Podocyte-specific expression of a novel trans-Golgi protein Vear in human kidney.
BACKGROUND
Vear is a recently identified Golgi apparatus-associated protein. It has been suggested to be involved in vesicular trafficking between the Golgi and the vacuolar/lysosomal system. Proteins similar to Vear have also been shown to interact with activated ARF proteins (ADP ribosylation factor), and they are probably involved in membrane trafficking from the trans-Golgi network (TGN). We have previously shown that Vear is widely distributed in human tissues, with an especially high level of mRNA in the kidney. This study further characterizes the distribution and subcellular localization of Vear in normal adult kidney and shows its association with glomerulogenesis in fetal kidney.


METHODS
Immunofluorescence and immunoelectron microscopy were used to study the expression of Vear in fetal and adult kidney. The expression of Vear in isolated glomeruli was shown by immunoblotting. The distribution of its mRNA was analyzed by using in situ and Northern hybridization.


RESULTS
In situ hybridization and immunofluorescence microscopy showed that in the kidney, Vear is present in glomerular structures. By fluorescence microscopy, the immunoreactivity for Vear was found only in podocytes, as judged by its distinct colocalization with podocalyxin and vimentin, well-established marker proteins of podocytes. Its specific expression in the glomeruli versus other compartments of the kidney was also verified by Western blotting. By using immunogold electron microscopy, Vear was seen in the Golgi apparatus, tubulovesicular structures, and membranes adjacent to the Golgi complex. In fetal kidney, expression of Vear coincided with the formation of segmental structures of the glomeruli. It was first seen close to the undifferentiated luminal cells at the vesicular stage and increasingly in the differentiating podocytes at the more advanced stages of glomerulogenesis.


CONCLUSIONS
In the kidney, Vear shows a distinct, specific, and developmentally regulated expression in glomerular podocytes. This suggests that Vear has a specific function in podocytes. It could be associated with the known high secretory and synthetic activity of the podocytes, especially the production of the basement membrane components, which are critically involved in the glomerulogenesis and the maintenance of the glomerular function.
Poussu
A M
AM
Department of Pathology, University of Oulu, Oulu, Finland.
Virtanen
I
I
Autio-Harmainen
H
H
Lehto
V P
VP
D033942:Adaptor Proteins, Vesicular Transport; D000328:Adult; D000818:Animals; D001485:Basement Membrane / Q000737:chemistry / Q000502:physiology / Q000648:ultrastructure; D015152:Blotting, Northern; D015153:Blotting, Western; D019556:COS Cells; D002352:Carrier Proteins; D005333:Fetus / Q000166:cytology; D005455:Fluorescent Antibody Technique; D015870:Gene Expression / Q000502:physiology; D006801:Humans; D017403:In Situ Hybridization; D007678:Kidney Glomerulus / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D016253:Microscopy, Immunoelectron; D021381:Protein Transport / Q000502:physiology; D011506:Proteins / Q000032:analysis / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis; D012795:Sialoglycoproteins / Q000032:analysis; D014746:Vimentin / Q000032:analysis; D021601:trans-Golgi Network / Q000737:chemistry / Q000502:physiology* / Q000648:ultrastructure
D033942:Adaptor Proteins, Vesicular Transport; D002352:Carrier Proteins; C405944:GGA2 protein, human; D011506:Proteins; D012333:RNA, Messenger; D012795:Sialoglycoproteins; D014746:Vimentin; C001643:podocalyxin