Social discrimination by quantitative assessment of immunogenetic similarity

Genes of the major histocompatibility complex (MHC) that underlie the adaptive immune system may allow vertebrates to recognize their kin. True kin-recognition genes should produce signalling products to which organisms can respond. Allelic variation in the peptide-binding region (PBR) of MHC molecules determines the pool of peptides that can be presented to trigger an immune response. To examine whether these MHC peptides also might underlie assessments of genetic similarity, we tested whether Xenopus laevis tadpoles socially discriminate between pairs of siblings with which they differed in PBR amino acid sequences. We found that tadpoles (four sibships, n = 854) associated preferentially with siblings with which they were more similar in PBR amino acid sequence. Moreover, the strength of their preference for a conspecific was directly proportional to the sequence similarity between them. Discrimination was graded, and correlated more closely with functional sequence differences encoded by MHC class I and class II alleles than with numbers of shared haplotypes. Our results thus suggest that haplotype analyses may fail to reveal fine-scale behavioural responses to divergence in functionally expressed sequences. We conclude that MHC–PBR gene products mediate quantitative social assessment of immunogenetic similarity that may facilitate kin recognition in vertebrates.     

The ontogeny of kin recognition in tadpoles of the toadBufo melanostictus (Anura; Bufonidae)

The ontogeny of kin recognition and influence of social environment on the development of kin recognition behaviour was experimentally investigated in tadpoles of Bufo melanostictus that lived in aggregations and showed low larval dispersion. Embryos and tadpoles of the toad were reared as (i) kin only, (ii) with kin and non-kin (separated by a mesh screen), and (iii) in isolation. They were tested for the ability to discriminate between (i) familiar siblings and unfamiliar non-siblings, (ii) familiar siblings and familiar non-siblings and, (iii) unfamiliar siblings and unfamiliar non-siblings. All tadpoles were fed on boiled spinach before conducting trials. Preference of test tadpoles to associate near the end compartments whether empty or containing members of specific stimulus groups was assessed using a rectangular choice tank. When tested in tanks with empty end compartments, the test tadpoles showed random distribution and thus no bias for the apparatus or the procedure. In the presence of kin/non-kin in the end compartments a significantly greater number of test tadpoles spent the majority of the time near familiar or unfamiliar kin rather than near familiar or unfamiliar non-kin. Kin discrimination ability persisted throughout larval development. Familiarity with siblings is not required for discriminating kin from non-kin, and kin discrimination ability is not modified following exposure to non-kin. Also, involvement of dietary cues is unlikely to be the prime mechanism of kin recognition inB. melanostictus unlike in some other anurans.     

Discrimination between full-sibling spiny mice (Acomys cahirinus) by olfactory signatures

Discrimination between individual full siblings was tested in spiny mice (Acomys cahirinus). At weaning, intact litters were separated into two pairs so that animals only had social contact with one of their full siblings. Following 10 days of being housed in pairs, the four weanlings from a given litter were reunited in an observation terrarium. Pairings by familiar siblings were more frequent than unfamiliar-sibling pairings, indicating that interactions among close kin are mediated by the recognition of individual signatures rather than general phenotypic traits shared by all members of a kin class. Animals rendered anosmic by intranasal injections of zinc sulphate displayed no evidence of discriminating between familiar and unfamiliar siblings. Olfactory cues therefore appear to function as the signatures by which individual siblings are recognized. Since all animals were fed identical diets and otherwise maintained in similar environments, the differences in olfactory signatures between full siblings are most likely a function of genetic variability within this class of kin.     

