Establishment of high-resolution FISH mapping system and its application for molecular cytogenetic characterization of chromosomes in newt, Cynops pyrrhogaster (Urodela, Amphibia).

Takahiro Murakami, Nobuyasu Maki, Chizuko Nishida-Umehara, Yoichi Matsuda, Kiyokazu Agata
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引用次数: 10

Abstract

Urodele amphibians (newts and salamanders) are important animal models for understanding regeneration mechanisms and genome evolution. We constructed ideograms of BrdU/dT- and C-banded karyotypes in the Japanese fire-belly newt, Cynops pyrrhogaster, which is useful as a model animal with extremely high ability of regeneration. We also established a high-resolution FISH mapping system for newts, and localized satellite DNA sequences, 18S rDNAs, telomeric (TTAGGG)n repeats and seven functional genes, including genes associated with lens regeneration, tyrosinase and two types of gamma crystallins, to chromosomes of the newt. The 18S rDNAs were localized to three chromosomal pairs in males, whereas the chromosomal locations were highly variable in females. No hybridization signals were detected for the telomeric (TTAGGG)n sequence. All three lens regeneration-related genes were mapped on the short arm of chromosome 7, suggesting that the location of the genes in the same linkage group may be correlated with the regulation of gene expression associated with chromatin dynamics in interphase nuclei during regeneration. The chromosomal distribution and nucleotide sequences of pericentric satellite DNA sequences were well conserved between C. pyrrhogaster and European newts; in contrast, there was species specificity of nucleotide sequences for centromere-specific satellite DNAs.

高分辨率FISH定位系统的建立及其在蝾螈(尾纲,两栖纲)染色体分子细胞遗传学鉴定中的应用
尾纲两栖动物(蝾螈和蝾螈)是了解再生机制和基因组进化的重要动物模型。构建了再生能力极高的日本火腹蝾螈(Cynops pyrrhogaster) BrdU/dT-和c -带核型表意图。我们还建立了一个高分辨率的蝾螈FISH定位系统,并将卫星DNA序列、18S rnas、端粒(TTAGGG)n重复序列和7个功能基因定位到蝾螈的染色体上,包括与晶体再生、酪氨酸酶和两种γ晶体蛋白相关的基因。在雄性中,18S rnas定位在3对染色体上,而在雌性中,染色体位置变化很大。端粒(TTAGGG)n序列未检测到杂交信号。所有三个晶状体再生相关基因都被定位在7号染色体的短臂上,这表明基因在同一连锁组中的位置可能与再生过程中间期核染色质动力学相关基因的表达调控有关。金舌螈与欧洲蝾螈的染色体分布和近心卫星DNA序列具有较好的保守性;相反,着丝粒特异性卫星dna的核苷酸序列具有物种特异性。
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