N.A. de Haan

The PiGMaP consortium cytogenetic map of the domestic pig (Sus scrofa domestica)

llNRA, Laboratoire de Grnrtique Cellulaire, BP27, 31326 Castanet-Tolosan, France 2Department of Functional Morphology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands 3Department of Animal Breeding and Genetics, Swedish University of Agricultural Sciences, Uppsala, Sweden 4Division of Anatomy, Department of Anatomy and Physiology, The Royal Veterinary and Agricultural University, Copenhagen, Denmark 5Division of Animal Genetics, Department of Animal Science and Animal Health, The Royal Veterinary and Agricultural University, Copenhagen, Denmark 6Department of Clinical Genetics, University of Ulm, Ulm, Germany 7Laboratoire de Cytomrtrie, CEA, Fontenay-aux Roses, France

Establishment of an R-banded rabbit karyotype nomenclature by FISH localization of 23 chromosome-specific genes on both G- and R-banded chromosomes

Direct detection of fluorescent in situ hybridization signals on R-banded chromosomes stained with propidium iodide is a rapid and efficient method for constructing cytogenetic maps for species with R-banded standard karyotypes. In this paper, our aim is to establish an R-banded rabbit karyotype nomenclature that is in total agreement with the 1981 G-banded standard nomenclature. For this purpose, we have produced new GTG- and RBG-banded mid-metaphase karyotypes and an updated version of ideograms of R-banded rabbit chromosomes. In addition, to confirm correlations between G- and R-banded chromosomes, we have defined a set of 23 rabbit BAC clones, each containing a specific gene, one marker gene per rabbit chromosome, and we have localized precisely each BAC clone by FISH on both G- and R-banded chromosomes.

Localization of 18S + 28S and 5S ribosomal RNA genes in the dog by fluorescence in situ hybridization.

The gene clusters encoding 18S + 28S and 5S rRNA in the dog (Canis familiaris) have been localized by using GTG-banding and fluorescence in situ hybridization. The 18S + 28S rDNA maps to chromosome regions 7q2.5-->q2.7, 17q1.7, qter of a medium-sized, not yet numbered autosome, and Yq1.2-->q1.3. Our data show that there is one cluster of 5S rDNA in the dog, which maps to chromosome region 4q1.4.

Localization of the 18S, 5.8S and 28S rRNA genes and the 5S rRNA genes in the babirusa and the white-lipped peccary

The locations of the genes encoding 18S, 5.8S and 28S rRNA and 5S rRNA were studied in two relatives of the domestic pig, the babirusa (Babyrousa babyrussa) and the white-lipped peccary (Tayassu pecari). In the babirusa, the 18S, 5.8S and 28S rDNA is located on chromosomes 6, 8 and 10. The genes on chromosomes 8 and 10 are actively transcribed, in contrast to those on chromosomes 6. In the white-lipped peccary, this rDNA was found to be located on chromosomes 4 and 8. The genes on both of these pairs of chromosomes are actively transcribed. The 5S rDNA was physically mapped to chromosome 16 in the babirusa, and to chromosome 11 in the white-lipped peccary. These data are compared to similar data obtained for the domestic pig, and confirm previously recognized chromosome homologies.

Comparative chromosome painting between the domestic pig (Sus scrofa) and two species of peccary, the collared peccary (Tayassu tajacu) and the white-lipped peccary (T. pecari): a phylogenetic perspective

The Suidae and the Dicotylidae (or Tayassuidae) are related mammalian families, both belonging to the artiodactyl suborder Suiformes, which diverged more than 37 million years ago. Cross-species chromosome painting was performed between the domestic pig (Sus scrofa; 2n = 38), a representative of the Suidae, and two species of the Dicotylidae: the collared peccary (Tayassu tajacu; 2n = 30) and the white-lipped peccary (T. pecari; 2n = 26). G-banded metaphase chromosomes of the two peccaries were hybridized with whole chromosome painting probes derived from domestic pig chromosomes 1–18 and X. For both peccary species, a total of 31 autosomal segments that are conserved between pig and peccary could be identified. The painting results confirm conclusions inferred from G-band analyses that the karyotypes of the collared peccary and the white-lipped peccary are largely different. The karyotypic heterogeneity of the Dicotylidae contrasts with the relative homogeneity among the karyotypes of the Suidae. For this difference between the Dicotylidae and the Suidae, a number of explanations are being postulated: 1) the extant peccaries are phylogenetically less closely related than is usually assumed; 2) the peccary genome is less stable than the genome of the pigs; and 3) special (e.g. biogeographical or biosocial) circumstances have facilitated the fixation of chromosome rearrangements in ancestral dicotylid populations.

