It has long been hypothesized that chromosomal rearrangements play a central role in different evolutionary processes, particularly in speciation and adaptation. Interchromosomal rearrangements have been extensively mapped using chromosome painting. However, intrachromosomal rearrangements have only been described using molecular cytogenetics in a limited number of mammals, including a few rodent species. This situation is unfortunate because intrachromosomal rearrangements are more abundant than interchromosomal rearrangements and probably contain essential phylogenomic information. Significant progress in the detection of intrachromosomal rearrangement is now possible, due to recent advances in molecular biology and bioinformatics. We investigated the level of intrachromosomal rearrangement in the Arvicolinae subfamily, a species-rich taxon characterized by very high rate of karyotype evolution. We made a set of region specific probes by microdissection for a single syntenic region represented by the p-arm of chromosome 1 of Alexandromys oeconomus, and hybridized the probes onto the chromosomes of four arvicolines (Microtus agrestis, Microtus arvalis, Myodes rutilus, and Dicrostonyx torquatus). These experiments allowed us to show the intrachromosomal rearrangements in the subfamily at a significantly higher level of resolution than previously described. We found a number of paracentric inversions in the karyotypes of M. agrestis and M. rutilus, as well as multiple inversions and a centromere shift in the karyotype of M. arvalis. We propose that during karyotype evolution, arvicolines underwent a significant number of complex intrachromosomal rearrangements that were not previously detected.

An integrative analysis of an isolated population of Harting’s vole (Microtus hartingi) from the Eastern Rhodope Mountains (Bulgaria) was carried out by morphological and morphometric methods, computed tomography, Cytb variation data, and experimental hybridization. Substantial changes in the development of the skull and teeth were found. Nevertheless, those voles can live to the senex stage. A phylogenetic reconstruction based on Cytb sequences showed that M. hartingi from the Eastern Rhodopes forms a separate clade, which is a sister clade to the voles from Northeastern Greece (also from the foothills of the Rhodopes). M. hartingi from the Rhodopes is mostly isolated reproductively from M. h. ankaraensis from Anatolia (Turkey), as evidenced by a decrease in the proportion of pairs that started breeding, relatively high mortality of pups, and increased sterility of hybrid males. Possible time of isolation of the Rhodopean population and consequences of the “bottleneck” effect on its current state are discussed. At the same time, we believe that, at present, it is impossible to determine its taxonomic status. It is necessary to recognize M. hartingi from the Bulgarian Rhodopes as an endangered population and to include it in regional Red Lists of Bulgaria.

The taxonomy of the genus Calomyscus remains controversial. According to the latest systematics the genus includes eight species with great karyotypic variation. Here, we studied karyotypes of 14 Calomyscus individuals from different regions of Iran and Turkmenistan using a new set of chromosome painting probes from a Calomyscus sp. male (2n = 46, XY; Shahr-e-Kord-Soreshjan-Cheshme Maiak Province). We showed the retention of large syntenic blocks in karyotypes of individuals with identical chromosome numbers. The only rearrangement (fusion 2/21) differentiated Calomyscus elburzensis, Calomyscus mystax mystax, and Calomyscus sp. from Isfahan Province with 2n = 44 from karyotypes of C. bailwardi, Calomyscus sp. from Shahr-e-Kord, Chahar Mahal and Bakhtiari-Aloni, and Khuzestan-Izeh Provinces with 2n = 46. The individuals from Shahdad tunnel, Kerman Province with 2n = 51–52 demonstrated non-centric fissions of chromosomes 4, 5, and 6 of the 46-chromosomal form with the formation of separate small acrocentrics. A heteromorphic pair of chromosomes in a specimen with 2n = 51 resulted from a fusion of two autosomes. C-banding and chromomycin A3-DAPI staining after G-banding showed extensive heterochromatin variation between individuals.

The mandarin vole, Lasiopodomys mandarinus, is one of the most intriguing species among mammals with non-XX/XY sex chromosome system. It combines polymorphism in diploid chromosome numbers, variation in the morphology of autosomes, heteromorphism of X chromosomes, and several sex chromosome systems the origin of which remains unexplained. Here we elucidate the sex determination system in Lasiopodomys mandarinus vinogradovi using extensive karyotyping, crossbreeding experiments, molecular cytogenetic methods, and single chromosome DNA sequencing. Among 205 karyotyped voles, one male and three female combinations of sex chromosomes were revealed. The chromosome segregation pattern and karyomorph-related reproductive performances suggested an aberrant sex determination with almost half of the females carrying neo-X/neo-Y combination. The comparative chromosome painting strongly supported this proposition and revealed the mandarin vole sex chromosome systems originated due to at least two de novo autosomal translocations onto the ancestral X chromosome. The polymorphism in autosome 2 was not related to sex chromosome variability and was proved to result from pericentric inversions. Sequencing of microdissection derived of sex chromosomes allowed the determination of the coordinates for syntenic regions but did not reveal any Y-specific sequences. Several possible sex determination mechanisms as well as interpopulation karyological differences are discussed.

We investigate the acoustic structure of alarm calls in a highly social rodent, the Harting’s vole (Microtus hartingi) and describe the phenomenon of collective shouting of bursts of alarm calls which could be produced in synchronized series. The alarm calls of Harting’s voles were recorded using an automatic device from 10 different family groups, containing from 4 to 15 potential callers, released to outdoor enclosures. Natural predators and humans served as call-eliciting stimuli. We reveal acoustic differences between urgent alarm calls, evoked by close-vicinity predators and identified from recordings by their calls and by rustle noise from the caller’s escape to burrow, and other alarm calls. We also reveal acoustic parameters between alarm calls produced in bursts and other alarm calls (produced in individual series or by a few non-synchronized callers). We discuss why the alarm calls of Harting’s voles are unusually high-frequency (about 17 kHz on average in the maximum fundamental frequency) among other vole species producing high-frequency alarms. Adaptive significance of producing the collective synchronized bursts of alarm calls by Harting’s voles remains unclear without direct observations of vocal vigilance in this species under natural conditions.

We analyzed the cranium dorsal projection and the mandible lateral projection in bone specimens from five Microtus guentheri and Microtus hartingi forms by geometric morphometrics (GM) methods (generalized Procrustes analysis, principal component analysis, canonical variance analysis, and discriminant function analysis). Analyses of the linear size and shapes of the cranium and lower jaw showed clear-cut differentiation among the forms into an eastern cluster and western cluster, matching M. guentheri and M. hartingi, respectively. Differences were revealed both between two subspecies of M. guentheri and between the subspecies M. h. strandzensis and Rhodopean M. hartingi, whose subspecies status has not yet been determined. M. h. ankaraensis bone specimens differ in many parameters of GM from the studied European specimens and to a lesser extent from M. g. guentheri and M. g. philistinus. Calculated morpho-ecological indices of the lower jaw revealed significant differences among all these forms, thereby possibly indicating adaptation of each to a specific habitat and dietary habits. Because of the emergence of impenetrable barriers for voles (the Anatolian Diagonal in the east and the Dardanelles and Bosporus in the west), the resultant vole groups have evolved independently.