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2017 ; 7
(1
): 11696
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Ultrastructural Characterization of the Glomerulopathy in Alport Mice by Helium
Ion Scanning Microscopy (HIM)
#MMPMID28916834
Tsuji K
; Suleiman H
; Miner JH
; Daley JM
; Capen DE
; P?unescu TG
; Lu HAJ
Sci Rep
2017[Sep]; 7
(1
): 11696
PMID28916834
show ga
The glomerulus exercises its filtration barrier function by establishing a
complex filtration apparatus consisting of podocyte foot processes, glomerular
basement membrane and endothelial cells. Disruption of any component of the
glomerular filtration barrier leads to glomerular dysfunction, frequently
manifested as proteinuria. Ultrastructural studies of the glomerulus by
transmission electron microscopy (TEM) and conventional scanning electron
microscopy (SEM) have been routinely used to identify and classify various
glomerular diseases. Here we report the application of newly developed helium ion
scanning microscopy (HIM) to examine the glomerulopathy in a Col4a3 mutant/Alport
syndrome mouse model. Our study revealed unprecedented details of glomerular
abnormalities in Col4a3 mutants including distorted podocyte cell bodies and
disorganized primary processes. Strikingly, we observed abundant filamentous
microprojections arising from podocyte cell bodies and processes, and presence of
unique bridging processes that connect the primary processes and foot processes
in Alport mice. Furthermore, we detected an altered glomerular endothelium with
disrupted sub-endothelial integrity. More importantly, we were able to clearly
visualize the complex, three-dimensional podocyte and endothelial interface by
HIM. Our study demonstrates that HIM provides nanometer resolution to uncover and
rediscover critical ultrastructural characteristics of the glomerulopathy in
Col4a3 mutant mice.