Bombesin and other growth factors activate cell proliferation in chick embryo otic vesicles in culture.

Abstract

The ability of the mitogenic peptide bombesin and other growth factors to trigger and support early development of the inner ear was studied on chick embryo otocysts in culture. The normal pattern of development was preserved in cultured otic vesicles in the presence of 20% fetal calf serum in the medium. Differentiation proceeded from stage 18 to 22 during the first 24 h and further to stage 24 in 48 h. Estimates of cell number and mitotic rates revealed a distinct period of proliferative growth which was maximum at the 24 h period of incubation. This was coincident with a high rate of DNA synthesis as measured by the acid-precipitable incorporation of [3H]thymidine. Development could be arrested by deprivation of serum during 24h. It could then be reactivated by readmission of serum to proceed with the normal pattern of morphological differentiation and cell proliferation. Bombesin (100 nM) was able to reactivate development in growth-arrested vesicles. Its effect was dose-dependent, saturable and potentiated by insulin (5 micrograms ml-1) which was ineffective if used alone. When associated with insulin, bombesin carried differentiation to stage 21 and stimulated mitotic activity above the level of serum as judged from estimates of cell number and [3H]thymidine uptake. EGF and PDGF were also effective in reinitiating development although their potency was smaller than bombesin. The reactivation by serum or bombesin was blocked by amiloride. The results show that (1) the otic vesicle can provide a useful model for studying the mechanisms that control proliferative growth and differentiation during normal development and (2) bombesin and other growth factors are able to activate growth in embryonic developing tissues.