An Essential Role of the Arginine Vasotocin System in Mate-Guarding Behaviors in Triadic Relationships of Medaka Fish (Oryzias latipes)

To increase individual male fitness, males of various species remain near a (potential) mating partner and repel their rivals (mate-guarding). Mate-guarding is assumed to be mediated by two different types of motivation: sexual motivation toward the opposite sex and competitive motivation toward the same sex. The genetic/molecular mechanisms underlying how mate presence affects male competitive motivation in a triadic relationship has remained largely unknown. Here we showed that male medaka fish prominently exhibit mate-guarding behavior. The presence of a female robustly triggers male-male competition for the female in a triadic relationship (2 males and 1 female). The male-male competition resulted in one male occupying a dominant position near the female while interfering with the other male's approach of the female. Paternity testing revealed that the dominant male had a significantly higher mating success rate than the other male in a triadic relationship. We next generated medaka mutants of arginine-vasotocin (avt) and its receptors (V1a1, V1a2) and revealed that two genes, avt and V1a2, are required for normal mate-guarding behavior. In addition, behavioral analysis of courtship behaviors in a dyadic relationship and aggressive behaviors within a male group revealed that avt mutant males displayed decreased sexual motivation but showed normal aggression. In contrast, heterozygote V1a2 mutant males displayed decreased aggression, but normal mate-guarding and courtship behavior. Thus, impaired mate-guarding in avt and V1a2 homozygote mutants may be due to the loss of sexual motivation toward the opposite sex, and not to the loss of competitive motivation toward rival males. The different behavioral phenotypes between avt, V1a2 heterozygote, and V1a2 homozygote mutants suggest that there are redundant systems to activate V1a2 and that endogenous ligands activating the receptor may differ according to the social context.

[1]  I. Khan,et al.  The distribution of an AVT V1a receptor in the brain of a sex changing fish, Epinephelus adscensionis , 2011, Journal of Chemical Neuroanatomy.

[2]  H. Ariga,et al.  Efficient Targeted Mutagenesis in Medaka Using Custom-Designed Transcription Activator-Like Effector Nucleases , 2013, Genetics.

[3]  美香 小井土,et al.  屋外池における野生メダカ Oryzias latipes の繁殖行動 , 2012 .

[4]  F. Hinz,et al.  Arginine vasotocin regulates social ascent in the African cichlid fish Astatotilapia burtoni. , 2015, General and comparative endocrinology.

[5]  A. Bass,et al.  New insights into neuropeptide modulation of aggression: field studies of arginine vasotocin in a territorial tropical damselfish , 2006, Proceedings of the Royal Society B: Biological Sciences.

[6]  Y. Kamei,et al.  Controlled Cre/loxP Site-Specific Recombination in the Developing Brain in Medaka Fish, Oryzias latipes , 2013, PLoS ONE.

[7]  K. Ewen,et al.  DNA fingerprinting in relation to male dominance and paternity in a captive colony of tammar wallabies (Macropus eugenii). , 1993, Journal of reproduction and fertility.

[8]  Yasuhiro Kamei,et al.  A Neural Mechanism Underlying Mating Preferences for Familiar Individuals in Medaka Fish , 2014, Science.

[9]  W. F. Dongen.  Mate guarding and territorial aggression vary with breeding synchrony in golden whistlers (Pachycephala pectoralis) , 2008, Naturwissenschaften.

[10]  J. Haller.  The neurobiology of abnormal manifestations of aggression—A review of hypothalamic mechanisms in cats, rodents, and humans , 2013, Brain Research Bulletin.

[11]  J. Godwin,et al.  Manipulations of the AVT System Shift Social Status and Related Courtship and Aggressive Behavior in the Bluehead Wrasse , 2001, Hormones and Behavior.

[12]  N. Seddon,et al.  Yelling for sex: harem males compete for female access in bronze-winged jacanas , 1999, Animal Behaviour.

[13]  H. Takeuchi,et al.  Mass spectrometric map of neuropeptide expression and analysis of the gamma-prepro-tachykinin gene expression in the medaka (Oryzias latipes) brain. , 2009, General and comparative endocrinology.

[14]  N. Kagawa.  Social rank-dependent expression of arginine vasotocin in distinct preoptic regions in male Oryzias latipes. , 2013, Journal of fish biology.

