Spatio-temporal expression of membrane transporters, Megalin and Cubilin, in renal development of C57BL/6 mice

Mei LUO; Xiaoyue ZHAI; Yunsheng MA

doi:10.16016/j.2097-0927.202306091

您当前的位置：

首页 >

文章列表页 >

Spatio-temporal expression of membrane transporters, Megalin and Cubilin, in renal development of C57BL/6 mice

Basic Medicine | 更新时间：2026-01-09

- Spatio-temporal expression of membrane transporters, Megalin and Cubilin, in renal development of C57BL/6 mice
- Journal of Army Medical University Vol. 46, Issue 5, Pages: 434-441(2024)
- 作者机构：
  
  1.121001 辽宁锦州，锦州医科大学生科院电镜室
  2.110001 沈阳，中国医科大学组织学与胚胎学教研室
  3.121001 辽宁锦州，锦州医科大学医疗学院形态学教研室
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(30871313);the Basic Research Project of Colleges and Universities of Liaoning Provincial Department of Education(LJKMZ20222086)
- DOI：10.16016/j.2097-0927.202306091
  CLC： R322.61;R329.26;R341
- Received：25 June 2023，
  
  Revised：2023-09-07，
  
  Published：15 March 2024
- 稿件说明：
移动端阅览
Mei LUO, Xiaoyue ZHAI, Yunsheng MA. Spatio-temporal expression of membrane transporters, Megalin and Cubilin, in renal development of C57BL/6 mice[J]. Journal of Army Medical University, 2024, 46(5): 434-441.
DOI：

Mei LUO, Xiaoyue ZHAI, Yunsheng MA. Spatio-temporal expression of membrane transporters, Megalin and Cubilin, in renal development of C57BL/6 mice[J]. Journal of Army Medical University, 2024, 46(5): 434-441. DOI： 10.16016/j.2097-0927.202306091.

摘要

目的

观察膜转运体Megalin、Cubilin在发生发育小鼠肾内的分布，从而揭示它们在肾发生发育中相应功能分化的时空规律。

方法

C57BL/6小鼠按雌雄1∶1合笼饲养，并于胚(胎)龄(Ed)E10、12、14、16、18 d，及生后日龄(Pd)P1、3、7、14、28、42 d，分离胎鼠、仔鼠或成鼠的肾脏。应用改良的块染技术对不同发育阶段的肾脏组织进行整体固定及染色，分别进行常规石蜡包埋，制成连续的石蜡切片(5 μm)。选取连续切片中目标切片，采用免疫组织化学技术和免疫荧光技术对不同发育阶段的小鼠肾脏膜转运体Megalin和Cubilin进行定位，观察其在不同发育阶段的表达情况。

结果

Megalin和Cubilin在E10 d的中肾管上皮细胞开始有表达，E12 d时表达于肾脏输尿管芽上皮细胞的近腔面游离缘，二者主要以共表达模式存在。E14 d以后在发生发育小鼠肾脏的“逗号”小体、“S”型小体上皮细胞的游离缘呈弱表达，于肾脏近端小管的上皮细胞游离缘表达明显，偶尔可见在未发育成熟的血管球有弱表达。Megalin和Cubilin在细段、远端小管、集合管系统以及间质部分都未见表达。

结论

膜转运体Megalin和Cubilin在中肾管、输尿管芽、“逗号”小体、“S”型小体，以及近端小管的依次表达规律提示小鼠肾脏在中肾形成时期就开始有蛋白或多肽的转运功能。此外，二者稳定地出现在较为成熟的近端小管，较弱地出现在未成熟近端小管，表明其重吸收蛋白质的功能与近端小管结构的成熟度有关。

Abstract

Objective

To observe the distribution of membrane transporters

Megalin and Cubilin

in the renal development of mice

and to reveal the temporal and spatial rules of their functional differentiation during renal development.

Methods

C57BL/6 mice were housed in a female/male ratio of 1∶1

and the kidneys were isolated at embryonic (Ed) ages of E10

E12

E14

E16 and E18 d

and postnatal ages of P1

P14

P28 and P42 d. A modified block staining technique was applied to fix and stain the whole kidney tissues at different developmental stages

and conventional paraffin and resin 812 embedding were performed to make continuous paraffin sections (5 μm). The target sections in the continuous sections were selected

and immunohistochemical and immunofluorescence assays were used to localize the mouse kidney membrane transporters Megalin and Cubilin at different developmental stages

and to observe their expression levels.

Results

Megalin and Cubilin were expressed in the epithelial cells of the mesonephric tubules at E10d

and in the proximal luminal surface on the epithelial cells in the ureteric buds of the kidneys at E12d

with co-expression in the proximal luminal surface on the epithelial cells in the ureteric buds of the kidneys

and were expressed weakly in the free margin of the epithelial cells of comma cells and S-shaped cells in the kidneys of the mice at the onset of the developmental process from E14d

and were expressed in the free margins of epithelial cells of proximal tubules of the kidneys. Megalin and Cubilin were not expressed in the fine segments

the distal tubules

the collecting duct system

or the mesangial portion of the kidney.

Conclusion

The sequential expression pattern of the membrane transporters Megalin and Cubilin in the mesonephric tubules

ureteric buds

comma vesicles

S-shaped vesicles

and proximal tubules suggests that the mouse kidney begins to have a protein or polypeptide transport function during the period of mesonephric formation. In addition

both proteins appear stably in the more mature proximal tubules and more weakly in the immature proximal tubules

suggesting that their protein reabsorption function is related to the maturity of the proximal tubular structure.

