1. Cell culture-based karyotyping of orectolobiform sharks for chromosome-scale genome analysis
- Author
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Keiichi Sato, Yoshinobu Uno, Shigehiro Kuraku, Ryo Nozu, Nobuyuki Higashiguchi, Itsuki Kiyatake, Kiyomi Murakumo, and Shuji Sodeyama
- Subjects
Male ,0106 biological sciences ,0301 basic medicine ,Chiloscyllium punctatum ,Cell Culture Techniques ,Medicine (miscellaneous) ,Evolutionary biology ,chemical and pharmacologic phenomena ,Biology ,Whale shark ,010603 evolutionary biology ,01 natural sciences ,Genome ,Article ,Chromosomes ,General Biochemistry, Genetics and Molecular Biology ,Cytogenetics ,03 medical and health sciences ,biology.animal ,medicine ,Animals ,Carpet shark ,Lymphocytes ,lcsh:QH301-705.5 ,Cells, Cultured ,In Situ Hybridization, Fluorescence ,Comparative Genomic Hybridization ,medicine.diagnostic_test ,Chromosome ,Vertebrate ,Karyotype ,Fibroblasts ,biology.organism_classification ,Genome evolution ,030104 developmental biology ,lcsh:Biology (General) ,Karyotyping ,Sharks ,Female ,General Agricultural and Biological Sciences ,human activities ,Fluorescence in situ hybridization - Abstract
Karyotyping, traditionally performed using cytogenetic techniques, is indispensable for validating genome assemblies whose sequence lengths can be scaled up to chromosome sizes using modern methods. Karyotype reports of chondrichthyans are scarce because of the difficulty in cell culture. Here, we focused on carpet shark species and the culture conditions for fibroblasts and lymphocytes. The utility of the cultured cells enabled the high-fidelity characterization of their karyotypes, namely 2n = 102 for the whale shark (Rhincodon typus) and zebra shark (Stegostoma fasciatum), and 2n = 106 for the brownbanded bamboo shark (Chiloscyllium punctatum) and whitespotted bamboo shark (C. plagiosum). We identified heteromorphic XX/XY sex chromosomes for the two latter species and demonstrated the first-ever fluorescence in situ hybridization of shark chromosomes prepared from cultured cells. Our protocols are applicable to diverse chondrichthyan species and will deepen the understanding of early vertebrate evolution at the molecular level., Shark cells are difficult to culture in the laboratory as the high osmolality physiological conditions do not reflect those of commonly cultured mammalian or teleost fish cell lines. Uno et al. present a new method of culturing shark fibroblasts and lymphocytes, and characterized the karyotypes of four carpet shark species, enabling whole genome analysis of these species.
- Published
- 2020
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