Animal Cytogenomics - Jiří Rubeš

Research areas

• Comparative molecular cytogenetics and genetics of reproduction

Main objectives

• To obtain novel information on chromosomal rearrangements that have taken place during the process of evolution in the family Bovidae and Equidae by using comparative FISH.
• Determination of associations between the frequencies of chromosomally abnormal sperm, semen parameters and the reproductive outcome of the carriers of chromosomal translocations.
• Meiotic process including homologous synapsis, and frequency and distribution of recombination events will be studied on pachytene spermatocytes in animal models to elucidate the progress of meiosis in individuals with normal or abnormal karyotypes.

Content of research

Comparative cytogenomics and the genetics of reproduction

The conservation of selected chromosome regions in different species of Equidae and Bovidae will be analysed. Changes in chromosome sizes and morphology are characteristic of the evolutionary process. Different karyotypes are usually detected even among closely related species. Many pathways have been described by which chromosomes and whole genomes change during evolution. The objectives are to obtain novel information on the chromosomal rearrangements that have taken place during the process of evolution in the Bovidae and Equidae families by using comparative FISH with painting and BAC probes to provide further insights into the evolution of the karyotypes in these families. Whole chromosome and region-specific painting probes will be prepared by laser microdissection, flow-sorting and DOP PCR. On the basis of the homologies and cross hybridisations of these sequences within the various subfamilies, phylogenetic associations between the investigated species will be specified.

Meiotic cell division is a complex process including recombination and equal distribution of chromosomes into gametes. The objective of this work package is to obtain information on the similarities and differences in the meiotic behaviour of chromosomes which are separate in one and fused in other related species of the Bovidae family using immunofluorescence and FISH methods. Special attention will be paid to the synapsis and recombination of sex chromosomes, especially in those species, where the fusion of one or both sex chromosomes with some autosome occurred during karyotypic evolution. The functional isolation of the autosomal and the ancestral sexual segment by intercalary heterochromatic block is necessary for the proper progression of meiosis in X-autosome translocated species. Pachytene spermatocytes from testicular samples obtained from captive-bred animals belonging to species of the Bovidae family will be analysed using immunofluorescence and FISH methods. The results obtained in species with related karyotypes will be compared. We have tests of the following species for the study: Aepyceros melampus, Kobus megaceros, Gazella dama ruficollis, Oryx gazella, Gazella leptoceros, Connochaetes taurinus taurinus, Connochaetes gnou, Hippotragus niger, Tragelaphus imberbis and Taurotragus derbianus.

The primary cause of decreased reproductive potential in carriers of translocation is incorrect meiotic segregation of chromosomal pairs included in translocation, which form a trivalent in Robertsonian translocations or quadrivalent in reciprocal translocations during the first meiotic division. Our aim is to obtain and draw a general conclusion from the information on the associations between the frequencies of chromosomally abnormal sperm and semen parameters and the reproductive outcome of the carriers of balanced translocations. The anticipated achievements will be novel information on the relationships between the frequency of chromosomally aberrant spermatozoa, the quality of spermatogenesis and the entire reproductive potential of males who are carriers of translocations. Fluorescence in situ hybridisation will be used for the investigation of meiotic segregation of normal and translocated chromosomes and the interchromosomal effect. Samples will be continuously examined by sperm analysis, FISH method and flow cytometry.

list / cards

SELECTED PUBLICATIONS

2017

• FROHLICH, J; KUBICKOVA, S; MUSILOVA, P; CERNOHORSKA, H; MUSKOVA, H; RUBES, J, 2017:A Comparative Study of Pygmy Hippopotamus (Choeropsis liberiensis) Karyotype by Cross-Species Chromosome Painting. JOURNAL OF MAMMALIAN EVOLUTION 24 (4), p. 465 - 474.
• FROHLICH, J; KUBICKOVA, S; MUSILOVA, P; CERNOHORSKA, H; MUSKOVA, H; VODICKA, R; RUBES, J, 2017:Karyotype relationships among selected deer species and cattle revealed by bovine FISH probes. PLOS ONE 12 (11)

2016

• PLASIL, M; MOHANDESAN, E; FITAK, RR; MUSILOVA, P; KUBICKOVA, S; BURGER, PA; HORIN, P, 2016:The major histocompatibility complex in Old World camelids and low polymorphism of its class II genes. BMC GENOMICS 17
• SEBESTOVA, H; VOZDOVA, M; KUBICKOVA, S; CERNOHORSKA, H; KOTRBA, R; RUBES, J, 2016:Effect of species-specific differences in chromosome morphology on chromatin compaction and the frequency and distribution of RAD51 and MLH1 foci in two bovid species: cattle (Bos taurus) and the common eland (Taurotragus oryx). CHROMOSOMA 125 (1), p. 137 - 149.

2015

• CERNOHORSKA, H; KUBICKOVA, S; KOPECNA, O; VOZDOVA, M; MATTHEE, CA; ROBINSON, TJ; RUBES, J, 2015:Nanger, Eudorcas, Gazella, and Antilope form a well-supported chromosomal clade within Antilopini (Bovidae, Cetartiodactyla). CHROMOSOMA 124 (2), p. 235 - 247.

2012

• VOZDOVA, M; KASIKOVA, K; ORACOVA, E; PRINOSILOVA, P; RYBAR, R; HORINOVA, V; GAILLYOVA, R; RUBES, J, 2012:The effect of the swim-up and hyaluronan-binding methods on the frequency of abnormal spermatozoa detected by FISH and SCSA in carriers of balanced chromosomal translocations. HUMAN REPRODUCTION 27 (3), p. 930 - 937.

2011

• HORNAK, M; JESETA, M; MUSILOVA, P; PAVLOK, A; KUBELKA, M; MOTLIK, J; RUBES, J; ANGER, M, 2011:Frequency of Aneuploidy Related to Age in Porcine Oocytes. PLOS ONE 6 (4)

GRANTY

• The application of modern molecular genetic methods for the determination of genotoxic changes in at-risk populations (15-33968A), Ministry of Health - National Programme of Research, 2015 - 2018
• Analysis of sex chromosomes in Ruminantia by molecular cytogenetic methods (15-11299S), Czech Science Foundation - Standard Grants, 2015 - 2018
• Cytogenetic and mutational analysis of mast cell tumours in dogs (15-04197S), Czech Science Foundation - Standard Grants, 2015 - 2018
• Fylogenetic relationships in the family Bovidae studied by analysis of subfamily specific DNA repeats and karyotype (GAP506/10/0421), Czech Science Foundation - Standard Grants, 2010 - 2013
• Study of meiotic disturbances in men – carriers of congenital balanced translocations and their effect on the outcome of assisted reproduction (NS9842), Ministry of Health - Departmental Programme of Research and Development (2008-2011), 2009 - 2011
• Comparative study of male meiosis in members of the family Bovidae (GAP502/11/0719), Czech Science Foundation - Standard Grants, 2011 - 2015

PLANNED RESEARCH INFRASTRUCTURE

Technology Units

Advance microscopic study of mammalian chromosomes

CURRENT RESEARCH INFRASTRUCTURE

The research group has a 10-year experience with the use of microdissection and includes leading experts in this technology. Department of Genetics and Reproduction of the Veterinary Research Institute is equipped with the necessary instruments and devices for cell culture, FISH and the follow-up molecular-biological methods. The staff of the department is experienced in preparation of DNA probes for FISH on chromosomes of various species of the Bovidae and Equidae as well as in isolation of chromosome-specific centromeric repeats and germ cell analysis.