Kim Hee Jeong, Lee Kina, Yoo Kiseo, Kim Jeong Eun, Kim Heeju, Lim Chae-Seok, Park Young Seok, Kim Hyong Kyu
Graduate Program in Neuroscience, Department of Medicine and Microbiology, Chungbuk National University, Rm #421, 194-15, Osongsaengmyeong 1-Ro, Heungdeok-Gu, Cheongju, 28160, Republic of Korea.
Department of Pharmacology, Wonkwang University School of Medicine, Iksan, 54538, Republic of Korea.
Mol Neurobiol. 2025 Jun;62(6):7647-7663. doi: 10.1007/s12035-025-04734-7. Epub 2025 Feb 8.
Although Rho guanine nucleotide exchange factor 4 (Arhgef4) is highly expressed in the brain, its function remains poorly understood. Our previous study showed that Arhgef4 negatively regulates excitatory postsynaptic regional activity. This study investigated the effects of Arhgef4 deletion in postnatal forebrain-specific knockout mice on brain function, synaptic proteins, and behaviors. We generated a knockout mouse with Arhgef4 deleted from the forebrain and analyzed gene expression and protein levels by RT-PCR and western blot. Synaptic function was assessed through electrophysiological recordings, and behavioral tests evaluated memory and anxiety. In these conditional knockout (cKO) mice, we observed a significant decrease in the expression of a 75-kDa brain-enriched isoform of Arhgef4 in the forebrain. In KO mice, pre- and post-synaptic protein levels were unchanged. However, in cultured hippocampal neurons from KO mice, the levels of postsynaptic density protein 95 (PSD-95) in the postsynaptic regions were significantly increased from the pre-mature stage to the fully mature stage during neuronal development. In contrast, the number of dendritic protrusions decreased during the early mature stage of the cultured neurons. Electrophysiological recordings of hippocampal neurons from KO mice showed a significant increase in miniature excitatory postsynaptic currents (mEPSC) frequency. Furthermore, Arhgef4 KO mice exhibited enhanced long-term memory and reduced anxiety-related behaviors. These findings suggest that Arhgef4 plays a role in regulating brain functions such as learning, memory, and anxiety.
尽管Rho鸟嘌呤核苷酸交换因子4(Arhgef4)在大脑中高度表达,但其功能仍知之甚少。我们之前的研究表明,Arhgef4对兴奋性突触后区域活动起负调节作用。本研究调查了出生后前脑特异性敲除小鼠中Arhgef4缺失对脑功能、突触蛋白和行为的影响。我们构建了前脑缺失Arhgef4的敲除小鼠,并通过RT-PCR和蛋白质免疫印迹分析基因表达和蛋白质水平。通过电生理记录评估突触功能,通过行为测试评估记忆和焦虑。在这些条件性敲除(cKO)小鼠中,我们观察到前脑中75 kDa富含脑的Arhgef4亚型的表达显著降低。在敲除小鼠中,突触前和突触后蛋白水平未发生变化。然而,在敲除小鼠培养的海马神经元中,突触后密度蛋白95(PSD-95)在神经元发育过程中从早熟阶段到完全成熟阶段,其在突触后区域的水平显著增加。相反,在培养神经元的早期成熟阶段,树突突起的数量减少。对敲除小鼠海马神经元的电生理记录显示,微小兴奋性突触后电流(mEPSC)频率显著增加。此外,Arhgef4敲除小鼠表现出增强的长期记忆和减少的焦虑相关行为。这些发现表明,Arhgef4在调节学习、记忆和焦虑等脑功能中发挥作用。