Publication: High resolution SEM analysis of acellular glomerular basement membrane following pepsin digestion, intrinsic fibrillar structures
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Date
1990
Authors
Berger, Walter J. ; Carlson, E. C.
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Publisher
Murcia : F. Hernández
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DOI
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info:eu-repo/semantics/article
Description
Abstract
Microdissection of acellular rat renal cortex
with pepsin was carried out to investigate the morphological
substructure of glomerular basement membrane
(GBM) by high resolution SEM. Renal cortical blocks
(< 5 mm ) from adult male Sprague Dawley rats were
rendered acellular by sequential detergent extraction
and digested up to 184 hrs with 5 mglml pepsin (185 U/
mg) in 0.5 M acetic acid (pH 2) at 10-15°C. Samples were
conventionally prepared for SEM, and observed at original
magnifications of 500-100,000 diameters.
At low magnifications (500-5,00Ox), acellular GBM
surfaces appeared smooth at al1 digestion times. At
higher magnifications (50,000-100,00Ox), control GBM
surfaces were finely granular. Granule diameter ranged
from 20-80 nm, with most between 30-40 nm. Pepsin digestion
did not affect average granule size.
Beginning at 44 hrs of digestion, intrinsic fibrillar
structures comprised of linear arrays of 20-40 nm granules
were observed onlin GBM surfaces. At later incubation
times, this component of GBM became more extensive.
At 160 hrs, the fibrillar arrays frequently bifurcated
and showed distinctive «forked» termini, some of which
comprised two sides of a triangle (120-150 nm on a side).
Fork «handles» (310-350 nm in length) radiated from
each angle of the triangle. These sometimes terminated
in large granules (approximately 100 nm in diameter),
two of which appeared to connect fibrillar arrays end-toend.
Together with other arrays, the interconnected
triangles appeared to comprise a three-dimensional
meshwork extending into the GBM and possibly providing
support for, its granular components.
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