关键词

Keywords

references

AGRE P. Molecular physiology of water transport: aquaporin nomenclature workshop. Mammalian aquaporins[J]. Biol Cell, 1997, 89(5/6): 255-257.

SHIPMAN K E, LONG K R, COWAN I A, et al . An adaptable physiological model of endocytic megalin trafficking in opossum kidney cells and mouse kidney proximal tubule[J ] . Function (Oxf), 2022, 3(6): zqac046. DOI: 10.1093/function/zqac046.

DE S, KUWAHARA S, SAITO A. The endocytic receptor megalin and its associated proteins in proximal tubule epithelial cells[J]. Membranes (Basel), 2014, 4(3): 333-355. DOI: 10.3390/membranes4030333.

SUN Y, LU X F, DANSER A. Megalin: a novel determinant of renin-angiotensin system activity in the kidney?[J]. Curr Hypertens Rep, 2020, 22(4): 30. DOI: 10.1007/s11906-020-01037-1.

许平庆. 916塑料包埋剂在病理技术中应用的体会[J]. 第三军医大学学报, 1993, 15(5): 452. DOI: 10.16016/j.1000-5404.193.05.020.

XU P Q. 916 experience with the use of plastic embedding agents in pathologic techniques[J]. Acta Acad Med Mil Tertiae, 1993, 15(5): 452. DOI: 10.16016/j.1000-5404.193.05.020.

CHRISTENSEN E I, BIRN H. Megalin and cubilin: synergistic endocytic receptors in renal proximal tubule[J]. Am J Physiol Ren Physiol, 2001, 280(4): F562-F573. DOI: 10.1152/ajprenal.2001.280.4.f562.

MIAO X M, ZHAI X Y, GUO M, et al . Expression of endocytic receptor megalin/cubilin in embryonic mouse kidneys[J ] . Acta Anatomica Sinica, 2009, 40(4): 671-674. DOI: 10.3969/j.issn.0529-1356.2009.04.030.

ZOU Z Y, CHUNG B, NGUYEN T, et al . Linking receptor-mediated endocytosis and cell signaling[J ] . J Biol Chem, 2004, 279(33): 34302-34310. DOI: 10.1074/jbc.m405608200.

ZHAI X Y, NIELSEN R, BIRN H, et al . Cubilin- and megalin-mediated uptake of albumin in cultured proximal tubule cells of opossum kidney[J ] . Kidney Int, 2000, 58(4): 1523-1533. DOI: 10.1046/j.1523-1755.2000.00314.x.

DRAKE C J, FLEMING P A, LARUE A C, et al . Differential distribution of cubilinand megalin expression in the mouse embryo[J ] . Anat Rec, 2004, 277A(1): 163-170. DOI: 10.1002/ar.a.10123.

CHRISTENSEN E I, KRISTOFFERSEN I B, GRANN B, et al . A well-developed endolysosomal system reflects protein reabsorption in segment 1 and 2 of rat proximal tubules[J ] . Kidney Int, 2021, 99(4): 841-853. DOI: 10.1016/j.kint.2020.11.015.

SUN Y, MARTINI A G, JANSSEN M J, et al . Megalin: A novel endocytic receptor for prorenin and renin[J ] . Hypertension, 2020, 75(5). DOI: 10.1161/HYPERTENSIONAHA.120.14845.

YAMMANI R R, SEETHARAM S, SEETHARAM B. Cubilin and megalin expression and their interaction in the rat intestine: effect of thyroidectomy[J]. Am J Physiol Endocrinol Metab, 2001, 281(5): E900-E907. DOI: 10.1152/ajpendo.2001.281.5.E900.

SMITH C P, LEE W K, HALEY M, et al . Proximal tubule transferrin uptake is modulated by cellular iron and mediated by apical membrane megalin-cubilin complex and transferrin receptor 1[J ] . J Biol Chem, 2019, 294(17): 7025-7036. DOI: 10.1074/jbc.RA118.006390.

CARRARO-LACROIX L R, MALNIC G. Acid-base transport by the renal distal nephron[J]. J Nephrol, 2010, 23(Suppl 16): S19-S27.

MOESTRUP S K, VERROUST P J. Megalin- and cubilin-mediated endocytosis of protein-bound vitamins, lipids, and hormones in polarized epithelia[J]. Annu Rev Nutr, 2001, 21(1): 407-428.

NIELSEN R, CHRISTENSEN E I, BIRN H. Megalin and cubilin in proximal tubule protein reabsorption: from experimental models to humandisease[J]. Kidney Int, 2016, 89(1): 58-67. DOI: 10.1016/j.kint.2015.11.007.

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Renal protective effect of equol on type 2 diabetic rats

Effect and mechanism of Dickkopf-related protein 1 on urinary sodium excretion in spontaneously hypertensive rats

Recombinant human erythropoietin attenuates renal ischemia reperfusion injury in rats through inhibiting TLR4/NF-κB signal transduction pathway

hBMP-7-modified bone marrow mesenchymal stem cells attenuate kidney ischemia-reperfusion injury in rabbits

Effect of vitamin A deficiency on ABCA1-mediated lipid efflux pathway in rat kidneys

Related Author

Jian WANG

Hanqiang CUI

Guiming ZHANG

Zhe XU

Xiangmin NI

Shuo LI

Yongjian YANG

Hongyong WANG

Related Institution

Department of Nutrition, Second Affiliated Hospital, Army Medical University (Third Military Medical University)

⁰