Russian Journal of PhysiologyRussian Journal of PhysiologyРоссийский физиологический журнал им. И.М. Сеченова0869-81392658-655XThe Russian Academy of Sciences65167510.31857/S0869813924020071DJINUEEXPERIMENTAL ARTICLESЭКСПЕРИМЕНТАЛЬНЫЕ СТАТЬИResearch ArticleInfluence of long-term social isolation on the behavior and dopamine system of the brain in miceВлияние длительной социальной изоляции на поведение и дофаминовую систему мозга у мышейBazovkinaD. V.БазовкинаД. В.
Russian Federation
daryabazovkina@gmail.comUstinovaU. S.УстиноваУ. С.
Russian Federation
daryabazovkina@gmail.comAdoninaS. N.АдонинаС. Н.
Russian Federation
daryabazovkina@gmail.comKomlevaP. D.КомлеваП. Д.
Russian Federation
daryabazovkina@gmail.comArefievaА. B.АрефьеваА. Б.
Russian Federation
daryabazovkina@gmail.comKulikovaЕ. А.КуликоваЕ. А.
Russian Federation
daryabazovkina@gmail.comInstitute of Cytology and Genetics, Siberian Branch of the Russian Academy of SciencesИнститут цитологии и генетики СО РАНNovosibirsk National Research State UniversityНовосибирский национальный исследовательский государственный университет15022024110223825301022025Copyright ©; 2024, Russian Academy of SciencesCopyright ©; 2024, Российская академия наук2024Russian Academy of SciencesРоссийская академия наукhttps://journals.eco-vector.com/0869-8139/article/view/651675

The dopamine brain system is involved in the regulation of various physiological processes and behavioral responses, including social behavior. It was shown in rodents that prolonged social isolation can lead to changes in behavioral parameters and neurochemical parameters, but these data remain unclear. In this work, we studied the effects of prolonged social isolation on the behavior and state of the brain dopamine system in male C57Bl/6 mice. Animals of the experimental group from the age of 40 - 42 days were kept individually in separate cages for six weeks, while the mice of the control group were kept in-group conditions. Isolation did not affect locomotor and exploratory activity in the “open field” test compared to the content in the group. At the same time, animals subjected to prolonged isolation demonstrated a longer duration of social contacts in the “resident-intruder” model and a weakening of stereotypical behavior in the “marbles burying” test compared to the control group. These behavioral changes were accompanied by an increase in mRNA levels of the genes encoding dopamine D1 and D2 receptors in the striatum of mice. In addition, the level of the dopamine metabolite DOPAC decreased in the hypothalamus and increased in the frontal cortex of mice due to the effect of social isolation compared with the control group. The results contribute to the understanding of the effects of prolonged social isolation on the behavior and dopamine brain system in mice.

Дофаминовая система мозга участвует в регуляции различных физиологических процессов и поведенческих реакций, включая социальное поведение. На грызунах было показано, что длительное одиночное содержание может приводить к изменениям параметров поведения и нейрохимических показателей, однако не всегда эти данные можно однозначно интерпретировать. В настоящей работе исследовали эффекты продолжительной социальной изоляции на поведение и состояние дофаминовой системы мозга у самцов мышей линии C57Bl/6. Животных опытной группы с 40–42-дневного возраста содержали индивидуально в отдельных клетках в течение шести недель, в то время как мышей контрольной группы – в групповых условиях. Изоляция не повлияла на двигательную и исследовательскую активность в тесте «открытое поле» по сравнению с групповым содержанием. В то же время животные, подвергавшиеся продолжительной изоляции, демонстрировали большую длительность социальных контактов в модели «резидент – интрудер» и ослабление стереотипного поведения в тесте «закапывание шариков» по сравнению с контрольной группой. Эти изменения в поведении сопровождались повышением уровней мРНК генов, кодирующих дофаминовые рецепторы D1 и D2, в стриатуме мышей. Кроме этого, уровень метаболита дофамина DOPAC снижался в гипоталамусе и повышался во фронтальной коре мышей вследствие действия социальной изоляции по сравнению с контрольной группой. Результаты вносят вклад в понимание эффектов длительной социальной изоляции на поведение и дофаминовую систему мозга у мышей.

