Williams S (2016) Optogenetic activation of septal glutamatergic neurons drive hippocampal theta rhythms. J Neurosci 36:3016–3023. doi:10.1523/JNEUROSCI.2141-15.2016

    Article  CAS  PubMed  Google Scholar 

  • Roland JJ, Stewart AL, Janke KL, Gielow MR, Kostek JA, Savage LM, Servatius RJ, Pang KC (2014) Medial septum-diagonal band of Broca (MSDB) GABAergic regulation of hippocampal acetylcholine efflux is dependent on cognitive demands. J Neurosci 34:506–514. doi:10.1523/JNEUROSCI.2352-13.2014

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ryan PJ, Ma S, Olucha-Bordonau FE, Gundlach AL (2011) Nucleus incertus-an emerging modulatory role in arousal, stress and memory. Neurosci Biobehav Rev 35:1326–1341. doi:10.1016/j.neubiorev.2011.02.004

    Article  PubMed  Google Scholar 

  • Sanchez-Perez AM, Arnal-Vicente I, Santos FN, Pereira CW, ElMlili N, Sanjuan J, Ma S, Gundlach AL, Olucha-Bordonau FE (2015) Septal projections to nucleus incertus in the rat: bidirectional pathways for modulation of hippocampal function. J Comp Neurol 523:565–588. doi:10.1002/cne.23687

    Article  CAS  PubMed  Google Scholar 

  • Schindelin J, Arganda-Carreras I, Frise E et al (2012) Fiji: an open-source platform for biological-image analysis. Nat Methods 9:676–682. doi:10.1038/nmeth.2019

    Article  CAS  PubMed  Google Scholar 

  • Shinder ME, Taube JS (2011) Active and passive movement are encoded equally by head direction cells in the anterodorsal thalamus. J Neurophysiol 106:788–800. doi:10.1152/jn.01098.2010

    Article  PubMed  PubMed Central  Google Scholar 

  • Smith CM, Shen PJ, Banerjee A, Bonaventure P, Ma S, Bathgate RA, Sutton SW, Gundlach AL (2010) Distribution of relaxin-3 and RXFP3 within arousal, stress, affective, and cognitive circuits of mouse brain. J Comp Neurol 518:4016–4045. doi:10.1002/cne.22442

    Article  CAS  PubMed  Google Scholar 

  • Sotres-Bayon F, Diaz-Mataix L, Bush DE, LeDoux JE (2009) Dissociable roles for the ventromedial prefrontal cortex and amygdala in fear extinction: NR2B contribution. Cereb Cortex 19:474–482. doi:10.1093/cercor/bhn099

    Article  PubMed  Google Scholar 

  • Streeter GL (1903) Anatomy of the floor of the fourth ventricle. Am J Anat 2:U297–U299. doi:10.1002/aja.1000020303

    Article  Google Scholar 

  • Sutin EL, Jacobowitz DM (1988) Immunocytochemical localization of peptides and other neurochemicals in the rat laterodorsal tegmental nucleus and adjacent area. J Comp Neurol 270:243–270. doi:10.1002/cne.902700206

    Article  CAS  PubMed  Google Scholar 

  • Tanaka M, Iijima N, Miyamoto Y, Fukusumi S, Itoh Y, Ozawa H, Ibata Y (2005) Neurons expressing relaxin 3/INSL 7 in the nucleus incertus respond to stress. Eur J Neurosci 21:1659–1670. doi:10.1111/j.1460-9568.2005.03980.x

    Article  PubMed  Google Scholar 

  • Urban DJ, Roth BL (2015) DREADDs (designer receptors exclusively activated by designer drugs): chemogenetic tools with therapeutic utility. Ann Rev Pharmacol Toxicol 55:399–417. doi:10.1146/annurev-pharmtox-010814-124803

    Article  CAS  Google Scholar 

  • Wang XD, Chen C, Zhang D, Yao H (2014) Cumulative latency advance underlies fast visual processing in desynchronized brain state. Proc Natl Acad Sci USA 111:515–520. doi:10.1073/pnas.1316166111

    Article  CAS  PubMed  Google Scholar 

  • Watanabe Y, Tsujimura A, Takao K, Nishi K, Ito Y, Yasuhara Y, Nakatomi Y, Yokoyama C, Fukui K, Miyakawa T, Tanaka M (2011) Relaxin-3-deficient mice showed slight alteration in anxiety-related behavior. Front Behav Neurosci 5:50. doi:10.3389/fnbeh.2011.00050

