Department of Anatomy, School of Medicine, University of Mostar, 88000 Mostar, Bosnia and Herzegovina.
Department of Anatomy, Histology and Embryology, University of Split School of Medicine, 21000 Split, Croatia.
Biomolecules. 2023 Feb 21;13(3):402. doi: 10.3390/biom13030402.
The purpose of this study was to compare the immunofluorescence patterns of autophagic markers: Light chain 3 beta (LC3B), Glucose regulating protein 78 (GRP78), Heat shock cognate 71 (HSC70) and Lysosomal-associated membrane protein 2A (LAMP2A) in the developing and postnatal kidneys of () mice to those of wild-type samples. Embryos were obtained on gestation days 13.5 and 15.5 (E13.5 and E15.5), and adult animals were sacrificed at postnatal days 4, 11 and 14 (P4, P11, and P14). After fixation and dehydration, paraffin-embedded kidney tissues were sectioned and incubated with specific antibodies. Using an immunofluorescence microscope, sections were analyzed. For statistical analysis, a two-way ANOVA test and a Tukey's multiple comparison test were performed with a probability level of < 0.05. A significant increase in GRP78 and LAMP2A expression was observed in the renal vesicles and convoluted tubules of in embryonic stages. In postnatal kidneys, all observed proteins showed higher signal intensities in proximal and distal convoluted tubules of , while a higher percentage of LC3B-positive cells was also observed in glomeruli. Our findings suggest that all of the examined autophagic markers play an important role in normal kidney development, as well as the potential importance of these proteins in renal pathology, where they primarily serve a protective function and thus may be used as diagnostic and therapeutic targets.
微管相关蛋白轻链 3β(LC3B)、葡萄糖调节蛋白 78(GRP78)、热休克同源蛋白 71(HSC70)和溶酶体相关膜蛋白 2A(LAMP2A)在()小鼠发育和出生后肾脏中的表达模式,并与野生型样本进行比较。胚胎分别于妊娠第 13.5 天和第 15.5 天(E13.5 和 E15.5)获得,成年动物分别于出生后第 4、11 和 14 天(P4、P11 和 P14)处死。组织固定和脱水后,用石蜡包埋切片并用特异性抗体孵育。使用免疫荧光显微镜分析切片。采用双因素方差分析和 Tukey 多重比较检验,概率水平设为<0.05。结果显示,在胚胎期,GRP78 和 LAMP2A 在肾小囊和曲管中的表达显著增加。在出生后的肾脏中,所有观察到的蛋白在近端和远端曲管中的信号强度均较高,而 LC3B 阳性细胞的比例也较高。这些发现表明,所有检测到的自噬标志物在正常肾脏发育中均发挥重要作用,这些蛋白在肾脏病理中可能具有重要作用,主要起保护作用,因此可能作为诊断和治疗靶点。