Laboratory of Molecular Neurobiology, Graduate School of Biostudies, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan.
Mol Biol Cell. 2013 May;24(10):1602-13. doi: 10.1091/mbc.E12-11-0782. Epub 2013 Mar 27.
In neuronal development, dendritic spine formation is important for the establishment of excitatory synaptic connectivity and functional neural circuits. Developmental deficiency in spine formation results in multiple neuropsychiatric disorders. Dock4, a guanine nucleotide exchange factor (GEF) for Rac, has been reported as a candidate genetic risk factor for autism, dyslexia, and schizophrenia. We previously showed that Dock4 is expressed in hippocampal neurons. However, the functions of Dock4 in hippocampal neurons and the underlying molecular mechanisms are poorly understood. Here we show that Dock4 is highly concentrated in dendritic spines and implicated in spine formation via interaction with the actin-binding protein cortactin. In cultured neurons, short hairpin RNA (shRNA)-mediated knockdown of Dock4 reduces dendritic spine density, which is rescued by coexpression of shRNA-resistant wild-type Dock4 but not by a GEF-deficient mutant of Dock4 or a truncated mutant lacking the cortactin-binding region. On the other hand, knockdown of cortactin suppresses Dock4-mediated spine formation. Taken together, the results show a novel and functionally important interaction between Dock4 and cortactin for regulating dendritic spine formation via activation of Rac.
在神经元发育过程中,树突棘的形成对于兴奋性突触连接和功能神经回路的建立非常重要。树突棘形成的发育缺陷会导致多种神经精神疾病。Dock4 是 Rac 的鸟嘌呤核苷酸交换因子 (GEF),已被报道为自闭症、阅读障碍和精神分裂症的候选遗传风险因素。我们之前曾表明,Dock4 在海马神经元中表达。然而,Dock4 在海马神经元中的功能及其潜在的分子机制仍知之甚少。在这里,我们表明 Dock4 高度集中在树突棘中,并通过与肌动蛋白结合蛋白 cortactin 的相互作用参与形成树突棘。在培养的神经元中,短发夹 RNA (shRNA)介导的 Dock4 敲低会降低树突棘密度,而 shRNA 抗性野生型 Dock4 的共表达可挽救这种情况,但 Dock4 的 GEF 缺陷突变体或缺乏 cortactin 结合区的截断突变体则不能挽救。另一方面,cortactin 的敲低会抑制 Dock4 介导的树突棘形成。总之,这些结果表明 Dock4 和 cortactin 之间存在一种新颖的、功能重要的相互作用,通过激活 Rac 来调节树突棘的形成。