Kato Takashi, Mizuno Shinya
Department of Biochemistry and Molecular Biology, Graduate School of Medicine, Osaka University, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan.
Department of Pathology, Faculty of Medicine, Kindai University, 377-2 Ohno-higashi, Osaka-sayama, Osaka 589-8511, Japan.
Exp Anim. 2017 Aug 5;66(3):183-189. doi: 10.1538/expanim.16-0101. Epub 2017 Feb 7.
Newborn mouse glomeruli are still immature with a morphological feature of an early capillary loop stage, but infant mice do not manifest proteinuria. Little is known about the molecular mechanism whereby infant mice are resistant to proteinuria. Nephrin and synaptopodin are crucial for slit diaphragm and foot process (FP) formation for avoiding proteinuria. Nephrin tyrosine phosphorylation means a transient biological signaling required for FP repair or extension during nephrotic disease. Using an immunohistochemical technique, we examined the natural course of nephrin, Wilms' tumor-1 (WT1) and synaptopodin at 16.5 days of embryonic age (E16.5d) and E19.5d, 7 days of post-neonatal age (P7d) and P42d during renal development of mice. As a result, nephrin and synaptopodin were detected at E19.5d in S-shaped bodies. WT1, a transcriptional factor for nephrin, was detected in nucleus in podocyte-like cells in all stages. Nephrin tyrosine phosphorylation was evident in glomeruli at P7d, and this was associated with an early-stage of FP extension. Inversely, nephrin phosphorylation became faint at P42d, along with maturated FP. Based on the present results, we suggest the sequential molecular mechanism to protect growing mice from proteinuria: (i) WT1-induced nephrin production by podocytes in S-shaped bodies at E19.5d; (ii) Synchronized induction of synaptopodin at the same period; and (iii) FP extension is initiated at a milk-suckling stage under a nephrin tyrosine-phosphorylated condition, while it is arrested at an adult stage, associated with a loss of nephrin-based signaling.
新生小鼠的肾小球仍不成熟,具有早期毛细血管袢阶段的形态特征,但幼鼠并不表现出蛋白尿。关于幼鼠对蛋白尿具有抗性的分子机制,目前了解甚少。Nephrin和突触足蛋白对于防止蛋白尿的裂孔隔膜和足突(FP)形成至关重要。Nephrin酪氨酸磷酸化意味着肾病期间FP修复或延伸所需的短暂生物信号。我们使用免疫组织化学技术,检测了小鼠肾脏发育过程中胚胎期16.5天(E16.5d)和E19.5d、出生后7天(P7d)和P42d时Nephrin、肾母细胞瘤-1(WT1)和突触足蛋白的自然进程。结果显示,在E19.5d的S形体中检测到了Nephrin和突触足蛋白。WT1作为Nephrin的转录因子,在所有阶段的足细胞样细胞核中均有检测到。Nephrin酪氨酸磷酸化在P7d的肾小球中很明显,这与FP延伸的早期阶段相关。相反,在P42d时,随着FP成熟,Nephrin磷酸化变得微弱。基于目前的结果,我们提出了保护生长中小鼠免受蛋白尿影响的顺序分子机制:(i)在E19.5d时,WT1诱导S形体中的足细胞产生Nephrin;(ii)同期同步诱导突触足蛋白;(iii)FP延伸在吸乳阶段在Nephrin酪氨酸磷酸化的条件下启动,而在成年阶段停止,这与基于Nephrin的信号丧失有关。