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.
The extracellular matrix (ECM) is an important regulator of the differentiated phenotype of mammary epithelial cells in culture. Despite the fact that ECM-degrading enzymes have been implicated in morphogenesis and tissue remodeling, there is little evidence for a direct role for such regulation in vivo. We generated transgenic mice that express autoactivated isoforms of the matrix metalloproteinase stromelysin-1, under the control of the whey acidic protein gene promoter, to examine the effect of inappropriate expression of this enzyme. Stromelysin-1 is implicated as the primary player in the loss of basement membrane and loss of function in the mammary gland during involution. The transgene was expressed at low levels in mammary glands of virgin female mice, leading to an unexpected phenotype: The primary ducts had supernumerary branches and showed precocious development of alveoli that expressed beta-casein at levels similar to that of an early- to mid-pregnant gland. Lactating glands showed high levels of transgene expression, with accumulation at the basement membrane, and a decrease in laminin and collagen IV, resulting in a loss of basement membrane integrity; this was accompanied by a dramatic alteration of alveolar morphology, with decreased size and shrunken lumina containing little beta-casein. During pregnancy, expression of endogenous whey acidic protein and beta-casein was reduced in transgenic glands, confirming the observed dependence of milk protein transcription of ECM in mammary epithelial cells in culture. These data provide direct evidence that stromelysin-1 activity can be morphogenic for mammary epithelial cells, inducing hyperproliferation and differentiation in virgin animals, and that its lytic activity can, indeed, disrupt membrane integrity and reduce mammary-specific function. We conclude that the balance of ECM-degrading enzymes with their inhibitors, and the associated regulation of ECM structure, is crucial for tissue-specific gene expression and morphogenesis in vivo.
Sympson
C J
CJ
Life Sciences Division, Lawrence Berkeley Laboratory, Berkeley, California 94720.
Talhouk
R S
RS
Alexander
C M
CM
Chin
J R
JR
Clift
S M
SM
Bissell
M J
MJ
Werb
Z
Z
D000818:Animals; D001483:Base Sequence; D001485:Basement Membrane / Q000648:ultrastructure; D002364:Caseins / Q000235:genetics; D002454:Cell Differentiation; D003094:Collagen / Q000235:genetics; D017931:DNA Primers / Q000737:chemistry; D005260:Female; D005786:Gene Expression Regulation; D005796:Genes; D007774:Lactation; D007797:Laminin / Q000235:genetics; D008321:Mammary Glands, Animal / Q000166:cytology* / Q000201:enzymology*; D019278:Matrix Metalloproteinase 3; D008666:Metalloendopeptidases / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic; D008894:Milk Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D009024:Morphogenesis; D011247:Pregnancy; D011401:Promoter Regions, Genetic; D015183:Restriction Mapping; D000067816:Whey Proteins
D002364:Caseins; D017931:DNA Primers; D007797:Laminin; D008894:Milk Proteins; D000067816:Whey Proteins; D003094:Collagen; D008666:Metalloendopeptidases; D019278:Matrix Metalloproteinase 3