The genetic basis of inbreeding avoidance in house mice

Animals might be able to use highly polymorphic genetic markers to recognize very close relatives and avoid inbreeding. The major histocompatibility complex (MHC) is thought to provide such a marker because it influences individual scent in a broad range of vertebrates. However, direct evidence is very limited. In house mice (Mus musculus domesticus), the major urinary protein (MUP) gene cluster provides another highly polymorphic scent signal of genetic identity that could underlie kin recognition. We demonstrate that wild mice breeding freely in seminatural enclosures show no avoidance of mates with the same MHC genotype when genome-wide similarity is controlled. Instead, inbreeding avoidance is fully explained by a strong deficit in successful matings between mice sharing both MUP haplotypes. Single haplotype sharing is not a good guide to the identification of full sibs, and there was no evidence of behavioral imprinting on maternal MHC or MUP haplotypes. This study, the first to examine wild animals with normal variation in MHC, MUP, and genetic background, demonstrates that mice use self-referent matching of a species-specific polymorphic signal to avoid inbreeding. Recognition of close kin as unsuitable mates might be more variable across species than a generic vertebrate-wide ability to avoid inbreeding based on MHC.     

MHC odours are not required or sufficient for recognition of individual scent owners

To provide information about specific depositors, scent marks need to encode a stable signal of individual ownership. The highly polymorphic major histocompatibility complex (MHC) influences scents and contributes to the recognition of close kin and avoidance of inbreeding when MHC haplotypes are shared. MHC diversity between individuals has also been proposed as a primary source of scents used in individual recognition. We tested this in the context of scent owner recognition among male mice, which scent mark their territories and countermark scents from other males. We examined responses towards urine scent according to the scent owner's genetic difference to the territory owner (MHC, genetic background, both and neither) or genetic match to a familiar neighbour. While urine of a different genetic background from the subject always stimulated greater scent marking than own, regardless of familiarity, MHC-associated odours were neither necessary nor sufficient for scent owner recognition and failed to stimulate countermarking. Urine of a different MHC type to the subject stimulated increased investigation only when this matched both the MHC and genetic background of a familiar neighbour. We propose an associative model of scent owner recognition in which volatile scent profiles, contributed by both fixed genetic and varying non-genetic factors, are learnt in association with a stable involatile ownership signal provided by other highly polymorphic urine components.     

Diverse sensory cues for individual recognition

To be social, the ability to recognize and discriminate conspecific individuals is indispensable in social animals, including primates, rodents, birds, fish, and social insects which live in societies or groups. Recent studies using molecular biology, genetics, in vivo and in vitro physiology, and behavioral neuroscientific approaches have provided detailed insights into how animals process and recognize the information of individuals. Here, we review the most distinct sensory modalities for individual recognition in animals, namely, olfaction and vision. In the case of rodents, two polymorphic gene complexes have been identified in their urine as the key and essential pheromonal components for individual recognition: the major histocompatibility complex (MHC) and the major urinary protein (MUP). Animals flexibly utilize MHC and/or MUP, which are detected by the main olfactory epithelium (MOE) and/or the vomeronasal organ (VNO) for various types of social recognition, such as strain recognition, kin recognition, and individual recognition. In contrast, primates, including humans, primarily use facial appearance to identify others. Face recognition in humans and other animals is naturally unique from genetic, cognitive, developmental, and functional points of view. Importantly note that nurture effects during growth phase such as social experience and environment can also shape and tune this special cognitive ability, in order to distinguish subtle differences between individuals. In this review, we address such unique nature and nurture mechanisms for individual recognition.     

Olfactory Communication and Neighbor Recognition in Giant Kangaroo Rats

We hypothesized that olfactory communication facilitates neighbor recognition in the giant kangaroo rat, Dipodomys ingens , and is therefore influential in coordinating social interactions in this solitary, desert rodent. We tested whether (i) D. ingens can discriminate between odors of same- and opposite-sex conspecifics; and (ii) the kangaroo rats exhibit scent preferences based on familiarity. In habituation-discrimination tests, we found that both genders distinguish differences between the scent of individuals of the same- and opposite-sex. In olfactory preference tests, both males and females spent significantly more time investigating the scent of their familiar cagemate than the scent of an unfamiliar conspecific. Giant kangaroo rats may be able to recognize familiar neighbors from olfactory cues, thus supporting a hypothesis of neighbor recognition. Neighbor recognition may be an important mechanism of social interactions in this endangered species.     

