Expression of coxsackievirus and adenovirus receptor isoforms in developing mouse bladder uroepithelium.
OBJECTIVE
Tight junctions are important for uroepithelial paracellular permeability barriers. In the present study, we examined the developmental changes in the expression of coxsackievirus and adenovirus receptor (CAR) isoforms in mouse bladder uroepithelium.


METHODS
Multiplex reverse transcriptase polymerase chain reaction using CAR isoform-specific primer sets and Western blotting were conducted on gestational day 19 and postnatal days 1, 7, and 55. Subcellular localization of CAR was examined, together with occludin and zonula occludens-1, in neonatal and adult bladder using light microscopy and immunofluorescence microscopy.


RESULTS
The total CAR and short CAR isoform mRNA were significantly increased from gestational day 19 to birth. Long CAR isoform mRNA was transiently decreased on postnatal day 7 and had recovered during adulthood. On Western blotting, molecular weight 46-kDa CAR was abundant in the mucosa and increased postnatally. In the neonatal, as well as the adult, bladder uroepithelium, CAR immunoreactivity was observed, together with occludin and zonula occludens-1 at the apical tight junctions and basolateral contacts between the adjacent uroepithelial cells. In adult bladder uroepithelium, CAR was increased at the interface between the basal cells and basal lamina.


CONCLUSIONS
The expression patterns of CAR isoforms changed during the late fetal to adult development of the mouse bladder. CAR at the apical tight junctions and cellular adhesions between the uroepithelial cells and the interfaces between the basal cells and basal lamina might support the paracellular permeability barrier and structural integrity of the uroepithelium in the mouse bladder. The expression of CAR in the uroepithelial cells can be integrated as a part of the strategy for virus-mediated gene therapy in the bladder uroepithelium.
Department of Life Science, Hanyang University College of Natural Sciences, Seoul Korea.
Myung Chan
MC
Gye
Yeong Seok
YS
Oh
Jae Eun
JE
Lee
Sarah
S
Shim
Kyung Jin
KJ
Choi
Hyun Soo
HS
Ahn
D000256:Adenoviridae; D000818:Animals; D001485:Basement Membrane / Q000502:physiology; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D004591:Electrophoresis, Polyacrylamide Gel; D004848:Epithelium / Q000196:embryology; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D020033:Protein Isoforms; D011991:Receptors, Virus / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019108:Tight Junctions / Q000502:physiology* / Q000821:virology; D001743:Urinary Bladder / Q000196:embryology*
C568772:CLMP protein, mouse; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D020033:Protein Isoforms; D011991:Receptors, Virus; C105941:coxsackievirus B receptor