Bairstow B M, Harvey A R
Department of Anatomy and Human Biology, University of Western Australia, Nedlands, Perth.
Exp Brain Res. 1992;89(3):560-70. doi: 10.1007/BF00229881.
Tectal tissue from E15 or E16 Wistar rat embryos was dissociated and reaggregated (DR) prior to transplantation on to the midbrain of newborn host rats. We wished to determine how complete disruption of the donor tissue (i) affected the subsequent morphological development of the grafts in the host brain, and (ii) whether this procedure affected the selectivity with which host retinal axons innervated target regions in the tectal transplants. Forty-three to 135 days after transplantation, host rats received binocular injections of wheatgerm agglutinin-conjugated horseradish peroxidase. After perfusion, frozen sections of the grafts and underlying host brainstem were cut and reacted with tetramethylbenzidine to identify retinal projections, or stained for either acetylcholinesterase (AChE), Nissl or neurofibrils. All host brains contained identifiable DR grafts. Each brain contained at least one large transplant and numerous smaller pieces of graft tissue. The fragmentation of DR grafts was greater than that seen in direct, undissociated tectal transplants; however the morphology of individual DR grafts was markedly similar to direct grafts. Of particular interest was the presence in DR grafts of localized, often oval or circular regions, that possessed high AChE activity and contained mostly small (5 to 10 microns) close-packed neurons. AChE-dense patches were found both superficially and deep within DR grafts and appeared identical to those seen in direct transplants. These regions are thought to be homologous to the superficial, retinorecipient layers of normal superior colliculus (SC) and it is likely that the formation of these localized areas resulted from the selective association of presumptive SGS neurons within the reaggregating neuropil. In almost all cases, host retinal input to DR grafts was confined to the localized AChE-dense patches, suggesting that despite the dissociation procedure, specific retinal innervation of regions containing at least some of the appropriate target cells was maintained in DR tectal grafts.
将E15或E16期Wistar大鼠胚胎的顶盖组织解离并重新聚集(DR),然后移植到新生宿主大鼠的中脑上。我们希望确定供体组织的完全破坏如何(i)影响宿主脑中移植物随后的形态发育,以及(ii)该程序是否影响宿主视网膜轴突支配顶盖移植中靶区域的选择性。移植后43至135天,宿主大鼠接受双目注射麦胚凝集素结合辣根过氧化物酶。灌注后,将移植物和下方宿主脑干的冰冻切片切开,与四甲基联苯胺反应以鉴定视网膜投射,或进行乙酰胆碱酯酶(AChE)、尼氏或神经原纤维染色。所有宿主脑均含有可识别的DR移植物。每个脑至少包含一个大的移植物和许多较小的移植物组织碎片。DR移植物的碎片化程度大于直接的、未解离的顶盖移植物;然而,单个DR移植物的形态与直接移植物明显相似。特别令人感兴趣的是,DR移植物中存在局部的、通常为椭圆形或圆形的区域,这些区域具有高AChE活性,并且大多包含紧密排列的小(5至10微米)神经元。在DR移植物的表面和深部均发现了AChE密集斑块,其外观与直接移植中所见的斑块相同。这些区域被认为与正常上丘(SC)的浅表视网膜接受层同源,并且这些局部区域的形成可能是由于重新聚集的神经毡内假定的SGS神经元的选择性结合所致。在几乎所有情况下,宿主视网膜对DR移植物的输入都局限于局部AChE密集斑块,这表明尽管经过了解离程序,但DR顶盖移植物中含有至少一些合适靶细胞的区域仍维持了特定的视网膜神经支配。