Involvement of estrogen receptor alpha, beta and oxytocin in social discrimination: A detailed behavioral analysis with knockout female mice

Social recognition, processing, and retaining information about conspecific individuals is crucial for the development of normal social relationships. The neuropeptide oxytocin (OT) is necessary for social recognition in male and female mice, with its effects being modulated by estrogens in females. In previous studies, mice whose genes for the estrogen receptor-alpha (alpha-ERKO) and estrogen receptor-beta (beta-ERKO) as well as OTKO were knocked out failed to habituate to a repeatedly presented conspecific and to dishabituate when the familiar mouse is replaced by a novel animal (Choleris et al. 2003, Proc Natl Acad Sci USA 100, 6192-6197). However, a binary social discrimination assay, where animals are given a simultaneous choice between a familiar and a previously unknown individual, offers a more direct test of social recognition. Here, we used alpha-ERKO, beta-ERKO, and OTKO female mice in the binary social discrimination paradigm. Differently from their wild-type controls, when given a choice, the KO mice showed either reduced (beta-ERKO) or completely impaired (OTKO and alpha-ERKO) social discrimination. Detailed behavioral analyses indicate that all of the KO mice have reduced anxiety-related stretched approaches to the social stimulus with no overall impairment in horizontal and vertical activity, non-social investigation, and various other behaviors such as, self-grooming, digging, and inactivity. Therefore, the OT, ER-alpha, and ER-beta genes are necessary, to different degrees, for social discrimination and, thus, for the modulation of social behavior (e.g. aggression, affiliation).     

基于交叉抚育的雄性根田鼠对异性同胞尿气味的识别

通过交叉抚育建立室内繁殖种群,在断奶后（80日龄）分别取这些供体的新鲜尿气味作刺激物,在行为观察箱中观察和记录雄性根田鼠对雌鼠气味的行为反应,以研究根田鼠同胞识别的化学通讯机制。结果表明：①在不同的发育时期（2～70日龄）,雄性同巢同胞与异巢同胞的体重没有显著差异。②雄性根田鼠对雌性同巢非同胞气味的接近潜伏期显著长于对异巢非同胞的接近潜伏期（P〈0．05）,其对异巢非同胞气味的访问时间和嗅舔时间都显著高于同巢非同胞气味（P〈0．05）。③雄性根田鼠对雌性异巢同胞和异巢非同胞气味的不存在明显偏好。其对两者的接近潜伏期、访问频次、访问时间、嗅舔频次和嗅舔时间等行为响应均无显著差异（P〈0．05）。这些结果表明,80日龄时,雄性根田鼠能够识别熟悉和陌生的无亲属关系雌性尿气味,但不能区分陌生的亲属和非亲属,因此,其异性同胞识别的机制为共生熟悉模式。     

Genetic Basis of Mating Preferences in Wild House Mice

This paper reviews work conducted over the last several yearson the effect of genetic differences within the t-complex ofwild house mice on female mating preference. Wild mice are polymorphicfor a mutation within the t complex on chromosome 17. About25% of wild mice are heterozygous (+/t) for a t-haplotype andthe remainder are +/+. These t-haplotypes have a number of deleteriouseffects when homozygous and hence t/t individuals are rarelyfound in wild populations. We have examined preferences of +/+and +/t females for males of both genotypes. We have found that+/t, but not +/+ females have strong preferences for +/+ males.These preferences can be modified by a variety of factors includingestrous condition of the female (the preferences are strongeramong estrous than diestrous females) and the dominance statusof the male (when forced to choose, females give priority tomale dominance status over t complex genotype in choosing males).The restiction of preference to +/t females indicates that geneson t haplotypes modulate these preferences. Because t haplotypesinclude the major histocompatibility complex (MHC) of the mousewe designed a study to ascertain whether the preferences of+/t females were associated with the MHC. Results of the studyindicate that the preferences are independent of the MHC. Furtherwork testing females carrying a partial t-haplotype (t^w18) indicatesthat the genes for mating preference are localized in the regionof the t complex distal to the MHC. A large number of t haplotypesare found in wild mouse populations. Females that are themselves+/t when forced to choose between 2 +/t males (one carryinga haplotype that is the same as their own and one carrying ahaplotype that is different) prefer males carrying t-haplotypesthat differ from their own. Finally, we conclude that matingpreference may only be a weak force regulating the frequencyof t-mutations in wild mouse populations. The impact of matingpreference on population genetics of genes within this regionis muted because of the great importance of male dominance rankin determining mating patterns within interacting social groups.     

