Hayashi Kanehiro, Furuya Asako, Sakamaki Yuriko, Akagi Takumi, Shinoda Yo, Sadakata Tetsushi, Hashikawa Tsutomu, Shimizu Kazuki, Minami Haruka, Sano Yoshitake, Nakayama Manabu, Furuichi Teiichi
Laboratory for Molecular Neurogenesis, RIKEN Brain Science Institute, Wako, Saitama, Japan.
Department of Anatomy, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan.
PLoS One. 2017 Mar 6;12(3):e0173175. doi: 10.1371/journal.pone.0173175. eCollection 2017.
Very-KIND/Kndc1/KIAA1768 (v-KIND) is a brain-specific Ras guanine nucleotide exchange factor carrying two sets of the kinase non-catalytic C-lobe domain (KIND), and is predominantly expressed in cerebellar granule cells. Here, we report the impact of v-KIND deficiency on dendritic and synaptic growth in cerebellar granule cells in v-KIND knockout (KO) mice. Furthermore, we evaluate motor function in these animals. The gross anatomy of the cerebellum, including the cerebellar lobules, layered cerebellar cortex and densely-packed granule cell layer, in KO mice appeared normal, and was similar to wild-type (WT) mice. However, KO mice displayed an overgrowth of cerebellar granule cell dendrites, compared with WT mice, resulting in an increased number of dendrites, dendritic branches and terminals. Immunoreactivity for vGluT2 (a marker for excitatory presynapses of mossy fiber terminals) was increased in the cerebellar glomeruli of KO mice, compared with WT mice. The postsynaptic density around the terminals of mossy fibers was also increased in KO mice. Although there were no significant differences in locomotor ability between KO and WT animals in their home cages or in the open field, young adult KO mice had an increased grip strength and a tendency to exhibit better motor performance in balance-related tests compared with WT animals. Taken together, our results suggest that v-KIND is required for compact dendritic growth and proper excitatory synaptic connections in cerebellar granule cells, which are necessary for normal motor coordination and balance.
极亲和蛋白/Kindlin-3/KIAA1768(v-KIND)是一种大脑特异性Ras鸟嘌呤核苷酸交换因子,带有两组激酶非催化C叶结构域(KIND),主要在小脑颗粒细胞中表达。在此,我们报告了v-KIND基因敲除(KO)小鼠中v-KIND缺乏对小脑颗粒细胞树突和突触生长的影响。此外,我们评估了这些动物的运动功能。KO小鼠小脑的大体解剖结构,包括小脑小叶、分层的小脑皮质和密集排列的颗粒细胞层,看起来正常,与野生型(WT)小鼠相似。然而,与WT小鼠相比,KO小鼠的小脑颗粒细胞树突过度生长,导致树突、树突分支和末端数量增加。与WT小鼠相比,KO小鼠小脑小球中vGluT2(苔藓纤维末梢兴奋性突触前标记物)的免疫反应性增加。KO小鼠苔藓纤维末梢周围的突触后密度也增加。尽管在饲养笼或旷场中,KO和WT动物的运动能力没有显著差异,但与WT动物相比,年轻成年KO小鼠握力增加,并且在平衡相关测试中表现出更好运动性能的趋势。综上所述,我们的结果表明,v-KIND是小脑颗粒细胞中紧密树突生长和适当兴奋性突触连接所必需的,而这对于正常的运动协调和平衡是必要的。