Gene Mapping in Fishes and Other Vertebrates

Morizot, Donald C.; Siciliano, Michael J.

doi:10.1007/978-1-4684-4652-4_4

Cited by 25 publications

(14 citation statements)

References 83 publications

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“…Also, neither hatching success of the clutch spawned by the amplectant female nor mean There is no evidence of linkage disequilibrium among the loci examined in this study. This result is consistent with other data on conserved linkage associations in vertebrate species (Morizot & Siciliano, 1984).…”

Section: Resultssupporting

confidence: 93%

Genetic heterozygosity and reproductive success in the green treefrog, Hyla cinerea

McAlpine

1993

Heredity

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The relationship between genetic variation and fitness characteristics is important because reduced variation at the population level is usually associated with reduced viability of individuals. Adult green treefrogs from a single population were examined at eight polymorphic allozyme loci to test the prediction that a positive correlation exists between reproductive success and number of heterozygous loci. In females, clutch size, corrected for snout-vent length, and total number of surviving offspring were significantly correlated with number of heterozygous loci. The relationship with percentage hatched showed the same correlation, but it was not significant. Body size as measured by snout-vent length was not associated with number of heterozygous loci, although larger females had significantly more hatched offspring. In males, heterozygosity was not associated with body size or hatching success of the clutch. Total reproductive success in females was positively correlated with number of heterozygous loci, while factors such as body size, which may affect fitness, were not.

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Section: Resultssupporting

confidence: 93%

Genetic heterozygosity and reproductive success in the green treefrog, Hyla cinerea

McAlpine

1993

Heredity

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“…Dissection and tissue preparation for starch gel electrophoresis of proteins followed described methods (30). A listing of polymorphic proteins analyzed in this study, with conditions for electrophoresis and histochemical staining, may be found in Morizot and Schmidt (31).…”

Section: Methodsmentioning

confidence: 99%

A CDKN2 -like polymorphism in Xiphophorus LG V is associated with UV-B-induced melanoma formation in platyfish–swordtail hybrids

Nairn

Kazianis

McEntire

et al. 1996

Proc. Natl. Acad. Sci. U.S.A.

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The genetic basis of spontaneous melanoma formation in spotted dorsal (Sd) Xiphophorus platyfishswordtail hybrids has been studied for decades, and is adequately explained by a two-gene inheritance model involving a sex-linked oncogene, Xmrk, and an autosomal tumor suppressor, DIFF. The Xmrk oncogene encodes a receptor tyrosine kinase related to EGFR; the nature of the DIFF tumor suppressor gene is unknown. We analyzed the genetic basis of UV-B-induced melanoma formation in closely related, spotted side platyfish-swordtail hybrids, which carry a different sex-linked pigment pattern locus, Sp. We UV-irradiated spotted side Xiphophorus platyfish-swordtail backcross hybrids to induce melanomas at frequencies 6-fold higher than occur spontaneously in unirradiated control animals. To identify genetic determinants of melanoma susceptibility in this UVinducible Xiphophorus model, we genotyped individual animals from control and UV-irradiated experimental regimes using allozyme and DNA restriction fragment length polymorphisms and tested for joint segregation of genetic markers with pigmentation phenotype and UV-induced melanoma formation. Joint segregation results show linkage of a CDKN2-like DNA polymorphism with UV-B-induced melanoma formation in these hybrids. The CDKN2-like polymorphism maps to Xiphophorus linkage group V and exhibits recombination fractions with ES1 and MDH2 allozyme markers consistent with previous localization of the DIFF tumor suppressor locus. Our results indicate that the CDKN2-like sequence we have cloned and mapped is a candidate for the DIFF tumor suppressor gene.Genetic hybrids between species of the genus Xiphophorus (Teleostei: Poeciliidae) exhibit spontaneous melanoma formation in several different cross types and have been used for decades to investigate genetic factors contributing to melanoma formation (1). The most studied and best understood Xiphophorus hybrid melanoma is the spotted dorsal GordonKosswig platyfish-swordtail model (2-5), represented by genetic hybrids derived from crossing F 1 hybrids between the platyfish Xiphophorus maculatus Jp 163 A and the swordtail Xiphophorus helleri back to X. helleri. Melanoma formation in this tumor model is genetically controlled by inheritance of a sex-linked receptor tyrosine kinase gene (Xmrk), associated with the spotted dorsal (Sd) pigment pattern locus, and segregation of an autosomal locus in Xiphophorus linkage group (LG) V, variously referred to in the literature as DIFF, R DIFF , and R (3-6). The DIFF locus is believed to regulate macromelanophore pigment cell differentiation (4, 6), and behaves in the Gordon-Kosswig model as a classical tumor suppressor for which loss of species-specific alleles in pigmented backcross hybrids results in melanoma formation according to simple, Mendelian inheritance (1, 3-5).Elegant experiments have shown that Xmrk is a duplicated gene, which has adventitiously acquired the promoter from another gene (7,8). It has been postulated that expression of the oncogenic Xmrk gene duplicate is re...