In search of the chemical basis for MHC odourtypes

Mice can discriminate between chemosignals of individuals based solely on genetic differences confined to the major histocompatibility complex (MHC). Two different sets of compounds have been suggested: volatile compounds and non-volatile peptides. Here, we focus on volatiles and review a number of publications that have identified MHC-regulated compounds in inbred laboratory mice. Surprisingly, there is little agreement among different studies as to the identity of these compounds. One recent approach to specifying MHC-regulated compounds is to study volatile urinary profiles in mouse strains with varying MHC types, genetic backgrounds and different diets. An unexpected finding from these studies is that the concentrations of numerous compounds are influenced by interactions among these variables. As a result, only a few compounds can be identified that are consistently regulated by MHC variation alone. Nevertheless, since trained animals are readily able to discriminate the MHC differences, it is apparent that chemical studies are somehow missing important information underlying mouse recognition of MHC odourtypes. To make progress in this area, we propose a focus on the search for behaviourally relevant odourants rather than a random search for volatiles that are regulated by MHC variation. Furthermore, there is a need to consider a ‘combinatorial odour recognition’ code whereby patterns of volatile metabolites (the basis for odours) specify MHC odourtypes.     

Social discrimination in lambs: the role of indirect familiarization and methods of assessment

Familiarization with individuals resulting from direct exposure is the primary mechanism mediating social recognition in many vertebrates. However, discernible phenotypic similarity among close kin may provide an additional basis for recognizing previously unencountered individuals. We investigated the existence of such a mechanism of indirect familiarity among artificially reared twin lambs, Ovis aries, that were separated shortly after birth. When they were 2-3 weeks old, we tested their recognition of familiar penmates, their unfamiliar twin and twins of familiar penmates in two-choice and paired tests. Lambs responded discriminatively to familiar penmates. When allowed to choose between their twins or twins of penmates versus an unfamiliar unrelated individual, lambs did not show any preference. Nevertheless, when paired with a twin or penmate’s twin, lambs bleated less than did those paired with an unfamiliar unrelated partner, which suggests that twins and twins of penmates were recognized. Discrimination between twin and unrelated individuals could be based either on early learning of the twin's phenotypic traits (template) before separation or on a process of indirect familiarization (comparison with familiar self cues). However, indirect familiarization was clearly implicated in the discrimination of twins of penmates; that is, lambs learned the characteristic phenotypic traits of (unrelated) penmates and detected a resemblance between those familiar individuals and their twins. Overall, the two-choice and paired tests were differentially effective procedures depending on the research question being addressed, that is, relatively simple or subtle social discrimination. Copyright 2003 Published by Elsevier Ltd on behalf of The Association for the Study of Animal Behaviour.      

Regulation of the NK cell alloreactivity to bone marrow cells by the combination of the host NK gene complex and MHC haplotypes

Host NK cells can reject MHC-incompatible (allogeneic) bone marrow cells (BMCs), suggesting their effective role for graft-vs leukemia effects in the clinical setting of bone marrow transplantation. NK cell-mediated rejection of allogeneic BMCs is dependent on donor and recipient MHC alleles and other factors that are not yet fully characterized. Whereas the molecular mechanisms of allogeneic MHC recognition by NK receptors have been well studied in vitro, guidelines to understand NK cell allogeneic reactivity under the control of multiple genetic components in vivo remain less well understood. In this study, we use congenic mice to show that BMC rejection is regulated by haplotypes of the NK gene complex (NKC) that encodes multiple NK cell receptors. Most importantly, host MHC differences modulated the NKC effect. Moreover, the NKC allelic differences also affected the outcome of hybrid resistance whereby F1 hybrid mice reject parental BMCs. Therefore, these data indicate that NK cell alloreactivity in vivo is dependent on the combination of the host NKC and MHC haplotypes. These data suggest that the NK cell self-tolerance process dynamically modulates the NK cell alloreactivity in vivo.     