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wells CE, Amos DP, Jeewajee A, Douchamps V, Rodgers J, O’Keefe J, Burgess N, Lever C (2013) Novelty and anxiolytic drugs dissociate two components of hippocampal theta in behaving rats. J Neurosci 33:8650–8667. doi:10.1523/JNEUROSCI.5040-12.2013

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • White MD, Milne RV, Nolan MF (2011) A molecular toolbox for rapid generation of viral vectors to up- or down-regulate neuronal gene expression in vivo. Front Mol Neurosci 4:8. doi:10.3389/fnmol.2011.00008

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zant JC, Kim T, Prokai L, Szarka S, McNally J, McKenna JT, Shukla C, Yang C, Kalinchuk AV, McCarley RW, Brown RE, Basheer R (2016) Cholinergic neurons in the basal forebrain promote wakefulness by actions on neighboring non-cholinergic neurons: an opto-dialysis study. J Neurosci 36:2057–2067. doi:10.1523/JNEUROSCI.3318-15.2016

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ziegler DR, Edwards MR, Ulrich-Lai YM, Herman JP, Cullinan WE (2012) Brainstem origins of glutamatergic innervation of the rat hypothalamic paraventricular nucleus. J Comp Neurol 520:2369–2394. doi:10.1002/cne.23043

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgments

The authors would like to thank Prof Bryan Roth (University of North Carolina, NC, USA) and Dr. Melanie White (ARMI, Monash University, VIC, Australia) for access to the DREADD construct and assistance in establishing the approach in our laboratory, respectively; and Prof Neil McNaughton (University of Otago, Dunedin, NZ) for valuable comments on the manuscript. This research was supported by project grants from the National Health and Medical Research Council (NHMRC) of Australia (1005988 and 1067522, A. L. G. and R. A. D. B.); a Grant from The Florey Institute of Neuroscience and Mental Health Foundation (A. L. G. and S. M.); a Grant from the Besen Family Foundation (A. L. G.); and by the Victorian Government Operational Infrastructure Support Program. S. J. W. is an Australian Research Council (ARC) Future Fellow. R. A. D. B. and A. L. G. are NHMRC (Australia) Senior Research Fellows. G.A. is the recipient of a Commonwealth of Australia International Postgraduate Research Scholarship (IPRS); and E. K. E. O-P. is the recipient of a University of Melbourne International Research Scholarship (MIFRS/MRS).

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Correspondence to Sherie Ma or Andrew L. Gundlach.

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429_2016_1230_MOESM1_ESM.mp4

Online resource 1. NI activation is associated with persistent locomotor activity. Concurrent video recording of 2 × NI-hM3Dq rats (upper chambers) and 2 × NI-mCherry rats (lower chambers) in automated locomotor cells during a 3 h trial immediately after injection of CNO (3 mg/kg, i.p.). Video speed is 2 × normal. Flashes of light are infrared beams activated automatically by the locomotor cell software that are not detected by the rats. Time-lapsed video recordings of activity illustrate the persistent ambulation and ‘interrupted’ attempts to rest of NI-hM3Dq rats. (MP4 3294 kb)

429_2016_1230_MOESM2_ESM.mp4

Online resource 2. NI activation is associated with increased risk-assessment and head-scanning behavior in response to learned potential threat. Concurrent video recording of 2 × NI-hM3Dq rats (lower chambers) and 2 × NI-mCherry rats (upper chambers) in operant chambers 24 h after fear conditioning where rats were trained to associate an audible tone (30 s, 80 dB, 5 kHz sine wave) with footshock (1 s, 0.7 mA). Video speed is 2 × normal. The video starts during the intertrial interval between tones 2-3, during which scanning behavior is exhibited by NI-hM3Dq rats, but infrequently by NI-mCherry rats that instead exhibit predominant freezing and immobility. A 30 s tone presentation (tone 3) is indicated by the presence of a small red LED on top of the chamber (not within view of the rats). During and after tone presentation, NI-hM3Dq rats exhibit high levels of head-scanning and risk assessment behavior. (MP4 2264 kb)

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Ma, S., Allocca, G., Ong-Pålsson, E.K.E. et al. Nucleus incertus promotes cortical desynchronization and behavioral arousal. Brain Struct Funct 222, 515–537 (2017). https://doi.org/10.1007/s00429-016-1230-0

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