Lee Sang-Eun, Kim Yoonju, Han Jeong-Kyu, Park Hoyong, Lee Unghwi, Na Myeongsu, Jeong Soomin, Chung ChiHye, Cestra Gianluca, Chang Sunghoe
Department of Physiology and Biomedical Sciences, Seoul National University College of Medicine, Seoul 01030, South Korea; Biomembrane Plasticity Research Center, Seoul National University College of Medicine, Seoul 01030, South Korea;
Department of Physiology and Biomedical Sciences, Seoul National University College of Medicine, Seoul 01030, South Korea; Biomembrane Plasticity Research Center, Seoul National University College of Medicine, Seoul 01030, South Korea; KM Convergence Research Division, Korea Institute of Oriental Medicine, Daejeon 34054, South Korea;
Proc Natl Acad Sci U S A. 2016 Jun 14;113(24):6749-54. doi: 10.1073/pnas.1600944113. Epub 2016 May 25.
Neural Abelson-related gene-binding protein 2 (nArgBP2) was originally identified as a protein that directly interacts with synapse-associated protein 90/postsynaptic density protein 95-associated protein 3 (SAPAP3), a postsynaptic scaffolding protein critical for the assembly of glutamatergic synapses. Although genetic deletion of nArgBP2 in mice leads to manic/bipolar-like behaviors resembling many aspects of symptoms in patients with bipolar disorder, the actual function of nArgBP2 at the synapse is completely unknown. Here, we found that the knockdown (KD) of nArgBP2 by specific small hairpin RNAs (shRNAs) resulted in a dramatic change in dendritic spine morphology. Reintroducing shRNA-resistant nArgBP2 reversed these defects. In particular, nArgBP2 KD impaired spine-synapse formation such that excitatory synapses terminated mostly at dendritic shafts instead of spine heads in spiny neurons, although inhibitory synapse formation was not affected. nArgBP2 KD further caused a marked increase of actin cytoskeleton dynamics in spines, which was associated with increased Wiskott-Aldrich syndrome protein-family verprolin homologous protein 1 (WAVE1)/p21-activated kinase (PAK) phosphorylation and reduced activity of cofilin. These effects of nArgBP2 KD in spines were rescued by inhibiting PAK or activating cofilin combined with sequestration of WAVE. Together, our results suggest that nArgBP2 functions to regulate spine morphogenesis and subsequent spine-synapse formation at glutamatergic synapses. They also raise the possibility that the aberrant regulation of synaptic actin filaments caused by reduced nArgBP2 expression may contribute to the manifestation of the synaptic dysfunction observed in manic/bipolar disorder.
神经阿贝尔森相关基因结合蛋白2(nArgBP2)最初被鉴定为一种能与突触相关蛋白90/突触后致密蛋白95相关蛋白3(SAPAP3)直接相互作用的蛋白质,SAPAP3是一种对谷氨酸能突触组装至关重要的突触后支架蛋白。尽管在小鼠中基因敲除nArgBP2会导致出现类似双相情感障碍患者许多症状的躁狂/双相样行为,但nArgBP2在突触处的确切功能却完全未知。在此,我们发现通过特异性小发夹RNA(shRNA)敲低(KD)nArgBP2会导致树突棘形态发生显著变化。重新引入对shRNA有抗性的nArgBP2可逆转这些缺陷。特别是,nArgBP2敲低损害了棘突-突触形成,使得兴奋性突触大多终止于有棘神经元的树突干而非棘突头部,尽管抑制性突触形成未受影响。nArgBP2敲低还进一步导致棘突中肌动蛋白细胞骨架动力学显著增加,这与威斯科特-奥尔德里奇综合征蛋白家族维普洛林同源蛋白1(WAVE1)/p21激活激酶(PAK)磷酸化增加以及丝切蛋白活性降低有关。通过抑制PAK或激活丝切蛋白并结合隔离WAVE可挽救nArgBP2敲低对棘突的这些影响。总之,我们的结果表明nArgBP2在谷氨酸能突触处发挥作用以调节棘突形态发生及随后的棘突-突触形成。它们还提出了一种可能性,即nArgBP2表达降低导致的突触肌动蛋白丝异常调节可能促成了在躁狂/双相情感障碍中观察到的突触功能障碍的表现。