Mouse fetal hemoglobin synthesis in murine erythroleukemic cells.
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Biomedical subjects
Publications and source records attributed to R M Zucker.
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Friend leukemia cells (GM86, clone 745) were induced to differentiate with dimethyl sulfoxide or butyric acid. The kinetics of induction were measured by cell growth, cell volume distributions, and [3H]thymidine incorporation. From the volume distributions, it was found that the rate of induction was both agent sensitive and concentration dependent. The changes in volume distributions occurred approximately 4 hr earlier with dimethyl sulfoxide induction relative to butyric acid induction. However, the changes with the butyric acid induction were more dependent on concentration. A decrease in labeling indices during the 12- to 20-hr time period was correlated to a decrease in mean cell volume and an increase in the proportion of G1 cells. After the 20-hr time period of induction, an increase in labeling indices and in the percentage of large cells was observed. The data suggest that a transient block of cells in G1 occurred between 12 to 20 hr, and that the early differentiation involved a volume decrease which was related to a redistribution of cell cycle stages. The study was also shown that the changes of cell volume are a rapid monitor to determine the early events of the differentiation process.
Using isoelectric focusing, a fetal hemoglobin was found in the peripheral blood of C57BL/6 fetal mouse during the 14 to 20 days gestational period. In acid-urea polyacrylamide gel the pattern of this fetal hemoglobin was different from that of the adult hemoglobin. The mouse fetal hemoglobin was differentiated from the mouse adult hemoglobin by immunodiffusion reaction. It suggests that there is a transient fetal hemoglobin in the C57BL/6 mouse during gestational age.
The thymus cells from spontaneous and 1st generation AKR leukemic mice were investigated, using electronic cell-volume distributions. The growth pattern of thymus cells in the mice with transplanted leukemic cells varied with the type of donor cells injected. After the transplantation of leukemic cells into 2 month-old recipients, the cortex of thymus decreased in size and medulla of thymus enlarged. The thymus underwent atrophy next, followed by a proliferation of the large-sized cells (greater than channel 40, 136 micrometer3). The electronic cell-volume distributions of the thymus cells from the spontaneous leukemic mice encompass the distributions of thymus cells during the growth of the transplanted leukemia.
The in vivo growth of EL-4 ascites tumor cells in peritoneal cavities of syngeneic (C57BL/6) and allogeneic (DBA/2) mice was monitored by electronic volume analysis. In the growth of EL-4 cells in the C57BL/6 mice, daily decrease in electronic modal cell volume and labeling indices was observed. After day 7, the rejection of EL-4 cells in DBA/2 mice was indicated by increase in the percentage of small cytotoxic lymphocytes and decrease in the labeling indices of tumor cells.
Multiple hemoglobins were found in the peripheral blood of hamsters when separated by analytical isoelectric focusing in polyacrylamide gel between pH 6.0 and pH 8.0. Five of these hemoglobins (I-V) were present in sufficient quantities for isolation by preparative isoelectric focusing and subsequent characterization. Cellulose acetate electrophoresis of the isolated hemoglobins resulted in producing as many as six different globin chains. The five hemoglobins were further characterized by their amino acid composition, N-terminal amino acid residue, and molecular weight.
Four animal transplantable solid tumors, composed of varying morphologic architecture and intercellular specializations, were studied by light and electron microscopy. These tumors were dissociated into viable single cell populations using a combination of mechanical and enzymatic methods. The conditions necessary for optimal dissociation consisted of (a) preparation of the tumor to maximize the tissue surface area, (b) enzymatic digestion with continuous agitation and (c) additional agitation to release loosely attached cells. Other factors that influenced the dissociation were optimized and discussed.
One hemaglobin was found in the adult rhesus monkey (Macaca mulatta) by the technique of isoelectric focusing. In addition to the adult hemoglobin, one fetal hemaglobin was observed in neonates of the same species. The alpha and non-alpha globins of rhesus monkey hemoglobins showed similar electrophoretic mobilities as those of human hemoglobin A by cellulose acetate membrane electrophoresis. It appeared that the rhesus monkey hemoglobin system is a good model system for the study of human hemoglobin development.
Rabbit and goat antibodies to monkey adult and fetal hemoglobin were prepared and purified to apparent monospecificity. After conjugation with fluorescein isothiocyanate, the antibodies were employed to identify the hemoglobin types within individual cells in peripheral erythrocyte smears. The percentage of neonatal monkey erythrocytes containing fetal hemoglobin was found to decrease with time. The existence of adult or fetal hemoglobin in the erythrocytes appeared to be mutually exclusive.
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The rapid analysis of in vivo chemotherapy on the L1210 ascites tumor grown in C57BL/6 X DBA/2F1 mice has been shown by means of an electronic volume analysis. The drugs were injected on the 4th day of tumor growth, and the cells in the peritoneal cavity were studied at 24-hr intervals on the 5th through 7th day. Using the electronic cell volume distributions, combined with labeling indices, cell morphology, and cell counts, it was found that the alkylating agents. 1,3-bis(2-chloroethyl)-1-nitrosourea and cyclophosphamide, at the dosages used, were more effective than the S-phase-specific drugs, palmitoyl ester of 1-beta-D-arabinofuranosylcytosine, vincristine, and methotrexate.
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