Grasbon-Frodl E M, Nakao N, Lindvall O, Brundin P
Department of Neurological Surgery, Wakayama Medical College, Japan.
Neuroscience. 1996 Jul;73(1):171-83. doi: 10.1016/0306-4522(96)00008-5.
Basic parameters which are crucial for the survival of human embryonic striatal grafts need to be investigated before initiating clinical trials in Huntington's disease. In order to define the dissection of human striatal-donor tissue which gives rise to the largest amount of striatal neurons after intrastriatal transplantation, we studied the lateral and medial ganglionic eminences of embryonic striatal primordia obtained from human embryos sized 17-30 mm in crown-to-rump length (corresponding to Carnegie stages 18-23). Anatomical landmarks that demarcated the lateral and medial ganglionic eminences from each other were present only in embryos with 20 mm crown-to-rump length or larger. In monolayer cultures, the lateral ganglionic eminence gave rise to a six-fold higher yield of dopamine- and cyclic AMP-regulated phosphoprotein 32-immunoreactive striatal neurons as compared to the medial ganglionic eminence. We also xenografted the lateral and medial ganglionic eminences from five embryos sized 21-30 mm in crown-to-rump length to the ibotenate lesioned striatum of immunosuppressed rats. The grafts were evaluated with respect to general morphology, survival and integration using (immuno-) histochemical stains for acetylcholinesterase/Cresyl Violet, nicotinamide adenine dinucleotide phosphate-diaphorase, dopamine- and cyclic AMP-regulated phosphoprotein-32, tyrosine hydroxylase and calbindin-D28KD. As assessed 9-25 weeks after implantation, 13 out of 16 and 8 out of 13 grafts, in the groups grafted with the medial and lateral ganglionic eminences, respectively, had survived. Previous studies with rat donor tissue have indicated that the functional efficacy of striatal grafts is related to the development of striatal-specific P-zone regions and that these are enriched in transplants derived from the lateral as opposed to the medial ganglionic eminence. Also in the human striatal xenografts of the present study, P-zones appeared more abundant when the donor tissue was derived from the lateral ganglionic eminence. However, the proportion of graft tissue that expressed P-zone properties was always very low (at most 30%) and never approached the 80-90% previously observed in transplants of rat lateral ganglionic eminence. We conclude that the relative yield of striatal neurons in grafts of the human embryonic striatal primordium has to be improved before neural transplantation should be applied in patients with Huntington's disease.
在针对亨廷顿舞蹈症开展临床试验之前，需要对人类胚胎纹状体移植存活的关键基础参数进行研究。为了确定纹状体内移植后能产生最多纹状体神经元的人类纹状体供体组织的解剖结构，我们研究了从冠臀长17 - 30毫米（对应卡内基分期18 - 23期）的人类胚胎获取的胚胎纹状体原基的外侧和内侧神经节隆起。将外侧和内侧神经节隆起彼此区分开的解剖学标志仅存在于冠臀长20毫米或更长的胚胎中。在单层培养中，与内侧神经节隆起相比，外侧神经节隆起产生的多巴胺和环磷酸腺苷调节磷蛋白32免疫反应性纹状体神经元产量高六倍。我们还将5个冠臀长21 - 30毫米胚胎的外侧和内侧神经节隆起移植到免疫抑制大鼠的鹅膏蕈氨酸损伤纹状体中。使用乙酰胆碱酯酶/甲酚紫、烟酰胺腺嘌呤二核苷酸磷酸黄递酶、多巴胺和环磷酸腺苷调节磷蛋白-32、酪氨酸羟化酶和钙结合蛋白-D28KD的（免疫）组织化学染色对移植体的一般形态、存活和整合情况进行评估。在植入后9 - 25周进行评估时，分别移植内侧和外侧神经节隆起的组中，16个移植体中有13个、13个移植体中有8个存活。先前对大鼠供体组织的研究表明，纹状体移植的功能效力与纹状体特异性P区的发育有关，并且这些区域在源自外侧而非内侧神经节隆起的移植体中更为丰富。在本研究的人类纹状体异种移植中，当供体组织源自外侧神经节隆起时，P区也显得更为丰富。然而，表达P区特性的移植组织比例始终非常低（最多30%），从未接近先前在大鼠外侧神经节隆起移植中观察到的80 - 90%。我们得出结论，在将神经移植应用于亨廷顿舞蹈症患者之前，必须提高人类胚胎纹状体原基移植中纹状体神经元的相对产量。