Chromosome homology between the domestic pig and the babirusa (family Suidae) elucidated with the use of porcine painting probes

Homology among three pairs of domestic pig (Sus scrofa) and five pairs of babirusa (Babyrousa babyrussa) autosomes has been demonstrated with the use of porcine painting probes. With the results of this study, in addition to data obtained earlier through the application of banding techniques, correspondence between all individual chromosomes of these two distantly related pigs has been identified.

Construction of a cytogenetically characterized porcine somatic cell hybrid panel and its use as a mapping tool

A new panel of cytogenetically characterized pigrodent somatic cell hybrids was constructed and tested for twelve microsatellite markers with PCR. Cytogenetic characterization of hybrids was accomplished by fluorescence painting and GTG-banding of metaphase chromosomes. The panel consists of 15 independent pig-hamster and 6 independent pig-mouse cell lines. In the panel, all pig autosomes and the X Chromosome (Chr) are represented, and it is informative for all chromosome pairs except 2–14, 2–15, 3–9, 14–15, 14–16, and 16–17. The microsatellites tested were S0022, S0023, S0084, S0098, SO112, SO113, SO114, S0115, S0117, S0118, S0119, and S0120. The PCR results obtained in the 21 hybrids were compared with the cytogenetic data and analyzed for concordancy and correlation. Eight microsatellites could be assigned to specific pig chromosomes, confirming seven assignments based on linkage analysis.

Gene mapping in the river buffalo (Bubalus bubalis L)

Somatic cell hybridization was used as a tool to examine synteny of genes in the buffalo. Parental cells were blood lymphocytes from two Egyptian river buffaloes, and cells of the HPRT(hypoxanthine phosphoribosyltransferase-negative) Chinese hamster cell line wg3h C12 (Echard et al, 1984). Hybrid cells were produced by polyethylene glycol (PEG)-mediated fusion, followed by hypoxanthine-aminopterinethymidine (HAT) selection. Twenty-one independent hybrid cell lines were established. For enzyme analysis, hybrid cells and hamster cells were lysed in lysis buffer (Meera Khan, 1971). Buffalo cardiac muscle, skeletal muscle and erythrocytes were homogenized in the same buffer, and were used as samples of reference for buffalo enzyme activity. Starch gel electrophoresis (Harris and Hopkinson, 1978) was carried out to analyze the following enzymes: glyceraldehyde-3-phosphate dehydrogenase (GAPD; EC 1.2.1.12); lactate dehydrogenase (LDH; EC 1.1.1.27); malic

Fourteen chromosomal localizations and an update of the cytogenetic map of the rabbit.

In order to improve the informativeness of the cytogenetic map of the rabbit genome, fourteen markers were regionally mapped to individual chromosomes. The localizations comprise eleven gene loci (PRLR, GHR, HK1, ACE, TF, 18S+28S rDNA, CYP2C4, PMP2, TCRB, ALOX15 and MT1) and three microsatellite loci (Sat13, Sol33 and D1Utr6). Five of the genes contain known microsatellite sequences. To achieve these localizations, homologous and heterologous small insert clones, and clones from a rabbit Bacterial Artificial Chromosome (BAC) library were used as probes for fluorescence in situ hybridization experiments. Results indicate that especially BAC clones are a valuable tool for cytogenetic mapping. Some of the genes were selected for mapping on the basis of human- rabbit comparative painting data, to achieve localizations on gene-poor rabbit chromosomes. Our data are, in general, in agreement with the human-rabbit comparative painting data. By mapping microsatellite sequences that have also been used in linkage studies, links are provided between the genetic and physical maps of the rabbit genome. Linkage groups I, VI and XI could be assigned to chromosomes 1, 5 and 3 respectively. Moreover, in this paper we give an overview of the current status of the rabbit cytogenetic map. This map now comprises 62 physically mapped genes, which are scattered over all autosomes, except chromosome 2, and the X chromosome.

Analysis of chromosome aberrations in a mammary carcinoma cell line from a dog by using canine painting probes

A cell line derived from a spleen metastasis of a mammary carcinoma in a female dog was analyzed by fluorescence in situ hybridization with canine chromosome-specific paints. The cell line showed a modal chromosome number of 77, with three (90% of the cells) or four (10% of the cells) biarmed chromosomes. Aberrations observed relate to chromosomes 8 or 11, 13 or 15, 37, 38, and X and include chromosome loss (X), formation of isochromosomes (8 or 11, 13 or 15), and centric fusion (37 and 38). In all aberrations, whole chromosomes are involved. None of the genes known to be related to breast cancer development in humans and that has mapped in the dog is located on one of the aberrant chromosomes. The results of this study show that chromosome painting is a most useful tool for the analysis of canine tumor cells.