Selective immunoreactivities of kidney basement membranes to monoclonal antibodies against laminin: Localization of the end of the long arm and the short arms to discrete microdomains

Research output: Contribution to journalJournal articleResearchpeer-review

Standard

Selective immunoreactivities of kidney basement membranes to monoclonal antibodies against laminin : Localization of the end of the long arm and the short arms to discrete microdomains. / Abrahamson, D. R.; Irwin, M. H.; St. John, P. L.; Perry, E. W.; Accavitti, M. A.; Heck, L. W.; Couchman, J. R.

In: Journal of Cell Biology, Vol. 109, No. 6 II, 01.12.1989, p. 3477-3491.

Research output: Contribution to journalJournal articleResearchpeer-review

Harvard

Abrahamson, DR, Irwin, MH, St. John, PL, Perry, EW, Accavitti, MA, Heck, LW & Couchman, JR 1989, 'Selective immunoreactivities of kidney basement membranes to monoclonal antibodies against laminin: Localization of the end of the long arm and the short arms to discrete microdomains', Journal of Cell Biology, vol. 109, no. 6 II, pp. 3477-3491. https://doi.org/10.1083/jcb.109.6.3477

APA

Abrahamson, D. R., Irwin, M. H., St. John, P. L., Perry, E. W., Accavitti, M. A., Heck, L. W., & Couchman, J. R. (1989). Selective immunoreactivities of kidney basement membranes to monoclonal antibodies against laminin: Localization of the end of the long arm and the short arms to discrete microdomains. Journal of Cell Biology, 109(6 II), 3477-3491. https://doi.org/10.1083/jcb.109.6.3477

Vancouver

Abrahamson DR, Irwin MH, St. John PL, Perry EW, Accavitti MA, Heck LW et al. Selective immunoreactivities of kidney basement membranes to monoclonal antibodies against laminin: Localization of the end of the long arm and the short arms to discrete microdomains. Journal of Cell Biology. 1989 Dec 1;109(6 II):3477-3491. https://doi.org/10.1083/jcb.109.6.3477

Author

Abrahamson, D. R. ; Irwin, M. H. ; St. John, P. L. ; Perry, E. W. ; Accavitti, M. A. ; Heck, L. W. ; Couchman, J. R. / Selective immunoreactivities of kidney basement membranes to monoclonal antibodies against laminin : Localization of the end of the long arm and the short arms to discrete microdomains. In: Journal of Cell Biology. 1989 ; Vol. 109, No. 6 II. pp. 3477-3491.

Bibtex

@article{96d747a5a95542a7a3654ddd0feba9e4,
title = "Selective immunoreactivities of kidney basement membranes to monoclonal antibodies against laminin: Localization of the end of the long arm and the short arms to discrete microdomains",
abstract = "To examine the ultrastructural distribution of laminin within kidney basement membranes, we prepared rat anti-mouse laminin mAbs to use in immunolocalization experiments. Epitope domains for these mAbs were established by immunoprecipitation, immunoblotting, affinity chromatography, and rotary shadow EM. One mAb bound to the laminin A and B chains on blots and was located to a site ~15 nm from the long arm-terminal globular domain as shown by rotary shadowing. Conjugates of this long arm-specific mAb were coupled to horseradish peroxidase (HRP) and intravenously injected into mice. Kidney cortices were fixed for microscopy 3 h after injection. HRP reaction product was localized irregularly within the renal glomerular basement membrane (GBM) and throughout mesangial matrices. In addition, this mAb bound in linear patterns specifically to the laminae rarae of basement membranes of Bowman's capsule and proximal tubule. This indicates the presence of the long arm immediately beneath epithelial cells in these sites. The laminae densae of these basement membranes were negative by this protocol. In contrast, the lamina rara and densa of distal tubular basement membranes (TBM) were both heavily labeled with this mAb. A different ultrastructural binding patern was seen with eight other mAbs, including two that mapped to different sites on the short arms by rotary shadowing and five that blotted to a large pepsin-resistant laminin fragment (P1). These latter mAbs bound weakly or not at all to GBM but all bound throughout mesangial matrices. In contrast, discrete spots of HRP reaction product were seen across all layers of Bowman's capsule BM and proximal TBM. These same mAbs, however, bound densely across the full width of distal TBM. Our findings therefore show that separate strata of different basement membranes are variably immunoreactive to these laminin mAbs. The molecular orientation or integration of laminin into the three dimensional BM meshwork therefore varies with location. Alternatively, there may be a family of distinct laminin-like molecules distributed within basement membranes.",
author = "Abrahamson, {D. R.} and Irwin, {M. H.} and {St. John}, {P. L.} and Perry, {E. W.} and Accavitti, {M. A.} and Heck, {L. W.} and Couchman, {J. R.}",
year = "1989",
month = "12",
day = "1",
doi = "10.1083/jcb.109.6.3477",
language = "English",
volume = "109",
pages = "3477--3491",
journal = "Journal of Cell Biology",
issn = "0021-9525",
publisher = "Rockefeller University Press",
number = "6 II",

}

RIS

TY - JOUR

T1 - Selective immunoreactivities of kidney basement membranes to monoclonal antibodies against laminin

T2 - Localization of the end of the long arm and the short arms to discrete microdomains

AU - Abrahamson, D. R.

