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H Scherthan

Publications and source records attributed to H Scherthan.

79 records · Page 5Linked to original sources

Characterisation of a tandem repetitive sequence cloned from the deer Capreolus capreolus and its chromosomal localisation in two muntjac species.

The isolation and characterisation of a highly repetitive DNA sequence from the genome of the Roe deer Capreolus capreolus is reported. This sequence is characterised by tandem repetition and located within centric heterochromatin as demonstrated by non isotopic in situ hybridisation to the karyotypes of the Indian and Chinese muntjacs. Amplification and/or clustering of these sequences during the drastic karyotype evolution of the genus Muntiacus was noted in the large centromere of the X chromosome of the Indian muntjac. Partial sequence analysis revealed a 62% sequence homology with the sat 1A sequences of Muntiacus muntjak vaginalis.

Animals↗

Use of a centromere-specific DNA probe (p82H) in nonisotopic in situ hybridization for classification of micronuclei.

The DNA probe p82H was used to visualize centromeric DNA in micronuclei (MN) of human cells. Slides prepared from cultures treated by the aneugen (causing aneuploidy) colcemid showed significantly more MN with centromeric signals than those treated by the clastogen (causing chromosome breakage) bleomycin. These results indicate that in situ hybridization with the alphoid p82H DNA probe is a suitable method with which to distinguish between MN containing whole chromosomes and acentric fragments, and hence allows one to discriminate between the clastogenic and aneugenic effects in MN formation.

Aneuploidy↗

Localization of cloned, repetitive DNA sequences in deer species and its implications for maintenance of gene territory.

The deer family shows the largest variation in chromosome number known in mammals (2n = 6 to 2n = 70). The drastic rearrangement of the chromosomes allows to test the prediction, based on the chromosome field theory, according to which DNA sequences tend to occupy specific territories within the eukaryotic chromosome. Nuclear DNAs were isolated from eight Deer and two Bovidae species. These DNAs were cleaved with the restriction enzymes Eco RI and Alu I. Following Eco RI digestion highly repetitive sequences formed two bands which were common to all deer species. These bands had about 1520 and 2240 base pairs and were particularly clear in Capreolus capreolus (2n = 70, roe deer). The 1520 band DNA was cloned in plasmid pUC9, nick translated, and hybridized with the DNAs of all ten species. The Capreolus DNA showed a high homology with the DNAs of all the species including the Indian muntjac (Muntiacus muntjak, 2n = 7, male) and the Chinese muntjac (Muntiacus reevesi, 2n = 46). 3H hybridization was carried out in situ with metaphase chromosomes of Cervus elaphus (2n = 68, red deer), M. muntjak, and M. reevesi. In C. elaphus all the chromosomes are heavily labeled except the regions of the arms near the centromere. In the muntjacs all chromosomes and most regions appear to be labeled. These results were checked with a different technique that involved the use of two antibodies and biotin labeling of the DNA. The hybridization picture was essentially the same as obtained with 3H. These results disclose that the 1520 bp DNA piece maintains its main territory independently of the drastic changes in chromosome number.

Animals↗

Localization of the repetitive telomeric sequence (TTAGGG)n in two muntjac species and implications for their karyotypic evolution.

It has been suggested that the chromosome set of the Indian muntjac, Muntiacus muntjak vaginalis (female, 2n = 6; male, 2n = 7), evolved from small acrocentric chromosomes, such as those found in the complement of the Chinese muntjac, M. reevesi (2n = 46), by a series of tandem fusions and other rearrangements. The location of the highly conserved human telomeric sequence (TTAGGG)n in the metaphase chromosomes of M.m. vaginalis and its close relative, M. reevesi, was investigated by non-radioactive in situ hybridization. The (TTAGGG)n repeat was found adjacent to the centromeres in the short arm and at the telomeres in the long arm of M. reevesi acrocentric metaphase chromosomes. Tandem fusions present in the karyotype of M.m. vaginalis chromosomes were not reflected by interstitial signals of the telomere repeat, as these chromosomes displayed hybridization signals only at the ends of the chromatids. Mechanisms that might have played a role in the evolution of the reduced karyotype of the Indian muntjac are discussed.

Animals↗

Interspecies relationship of a repetitive chromosome-specific DNA.

A repetitive DNA motif, consisting of 48 bp units (D22Z3) was shown to reside in the pericentric region of the human chromosome 22. In genomic blots it was displaying cross-homology only to great ape species, the restriction patterns being similar but specific for each species investigated. Non-radioactive in situ hybridization revealed a minor homology to human chromosomes 14/15 when less stringent conditions were applied. In gorilla, only chromosomes corresponding to human G-group autosomes showed a distinct cross-hybridization, whereas in chimpanzee, homology to this sequence was observed for all acrocentric chromosomes, even with reduced signal accumulation for the human 22 equivalent at higher stringencies.

Animals↗

The p16/Cdkn2a/Ink4a gene is frequently deleted in nitrosourea-induced rat glial tumors.

The present study investigates nitrosourea-induced rat (Rattus norvegicus) glioma cell lines for the functional status of the p16/Cdkn2a/Ink4a gene, which encodes the p16 cdk4 inhibitor and the alternative reading frame protein, p19ARF. We detected homozygous deletions of the p16/Cdkn2a/Ink4a gene locus in 4 of 5 glioma cell lines (C6, F98, RG2, and RGL.3), but not in the 9L gliosarcoma cell line or in a rat primary fibroblast cell line. RT-PCR demonstrated expression of the p16 and p19ARF mRNAs only in 9L cells and in rat fibroblasts. Comparative genomic in situ hybridization showed that the copy number of rat chromosome RNO5 was not altered in any of the glioma cell lines investigated, indicating that the deletions result from a discrete loss in the region of the p16/Cdkn2a/Ink4a locus. This is the first report of p16/Cdkn2a/Ink4a deletions present in nitrosourea-induced rat glioma cell lines. Since this genetic alteration is also commonly observed in human malignant glial tumors, our results validate the use of chemically induced rat glioma cell lines as an experimental model in the development of gene therapy strategies.

Animals↗