Neuronotrophic effects of skeletal muscle fractions on neurite development.

Soluble fractions of chick embryonic skeletal muscle stimulated radial outgrowth of long neurites from peripheral ganglia. Dorsal root ganglia were more responsive to the growth stimulus of muscle fractions than sympathetic ganglia. Muscle fractions from chicks immobilized with d-tubocurarine chloride (dtc) were significantly more effective in stimulating growth than normal muscle fractions. Neuritic outgrowth stimulated by muscle fractions was not blocked by antisera to the Nerve Growth Factor (NGF) indicating that these neuronotrophic effects were not due to NGF.

Care and breeding of the gray, short-tailed opossum (Monodelphis domestica)

Husbandry procedures were developed for the gray short-tailed opossum (Monodelphis domestica) based on experience with several colonies that were self-sustaining for up to 4 years (five generations). Individual adult animals (80-155 g) were maintained in modified single rat or guinea pig cages, although larger cages with floor areas of at least 2,000 cm2 were required for breeding. Diets consisted primarily of ground meat, dried milk powder, wheat germ, and vitamin supplements. Switching males in the breeding pairs stimulated reproductive activity. Monodelphis domestica was found to be hardy, tractable, and able to reproduce throughout the year under laboratory conditions. This species is proposed as a practical marsupial model for biomedical research.

An autoradiographic study of the role of satellite cells and myonuclei during myogenesis in vitro.

Satellite cells and myonuclei of neonatal rat muscles were differentially labeled with 3H-thymidine according to the procedure of Moss and Leblond (1971). Minced muscles fragments containing either labeled satellite cells or labeled myonuclei were cultured until multinucleated myotubes grew out from the explants. Reutilzation of isotope released from degenerating nuclei was competitively inhibited by using a culture medium containing excess (0.32-0.41 mM) cold thymidine. after an 8-10 day growth period, the explants were fixed and prepared for autoradiographic (ARG) examination to determine whether labeled satellite cells or myonuclei had contributed to the myonuclear population of the developing myotubes. Counts were made of the number of labeled myotubes in the explants and compared with the number of labeled satellite cells and myonuclei in samples of the original muscle tissues fixed at the time of explantation. The original muscles showed a mean satellite cell labeling index of 51.7% and gave rise to myotubes with a mean labeling incidence of 40%. In contrast, myonuclear labeling in the original muscle tissues showed no correlation with subsequent myotube labeling. Only 3.4% myotube labeling was found in explants in which over 30% of the original tissue myonuclei had been labeled. Under conditions controlled for isotope reutilization, these observations confirm results of in vivo ARG studies indicating that satellite cells are the only significant source of regenerating myoblasts in injured muscle tissue.

The in vivo differentiation of murine neuroblastoma.

C1300 neuroblastoma was implanted with regenerating skeletal muscle to study the role of tissue interactions during tumor cell differentiation. Combined tumor-muscle implants, placed subcutaneously or within diffusion chambers were compared with control tumors implanted without muscle. Neuroblastoma implanted with injured muscle undergoes a partial neuronal differentiation. The tumor cells lose their normal round cell configuration and develop numerous cytoplasmic processes. Accompanying these outward changes are an increased content of microtubules in the neuritic processes; the appearance of glial-like processes containing abundant microfilaments; and the occurrence of growth vesicles identical to those of the growth cones of normal neurites. Although the implants usually contain large numbers of regenerated myofibers, tumor cell differentiation is not dependent upon the presence of these newly formed fibers. Tumor differentiation occurs equally well on the surfaces of degenerating muscle fragments, fibrin deposits and on the membrane surfaces of the diffusion chambers. These observations suggest that non-specific cell surface phenomena, rather than neuromuscular interactions were primarily responsible for the tumor cell differentiation in vivo.