Fetal liver cells of DBA/2 mice were infected with the anemic strain of Friend leukemia virus (FLV-A), which has no spleen focus-forming virus (SFFV) activity. The infected cells were grown in medium with or without erythropoietin. Transformed lines were isolated only from the infected cultures that had been treated with erythropoietin at the time of their initiation. The properties of three permanent cell lines in serial passage for over 2 years are described. Each has an aneuploid karyotype. Only the immature hematopoietic cells of the first line have metacentric chromosomes. They grow in suspension, as do the erythroleukemic lines derived from leukemic spleens of FLV-infected mice, and clone on agar. They produce tumors resembling reticulum cell sarcomas upon subcutaneous inoculation into syngeneic hosts. Stimulation of differentiation induced after treatment with dimethyl sulfoxide identifies the cells of the first line as being erythroid in origin. The two other lines are adherent and epithelioid in appearance. These lines may have originated from the nonhematopoietic cells present in fetal liver. No tumors were produced after the subcutaneous inoculation of 10(6) cells. All three lines synthesize virus. The virus is attenuated for leukemogenicity and has no SFFV activity. The transforming event appears to be specific, because fetal liver cells from C57BL/6 mice, which are resistant to the induction of leukemia by FLV, were not affected by the virus. Malignant transformation of erythroid cells by FLV-A in vitro confirms the in vivo findings that SFFV may not be a necessary prerequisite for the induction of erythroleukemia in susceptible hosts.

From day 12 to 16 of gestation a highly circumscribed zone of cellular degeneration has been observed in the otocyst of more than 50 normal rats. This zone appears to be produced by selective or programmed cell death, and results in the elimination of unneeded cellular elements. That cells in this zone are also selectively destroying subcellular organelles as a first step in specialization is also discussed. Programmed cellular death has long been known in other systems and is often an obligatory companion to cellular specialization. The ear now joins the long list of organs which demonstrate this phenomenon.

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The in vitro uptake of tritiated serotonin (3H-5HT) into hippocampal slices was measured in Ringer's solution (37#{176}C) containing pargyline, ascorbic acid, and dextrose. The specific uptake of 3H-5HT rose asymptotically as the 3H-5HT molarity was increased from 5 x 1O #{176}to 1.5 x lO M. Linear regression analysis gave a Km value for the specific uptake of 1.4 x 1O M. The nonspecific binding (NSB) was the amount of 3H-5HT retained by the slices following incubation in a medium with a very large excess of unlabeled 5-HT added to dilute the specific uptake of 3H-5HT. This NSB increased with increasing molarity of 3H-5HT, and was linearly related to 3H-5HT concentrations between 5 x 1O and 1.5 x 1O M. The ratio of specific uptake to NSB was highest at 5 x 1O M (2.75) and lowest at 1.5 x 1O M of3H-5HT (0.54). Competition studies with noradrenaline, desipramine (a noradrenergic uptake blocker), fluoxetine (a 5-HT uptake blocker), and tryptophan confirmed the specificity of the 3H-5HT uptake mechanism. Radioautographic studies of in vitro incubated hippocampal slices showed silver grain aggregates at 3H-5HT specific uptake sites. Addition of an excess of unlabeled 5-HT to the slices, or the use of 1 19.5 boutons/ 10,000 .tm2, in the dentate gyrus it was 67.4 boutons/ 10,000 m2, and in the subicular cortex it was 79.2 boutons/1O,000 am2. These results are consistent with previous quantitative results.

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