Molecular pathology of neuronal differentiation, migration and death in developmental disorders.

A.神经元死亡我们制备了一种抗Bak的多克隆抗体，Bak是一种促进神经元凋亡的蛋白质，因此我们用Western blotting和免疫染色方法研究了它在人脑中的表达。1996年，我们研究了与发育和衰老相关的变化，并证明Bak在胎儿和老年人的脑中高表达。1997年，我们比较了唐氏综合征患者和正常对照患者的Bak免疫反应。在唐氏综合症患者的大脑中，与衰老相关的Bak表达上调过早发生。脑神经元在发生神经纤维变化之前就呈Bak阳性。B.神经元分化我们制造了针对Tuberin N端和C端的兔抗体，Tuberin是导致结节性硬化症的TSC2基因的产物。1996年，我们展示了Tuberin在对照大脑中的表达。在发育过程中，块茎蛋白含量随着年龄的增长而增加。相比之下，结节性硬化症的大脑显示出结节蛋白的丢失，这在错构瘤病变(皮质结节和室管膜下巨细胞瘤)和组织学正常的皮质中都是严重的。肾错构瘤和心脏错构瘤的Tuberin免疫反应也丢失。1997年，我们在局灶性皮质发育不良中观察到了正常水平的tuberin表达，从而表明结节性硬化症和皮质发育不良之间的病理生理差异。C.神经元迁移我们扩大了对Lis1基因产物(PAF乙酰水解酶的45k亚基)的免疫组织化学研究，该基因的缺陷导致了Miller-Dieker裂脑综合征。1996年，我们研究了Lis1在各种迁移性疾病中的表达，并证明了Lis1的缺失是该综合征特有的。1997年，我们对人胎脑进行了免疫组织化学染色，观察到脑室神经上皮和Cajal-Retzius细胞的强标记。

A.Neuronal deathWe produced a polyclonal antibody against Bak, a protein promoting neuronal apoptosis, and thereby studied its expression in human brains by Western blotting and immunostaining. In 1996, we investigated the changes associated with development and aging, and demonstrated that the expression of Bak is high in the fetal and aged brains. In 1997, we comapared Bak immunoreactivity between Down syndrome and control patients. In Down syndrome brains, the aging-related upregulation of Bak occurred prematurely. Cerebral neurons became Bak-positive prior to the development of neurofibrillary changes.B.Neuronal differentiationWe produced rabbit antibodies against the N-and C-terminal of tuberin, the product of the TSC2 gene responsible for tuberous sclerosis. In 1996, we demonstrated the expression of tuberin in control cerebra. During development, tuberin content increased with age. Tuberous sclerosis brains by contrast showed loss of tuberin, which was severe in both the hamartomatous lesions (cortical tuber and subependymal giant cell tumor) and histologically normal cortices. Tuberin immunoreactivity was also lost from the renal and cardiac hamartomas. In 1997, we observed a normal level of tuberin expression in focal cortical dysplasia, thereby indicating pathophysiological difference between tuberous sclerosis and cortical dysplasia.C.Neuronal migrationWe extended immunohistochemical studies of the LIS1 gene product (a 45k subunit of PAF acetylhydrolase), the defect of which being responsible for the Miller-Dieker lissencephaly syndrome. In 1996, we studied the expression of LIS1 in various migration disorders, and demonstrated that the loss of LIS1 is specific to the syndrome. In 1997, we immunostained human fetal brains and observed strong labeling of the ventricular neuroepithlium and Cajal-Retzius cells.