PMID- 12399632
OWN - NLM
STAT- MEDLINE
DCOM- 20030717
LR  - 20171101
IS  - 1660-8151 (Print)
IS  - 1660-8151 (Linking)
VI  - 92
IP  - 4
DP  - 2002 Dec
TI  - Detection of mRNA for alpha-3 chain of type IV collagen in the glomerular 
      epithelium, and the effect of perfused elastase on its expression.
PG  - 853-9
AB  - BACKGROUND: The type IV collagen is a complex of trimetric molecule composed of 
      six genetically distinct polypeptide chains; alpha1-6(IV). Since alpha3(IV) 
      distribute specifically in the glomerular basement membrane (GBM) of glomerular 
      capillary, we tried to develop the detection methods for the transcripts of 
      alpha3(IV) in glomerular epithelial cells (GEC) which produce most of the 
      components for GBM. Then, using these molecular techniques, the influence of 
      elastase, one of the proteases released from activated polymorphonuclear 
      leukocytes at the site of inflammation, on GEC was determined as manifested by 
      expressional alteration of alpha3(IV) mRNA. METHODS: DIG-labeled oligo-DNA probe 
      designating non-collagenous region of alpha3(IV) was used for in situ 
      hybridization. Semiquantitative measurement of alpha3(IV) in the renal cortex was 
      performed by PCR reactions, each reaction being normalized by that for GAPDH. 
      Then, the femoral artery of each of 18 Sprague-Dawley rats was catheterized and 
      the left kidney was perfused with saline alone (0.5 ml) or saline containing 100 
      microg/ml elastase. After collection of urine for 24 h, the left kidney was 
      harvested for analysis of mRNA (4 for in situ hybridization and 5 kidneys for PCR 
      analysis). RESULTS: Antisense cDNA probe and PCR reaction well identified 
      alpha3(IV) mRNA in the cytoplasm of GEC and in the renal cortex, respectively. 
      Urinary protein excreted by rats with elastase perfusion was 47.2 +/- 3.8 mg/24 h 
      but this was only 13.9 +/- 1.1 mg/24 h in control rats (mean +/- SEM, p < 0.05). 
      In situ hybridization demonstrated that expression of alpha3(IV) mRNA in GEC was 
      focally or diffusely reduced in the glomeruli of rats with elastase perfusion, 
      whereas the transcripts were well stained in GEC of controls. PCR analysis showed 
      about 25% decrease in transcripts of alpha3(IV) in the renal cortex of rats with 
      elastase perfusion compared to those of control rats. CONCLUSIONS: alpha3(IV) 
      mRNA was identified specifically in the GEC in the glomeruli. Co-incidence of 
      proteinuria and reduced alpha3(IV) expression by elastase suggests adverse 
      effects of elastase on GEC and close association between proteinuria and GEC 
      injury.
CI  - Copyright 2002 S. Karger AG, Basel
FAU - Nogae, Shoji
AU  - Nogae S
AD  - Department of Nephrology, Endocrinology and Vascular Medicine, Tohoku University 
      School of Medicine, Sendai, Japan.
FAU - Michimata, Mari
AU  - Michimata M
FAU - Araki, Tsutomu
AU  - Araki T
FAU - Suzuki, Michiko
AU  - Suzuki M
FAU - Kazama, Itsuro
AU  - Kazama I
FAU - Ito, Sadayoshi
AU  - Ito S
FAU - Imai, Yutaka
AU  - Imai Y
FAU - Matsubara, Mitsunobu
AU  - Matsubara M
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Switzerland
TA  - Nephron
JT  - Nephron
JID - 0331777
RN  - 0 (Autoantigens)
RN  - 0 (Collagen Type IV)
RN  - 0 (Oligonucleotides, Antisense)
RN  - 0 (RNA, Messenger)
RN  - 0 (Serum Albumin)
RN  - 0 (type IV collagen alpha3 chain)
RN  - EC 3.4.21.36 (Pancreatic Elastase)
SB  - IM
MH  - Animals
MH  - Autoantigens/*genetics/*metabolism
MH  - Collagen Type IV/*genetics/*metabolism
MH  - Epithelial Cells/cytology/metabolism
MH  - In Situ Hybridization
MH  - Kidney Glomerulus/cytology/*drug effects/*metabolism
MH  - Oligonucleotides, Antisense/metabolism
MH  - Pancreatic Elastase/*pharmacology
MH  - RNA, Messenger/genetics/*metabolism
MH  - Rats
MH  - Rats, Sprague-Dawley
MH  - Serum Albumin/analysis
EDAT- 2002/10/26 04:00
MHDA- 2003/07/18 05:00
CRDT- 2002/10/26 04:00
PHST- 2002/10/26 04:00 [pubmed]
PHST- 2003/07/18 05:00 [medline]
PHST- 2002/10/26 04:00 [entrez]
AID - 65440 [pii]
AID - 10.1159/000065440 [doi]
PST - ppublish
SO  - Nephron. 2002 Dec;92(4):853-9. doi: 10.1159/000065440.