Isenmann S, Brandner S, Sure U, Aguzzi A
Department of Pathology, University Hospital, Zürich, Switzerland.
Neuropathol Appl Neurobiol. 1996 Apr;22(2):108-17.
Telencephalic grafting represents a powerful tool for developmental studies and for the investigation of biological features of transgenic brain tissue. The interpretation of grafting experiments, however, requires detailed knowledge of graft biology. Therefore, we have characterized growth rates, graft size, and differentiation of embryonic telencephalic tissue harvested at various developmental stages and grafted into the caudoputamen and lateral ventricles of histocompatible mice. A total of 164 grafts were analysed up to 500 days after transplantation. Of all transplants, 79.3% resulted in the formation of solid neural grafts. Grafted cells were identified by 3H-thymidine labelling and autoradiography. Proliferation was studied by bromodeoxyuridine incorporation and decreased from an initial 35% at 1-3 d after grafting to less than 1.6% after 40 days. The graft size was measured as a function of the embryonic age of the transplanted tissue. Our data indicate that telencephalic tissue harvested at embryonic day E 12.5 reproducibly yields large, fully differentiated neuroectodermal grafts. The parameters defined in this study will be useful for detailed analysis of neuroectodermal tissue from mice undergoing fatal neurodegeneration, such as knockout mice bearing lethal mutations.
端脑移植是发育研究以及转基因脑组织生物学特性研究的有力工具。然而,移植实验的解读需要对移植生物学有详细的了解。因此,我们对在不同发育阶段采集并移植到组织相容性小鼠的尾壳核和侧脑室中的胚胎端脑组织的生长速率、移植块大小及分化情况进行了表征。在移植后长达500天的时间里,共分析了164个移植块。在所有移植中,79.3%形成了实体神经移植块。通过3H-胸腺嘧啶核苷标记和放射自显影鉴定移植细胞。通过溴脱氧尿苷掺入研究增殖情况,增殖率从移植后1 - 3天的初始35%下降到40天后的不足l.6%。移植块大小作为移植组织胚胎年龄的函数进行测量。我们的数据表明,在胚胎第12.5天采集的端脑组织可重复性地产生大的、完全分化的神经外胚层移植块。本研究中定义的参数将有助于对患有致命性神经退行性变的小鼠(如携带致死突变的基因敲除小鼠)的神经外胚层组织进行详细分析。