[15]  G. Nevitt,et al.  Exogenous vasotocin alters aggression during agonistic exchanges in male Amargosa River pupfish (Cyprinodon nevadensis amargosae) , 2004, Hormones and Behavior.

[16]  A. Takeuchi,et al.  Sex differences in the expression of vasotocin/isotocin, gonadotropin-releasing hormone, and tyrosine and tryptophan hydroxylase family genes in the medaka brain , 2012, Neuroscience.

[17]  P. Sherman.  Mate guarding as paternity insurance in Idaho ground squirrels , 1989, Nature.

[18]  Daisuke Kurabayashi,et al.  Coordinated and Cohesive Movement of Two Small Conspecific Fish Induced by Eliciting a Simultaneous Optomotor Response , 2010, PloS one.

[19]  J. Y. Kuwada,et al.  Expression of arginine vasotocin receptors in the developing zebrafish CNS. , 2013, Gene expression patterns : GEP.

[20]  J. Komdeur.  Mate guarding in the Seychelles warbler is energetically costly and adjusted to paternity risk , 2001, Proceedings of the Royal Society of London. Series B: Biological Sciences.

[21]  David J. Anderson,et al.  A Framework for Studying Emotions across Species , 2014, Cell.

[22]  H. Drummond,et al.  Male whiptail lizards adjust energetically costly mate guarding to male–male competition and female reproductive value , 2010, Animal Behaviour.

[23]  Larry J. Young,et al.  The prairie vole: an emerging model organism for understanding the social brain , 2010, Trends in Neurosciences.

[24]  D. Buss.  Human mate guarding. , 2002, Neuro endocrinology letters.

[25]  J. Godwin,et al.  Variation in gene transcript profiles of two V1a-type arginine vasotocin receptors among sexual phases of bluehead wrasse (Thalassoma bifasciatum). , 2012, General and comparative endocrinology.

[26]  B. Sinervo,et al.  Polygyny, mate-guarding, and posthumous fertilization as alternative male mating strategies. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[27]  L. Simmons,et al.  Female crickets assess relatedness during mate guarding and bias storage of sperm towards unrelated males , 2013, Journal of evolutionary biology.

[28]  H. Takeda,et al.  Transient and permanent gene transfer into the brain of the teleost fish medaka (Oryzias latipes) using human adenovirus and the Cre‐loxP system , 2010, FEBS letters.

[29]  S. Lema.  Identification of multiple vasotocin receptor cDNAs in teleost fish: Sequences, phylogenetic analysis, sites of expression, and regulation in the hypothalamus and gill in response to hyperosmotic challenge , 2010, Molecular and Cellular Endocrinology.

[30]  Nobuaki Takemori,et al.  Proteome mapping of the Drosophila melanogaster male reproductive system , 2009, Proteomics.

[31]  G. Guillon,et al.  Peptide and non-peptide agonists and antagonists for the vasopressin and oxytocin V1a, V1b, V2 and OT receptors: research tools and potential therapeutic agents. , 2008, Progress in brain research.

[32]  T. Todo,et al.  High-resolution melting curve analysis for rapid detection of mutations in a Medaka TILLING library , 2010, BMC Molecular Biology.

[33]  J. Goodson,et al.  To flock or fight: Neurochemical signatures of divergent life histories in sparrows , 2012, Proceedings of the National Academy of Sciences.

[34]  T. Birkhead,et al.  Extra-pair copulation and sperm competition in the zebra finch , 1988, Nature.

[35]  Thomas R. Insel,et al.  A role for central vasopressin in pair bonding in monogamous prairie voles , 1993, Nature.

[36]  R. R. Capranica,et al.  Hormone-induced vocal behavior and midbrain auditory sensitivity in the green treefrog, Hyla cinerea , 2004, Journal of Comparative Physiology A.

[37]  H. Albers,et al.  Oxytocin induces social communication by activating arginine-vasopressin V1a receptors and not oxytocin receptors , 2014, Psychoneuroendocrinology.

[38]  A. Bass,et al.  Individual Behavioral and Neuronal Phenotypes for Arginine Vasotocin Mediated Courtship and Aggression in a Territorial Teleost , 2010, Brain, Behavior and Evolution.

