No abstract

A combination of cytogenetic and molecular biology techniques were used to study the molecular composition and organisation of the pericentromeric regions of house mouse metacentric chromosomes, the products of Robertsonian (Rb) translocations between telocentrics. Regardless of whether mitotic or meiotic preparations were used, in situ hybridisation failed to reveal pericentromeric telomeric sequences on any of the Rb chromosomes, while all metacentrics retained detectable, although reduced (average 50 kb), amounts of minor satellite DNA in the vicinity of their centromeres. These results were supported by slot blot hybridisation which indicated that mice with 2n=22 Rb chromosomes have 65% of telomeric sequences (which are allocated to the distal telomeres of both Rb and telocentric chromosomes and to the proximal telomeres of telocentrics) and 15% the amount of minor satellite, compared with mice with 2n=40 all-telocentric chromosomes. Pulsed field gel electrophoresis and Southern analysis of DNA from Rb mice showed that the size of the telomeric arrays is similar to that of mice with all-telocentric chromosomes and that the minor satellite sequences were hybridising to larger fragments incorporating major satellite DNA. Since the telomeric sequences are closer to the physical end of the chromosome than the minor satellite sequences, the absence of telomeric sequences and the reduced amount of minor satellite sequences at the pericentromeric region of the Rb metacentrics suggest that the breakpoints for the Rb translocation occur very close to the minor satellite-major satellite border. Moreover, it is likely that the minor satellite is required for centromeric function, 50-67 kb being enough DNA to organise one centromere with a functionally active kinetochore.

A survey is given on the occurrence, the geographic origin and the arm composition of 27 Robertsonian fusion metacentric chromosomes of wild populations of the mouse. Their study is of twofold interest: a) it is possible to introduce these naturally occuring metacentrics in laboratory strains for experimental use. At present, altogether 34 metacentric chromosomes of different composition are available including 7 cases of metacentrics known form laboratory strains of the mouse. b) With the search for metacentrics in the mouse and with their identification insights are permitted in the role of Robertsonian changes in the course of mammalian evolution--Several separate populations of the mouse with different sets of multiple (up to 9) metacentrics have been found in Switzerland and Italy. Some of the individual metacentrics may occur in different populations. The participation of an acrocentric autosome in the formation of metacentrics seem to be at random, but the sex chromosomes are never included in a metacentric.--Homology of the arms involved in metacentrics is conserved, so that in meiosis of interpopulation hybrids is due to mechanisms of segregational imbalance and subsequent prenatal elimination of fetal offspring, but it follows also the pattern of male limited hybrid sterility.--From an evolutionary view point, karyotype rearrangements of Robertsonian type may initiate reproductive isolation, which prepares the ground for further genetic diversification and, as in the case of the mouse, of incipient speciation.

In mammals, Robertsonian (Rb) translocation (the joining of two telo͞acrocentric chromosomes at their centromere to form a metacentric) is the most effective process in chromosomal evolution leading to speciation; its occurrence also affects human health (through the induction of trisomies) and the fertility of farm animals. To understand the mechanism of Rb translocation, we used the house mouse as a model system and studied the organization of pericentromeric satellite DNAs (satDNA) of telocentrics and Rb chromosomes, both minor and major satDNA. The chromosome-orientation fluorescence in situ hybridization (CO-FISH) technique was used to analyze the major satDNA. To detect the very small amount of minor satDNA, a procedure was developed that combines CO-FISH with primed in situ labeling and conventional FISH and is five times more sensitive than the CO-FISH procedure alone. It was found that both the major and the minor satDNA tandem repeats are oriented head-to-tail in telocentric and Rb chromosomes, and their polarity is always the same relative to the centromere. We suggest that all tandemly repetitive satDNAs in a species probably are locked into such a symmetry constraint as a universal consequence of chromosomal evolution. Rb translocation breakpoints were found localized within the minor satDNA of telocentrics, and these sequences contributed symmetrically to the formation of the centromeric region of the Rb chromosomes. These results are important for an understanding of the geometry of Rb translocations and suggest the study of DNA orientation as a new tool for investigating these rearrangements.