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目的:明确解剖和生理上与人非常接近的中国实验用小型猪肾脏发育过程中肾小管的形态学变化和肾小管各节段的特异性标志物。方法:采用高碘酸-希夫(PAS)染色和免疫荧光染色技术,系统观察中国实验用小型猪妊娠28～112d(E28d～E112d)和出生后1d、7d、14d、21d(P1d～21d),共17个不同时间点猪肾小管的发育及肾小管特异性标志物雪莲花凝集素(LTL)、水通道蛋白1(AQP1)、钙结合蛋白(calbindin)-D28k在肾小管不同节段的表达。结果:(1)中国实验用小型猪E28d可见后肾间充质和输尿管芽,即后肾已经开始发育;但这时还没有肾小管。E35d可见不同节段的肾小管,即肾小管已开始发育。从E35d～P14d(E112d仔猪出生),肾皮质均有生肾区存在,即不断有新的肾单位发生;P21d生肾区消失,即不再有新的肾单位产生。(2)①LTL在E28d表达在输尿管芽,E35d开始在近端小管表达,以刷状缘表达最为明显;表达由弱到强,由点状到线状。②AQP1在E28d未见表达,E35d开始表达;AQP1表达在近端小管和髓袢的降支细段,主要表达在细胞膜,尤其在管腔侧的表达更为明显。③Calbindin-D28k在E28d表达在输尿管芽,E35d开始表达在远端小管和集合管;Calbindin-D28k主要表达在细胞质,随着肾小管发育,表达逐渐增强。(3)发现集合管来源于输尿管芽,发源于输尿管芽的集合管从被膜下的生肾区一直延伸到肾髓质。结论:中国实验用小型猪妊娠35d可以见到不同节段肾小管。LTL、AQP1、Calbindin-D28k可以分别作为猪近端小管、髓袢、远端小管和集合管的标志物。
OBJECTIVE: To clarify the morphological changes of renal tubules and the specific markers of renal tubules during kidney development in Chinese experimental miniature pigs, which are very close to human anatomy and physiology. Methods: The effects of PAS staining and immunofluorescence staining on gestational age from 28 to 112 days (E28d to E112d) and 1d, 7d, 14d and 21d (P1d to 21d) , A total of 17 different time points pig kidney tubular development and tubular specific markers snow lotus lectin (LTL), aquaporin 1 (AQP1), calbindin-D28k in different segments of the renal tubules expression. Results: (1) In experimental piglets E28d, the posterior kidney mesenchyme and ureter buds were observed, that is, the posterior kidney had begun to develop; but at this time, there was no renal tubule. E35d shows the different segments of the renal tubules, tubules have begun to develop. From E35d to P14d (E112d piglets were born), renal cortex both have nephrotic areas, that there are new nephrons occur; P21d nephridial area disappears, that is no new nephron production. (2) ①LTL expression in the ureter at E28d bud, E35d began to express in the proximal tubule, brush border expression of the most obvious; expression from weak to strong, from punctate to linear. ② AQP1 was not expressed on E28d, and began to be expressed on E35d. AQP1 was expressed in the descending branch of proximal tubule and medulla oblongata and mainly expressed on the cell membrane, especially on the luminal side. The expression of Calbindin-D28k was mainly expressed in the cytoplasm of cytoplasm at E35d, and gradually increased at the beginning of E35d expression in the distal tubule and collecting duct. (3) The collection tube was found to originate from the ureter bud, and the collecting tube originating from the ureter bud extended from the nephric area under the capsule to the medulla. CONCLUSIONS: Different segments of tubules can be seen in 35-day pregnant mini-pigs in China. LTL, AQP1, Calbindin-D28k can be used as markers of pig proximal tubules, medulla oblongata, distal tubules and collecting duct, respectively.