[39]  Y. Sakaki,et al.  Generation of medaka gene knockout models by target-selected mutagenesis , 2006, Genome Biology.

[40]  J. Godwin,et al.  Nonapeptides and Social Behavior in Fishes , 2012, Hormones and Behavior.

[41]  I. Khan,et al.  Distribution of nonapeptide systems in the forebrain of an African cichlid fish, Astatotilapia burtoni , 2012, Journal of Chemical Neuroanatomy.

[42]  A. Meyer,et al.  Dual control by a single gene of secondary sexual characters and mating preferences in medaka , 2009, BMC Biology.

[43]  Larry J. Young,et al.  Enhanced partner preference in a promiscuous species by manipulating the expression of a single gene , 2004, Nature.

[44]  G. Hill,et al.  Fighting ability and motivation: determinants of dominance and contest strategies in females of a passerine bird , 2007, Animal Behaviour.

[45]  M. Mann,et al.  Stop and go extraction tips for matrix-assisted laser desorption/ionization, nanoelectrospray, and LC/MS sample pretreatment in proteomics. , 2003, Analytical chemistry.

[46]  N. Kagawa.  Comparison of Aggressive Behaviors Between Two Wild Populations of Japanese Medaka, Oryzias latipes and O. sakaizumii , 2014, Zoological science.

[47]  L. Young,et al.  The neurobiology of pair bonding , 2004, Nature Neuroscience.

[48]  J. Heierhorst,et al.  Structure, function, and phylogeny of [Arg8]vasotocin receptors from teleost fish and toad. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[49]  L. O’Connell,et al.  The Vertebrate mesolimbic reward system and social behavior network: A comparative synthesis , 2011, The Journal of comparative neurology.

[50]  M. Schartl,et al.  Design, evaluation, and screening methods for efficient targeted mutagenesis with transcription activator‐like effector nucleases in medaka , 2014, Development, growth & differentiation.

[51]  S. E. Walker,et al.  Information asymmetry and aggressive behaviour in male house crickets, Acheta domesticus , 2008, Animal Behaviour.

[52]  Caroline E. G. Tutin,et al.  Mating patterns and reproductive strategies in a community of wild chimpanzees (Pan troglodytes schweinfurthii) , 1979, Behavioral Ecology and Sociobiology.

[53]  J. Goodson.  Territorial Aggression and Dawn Song are Modulated by Septal Vasotocin and Vasoactive Intestinal Polypeptide in Male Field Sparrows (Spizella pusilla) , 1998, Hormones and Behavior.

[54]  K. A. Paczolt,et al.  Post-copulatory sexual selection and sexual conflict in the evolution of male pregnancy , 2010, Nature.

[55]  M. Eens,et al.  Copulation and mate-guarding patterns in polygynous European starlings , 1997, Animal Behaviour.

[56]  Yoshiaki Ono,et al.  Mating Ethogram in Oryzias latipes (With 1 Text-figure) , 1957 .

[57]  T. Sawasaki,et al.  High-throughput synthesis of stable isotope-labeled transmembrane proteins for targeted transmembrane proteomics using a wheat germ cell-free protein synthesis system. , 2015, Molecular bioSystems.

[58]  S. Goldberg,et al.  The effects of vasopressin deficiency on aggression and impulsiveness in male and female rats , 2014, Psychoneuroendocrinology.

[59]  M. Wells.  Effects of body size and resource value on fighting behaviour in a jumping spider , 1988, Animal Behaviour.

[60]  M. D. Beecher,et al.  Sociobiology of Bank Swallows: Reproductive Strategy of the Male , 1979, Science.

[61]  Zoe R. Donaldson,et al.  Central vasopressin V1a receptor activation is independently necessary for both partner preference formation and expression in socially monogamous male prairie voles. , 2010, Behavioral neuroscience.

[62]  T. Burke,et al.  Explicit experimental evidence for the effectiveness of proximity as mate‐guarding behaviour in reducing extra‐pair fertilization in the Seychelles warbler , 2007, Molecular ecology.

[63]  Hans A. Hofmann,et al.  Neuropeptide regulation of social behavior in a monogamous cichlid fish , 2011, Physiology & Behavior.