In vivo turnover of the basement membrane and other heparan sulfate proteoglycans of rat glomerulus

Journal Article · Wed Mar 01 04:00:00 EST 1989 · Arch. Biochem. Biophys.; (United States)

DOI:https://doi.org/10.1016/0003-9861(89)90143-4· OSTI ID:6279438

Beavan, L A; Davies, M; Couchman, J R; Williams, M A; Mason, R M

The metabolic turnover of rat glomerular proteoglycans in vivo was investigated. Newly synthesized proteoglycans were labeled during a 7-h period after injecting sodium (35S)sulfate intraperitoneally. At the end of the labeling period a chase dose of sodium sulfate was given. Subsequently at defined times (0-163 h) the kidneys were perfused in situ with 0.01% cetylpyridinium chloride in phosphate-buffered saline to maximize the recovery of 35S-proteoglycans. Glomeruli were isolated from the renal cortex and analyzed for 35S-proteoglycans by autoradiographic, biochemical, and immunochemical methods. Grain counting of autoradiographs revealed a complex turnover pattern of 35S-labeled macromolecules, commencing with a rapid phase followed by a slower phase. Biochemical analysis confirmed the biphasic pattern and showed that the total population of (35S)heparan sulfate proteoglycans had a metabolic half-life (t1/2) of 20 and 60 h in the early and late phases, respectively. Heparan sulfate proteoglycans accounted for 80% of total 35S-proteoglycans, the remainder being chondroitin/dermatan sulfate proteoglycans. Whole glomeruli were extracted with 4% 3-((cholamidopropyl)dimethy-lammonio)-1-propanesulfonate-4 M guanidine hydrochloride, a procedure which solubilized greater than 95% of the 35S-labeled macromolecules. Of these 11-13% was immunoprecipitated by an antiserum against heparan sulfate proteoglycan which, in immunolocalization experiments, showed specificity for staining the basement membrane of rat glomeruli. Autoradiographic analysis showed that 18% of total radioactivity present at the end of the labeling period was associated with the glomerular basement membrane.

• Research Organization: Charing Cross and Westminster Medical School, London (England)
• OSTI ID: 6279438
• Journal Information: Arch. Biochem. Biophys.; (United States), Journal Name: Arch. Biochem. Biophys.; (United States) Vol. 269:2; ISSN ABBIA
• Country of Publication: United States
• Language: English

Related Subjects

550501* -- Metabolism-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINES
ANIMALS
AUTORADIOGRAPHY
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOLOGICAL HALF-LIFE
BODY
CARBOHYDRATES
CELL CONSTITUENTS
CELL MEMBRANES
DAYS LIVING RADIOISOTOPES
EVEN-ODD NUCLEI
GLYCOPROTEINS
IN VIVO
INJECTION
INTAKE
INTRAPERITONEAL INJECTION
ISOTOPES
KIDNEYS
LIGHT NUCLEI
MAMMALS
MEMBRANES
METABOLISM
MUCOPOLYSACCHARIDES
NUCLEI
ORGANIC COMPOUNDS
ORGANS
OXYGEN COMPOUNDS
PERFUSED ORGANS
POLYSACCHARIDES
PROTEINS
RADIOISOTOPES
RATS
RODENTS
SACCHARIDES
SULFATES
SULFUR 35
SULFUR COMPOUNDS
SULFUR ISOTOPES
VERTEBRATES