Abstract
We used immunogold electron microscopic (IEM) techniques with periodate- lysine-paraformaldehyde-fixed and Lowicryl-embedded or cryopreserved tissues to study the distribution of α1(IV) and α3(IV) chains of Types IV and VI collagen in glomerular basement membrane (GBM) and mesangial matrix of glomeruli in normal human kidneys. Monoclonal antibodies to α1(IV) and α3(IV) collagen chains and Type VI collagen could be detected only with cryoultramicrotomy, whereas polyclonal anti-Type IV collagen antibody was detectable in Lowicryl-embedded tissue. Ultrastructural detail was better preserved in the Lowicryl-embedded tissue. IEM labeling provided more detailed information as to the site-specific array of these extracellular matrix molecules in glomeruli than did immunofluorescent microscopy. The labeling of α1(IV) collagen chain was distributed mainly along the endothelial side of glomerular basement membrane and the mesangial matrix. Mesangial GBM was relatively poorly labeled compared with that of mesangial matrix. In contrast, the α3(IV) chain was detected throughout the thickness of the GBM, but there was no labeling of mesangial matrix. Type IV collagen distribution was identical to that of the α1(IV) chain within the glomerulus but was also associated with interstitial collagen fibrils. This study documents and details the heterogeneous distribution of Type IV and VI collagen chains within the normal human glomerulus and provides the framework for the study of these matrix components in human glomerular diseases.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 577-584 |
| Number of pages | 8 |
| Journal | Journal of Histochemistry and Cytochemistry |
| Volume | 42 |
| Issue number | 5 |
| DOIs | |
| State | Published - Jan 1 1994 |
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Keywords
- Immunogold electron microscopy
- Kidney
- Type IV collagen
- Type VI collagen
Cite this
Application of electron microscopic immunocytochemistry to the human kidney : Distribution of type IV and type VI collagen in normal human kidney. / Zhu, D.; Kim, Youngki; Steffes, Michael W; Groppoli, T. J.; Butkowski, R. J.; Mauer, Michael.
In: Journal of Histochemistry and Cytochemistry, Vol. 42, No. 5, 01.01.1994, p. 577-584.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Application of electron microscopic immunocytochemistry to the human kidney
T2 - Distribution of type IV and type VI collagen in normal human kidney
AU - Zhu, D.
AU - Kim, Youngki
AU - Steffes, Michael W
AU - Groppoli, T. J.
AU - Butkowski, R. J.
AU - Mauer, Michael
PY - 1994/1/1
Y1 - 1994/1/1
N2 - We used immunogold electron microscopic (IEM) techniques with periodate- lysine-paraformaldehyde-fixed and Lowicryl-embedded or cryopreserved tissues to study the distribution of α1(IV) and α3(IV) chains of Types IV and VI collagen in glomerular basement membrane (GBM) and mesangial matrix of glomeruli in normal human kidneys. Monoclonal antibodies to α1(IV) and α3(IV) collagen chains and Type VI collagen could be detected only with cryoultramicrotomy, whereas polyclonal anti-Type IV collagen antibody was detectable in Lowicryl-embedded tissue. Ultrastructural detail was better preserved in the Lowicryl-embedded tissue. IEM labeling provided more detailed information as to the site-specific array of these extracellular matrix molecules in glomeruli than did immunofluorescent microscopy. The labeling of α1(IV) collagen chain was distributed mainly along the endothelial side of glomerular basement membrane and the mesangial matrix. Mesangial GBM was relatively poorly labeled compared with that of mesangial matrix. In contrast, the α3(IV) chain was detected throughout the thickness of the GBM, but there was no labeling of mesangial matrix. Type IV collagen distribution was identical to that of the α1(IV) chain within the glomerulus but was also associated with interstitial collagen fibrils. This study documents and details the heterogeneous distribution of Type IV and VI collagen chains within the normal human glomerulus and provides the framework for the study of these matrix components in human glomerular diseases.
AB - We used immunogold electron microscopic (IEM) techniques with periodate- lysine-paraformaldehyde-fixed and Lowicryl-embedded or cryopreserved tissues to study the distribution of α1(IV) and α3(IV) chains of Types IV and VI collagen in glomerular basement membrane (GBM) and mesangial matrix of glomeruli in normal human kidneys. Monoclonal antibodies to α1(IV) and α3(IV) collagen chains and Type VI collagen could be detected only with cryoultramicrotomy, whereas polyclonal anti-Type IV collagen antibody was detectable in Lowicryl-embedded tissue. Ultrastructural detail was better preserved in the Lowicryl-embedded tissue. IEM labeling provided more detailed information as to the site-specific array of these extracellular matrix molecules in glomeruli than did immunofluorescent microscopy. The labeling of α1(IV) collagen chain was distributed mainly along the endothelial side of glomerular basement membrane and the mesangial matrix. Mesangial GBM was relatively poorly labeled compared with that of mesangial matrix. In contrast, the α3(IV) chain was detected throughout the thickness of the GBM, but there was no labeling of mesangial matrix. Type IV collagen distribution was identical to that of the α1(IV) chain within the glomerulus but was also associated with interstitial collagen fibrils. This study documents and details the heterogeneous distribution of Type IV and VI collagen chains within the normal human glomerulus and provides the framework for the study of these matrix components in human glomerular diseases.
KW - Immunogold electron microscopy
KW - Kidney
KW - Type IV collagen
KW - Type VI collagen
UR - http://www.scopus.com/inward/record.url?scp=0028294097&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0028294097&partnerID=8YFLogxK
U2 - 10.1177/42.5.8157929
DO - 10.1177/42.5.8157929
M3 - Article
VL - 42
SP - 577
EP - 584
JO - Journal of Histochemistry and Cytochemistry
JF - Journal of Histochemistry and Cytochemistry
SN - 0022-1554
IS - 5
ER -