The expression profile of genes associated with behavior, stress, and adult neurogenesis along the hippocampal dorsoventral axis in tame and aggressive foxes.

The hippocampus plays the key role in stress response regulation, and stress response appears to be weakened in domesticated animals compared to their wild relatives. The hippocampus is functionally heterogeneous along its dorsoventral axis, with its ventral compartment being more closely involved in stress regulation. An earlier series of experiments was conducted with a unique breeding model of animal domestication, the farm silver fox (Vulpes vulpes), which included tame, aggressive, and unselected animals. A decrease in many indices of the hypothalamic-pituitary-adrenal activity was observed in tame animals. Also, adult hippocampal neurogenesis was more intense in tame foxes, and this fact may relate to reduced stress levels in this experimental population of foxes. Nevertheless, the molecular mechanisms responsible for the reduced stress response in tame animals remain obscure. In this study, serum cortisol levels and the mRNA levels of 13 genes in the dorsal and ventral hippocampus have been measured and compared in tame, aggressive, and unselected foxes. At the current stage of domestication, stress-induced cortisol levels in tame, aggressive, and unselected animals differ significantly from each other: tame foxes show the lowest levels, and aggressive ones, the highest. Twelve genes tested demonstrate significant gene expression differences between the dorsal and ventral hippocampi. These differences are mainly consistent with those found in rodents and humans. In tame foxes, significantly elevated mRNA levels were recorded for several genes: CYP26B1 for cytochrome P450 26B1 and ADRA1A for α1A adrenergic receptor in the dorsal hippocampus, whereas the level of NR3C2 mRNA for mineralocorticoid receptor was higher in the ventral. It is presumed that these genes constitute an important part of the mechanism reducing stress induced by contacts with humans and contribute to linking stress regulation with adult neurogenesis in tame foxes and domesticated animals in general.

Platinum coat color in red fox (Vulpes vulpes) is caused by a mutation in an autosomal copy of KIT.

The red fox (Vulpes vulpes) demonstrates a variety of coat colors including platinum, a common phenotype maintained in farm-bred fox populations. Foxes heterozygous for the platinum allele have a light silver coat and extensive white spotting, whereas homozygosity is embryonic lethal. Two KIT transcripts were identified in skin cDNA from platinum foxes. The long transcript was identical to the KIT transcript of silver foxes, whereas the short transcript, which lacks exon 17, was specific to platinum. The KIT gene has several copies in the fox genome: an autosomal copy on chromosome 2 and additional copies on the B chromosomes. To identify the platinum-specific KIT sequence, the genomes of one platinum and one silver fox were sequenced. A single nucleotide polymorphism (SNP) was identified at the first nucleotide of KIT intron 17 in the platinum fox. In platinum foxes, the A allele of the SNP disrupts the donor splice site and causes exon 17, which is part of a segment that encodes a conserved tyrosine kinase domain, to be skipped. Complete cosegregation of the A allele with the platinum phenotype was confirmed by linkage mapping (LOD 25.59). All genotyped farm-bred platinum foxes from Russia and the US were heterozygous for the SNP (A/G), whereas foxes with different coat colors were homozygous for the G allele. Identification of the platinum mutation suggests that other fox white-spotting phenotypes, which are allelic to platinum, would also be caused by mutations in the KIT gene.

[Does the selection of fox for tame and aggressive behavior changes their ability to focus attention, and training the motor skills?].

Foxes long time selected for tame and aggressive behavior were compared on ability to focus attention on the object of food reinforcement. Attenuation of this behavior and rate of training for motor skills also has been examined. Maximal duration of eye focusing was significantly higher in aggressive foxes, in comparison with tame ones. Our experiments allowed divide the group of tame foxes into two subgroups "calm" and "emotional", on the base of emotionality and motor activity during tests. Features of behavior of these two subgroups steadily differed in all tests. "Calm" tame foxes at the extinction test continued the trained skill longer than "emotional" and aggressive ones. Tame foxes were more successful than aggressive in the training for motor skills. The possible reasons of the data obtained are discussed.

Vocalization toward conspecifics in silver foxes (Vulpes vulpes) selected for tame or aggressive behavior toward humans.

