MICE LACKING ALKALINE PHOSPHATASE ISOZYMES.-MOLECULAR GENETICS AND PATHOLOGICAL INVESTIGATION

缺乏碱性磷酸酶同工酶的小鼠-分子遗传学和病理学研究

基本信息

批准号：
09044335
负责人：
WATANABE Keiichi
金额：
$ 4.03万
依托单位：
TOKAI UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for international Scientific Research
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09044335/
关键词：
ALKALINE PHOSPHATASE (AP)EMBRYONAL AP TISSUE NONSPECIFIC AP KOCK-OUT MICE alkaline phophatase(AP)

项目摘要

There are 3 to 4, depending on the difference of the animal species, isozymes of alkaline phosphatase (AP), and they are widely distributed in those animal bodies. However, little is known about their exact biological functions and left as a "mysterious enzyme". Attempts were thus made to disclose those biological functions and significances by establishing "knock-out mice" for the genes of mouse APs ((1)D embryonic (EAP), (2) tissue non-specific (TNAP) and (3) intestinal (IAP)) and by further investigations on genomic and phenotypic expression of those genes and diverse patho-physiological phenomena which may appear in those animals. Since the establishment of "IAP knock-out mice" is still in process and "EAP knock-out mice" did not show any significant phenotypic changes, the observations were centered to the changes appeared in the "TNAP knock-out mice" . (1) The majority of these "knock-out mice" died during the weaning period (averagely 2 weeks) after they exhibited characteristic epileptic seizure. Sine it is a well documented fact that pyridoxal (vitaminB^6=VB^6) deficient mice fell in the very similar epileptic seizure, we presume TNAP gene inactivation would cause in VB^6 depletion which is not experimentally proved yet. TNAP could be taking an important role in dephosphorylation of pyridoxal phosphates. (2) Another conspicuous pathological change observed in those "knock-out mice" was abnormal mineralization appeared in bones of almost whole bodies. The deficiency of TNAP in osteoblasts which show marked structural deformity may strongly be responsible for thatpathology. Concerning IAP knockout mouse preparation, ES cells transfected with the targeting vector described before. One of the clones was confirmed as a homologous recombinant by Southern blots, and this clone was injected into the bastocysts. We are currently breeding the four chimeric males in order to obtain germline heterozygote mice.

有3至4，具体取决于动物物种的差异，碱性磷酸酶（AP）的同工酶，它们被广泛分布在这些动物体内。然而，对它们的确切生物学功能知之甚少，并且是一种“神秘酶”。因此，通过建立对小鼠APS基因（（（1）D胚胎（EAP），（2）组织非特异性（TNAP）和（3）肠道（IAP）组织的“敲除小鼠”，试图通过建立“敲除小鼠”来披露这些生物学功能和意义的尝试，并通过对这些基因和多样性的进一步研究来揭示这些生物学功能和意义。由于建立“ IAP敲除小鼠”仍在过程中，并且“ EAP敲除小鼠”没有显示出任何显着的表型变化，因此观察结果集中在“ TNAP敲除小鼠”中出现的变化。（1）这些“淘汰小鼠”中的大多数在断奶时期（平均2周）死亡，它们表现出特征性的癫痫发作。正弦是一个有充分记载的事实，吡啶毒素（维生素^6 = vb^6）缺乏小鼠在非常相似的癫痫发作中，我们认为TNAP基因失活会在Vb^6耗尽中引起，这尚未在实验上证明。 TNAP可能在吡啶多磷酸盐的去磷酸化中发挥重要作用。（2）在那些“敲除小鼠”中观察到的另一种明显的病理变化是矿化异常，几乎全身的骨骼中出现。在成骨细胞中TNAP的缺乏表现出明显的结构畸形可能是该路径学的原因。关于IAP敲除小鼠的制备，ES细胞用前面描述的靶向载体转染。其中一个克隆被Southern印迹确认为同源重组，并且该克隆被注入了混蛋。我们目前正在繁殖这四个嵌合雄性，以获得种系杂合子小鼠。