DNA sequencing system from histopathological sections

本研究采用显微解剖方法从组织病理切片中建立DNA测序系统，并采用形态学和分子生物学方法对几种突变小鼠进行了研究。研究的突变小鼠如下：1)尿脲诱导肺肿瘤，2)肾素敲除小鼠，3)GM2激活蛋白过表达转基因小鼠，4)遗传性白内障小鼠，5)转基因雌性生殖肿瘤小鼠，6)雄性不育小鼠。此外，利用马立克病病毒感染细胞系，采用宿主基因组减法检测新的基因组多态性。在精子发生的研究中，MRL小鼠作为一种携带减数分裂中期特异性凋亡基因和抗热休克基因的新品系被建立。令人惊讶的是，这种菌株在产后的睾丸中暂时显示出卵母细胞样细胞。这些结果推测，MRL小鼠具有热休克抗性的精母细胞在Sertoli细胞的减数分裂检查点完成前分化为卵母细胞样细胞，并在完成后进入减数分裂中期的凋亡状态。为了鉴定减数分裂中期特异性凋亡和热休克抗性精母细胞相关基因，我们对C57BL/6和MRL小鼠的睾丸dna进行了差异展示，检测到几个新的克隆。特别是对200个回交后代的染色体连锁分析表明，携带中期特异性凋亡的基因定位在1号染色体上的100.0 cM处。

In the present study, DNA sequencing system was established from histopathological sections by using microdissection procedure, and several mutant mice were investigated by morphological and molecular biological approaches. Mutant mice investigated were the following ; 1) urethan-induced lung tumor, 2) renin knockout mice, 3) transgenic mice overexpressing GM2 activator protein, 4) hereditary cataract mice, 5) female genital tumor mice creating by transgenic strategy and 6) male sterile mice. Additionally, the novel genomic polymorphisms were detected by the host genome subtraction method using Marek's Disease virus infected cell lines.During the investigation of spermatogenesis, MRL mouse was established as a new strain carrying the genes of meiotic metaphase-specific apoptosis and heat-shock resistant spermatocytes. Surprisingly, this strain showed temporarily oocyte-like cells in testis during postnatal periods. These results assumed that spermatocyte possessing heat-shock resistance in MRL mice differentiates into oocytes-like cells before the completion of meiotic checkpoint in Sertoli cells, and then progresses to apoptotic figure of meiotic metaphase after it's completion. In order to identify the genes associating meiotic metaphase-specific apoptosis and heat-shock resistance spermatocytes, the differential display was performed between the testis cDNAs of C57BL/6 and MRL mice, detecting several novel clones. Especially, it was suggested by chromosomal linkage analysis of 200 backcross progenies that the gene carrying metaphase-specific apoptosis was mapped at 100.0 cM on chromosomal 1.