We examined the production of different vocalizations in three strains of silver fox (unselected, aggressive, and tame) attending three kinds of behavior (aggressive, affiliative, and neutral) in response to their same-strain conspecifics. This is a follow-up to previous experiments which demonstrated that in the presence of humans, tame foxes produced cackles and pants but never coughed or snorted, whilst aggressive foxes produced coughs and snorts but never cackled or panted. Thus, cackle/pant and cough/snort were indicative of the tame and aggressive fox strains respectively toward humans. Wild-type unselected foxes produced cough and snort toward humans similarly to aggressive foxes. Here, we found that vocal responses to conspecifics were similar in tame, aggressive and unselected fox strains. Both cackle/pant and cough/snort occurred in foxes of all strains. The difference in the use of cackle/pant and cough/snort among these strains toward humans and toward conspecifics suggest that silver foxes do not perceive humans as their conspecifics. We speculate that these vocalizations are produced in response to a triggering internal state, affiliative or aggressive, that is suppressed by default in these fox strains toward humans as a result of their strict selection for tame or aggressive behavior, whilst still remaining flexible toward conspecifics.

Measurement of segregating behaviors in experimental silver fox pedigrees.

Strains of silver foxes, selectively bred at the Institute of Cytology and Genetics of the Russian Academy of Sciences, are a well established, novel model for studying the genetic basis of behavior, and the processes involved in canine domestication. Here we describe a method to measure fox behavior as quantitative phenotypes which distinguish populations and resegregate in experimental pedigrees. We defined 50 binary observations that nonredundantly and accurately distinguished behaviors in reference populations and cross-bred pedigrees. Principal-component analysis dissected out the independent elements underlying these behaviors. PC1 accounted for >44% of the total variance in measured traits. This system clearly distinguished tame foxes from aggressive and wildtype foxes. F1 foxes yield intermediate values that extend into the ranges of both the tame and aggressive foxes, while the scores of the backcross generation resegregate. These measures can thus be used for QTL mapping to explore the genetic basis of tame and aggressive behavior in foxes, which should provide new insights into the mechanisms of mammalian behavior and canine domestication.

A marker set for construction of a genetic map of the silver fox (Vulpes vulpes).

The silver fox, a variant of the red fox (Vulpes vulpes), is a close relative of the dog (Canis familiaris). Cytogenetic differences and similarities between these species are well understood, but their genomic organizations have not been compared at higher resolution. Differences in their behavior also remain unexplained. Two silver fox strains demonstrating markedly different behavior have been generated at the Institute of Cytology and Genetics of the Russian Academy of Sciences. Foxes selected for tameness are friendly, like domestic dogs, while foxes selected for aggression resist human contact. To refine our understanding of the comparative genomic organization of dogs and foxes, and enable a study of the genetic basis of behavior in these fox strains, we need a meiotic linkage map of the fox. Towards this goal we generated a primary set of fox microsatellite markers. Four hundred canine microsatellites, evenly distributed throughout the canine genome, have been identified that amplify robustly from fox DNA. Polymorphism information content (PIC) values were calculated for a representative subset of these markers and population inbreeding coefficients were determined for tame and aggressive foxes. To begin to identify fox-specific single nucleotide polymorphisms (SNPs) in genes involved in the neurobiology of behavior, fox and dog orthologs of serotonin 5-HT1A and 5-HT1B receptor genes have been cloned. Sequence comparison of these genes from tame and aggressive foxes reveal several SNPs. The close relationship of the fox and dog enables canine genomic tools to be utilized in developing a fox meiotic map and mapping behavioral traits in the fox.

Directional left-sided asymmetry of adrenals in experimentally domesticated animals.

Directional left-sided asymmetry of the adrenals was typical of black and silver foxes, American minks, and gray rats selected by their behavior. In domesticated, but to a greater extent, in aggressive animals, the weight of the left adrenal and the width of its medulla and cortex markedly surpassed the corresponding parameters of the right adrenal. In aggressive animals enlargement of the left adrenal cortex was associated with widening of the zona reticularis, while in domesticated animals with enlargement of the zona fasciculata.

[Effect of selection for behavior on the cranial traits of the American mink (Mustela vison)]. / Vliianie selektsii po povedeniiu na kraniologicheskie priznaki Amerikanskoi norki (Mustela vison).

Cranial sizes of American mink selected for tame and aggressive behavior (towards humans) and control mink, which were not selected for behavior, were compared. Absolute sizes of the skull were demonstrated to change depending on the direction of selection. Sexual dimorphism was reduced in mink selected for aggressive behavior, because the cranial sizes of females increased and those of males decreased. Cranial traits were analyzed by the method of principal components. The groups studied differed from one another with respect to the first four components. Although the vector of selection determined the differences between the groups, some morphological changes were similar in the groups selected for tame and aggressive behavior.