social isolationbehaviordopaminedopamine receptorgene expressionbrainmiceсоциальная изоляцияповедениедофаминдофаминовый рецепторэкспрессия геновмозгмышиРоссийский научный фондRussian Science Foundation№ 21-15-00051This work was financed by the Russian Science Foundation (grant No. 21-15-00051). The mouse lines were maintained using funds from the budget project FWNR-2022-0023.Данная работа финансировалась за счет средств Российского Научного Фонда (грант № 21-15-00051). Содержание линий мышей осуществлялось за счет средств бюджетного проекта FWNR-2022-0023.Arakawa H (2018) Ethological approach to social isolation effects in behavioral studies of laboratory rodents. Behav Brain Res 341: 98–108. https://doi.org/10.1016/j.bbr.2017.12.022Brandt L, Liu S, Heim C, Heinz A (2022) The effects of social isolation stress and discrimination on mental health. Translational Psychiatry 12: 398. https://doi.org/10.1038/s41398-022-02178-4Nakama N, Usui N, Doi M, Shimada S (2023) Early life stress impairs brain and mental development during childhood increasing the risk of developing psychiatric disorders. Prog Neuropsychopharmacol Biol Psychiatry 126: 110783. https://doi.org/10.1016/j.pnpbp.2023.110783Kaneda Y, Kawata A, Suzuki K, Matsunaga D, Yasumatsu M, Ishiwata T (2021) Comparison of neurotransmitter levels, physiological conditions, and emotional behavior between isolation-housed rats with group-housed rats. Dev Psychobiol 63: 452–460. https://doi.org/10.1002/dev.22036Zhang X, Xun Y, Wang L, Zhang J, Hou W, Ma H, Cai W, Li L, Guo Q, Li Y, Lv Z, Jia R, Tai F, He Z (2021) Involvement of the dopamine system in the effect of chronic social isolation during adolescence on social behaviors in male C57 mice. Brain Res 1765: 147497. https://doi.org/10.1016/j.brainres.2021.147497Grigoryan GA, Pavlova IV, Zaichenko MI (2022) Effects of Social Isolation on the Development of Anxiety and Depression-Like Behavior in Model Experiments in Animals. Neurosci Behav Physiol 52: 722–738. https://doi.org/10.1007/s11055-022-01297-1Lukkes JL, Mokin MV, Scholl JL, Forster GL (2009) Adult rats exposed to early-life social isolation exhibit increased anxiety and conditioned fear behavior, and altered hormonal stress responses. Horm Behav 55: 248–256. https://doi.org/10.1016/j.yhbeh.2008.10.014Wang HT, Huang FL, Hu ZL, Zhang WJ, Qiao XQ, Huang YQ, Dai RP, Li F, Li CQ (2017) Early-Life Social Isolation-Induced Depressive-Like Behavior in Rats Results in Microglial Activation and Neuronal Histone Methylation that Are Mitigated by Minocycline. Neurotox Res 31: 505–520. https://doi.org/10.1007/s12640-016-9696-3Кoike H, Ibi D, Mizoguchi H, Nagai T, Nitta A, Takuma K, Nabeshima T, Yoneda Y, Yamada K (2009) Behavioral abnormality and pharmacologic response in social isolation-reared mice. Behav Brain Res 202: 114–121. https://doi.org/10.1016/j.bbr.2009.03.028Kercmar J, Büdefeld T, Grgurevic N, Tobet SA, Majdic G (2011) Adolescent social isolation changes social recognition in adult mice. Behav Brain Res 216: 647–651. https://doi.org/10.1016/j.bbr.2010.09.007Dankoski EC, Agster KL, Fox ME, Moy SS, Wightman RM (2014) Facilitation of serotonin signaling by SSRIs is attenuated by social isolation. Neuropsychopharmacology 39: 2928–2937. https://doi.org/10.1038/npp.2014.162Kinley BL, Kyne RF, Lawton-Stone TS, Walker DM, Paul MJ (2021) Long-term consequences of peri-adolescent social isolation on social preference, anxiety-like behaviour, and vasopressin neural circuitry of male and female rats. Eur J Neurosci 54: 7790–7804. https://doi.org/10.1111/ejn.15520.Perić I, Stanisavljević A, Gass P, Filipović D (2021) Fluoxetine exerts subregion/layer specific effects on parvalbumin/GAD67 protein expression in the dorsal hippocampus of male rats showing social isolation-induced depressive-like behaviour. Brain Res Bull 173: 174–183. https://doi.org/10.1016/j.brainresbull.2021Baskerville