Laminin chain expression by chick chondrocytes and mouse cartilaginous tissues in vivo and in vitro.
We have observed that laminins are expressed in the chondrocytes of chick embryo sternum, mouse limb bud, and adult mouse knee joint by the methods of in situ hybridization, immunohistochemistry, Western blotting, and immunoprecipitation. From in situ hybridization using similar sized RNA probes for different mouse laminin chains, mRNAs for the alpha 1, alpha 2, beta 1, beta 2, and gamma 1 chains were expressed in the chondrocytes of chick embryo sternum, mouse limb bud, and the articular cartilage cap and epiphyseal growth plate of adult mouse knee joint. Through the use of chain-specific antibodies, staining for laminins was observed in the cytoplasm of chondrocytes from chick embryo sternum, mouse limb bud, and adult mouse knee joint. Western blot analysis confirmed the presence of laminin chains in the cells and sternal tissues. Cultured chick embryonic sternal chondrocytes expressed laminin mRNAs in the proliferating stage (2-3 days of culture) but the level increased in the aggregated cells during the maturation stage (5-7 days of culture). Comparable data were also obtained after immunostaining the cells. Thus, laminins are expressed in significant amounts by chondrocytes and may have an important role in cartilage development.
Lee
S K
SK
Craniofacial Developmental Biology and Regeneration Branch, National Institute of Dental Research, National Institutes of Health, Bethesda, Maryland 20892-4370, USA.
Malpeli
M
M
Cancedda
R
R
Utani
A
A
Yamada
Y
Y
Kleinman
H K
HK
D000818:Animals; D015153:Blotting, Western; D002356:Cartilage / Q000166:cytology* / Q000196:embryology*; D002478:Cells, Cultured; D002642:Chick Embryo; D019902:Chondrocytes / Q000737:chemistry* / Q000166:cytology; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D007719:Knee Joint / Q000737:chemistry / Q000166:cytology / Q000196:embryology; D007797:Laminin / Q000032:analysis* / Q000235:genetics*; D018878:Limb Buds / Q000737:chemistry / Q000166:cytology / Q000196:embryology; D051379:Mice; D008815:Mice, Inbred Strains; D011233:Precipitin Tests; D012333:RNA, Messenger / Q000032:analysis; D013249:Sternum / Q000737:chemistry / Q000166:cytology / Q000196:embryology
D007797:Laminin; D012333:RNA, Messenger