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目的观察神经型一氧化氮合酶(nNOS)在胚胎小鼠肾发育过程中的表达,探讨nNOS在小鼠肾脏发育早期的作用。方法选取胚龄(E)12、14、16、18 d及新生组(P0 d)小鼠,应用免疫组织化学技术观察nNOS的表达部位。应用体视学检测体密度值方法和免疫印迹技术检测各组小鼠肾组织中nNOS含量。结果免疫组化染色显示,E12 d,nNOS在输尿管芽呈现阳性表达;E14 d,nNOS在皮质生肾区、近端小管和远端小管表达。E16 d,nNOS在输尿管芽、生肾区和近端小管的表达减弱,在远端小管的表达加强。至P0 d,nNOS在输尿管芽、生肾区和近端小管的表达进一步减弱,在远端小管尤其是致密斑处呈强表达。从E12 d至P0 d,除了在成熟肾小体的肾小囊偶见nNOS的阳性表达外,在发育各期的肾小体中均未见nNOS的阳性表达。在集合管中也无表达。体视学分析结果显示,从E12 d至P0 d,各组小鼠肾组织中nNOS的体密度值逐渐增加。免疫印迹结果显示,从E12 d至P0 d,nNOS的表达量逐渐增加。结论在胚胎小鼠肾发育过程中,nNOS表达部位主要在远端小管致密斑,可能对远端小管的发育起重要的调节作用。
Objective To observe the expression of neuronal nitric oxide synthase (nNOS) during embryonic kidney development and to explore the role of nNOS in the early development of kidney in mice. Methods The mice of embryonic day (E) 12, 14, 16 and 18 days and neonatal rats (P0 d) were selected for immunohistochemistry to observe the expression of nNOS. The volume of nNOS in the kidney of each group was detected by the methods of volume density and volumetric method. Results Immunohistochemical staining showed that the expression of nNOS was positive in ureteral buds on E12 d and nNOS in proximal cortex and distal tubule on E14 d. On day E16, the expression of nNOS in the ureter bud, nephrogenic zone and proximal tubule was weakened and the expression in the distal tubule was enhanced. At P0 d, the expression of nNOS in the ureter bud, nephrogenic zone and proximal tubule was further weakened, and was strongly expressed in distal tubules, especially in the dense plaque. From E12 d to P0 d, no positive expression of nNOS was found in all stages of nephrons, except for occasional positive expression of nNOS in renal cysts of mature kidney. No expression in collecting tube. Stereological analysis showed that from E12 d to P0 d, the body density of nNOS gradually increased in all groups of mice. Immunoblotting showed that the expression of nNOS gradually increased from E12 d to P0 d. Conclusion In the process of embryonic mouse kidney development, the nNOS expression site is mainly in distal tubule plaque, which may play an important regulatory role in the development of distal tubules.