[Pleiotropic effect of the Black crystal mutation on reproductive indicators in the American mink (Mustela vison)]. / Pleiotropnyi éffekt mutatsii Black crystal na reproduktivnye pokazateliu amerikanskoi norki (Mustela vison).

The parameters of reproduction were studied in American minks that were homo- and heterozygous for the Black crystal mutation (Cr). The given mutation that changes the hair pigmentation was obtained de novo duration selection of American minks for tame behavior. Apart from changed color, the following reproduction disorders were detected in the Black crystal mutants, as compared to standard animals: (1) delayed entry into the reproductive season in females; (2) greater percentage in both homo- and heterozygous animals of females bearing no offspring; (3) greater early postnatal lethality indices. The most marked suppressing effect on the development of offspring was detected in homozygous females (Cr/Cr), whose progeny perished within a few days after birth. An analysis of reproduction in mutants and standard animals is presented. An attempt was made to explain the conceivable mechanisms that form the basis of disorders detected.

Folliculogenesis, onset of puberty and fecundity of mink (Mustela vision Schreb) selectively bred for docility or aggressiveness.

It has been suggested that selective breeding of animals for docile behavior is correlated with early onset of puberty and improved fertility. We wished to test the hypothesis that mink bred for docility would show earlier onset of puberty and greater fecundity than mink bred for aggressiveness. We used farm-raised, 7-mo-old mink females that had been selectively bred for 7 to 10 generations on the basis of behavior towards humans. Onset of puberty was estimated once (between 15 and 20 December) by vaginal smears and was said to start wtih preponderance of cornified epithelial cells in the cytological specimen. Fecundity was measured by litter size and rate of folliculogenesis, with and without hCG stimulation, by histomorphometric examination of ovaries and uteri. A total of 43/100 (43%) docile females achieved proestrus and estrus as compared to 16/136 (12%) of the aggressive ones. Overall pregnancy rate, survival to 5 d after whelping and litter size did not differ between the docile and aggressive females. Docile females showed significantly higher numbers (P < 0.05 and < 0.001) of growing, maturing and atretic follicles than the aggressive ones, however the latter showed a highly significant (P < 0.001) folliculogenic response to hCG. The response of the ovary of aggressive females to hCG is particularly dramatic because in most aggressive females the ovaries contained none or only few follicles of any kind. The inhibition of folliculogenesis in aggressive mink is similar to that reported in Silverblue and Sapphire mink homozygous for the Stuart factor. The paradoxical response to hCG may be an indication that selective breeding for aggression may be correlated with the disturbance at the early stages of folliculogenesis which creates a deficiency of follicles that are sensitive to LH-type stimulation.

[The effect of the selection of Mustela vison mink for behavior on their folliculogenesis and reproductive function]. / Vliianie selektsii norok Mustela vison po povedeniiu na ikh follikulogenez i reproduktivnuiu funktsiiu.

The selection of minks for behaviour in two directions (aggressive and domesticated one) brought about the correlative changes in function of sexual system. The sexual maturing of domesticated mink controlled by vaginal smears began more earlier. The fertility of domesticated mink with signs of earlier oestrus was larger than one in the same females with later maturing. Such correlation was absent in the aggressive females. In ovary of the aggressive mink the number of follicles of all types was decreased in comparison with those of the control and tame type of females. The HCG treatment in doses of 50 U and 100 U brought about the stimulation of folliculogenesis with the sharp increasing in a number of well developed vesicular and atretic follicles in all groups of females. Such shifts of folliculogenesis were directly proportional to the treated doses in control and tame type females and inversely proportional to the treated doses in aggressive females.

[Adrenal function in the mink Mustela vison selected for the type of behavior]. / Funktsiia nadpochechnikov u norok Mustela vison, selektsioniruemykh po tipu povedeniia.

Glucocorticoid function of adrenals and its reactivity to stress caused by immobilization and to ACTH stimulation have been studied in the male minks selected both for domestic and aggressive behavior towards man. In autumn period the regression of cortisol content in adrenals was observed in domesticated males and particularly in aggressive ones compared with control. At the same period the increased level of cortisol in blood as the result of immobilization was noticed in aggressive minks compared with the domesticated and control ones. At ACTH stimulation in spring time the increase of cortisol level in blood was shown for animals of all the groups but it was more marked for the aggressive ones.