Glycosphingolipid biosynthesis in adult mouse kidney cultures
Primary kidney cultures from C57BL/6J mice, six weeks of age or older, were produced using D-valine medium to select for epithelial cell growth. After allowing cells to attach and proliferate for one week following plating, medium was changed once per week. Cells formed nearly confluent monolayers during the second week of culture. The cultured cells contained all of the glycosphingolipids seen in the adult kidney, as analyzed by HPLC as the perbenzoyl derivatives. Glucosylceramide, however, was highly predominant in the cultured cells, whereas dihexosyl- and trihexosylceramides predominate in the intact kidney. Sex differences in glycolipid contents found in the intact kidney were also apparent in these cultured cells: greater amounts of nonhydroxy fatty acid (Nfa) containing galactosylceramide and Nfa digalactosylceramide were present in the male cells. Neutral glycosphingolipids were labeled after two weeks in culture using /sup 3/H-labeled palmitate. The /sup 3/H-palmitate was incorporated into all of the glycolipids within two hours of labeling, and after three days of chase with unlabeled palmitate, some redistribution into higher glycosphingolipids was apparent. Hence, adult mouse kidney cells in D-valine medium retain their differentiated characteristics for a sufficient period of time to allow investigation of glycolipid syntheses in monolayer cultures of epithelial cells derived from this organ.
- Research Organization:
- Clark Univ., Worcester, MA
- OSTI ID:
- 7246765
- Report Number(s):
- CONF-8606151-; TRN: 86-039044
- Journal Information:
- Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Vol. 45:6; Conference: 76. annual meeting of the Federation of American Society for Experimental Biology, Washington, DC, USA, 8 Jun 1986
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
EPITHELIUM
CELL PROLIFERATION
GLYCOLIPIDS
BIOSYNTHESIS
LABELLING
AGE DEPENDENCE
CELL CULTURES
HEXADECANOIC ACID
KIDNEYS
LIQUID COLUMN CHROMATOGRAPHY
MICE
SEX DEPENDENCE
TRACER TECHNIQUES
TRITIUM COMPOUNDS
ANIMAL TISSUES
ANIMALS
BODY
CARBOHYDRATES
CARBOXYLIC ACIDS
CHROMATOGRAPHY
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
LIPIDS
MAMMALS
MONOCARBOXYLIC ACIDS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANS
RODENTS
SACCHARIDES
SEPARATION PROCESSES
SYNTHESIS
TISSUES
VERTEBRATES
550201* - Biochemistry- Tracer Techniques