【精选】Increased hyaluronate synthesis is required for fibroblast detachment.pdfVIP

Biochem. J. (1986) 239, 445-450 (Printed in Great Britain) 445 Increased hyaluronate synthesis is required for fibroblast detachment and mitosis Michael BRECHT, Ulrike MAYER, Eva SCHLOSSER and Peter PREHM* Max Planck Institut fur Biochemie, D-8033 Martinsried, Federal Republic of Germany Human-embryo fibroblasts were synchronized by means of colchicine and cytochalasin, and the production of hyaluronate was determined by [3H]glucosamine incorporation and ion-exchange chromatography. Cells arrested by colchicine synthesized small amounts ofhyaluronate, whereas cells blocked by cytochalasin were stimulated in hyaluronate production. When the colchicine block was released, there was an increased synthesis of hyaluronate, which appeared first in the cellular fraction and was then shed into the culture medium. After release of the cytochalasin block, the hyaluronate production declined to that found with unsynchronized cells. A comparable increase of hyaluronate synthase activity was observed during mitosis. When hyaluronate synthesis was blocked by periodate-oxidized UDP-glucuronic acid, the cells were arrested in mitosis before rounding of cells. These results suggest that hyaluronate synthesis is required for detachment and rounding of cells during mitosis. INTRODUCTION (Prehm, 1985) have been described previously. Cells were blocked by addition of colchicine (0.01,Ig/ml) or Hyaluronate synthesis has been related to many cytochalasin (2.0,ug/ml) to the growth medium before cellular functions, including migration (Toole, 1972), culture for 40 h. Cell synchrony was determined as invasion (Toole et al., 1979; Knudson et al.,