Good genes,complementary genes and human mate preferences

The past decade has witnessed a rapidly growing interest in the biological basis of human mate choice. Here we review recent studies that demonstrate preferences for traits which might reveal genetic quality to prospective mates, with potential but still largely unknown influence on offspring fitness. These include studies assessing visual, olfactory and auditory preferences for potential good-gene indicator traits, such as dominance or bilateral symmetry. Individual differences in these robust preferences mainly arise through within and between individual variation in condition and reproductive status. Another set of studies have revealed preferences for traits indicating complementary genes, focussing on discrimination of dissimilarity at genes in the major histocompatibility complex (MHC). As in animal studies, we are only just beginning to understand how preferences for specific traits vary and inter-relate, how consideration of good and compatible genes can lead to substantial variability in individual mate choice decisions and how preferences expressed in one sensory modality may reflect those in another. Humans may be an ideal model species in which to explore these interesting complexities.     

Good genes,complementary genes and human mate preferences

The past decade has witnessed a rapidly growing interest in the biological basis of human mate choice. Here we review recent studies that demonstrate preferences for traits which might reveal genetic quality to prospective mates, with potential but still largely unknown influence on offspring fitness. These include studies assessing visual, olfactory and auditory preferences for potential good-gene indicator traits, such as dominance or bilateral symmetry. Individual differences in these robust preferences mainly arise through within and between individual variation in condition and reproductive status. Another set of studies have revealed preferences for traits indicating complementary genes, focussing on discrimination of dissimilarity at genes in the major histocompatibility complex (MHC). As in animal studies, we are only just beginning to understand how preferences for specific traits vary and inter-relate, how consideration of good and compatible genes can lead to substantial variability in individual mate choice decisions and how preferences expressed in one sensory modality may reflect those in another. Humans may be an ideal model species in which to explore these interesting complexities.     

The use of olfactory and other cues for social recognition by juvenile pigs

Social recognition is essential for the maintenance of a stable group structure. Failure to recognise familiar conspecifics in social groups of juvenile pigs may initiate agonistic encounters that can compromise welfare and productivity. Current housing systems may allow build up of atmospheric ammonia that might, in turn, interfere with the olfactory system and compromise olfactory perception. In the present study, 16 juvenile pigs were housed in fresh air while another 16 pigs were kept in an ammoniated atmosphere (approximately 36ppm) for 1 week prior to test and another week during testing. We then assessed the role of olfaction in social recognition and determined whether chronic exposure to ammonia compromised discrimination based on olfactory perception by comparing the pigs' responses to selected cues from a familiar and an unfamiliar pig presented simultaneously in separate chambers of a modified Y-maze in each of two test situations (near, remote). Visual, auditory, olfactory, and tactile cues were all provided in the "near" test situation; here, the stimulus pigs were presented in two separate chambers behind clear perspex walls containing an aperture that allowed nose-to-nose contact between the test and stimulus pigs. On the other hand, the "remote" test provided only olfactory cues via air passed from the chambers containing the stimulus pigs into the test chamber. Each test lasted 5min and the pigs' behaviour was recorded via overhead video cameras; we then measured the accumulated times spent near and the numbers of visits made to the familiar and the unfamiliar stimulus pigs as well as the transitions between them. Overall, pigs made more visits to and spent significantly longer near both the stimulus pigs in the near test than in the remote one (ANOVA, P<0.001). They also made more transitions between the stimuli in the former test than the latter (P<0.001). Pigs from both the ammonia and the fresh-air treatment groups showed social discrimination. However, pigs that had received chronic exposure to ammonia visited the familiar pig more often and spent longer near it than the unfamiliar one regardless of the test situation (P<0.05) whereas those reared in fresh air spent longer near the unfamiliar animal (P<0.05). The present results suggest that pigs from both treatment groups employed olfactory cues in social recognition, but that chronic exposure to ammonia did not interfere with this ability. However, ammonia treatment seemingly affected social preferences, thus indicating an unknown and more fundamental effect of living in ammoniated atmospheres.     

