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Production of Sterile Mouse Model for Human and Domestic Animal

人和家畜无菌小鼠模型的制作

基本信息

批准号：
07558235
负责人：
WATANABE Tomomasa
金额：
$ 1.92万
依托单位：
Hokkaido University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1997
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07558235/
关键词：
mouse hybrid sterile cDNA subtraction sex chromosome spermatogenesis meiosis northern blot 流産外挿

项目摘要

In mice, the interspecific or intersubspecific hybrid sterility has been occasionally found in matings between laboratory mice and wild mice induced by Hybrid sterility (Hst) genes. One of them, Hst3, which controls the sterility of interspecific hybrids between laboratory mice and Mus spretus originated from Spanish wild mouse, is associated with X-Y chromosome dissociation accompanied by spermatogenic breakdown after metaphase I (MI). For detaild analysis of hybrid sterility caused by Hst3, C57BL/6 congenic mice, carring Hst3 derived from spretus, were created.Up to postnatal day 12, spermatogenesis progressed relatively normal in both type testis. At day 14, when early pachytene spermatocytes appeared in nomal testis, many degenerations weredetected on these cells in sterile testis. At day 20, many round spermatids appeared in nomal testis, while no round spermatids were found in stertile testis. At day 60 of sexually matured age, nomal testis involved in all kinds of germ cells, ho … More wever it was noted that few spermatids and spermatozoa were found in sterile testis throughout the histological sections.According to morphologial differeces between normal and sterile testis, the subtraction method between normal and sterile testis cDNA was performed using congenic mouse testis to analyze the normal spermatogenesis-associated genes. Transcripts of some subtracted cDNA clones were expressed higher in normal testis than in sterile testis by northern blot analysis. One of them, clone F77, showing significant unmatchies with entries in the demonstrated remarkable expression of apporoximately 4kb transcript in nomal testis, compared to those in sterile testis. Additionaly the transcript was not detected at any other organs except testis. Northern blot analysis using developing normal testis and germ cell-defficient W/Wv mutant testis, show that the transcript of cDNA clone F77 was expressed only in normal adult testis, in which it was detected in elongated spermatids and partially round spermatids by in situ hybridization. cDNA clone F77 was mapped at position between Mt-1 and Ctrb on mouse chromosome 8, in which no related genes were detected Less

在小鼠中，杂种不育（Hst）基因诱导的实验室小鼠与野生小鼠交配时偶尔会发现种间或亚种间杂种不育。其中之一，Hst3，控制实验室小鼠和源自西班牙野生小鼠的小家鼠之间种间杂交的不育性，与中期 I (MI) 后伴随着生精破坏的 X-Y 染色体解离有关。为了详细分析Hst3引起的杂交不育，创建了携带来自spretus的Hst3的C57BL/6同系小鼠。直到出生后第12天，两种类型睾丸的精子发生进展相对正常。第14天，当正常睾丸中出现早期粗线期精母细胞时，在无菌睾丸中的这些细胞上检测到许多变性。第20天，正常睾丸中出现许多圆形精子细胞，而不育睾丸中未发现圆形精子细胞。性成熟60天时，正常睾丸含有各种生殖细胞，但在整个组织学切片中发现，无菌睾丸中几乎没有精细胞和精子。根据正常睾丸与无菌睾丸形态的差异，利用同源小鼠睾丸进行正常睾丸cDNA与不育睾丸cDNA的差减法，对正常睾丸进行分析。精子发生相关基因。根据Northern印迹分析，一些消减的cDNA克隆的转录物在正常睾丸中的表达高于在无菌睾丸中的表达。其中之一，克隆 F77，与无菌睾丸中的相比，在正常睾丸中显示出大约 4kb 转录物的显着表达，显示出与条目的显着不匹配。此外，除睾丸外，在任何其他器官中均未检测到转录本。使用发育中的正常睾丸和生殖细胞缺陷的W/Wv突变体睾丸进行Northern印迹分析，表明cDNA克隆F77的转录物仅在正常成年睾丸中表达，其中通过原位杂交在细长精子细胞和部分圆形精子细胞中检测到该转录物。 cDNA克隆F77定位于小鼠8号染色体Mt-1和Ctrb之间，未检测到相关基因 Less