TA, Douglas AJ (2010) Dopamine and oxytocin interactions underlying behaviors: potential contributions to behavioral disorders. CNS Neurosci Ther 16: e92–e123. https://doi.org/10.1111/j.1755-5949.2010.00154.xBeaulieu JM, Espinoza S, Gainetdinov RR (2015) Dopamine receptors - IUPHAR Review 13. Br J Pharmacol 172: 1–23. https://doi.org/10.1111/bph.12906Villalba RM, Smith Y (2013) Differential striatal spine pathology in Parkinson’s disease and cocaine addiction: a key role of dopamine? Neuroscience 251: 2–20. https://doi.org/10.1016/j.neuroscience.2013.07.011Grace AA (2016) Dysregulation of the dopamine system in the pathophysiology of schizophrenia and depression. Nat Rev Neurosci 17: 524–532. https://doi.org/10.1038/nrn.2016.57Correia R, Coimbra B, Domingues AV, Wezik M, Vieitas-Gaspar N, Gaspar R, Sousa N, Pinto L, Rodrigues AJ, Soares-Cunha C (2023) Involvement of nucleus accumbens D2-medium spiny neurons projecting to the ventral pallidum in anxiety-like behaviour. J Psychiatry Neurosci 48: E267–E284. https://doi: 10.1503/jpn.220111Han X, Wang W, Shao F, Li N (2011) Isolation rearing alters social behaviors and monoamine neurotransmission in the medial prefrontal cortex and nucleus accumbens of adult rats. Brain Res 1385: 175–181. https://doi:10.1016/j.brainres.2011.02.035Trabace L, Zotti M, Colaianna M, Morgese MG, Schiavone S, Tucci P, Harvey BH, Wegener G, Cuomo V (2012) Neurochemical differences in two rat strains exposed to social isolation rearing. Acta Neuropsychiatr 24: 286–295. https://doi.org/10.1111/j.1601-5215.2011.00627.xBrenes JC, Fornaguera J (2009) The effect of chronic fluoxetine on social isolation-induced changes on sucrose consumption, immobility behavior, and on serotonin and dopamine function in hippocampus and ventral striatum. Behav Brain Res 198: 199–205. https://doi.org/10.1016/j.bbr.2008.10.036Hall FS (1998) Social deprivation of neonatal, adolescent, and adult rats has distinct neurochemical and behavioral consequences. Crit Rev Neurobiol 12: 129–162. https://doi.org/10.1615/critrevneurobiol.v12.i1-2.50Bean G, Lee T (1991) Social isolation and cohabitation with haloperidol-treated partners: effect on density of striatal dopamine D2 receptors in the developing rat brain. Psychiatry Res 36: 307–317. https://doi.org/10.1016/0165-1781(91) 90029-oFitzgerald ML, Mackie K, Pickel VM (2013) The impact of adolescent social isolation on dopamine D2 and cannabinoid CB1 receptors in the adult rat prefrontal cortex. Neuroscience 235: 40–50. https://doi.org/10.1016/j.neuroscience.2013.01.021King MV, Seeman P, Marsden CA, Fone KCF (2009). Increased dopamine D2Highreceptors in rats reared in social isolation. Synapse 63: 476–483. https://doi.org/10.1002/syn.20624Archer J (1969) Contrasting effects of group housing and isolation on subsequent open field exploration in laboratory rats. Psychon Sci 14: 234–235. https://doi.org/10.3758/BF03332812Науменко ЕВ, Попова НК, Старыгин АГ (1971) Гипофизарно-надпочечниковая система животных в группах и изоляции. Журн общей биол 32: 731–736. [Naumenko EV, Popova NK, Starygin AG (1971) Pituitary-adrenal system of animals in groups and isolation. J General Biol 32: 731–736. (In Russ)].Kulikov AV, Tikhonova MA, Kulikov VA (2008). Automated measurement of spatial preference in the open field test with transmitted lighting. J Neurosci Methods 170: 345–351. https://doi.org/10.1016/j.jneumeth.2008.01.024Deacon RM (2006) Digging and marble burying in mice: simple methods for in vivo identification of biological impacts. Nature Protocols 1: 122–124. https://doi.org/10/1038/nprot.2006.20Kulikova EA, Bazovkina DV, Antonov YV, Akulov AE, Kulikov AV, Kondaurova EM (2017) Alteration of the brain morphology and the response to the acute stress in the recombinant mouse lines with different predisposition to catalepsy. Neurosci Res 117: 