title: Deposition of collagen VI in the extracellular matrix during mouse embryogenesis correlates with expression of the alpha 3(VI) subunit gene.
abstract: Collagen VI is a microfibrillar component of the extracellular matrix that is predicted to have an important structural role in matrix organization and a biological function in mediating cell-matrix interactions. Secreted collagen VI molecules are composed of three distinct subunits, the alpha 1(VI), alpha 2(VI), and alpha 3(VI) chains. To determine when, and in which tissues, collagen VI is likely to have a role in embryonic processes, we have analyzed the expression patterns of the three subunit chains during postimplantation mouse development by reverse transcriptase-PCR (RT-PCR), in situ hybridization, and immunofluorescence. No collagen VI protein could be detected in the mouse embryo until Day 11.5 of gestation, when low levels were localized within the mesoderm layer of the visceral yolk sac, the subepidermal matrix of branchial arches, and the vessel wall of the dorsal aorta. Levels of collagen VI mRNA and protein increased during the period from Days 12.5 to 14.5 in the visceral yolk sac, subepidermal mesenchyme, lung, gut, meninges, muscle, perichondrium, and vertebral column. The cartilage matrix of ribs and developing long bones was not stained with collagen VI antisera, but pericellular staining of chondrocytes was seen in both tissues. Low levels of collagen VI mRNA and protein were seen in the fetal liver except for the connective tissue of the liver capsule, which was highly stained. Collagen VI was first detected at significant levels in the developing heart on Day 14.5. These data demonstrate a tissue-specific onset of collagen VI synthesis and deposition in the extracellular matrix of developing mouse embryos at a much later stage of development than that reported for fibronectin or collagen I. Sensitive RT-PCR assays showed that alpha 1(VI) and alpha 2(VI) mRNAs were amplified from extracts of embryonic tissues as early as Day 7.5, while alpha 3(VI) mRNA was not detected until Day 10.5. Expression of the alpha 3(VI) gene immediately preceded the appearance of collagen VI protein in embryonic tissues. This correlation is consistent with the proposal that expression of alpha 3(VI) chains regulates the formation and secretion of collagen VI trimers and collagen VI matrix deposition during development.
authors.affiliation: Institute of Reproduction and Development, Monash Medical Centre, Clayton, Victoria, Australia.
authors.forename: M
authors.initials: M
authors.lastname: Dziadek
authors.forename: P
authors.initials: P
authors.lastname: Darling
authors.forename: M
authors.initials: M
authors.lastname: Bakker
authors.forename: M
authors.initials: M
authors.lastname: Overall
authors.forename: R Z
authors.initials: RZ
authors.lastname: Zhang
authors.forename: T C
authors.initials: TC
authors.lastname: Pan
authors.forename: E
authors.initials: E
authors.lastname: Tillet
authors.forename: R
authors.initials: R
authors.lastname: Timpl
authors.forename: M L
authors.initials: ML
authors.lastname: Chu
mesh_terms: D000818:Animals; D001483:Base Sequence; D003094:Collagen / Q000032:analysis* / Q000096:biosynthesis / Q000235:genetics*; D004622:Embryo, Mammalian / Q000737:chemistry; D005314:Embryonic and Fetal Development; D005109:Extracellular Matrix / Q000737:chemistry*; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D051379:Mice; D008969:Molecular Sequence Data; D009928:Organ Specificity; D012333:RNA, Messenger / Q000032:analysis
chemical_list: D012333:RNA, Messenger; D003094:Collagen