Influence of a reconstituted basement membrane and its components on casein gene expression and secretion in mouse mammary epithelial cells
When primary mouse mammary epithelial cells are cultured on plastic, they rapidly lose their ability to synthesize and secrete most milk proteins even in the presence of lactogenic hormones, whereas cells cultured on release type I collagen gels show greatly enhanced mRNA levels and secretion rates of ..beta..-casein and of some other milk proteins. The authors show here that culture on a reconstituted basement membrane from Engelbreth-Holm-Swarm tumor (EHS) allows > 90% of cells to produce high levels of ..beta..-casein. By comparison, 30-40% of cells on released type 1 gels and only 2-10% of cells on plastic express ..beta..-casein after 6 days in culture. Because only 40% of cells from late pregnant gland produced ..beta..-casein before culture, the EHS matrix can both induce and maintain an increased level of casein gene expression. Individual basal lamina components were also evaluated. Type IV collagen and fibronectin had little effect on morphology and ..beta..-casein mRNA levels. In contrast, both laminin and heparan sulfate proteoglycan increased ..beta..-casein mRNA levels. Profound morphological differences were evident between cells cultured on plastic and on EHS matrix, the latter cells forming ducts, ductules, and lumina and resembling secretory alveoli. These results emphasize the vital role of the extracellular matrix in receiving and integrating structural and functional signals that can direct specific gene expression in differentiated tissues.
- Research Organization:
- Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
- DOE Contract Number:
- AC03-76SF00098
- OSTI ID:
- 5653199
- Journal Information:
- Proc. Natl. Acad. Sci. U.S.A.; (United States), Vol. 84:1
- Country of Publication:
- United States
- Language:
- English
Similar Records
Functional differentiation and alveolar morphogenesis of primary mammary cultures on reconstituted basement membrane
Targeted Expression of Stromelysin-1 in Mammary Gland Provides Evidence for a Role of Proteinases in Branching Morphogenesis and the Requirement for an Intact Basement Membrane for Tissue-specific Gene Expression
Related Subjects
CASEIN
SECRETION
EPITHELIUM
CYTOLOGICAL TECHNIQUES
TRANSMISSION ELECTRON MICROSCOPY
COLLAGEN
ELECTROPHORESIS
IN VITRO
MAMMARY GLANDS
MESSENGER-RNA
METHIONINE
SULFUR 35
AMINO ACIDS
ANIMAL TISSUES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BODY
CARBOXYLIC ACIDS
DAYS LIVING RADIOISOTOPES
DRUGS
ELECTRON MICROSCOPY
EVEN-ODD NUCLEI
GLANDS
ISOTOPES
LIGHT NUCLEI
LIPOTROPIC FACTORS
MICROSCOPY
NUCLEI
NUCLEIC ACIDS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC PHOSPHORUS COMPOUNDS
ORGANIC SULFUR COMPOUNDS
ORGANS
PROTEINS
RADIOISOTOPES
RNA
SCLEROPROTEINS
SULFUR ISOTOPES
TISSUES
550301* - Cytology- Tracer Techniques