Neph1 and nephrin interaction in the slit diaphragm is an important determinant of glomerular permeability.
Neph1-deficient mice develop nephrotic syndrome at birth, indicating the importance of this protein in the development of a normal glomerular filtration barrier. While the precise subcellular localization of Neph1 remains unknown, its relationship with other components of the glomerular filtration barrier is of great interest in this field. In this paper, we localize the expression of Neph1 to the glomerular slit diaphragm by immunogold electron microscopy in rodents and describe its direct interaction with two other components of the slit diaphragm, nephrin and ZO-1. Both native and recombinant Neph1 associate with each other as dimers and multimers and interact with nephrin via their extracellular segments. Disruption of the Neph1-nephrin interaction in vivo by injecting combinations of individual subnephritogenic doses of anti-Neph1 and anti-nephrin results in complement- and leukocyte-independent proteinuria with preserved foot processes. This disruption modestly reduces Neph1 and nephrin protein expression in podocytes and dramatically reduces ZO-1 protein expression via the interaction of ZO-1 PDZ domains with the cytoplasmic tail of Neph1, independent of changes in mRNA expression of all three genes. The interaction between nephrin and Neph1 is specific and not shared by either protein with P-cadherin, another integral slit diaphragm protein. The interaction between nephrin and Neph1 therefore appears to be an important determinant of glomerular permeability.
Liu
Gang
G
Department of Medicine, Feinberg School of Medicine, Northwestern University, Chicago, Illinois 60611, USA.
Kaw
Beenu
B
Kurfis
Jayson
J
Rahmanuddin
Syed
S
Kanwar
Yashpal S
YS
Chugh
Sumant S
SS
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D015820:Cadherins / Q000378:metabolism; D003001:Cloning, Molecular; D003593:Cytoplasm / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D003720:Densitometry; D019281:Dimerization; D005919:Glomerular Filtration Rate; D007150:Immunohistochemistry; D007668:Kidney / Q000166:cytology / Q000502:physiology*; D007678:Kidney Glomerulus / Q000502:physiology*; D008565:Membrane Proteins / Q000096:biosynthesis / Q000502:physiology*; D051379:Mice; D008856:Microscopy, Fluorescence; D008957:Models, Genetic; D008969:Molecular Sequence Data; D010750:Phosphoproteins / Q000096:biosynthesis; D011233:Precipitin Tests; D011506:Proteins / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D062826:Zonula Occludens-1 Protein
D015820:Cadherins; D018076:DNA, Complementary; C486606:Kirrel1 protein, mouse; C486607:Kirrel1 protein, rat; D008565:Membrane Proteins; D010750:Phosphoproteins; D011506:Proteins; D012333:RNA, Messenger; D011994:Recombinant Proteins; C569043:Tjp1 protein, mouse; C569044:Tjp1 protein, rat; D062826:Zonula Occludens-1 Protein; C113267:nephrin