Two candidate G1 polypeptides in chick embryo fibroblasts

Resting cultures of primary chick embryo cells have been labeled with ³H-leucine for the first 2 or 3 hours after feeding basal medium or basal medium supplemented with 10% dialyzed serum. The labeling patterns of the ³H-polypeptides of the soluble cell fraction have been compared by fluorography of two-dimensional gels. Large and consistent differences are seen in only three of the more than 900 spots that can visualized. This report concerns two of the three spots. The ³H contents of the two polypeptides (41,000 daltons, pI 7.1, designated 41–7.1, and 34,000 daltons, pI 6.2, designated 34–6.2) are increased by serum by about ten-fold. The highly selective effect of serum on the labeling of the two spots does not appear to be an artifact related to the extractability, solubility, or state of aggregation of the polypeptides. The radio-intensities of both polypeptides decrease markedly when the cells are labeled later than 3 hours after “shift-up”. Drugs that inhibit RNA synthesis and are known to stop the progression of the chick cells through the G1 period, camptothecin, cordycepin and 5,6-dichloro-β-D-ribofuranosylbenzimidazole, depress, with great specificity, the enhanced labeling of polypeptide 41–7.1 in the stimulated cells, and all but camptothecin have a similar action on polypeptide 34–6.2. A high level of actinomycin D (10 μg/ml), but neither a low level of the drug (0.02 μg/ml) nor 5-fluorouridine prevents the increased labeling of the two polypeptides in serum-fed cells. That 5-fluorouridine enters the chick cells and is converted to its active form is shown by the inhibition of the processing of pre-ribosomal RNA. The observations with the RNA inhibitors are at least consistent with the conclusions that the enhanced labeling of the two sports results from increased rates of synthesis of the polypeptïdes that depend upon mRNA production but not on the formation of ribosomal RNAs, and that the polypeptides play a role in the regulation of DNA replication in the chick cells.     

Artificial incubation,hand-rearing,behavior, and release of common 'Amakihi (Hemignathus virens virens): Surrogate research for restoration of endangered Hawaiian forest birds

In order to test the effectiveness of captive-rearing and release strategies for future restoration of birds in Hawai'i, this pilot study was conducted in forests where introduced avian disease and mammalian predators were present. Methodology used resulted in the first successful hatching of Drepanidinae eggs in an incubator and subsequent hand-rearing of chicks from hatch. Sixteen Common 'Amakihi (Hemignathus virens virens) (mean hatch weight = 1.4 g) were hand-reared. Two different reintroduction strategies were evaluated for small honeycreepers. Known mortality in the wild after release was due to mosquito-transmitted disease (avian malaria and pox). This pilot study shows that the techniques necessary to hatch, rear, and release endangered Hawaiian honeycreepers are available. However, restoration efforts will probably not succeed unless mosquito-free, predator-controlled reintroduction sites are available or strategies are developed to decrease mortality in naive honeycreepers exposed to disease after release. © 1996 Wiley-Liss, Inc.     

Distribution of binding sites for human chorionic gonadotropin in the preovulatory follicle of the rat

The distribution of binding sites for human chorionic gonadotropin (hCG) in the preovulatory follicle was studied by autoradiography. An ovulatory dose (10 IU/rat) of [125I]hCG (1.4 muCi/IU) was administered intravenously, and large Graafian follicles were isolated 3 h later by microdissection. Injection of excess unlabeled hCG (500 IU/rat) prevented uptake of radioactivity by the follicle, indicating that binding of iodinated hormone was confined to specific and saturable receptor sites. The density of bound hormone molecules was highest in the theca interna and in three to four layers of mural granulosa cells adjacent to the basement membrane; labeling was chiefly associated with the cell borders. No significant binding could be detected either on the oocyte or on the cumulus cells surrounding the oocyte. We therefore suggest that the induction of ovum maturation does not require attachment of the hormone to the oocyte itself or to follicle cells in its immediate vicinity.