Embryonic skin fibroblasts release TGF alpha and TGF beta able to influence synthesis and secretion of GAG.
Conditioned medium (CM), collected from 7 and 14 days-old chick embryo skin fibroblasts and added to the same cells, increases glycosaminoglycans (GAG) intra- and extracellular accumulation. The factors responsible for GAG enhancement are TGF alpha and TGF beta because they are trypsin and dithiothreitol sensitive, stable or enhanced by heat and transient acidification. Moreover, Sephadex G-75 fractions of CM active on GAG synthesis contain, when analysed on SDS-polyacrylamide gel electrophoresis, two bands that comigrate with TGF alpha and TGF beta and induce NRK cells clone 49F to form large colonies of mean size > 8.000 microns 2 in soft agar. Since both the factors must be present to induce the formation of large colonies we come to the conclusion that CM contains TGF alpha and TGF beta. The two growth factors have different effects on the accumulation of individual classes of GAG in the ECM. In particular, TGF beta stimulates a marked increase of CS and DS, TGF alpha of HA and DS in the medium. The contemporaneous addition of TGF alpha and TGF beta to 7 days-old fibroblasts produces a pattern of GAG response similar to CM. These embryonic fibroblasts may control their own GAG synthesis and secretion through autocrine TGF alpha and beta activity.
Locci
P
P
Dipartimento di Medicina Sperimentale e Scienze Biochimiche, Università degli Studi di Perugia, Italy.
Lilli
C
C
Marinucci
L
L
Baroni
T
T
Pezzetti
F
F
Becchetti
E
E
D000818:Animals; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D002642:Chick Embryo; D017077:Culture Media, Conditioned / Q000494:pharmacology; D004357:Drug Synergism; D004815:Epidermal Growth Factor / Q000494:pharmacology; D005109:Extracellular Matrix / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects*; D006025:Glycosaminoglycans / Q000096:biosynthesis*; D007424:Intracellular Fluid / Q000378:metabolism; D012867:Skin / Q000166:cytology* / Q000196:embryology; D016211:Transforming Growth Factor alpha / Q000378:metabolism* / Q000494:pharmacology; D016212:Transforming Growth Factor beta / Q000378:metabolism* / Q000494:pharmacology
D017077:Culture Media, Conditioned; D006025:Glycosaminoglycans; D016211:Transforming Growth Factor alpha; D016212:Transforming Growth Factor beta; D004815:Epidermal Growth Factor