Kin recognition in zebrafish: a 24-hour window for olfactory imprinting

Distinguishing kin from non-kin profoundly impacts the evolution of social behaviour. Individuals able to assess the genetic relatedness of conspecifics can preferentially allocate resources towards related individuals and avoid inbreeding. We have addressed the question of how animals acquire the ability to recognize kin by studying the development of olfactory kin preference in zebrafish (Danio rerio). Previously, we showed that zebrafish use an olfactory template to recognize even unfamiliar kin through phenotype matching. Here, we show for the first time that this phenotype matching is based on a learned olfactory imprinting process in which exposure to kin individuals on day 6 post fertilization (pf) is necessary and sufficient for imprinting. Larvae that were exposed to kin before or after but not on day 6 pf did not recognize kin. Larvae isolated from all contact with conspecifics did not imprint on their own chemical cues; therefore, we see no evidence for kin recognition through self-matching in this species. Surprisingly, exposure to non-kin odour during the sensitive phase of development did not result in imprinting on the odour cues of unrelated individuals, suggesting a genetic predisposition to kin odour. Urine-born peptides expressed by genes of the immune system (MHC) are important messengers carrying information about 'self' and 'other'. We suggest that phenotype matching is acquired through a time-sensitive learning process that, in zebrafish, includes a genetic predisposition potentially involving MHC genes expressed in the olfactory receptor neurons.     

Phylogeography and species discrimination in the Paracalliope fluviatilis species complex (Crustacea: Amphipoda): can morphologically similar heterospecifics identify compatible mates?

We examined the phylogeographic patterns of Paracalliope fluviatilis (Amphipoda) over its entire range and the prevalence of mate discrimination in laboratory mate choice tests using genetically distinct populations. We predicted that mate discrimination would increase as the level of genetic divergence increased between populations. Thirty different haplotypes were found with pairwise sequence divergences in the range 1–23.5% between locations. Individuals were selected from seven genetically distinct populations and males were presented with ‘local’ (same population) or ‘foreign’ (genetically divergent) females. Males were more likely to pair with local than foreign females but there was no evidence of a gradual increase in discrimination. Discrimination became most prominent (approximately 5 : 1 local : foreign) when genetic divergences exceeded approximately 20%; matings between divergent individuals also resulted in significantly fewer females producing eggs. We suggest that: (1) this abrupt shift in discrimination occurs because individuals from different, but similarly divergent, clades rely on different recognition cues (e.g. moulting pheromones with disparate chemical signatures) that trigger recognition and subsequent discrimination of incompatible mates; (2) geological history associated with sea level changes and a series of isolation events may be responsible for the patterns of discrimination that we observed; and (3) amphipods may be more genetically variable relative to other invertebrate taxa. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 99 , 196–205.     

How effective is recognition of siblings on the basis of genotype?

The ability to recognize kin based on genetic markers has been widely proposed as a mechanism to facilitate altruistic behaviour and inbreeding avoidance. Siblings are an important group of relatives to discriminate from unrelated individuals but present a problem, because siblings can share 0, 1 or 2 alleles at any single recognition locus. Here, we present a Bayesian model of kin recognition that defines the potential for genotypic information to convey kinship. Under the direct comparison model, where the signaller’s genotype is compared with that of the receiver, the odds ratio that a pair of individuals were siblings was substantially increased if they shared both alleles at a single locus, but only a minority of siblings were recognized; increasing the number of recognition loci used could not increase both the odds ratio and the proportion of siblings recognized. A maternal comparison model, where the signaller’s genotype is compared with that of the receiver’s mother, performed poorly when only a single recognition locus was considered, but became increasingly effective with more recognition loci. Nevertheless, incorporating partial‐matching information across multiple, independent loci are likely to be difficult. Further empirical work needs to establish the mechanistic basis of genetic kin recognition used by different taxa.