14–21. https://doi.org/10.1016/j.neures.2016.11.009Bazovkina D, Naumenko V, Bazhenova E, Kondaurova E (2021) Effect of Central Administration of Brain-Derived Neurotrophic Factor (BDNF) on Behavior and Brain Monoamine Metabolism in New Recombinant Mouse Lines Differing by 5-HT1A Receptor Functionality. Int J Mol Sci 22: 11987. https://doi.org/10.3390/ijms222111987Naumenko VS, Kulikov AV (2006) Quantitative assay of 5-HT(1A) serotonin receptor gene expression in the brain. Mol Biol (Mosk) 40: 37–44. https://doi.org/10.1134/s0026893306010079Naumenko VS, Osipova DV, Kostina EV, Kulikov AV (2008) Utilization of a two-standard system in real-time PCR for quantification of gene expression in the brain. J Neurosci Methods 170: 197–203. https://doi.org/10.1016/j.jneumeth.2008.01.008Du Preez A, Law T, Onorato D, Lim YM, Eiben P, Musaelyan K, Egeland M, Hye A, Zunszain PA, Thuret S, Pariante CM, Fernandes C (2020) The type of stress matters: repeated injection and permanent social isolation stress in male mice have a differential effect on anxiety- and depressive-like behaviours, and associated biological alterations. Transl Psychiatry 10: 325. https://doi.org/10.1038/s41398-020-01000-3Lander SS, Linder-Shacham D, Gaisler-Salomon I (2017) Differential effects of social isolation in adolescent and adult mice on behavior and cortical gene expression. Behav Brain Res 316: 245–254. https://doi.org/10.1016/j.bbr.2016.09.005Locci A, Geoffroy P, Miesch M, Mensah-Nyagan AG, Pinna G (2017) Social Isolation in Early versus Late Adolescent Mice Is Associated with Persistent Behavioral Deficits That Can Be Improved by Neurosteroid-Based Treatment. Front Cell Neurosci 11: 208. https://doi.org/10.3389/fncel.2017.00208Liu N, Wang Y, An AY, Banker C, Qian YH, O’Donnell JM (2020) Single housing-induced effects on cognitive impairment and depression-like behavior in male and female mice involve neuroplasticity-related signaling. Eur J Neurosci 52: 2694–2704. https://doi.org/10.1111/ejn.14565Oliver DK, Intson K, Sargin D, Power SK, McNabb J, Ramsey AJ, Lambe EK (2020) Chronic social isolation exerts opposing sex-specific consequences on serotonin neuronal excitability and behaviour. Neuropharmacology 168: 108015. https://doi.org/10.1016/j.neuropharm.2020.108015Amiri S, Haj-Mirzaian A, Rahimi-Balaei M, Razmi A, Kordjazy N, Shirzadian A, Ejtemaei Mehr S, Sianati H, Dehpour AR (2015) Co-occurrence of anxiety and depressive-like behaviors following adolescent social isolation in male mice; possible role of nitrergic system. Physiol Behav 145: 38–44. https://doi.org/10.1016/j.physbeh.2015.03.032Võikar V, Polus A, Vasar E, Rauvala H (2005) Long-term individual housing in C57BL/6J and DBA/2 mice: assessment of behavioral consequences. Genes Brain Behav 4: 240–252. https://doi.org/10.1111/j.1601-183X.2004.00106.xKraeuter AK, Guest PC, Sarnyai Z (2019) The Open Field Test for Measuring Locomotor Activity and Anxiety-Like Behavior. Methods Mol Biol 1916: 99–103. https://doi.org/10.1007/978-1-4939-8994-2_9Dixit PV, Sahu R, Mishra DK (2020) Marble-burying behavior test as a murine model of compulsive-like behavior. J Pharmacol Toxicol Methods 102: 106676. https://doi.org/10.1016/j.vascn.2020.106676Himanshu, Dharmila, Sarkar D, Nutan (2020) A Review of Behavioral Tests to Evaluate Different Types of Anxiety and Anti-anxiety Effects. Clin Psychopharmacol Neurosci 18: 341–351. https://doi.org/10.9758/cpn.2020.18.3.341Zlatković J, Todorović N, Bošković M, Pajović SB, Demajo M, Filipović D (2014) Different susceptibility of prefrontal cortex and hippocampus to oxidative stress following chronic social isolation stress. Mol Cell Biochem 393: 43–57. https://doi.org/10.1007/s11010-014-2045-zThomas A, Burant A, Bui N, Graham D, Yuva-Paylor LA, Paylor R (2009) Marble burying reflects a repetitive and perseverative behavior more than novelty-induced anxiety. Psychopharmacology (Berl) 204: 361–373. https://doi.org/10.1007/s00213-009-1466-yTulogdi A, Tóth M, Barsvári B, Biró L, Mikics E, Haller J (2014) Effects of resocialization on post-weaning social isolation-induced abnormal aggression and social deficits in rats. Dev Psychobiol 56: 49–57. https://doi.org/10.1002/dev.21090Biro L, Toth M, Sipos E, Bruzsik B, Tulogdi A, Bendahan S, Sandi C, Haller J (2017) Structural and functional alterations in the prefrontal cortex after post-weaning social isolation: relationship with species-typical and deviant aggression. Brain Struct Funct 222: 1861–1875. https://doi.org/10.1007/s00429-016-1312-zMa YK, Zeng PY, Chu YH, Lee CL, Cheng CC, Chen CH, Su YS, Lin KT, Kuo TH (2022) Lack of social touch alters anxiety-like and social behaviors in male mice. Stress 25: 134–144. https://doi.org/10.1080/10253890.2022.2047174Weele CMV, Siciliano CA, Tye KM (2019) Dopamine tunes prefrontal outputs to orchestrate aversive processing. Brain Res 1713: 16–31. https://doi.org/10.1016/j.brainres.2018.11.044Shirenova SD, Khlebnikova NN, Narkevich VB, Kudrin VS, Krupina NA (2023) Nine-month-long Social Isolation Changes the Levels of Monoamines in the Brain Structures of Rats: A Comparative Study of Neurochemistry and Behavior. Neurochem Res 48: 1755–1774. https://doi.org/10.1007/s11064-023-03858-3Ko C-Y, Liu Y-P (2016) Disruptions of sensorimotor gating, cytokines, glycemia, monoamines, and genes in both sexes of rats reared in social isolation can be ameliorated by oral chronic quetiapine administration. Brain Behav Immun 51: 119–130. https://doi.org/10.1016/j.bbi.2015.08.003Mncube K, Möller M, Harvey BH (2021) Post-weaning social isolated flinders sensitive line rats display bio-behavioural manifestations resistant to fluoxetine: a model of treatment-resistant depression. Front Psychiatry 12: 688150. https://doi.org/10.3389/ fpsyt.2021.688150Algamal M, Pearson AJ, Hahn-Townsend C, Burca I, Mullan M, Crawford F, Ojo JO (2021) Repeated unpredictable stress and social isolation induce chronic HPA axis dysfunction and persistent abnormal fear memory. Prog Neuropsychopharmacol Biol Psychiatry 104: 110035. https://doi.org/10.1016/j.pnpbp.2020.110035Watanabe T, Iba H, Moriyama H, Kubota K, Katsurabayashi S, Iwasaki K (2021) Sansoninto attenuates aggressive behavior and increases levels of homovanillic acid, a dopamine metabolite, in social isolation-reared mice. J Tradit Complement Med 12: 243–249. https://doi.org/10.1016/j.jtcme.2021.08.006Yun S, Yang B, Anair JD, Martin MM, Fleps SW, Pamukcu A, Yeh NH, Contractor A, Kennedy A, Parker JG (2023) Antipsychotic drug efficacy correlates with the modulation of D1 rather than D2 receptor-expressing striatal projection neurons. Nat Neurosci 26: 1417–1428. https://doi.org/10.1038/s41593-023-01390-9Bruins Slot LA, Bardin L, Auclair AL, Depoortere R, Newman-Tancredi A (2008) Effects of antipsychotics and reference monoaminergic ligands on marble burying behavior in mice. Behav Pharmacol 19: 145–152. https://doi.org/10.1097/FBP.0b013e3282f62cb2Egashira N, Kubota N, Goto Y, Watanabe T, Kubota K, Katsurabayashi S, Iwasaki K (2018) The antipsychotic trifluoperazine reduces marble-burying behavior in mice via D2 and 5-HT2A receptors: Implications for obsessive-compulsive disorder. Pharmacol Biochem Behav 165: 9–13. https://doi.org/10.1016/j.pbb.2017.12.006Del Arco A, Zhu S, Terasmaa A, Mohammed AH, Fuxe K (2004) Hyperactivity to novelty induced by social isolation is not correlated with changes in D2 receptor function and binding in striatum. Psychopharmacology (Berl) 171: 148–155. https://doi.org/10.1007/s00213-003-1578-8Yorgason JT, Calipari ES, Ferris MJ, Karkhanis AN, Fordahl SC, Weiner JL, Jones SR (2016) Social isolation rearing increases dopamine uptake and psychostimulant potency in the striatum. Neuropharmacology 101: 471–479. https://doi.org/10.1016/j.neuropharm.2015.10.025