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“…There is even evidence for the long-term stable gene linkage relations between groups as evolutionary divergent as fish and mammals although the data are not numerous, as summarized by Morizot and Siciliano (1984).…”

Section: No 3]mentioning

confidence: 99%

Fish gene mapping using mouse-fish cell hybrids.

Wang

Ojima

1988

Proceedings of the Japan Academy. Ser. B: Physical and Biologic

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In 1970, a nonreverting 8-azaguanine resistant mutant cell line (RAG), deficient in the HPRT gene, was isolated from cultured renal adenocarcinoma cells by Klebe et al. (1970). Since fish cells can usually grow only at temperatures lower than 30°C, a subline (RAG30) that can grow at 28 to 30°C was isolated from RAG to make mouse-fish hybridization easier. In an attempt to assign the fish HPRT gene, RAG30 cells were fused with fish cells derived from primarily cultured fin cells of the saddled weever (Parapercis sex f asciata, Perciformes) which is thought to have a relatively differentiated XY type sex chromosome (Ojima et al., 1984). Our results indicate evidence of nonconservation of gene linkage relations in the sex chromosomes of fish and mammals.Materials and methods, a) Isolation of RAG30. The culture temperature of RAG cells was gradually reduced to 30°C at a rate of 0.5°C day. During the procedure most cells became detached from the petri-dishes but some survived and proliferated.The growth rate of these cells became greater during cultivation at 30°C, and they acquired almost the same growth ability as the parent RAG cells. b) Cell fusion. About 10G RAG30 cells and 106 fish cells were mixed and cultured at 28°C overnight in HANKS' MEM containing double the amount of the Eagle's vitamins and amino acids, supplemented with 10% fetal calf serum.. One ml of 47% PEG (polyethylene glycol 4000, MERCK) solution containing 5% DMSO (dimethyl sulfoxide, Norwood et al., 1976) was added to the monolayer culture for one minute. The cells were thoroughly washed with BSS four times and cultured in the normal medium for two days for the expression time of fish genes. Two days after the fusion, GHAT was added to the medium and was changed every four to five days. Two weeks later, several developing colonies of hybrid cells were observed. c) Chromosome preparation. The cultures were exposed for one hour to 0.05 µg/ml of colchicine before harvesting. 0.8 µg/ml actinomycin D was added together with the colchicine to make the chromosomes elongated (Yu et al., 1981;Rybak et al., 1982;Liu, 1986). The cells were trypsinized and resuspended in 10 ml hypotonic solution (0.067 M KC1) for 20 min and fixed in methanol-acetic acid (3:1).G-banding and C-banding were carried out using the methods of Seabright (1971) and Sumner (1972), respectively. d) Primary culture. The method developed by 0 j ima (1982) was used for the primary fin culture.Results and discussion. About two weeks after the fusion, colonies of hybrid cells were observed. The karyotypes of the hybrid cells were analyzed one month after growth in GHAT medium.

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Gene Mapping in Fishes and Other Vertebrates

Cited by 25 publications

References 83 publications

Genetic heterozygosity and reproductive success in the green treefrog, Hyla cinerea

Genetic heterozygosity and reproductive success in the green treefrog, Hyla cinerea

A CDKN2 -like polymorphism in Xiphophorus LG V is associated with UV-B-induced melanoma formation in platyfish–swordtail hybrids

Fish gene mapping using mouse-fish cell hybrids.

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