AU - Irwin, M. H.

AU - St. John, P. L.

AU - Perry, E. W.

AU - Accavitti, M. A.

AU - Heck, L. W.

AU - Couchman, J. R.

PY - 1989/12/1

Y1 - 1989/12/1

N2 - To examine the ultrastructural distribution of laminin within kidney basement membranes, we prepared rat anti-mouse laminin mAbs to use in immunolocalization experiments. Epitope domains for these mAbs were established by immunoprecipitation, immunoblotting, affinity chromatography, and rotary shadow EM. One mAb bound to the laminin A and B chains on blots and was located to a site ~15 nm from the long arm-terminal globular domain as shown by rotary shadowing. Conjugates of this long arm-specific mAb were coupled to horseradish peroxidase (HRP) and intravenously injected into mice. Kidney cortices were fixed for microscopy 3 h after injection. HRP reaction product was localized irregularly within the renal glomerular basement membrane (GBM) and throughout mesangial matrices. In addition, this mAb bound in linear patterns specifically to the laminae rarae of basement membranes of Bowman's capsule and proximal tubule. This indicates the presence of the long arm immediately beneath epithelial cells in these sites. The laminae densae of these basement membranes were negative by this protocol. In contrast, the lamina rara and densa of distal tubular basement membranes (TBM) were both heavily labeled with this mAb. A different ultrastructural binding patern was seen with eight other mAbs, including two that mapped to different sites on the short arms by rotary shadowing and five that blotted to a large pepsin-resistant laminin fragment (P1). These latter mAbs bound weakly or not at all to GBM but all bound throughout mesangial matrices. In contrast, discrete spots of HRP reaction product were seen across all layers of Bowman's capsule BM and proximal TBM. These same mAbs, however, bound densely across the full width of distal TBM. Our findings therefore show that separate strata of different basement membranes are variably immunoreactive to these laminin mAbs. The molecular orientation or integration of laminin into the three dimensional BM meshwork therefore varies with location. Alternatively, there may be a family of distinct laminin-like molecules distributed within basement membranes.

AB - To examine the ultrastructural distribution of laminin within kidney basement membranes, we prepared rat anti-mouse laminin mAbs to use in immunolocalization experiments. Epitope domains for these mAbs were established by immunoprecipitation, immunoblotting, affinity chromatography, and rotary shadow EM. One mAb bound to the laminin A and B chains on blots and was located to a site ~15 nm from the long arm-terminal globular domain as shown by rotary shadowing. Conjugates of this long arm-specific mAb were coupled to horseradish peroxidase (HRP) and intravenously injected into mice. Kidney cortices were fixed for microscopy 3 h after injection. HRP reaction product was localized irregularly within the renal glomerular basement membrane (GBM) and throughout mesangial matrices. In addition, this mAb bound in linear patterns specifically to the laminae rarae of basement membranes of Bowman's capsule and proximal tubule. This indicates the presence of the long arm immediately beneath epithelial cells in these sites. The laminae densae of these basement membranes were negative by this protocol. In contrast, the lamina rara and densa of distal tubular basement membranes (TBM) were both heavily labeled with this mAb. A different ultrastructural binding patern was seen with eight other mAbs, including two that mapped to different sites on the short arms by rotary shadowing and five that blotted to a large pepsin-resistant laminin fragment (P1). These latter mAbs bound weakly or not at all to GBM but all bound throughout mesangial matrices. In contrast, discrete spots of HRP reaction product were seen across all layers of Bowman's capsule BM and proximal TBM. These same mAbs, however, bound densely across the full width of distal TBM. Our findings therefore show that separate strata of different basement membranes are variably immunoreactive to these laminin mAbs. The molecular orientation or integration of laminin into the three dimensional BM meshwork therefore varies with location. Alternatively, there may be a family of distinct laminin-like molecules distributed within basement membranes.

UR - http://www.scopus.com/inward/record.url?scp=0024788014&partnerID=8YFLogxK

U2 - 10.1083/jcb.109.6.3477

DO - 10.1083/jcb.109.6.3477

M3 - Journal article

C2 - 2480964

AN - SCOPUS:0024788014

VL - 109

SP - 3477

EP - 3491

JO - Journal of Cell Biology

JF - Journal of Cell Biology

SN - 0021-9525

IS - 6 II

ER -

ID: 225719801