Effects on Sperm Morphology by Alleles at the Pink-Eyed Dilution Locus in Mice

Sperm head morphology was analyzed in all genotypic combinations for alleles dark pink-eye (p^d) and p-sterile alleles, p^6H, p^bs (p -black-eyed sterile) and p^25H. Three of these, p^6H, p^bs and p^25H, were radiation induced; homozygotes and heterozygotes of these three alleles are male sterile, whereas p^d/— genotypes are fertile. Sperm heads were examined by light microscopy and assigned to one of five classes: A. normal and near-normal, B. triangulate and oblate, C. spatulate, D. elongate, and E. filamentous. Males of each sterile genotype had grossly abnormal sperm and each sterile genotype differed from all other sterile genotypes and from fertile genotypes in at least one class, except p^6H/p^6H compared to p^bs/p^bs.Frequency distribution profiles (1) revealed a complex pattern of allelic interaction and do not support a deletion-complementation hypothesis, (2) do not show simple bimodality, which might suggest post-meiotic (haploid) gene expression, and (3) together with unpublished breeding data, show that p^25H is not a remutation of p^6H.     

Genetic Modulation of RNA Metabolism in Drosophila. I. Increased Rate of Ribosomal RNA Synthesis

It has been suggested that a particular Y chromosome which is rDNA-deficient (YbbSuVar-5) may be associated with an increased utilization of rDNA template in adult testes (Shermoen and Kiefer 1975). To extend the observations on this chromosome, experiments were designed to determine if the chromosome has an effect on rRNA synthesis in bobbed adults and on classic bobbed phenotypes (shortened and thinner scutellar bristles and delayed development). Specific activity measurements were made on rRNA extracted from adult males of the genotypes car bb/YbbSuVar-5, which are rDNA-deficient to the same extent, and from Samarkand+ isogenic (Sam+ iso), which is a wild-type stock. The resulting data demonstrated that the presence of the YbbSuVar-5 chromosome increases the rate of ribosomal RNA synthesis in adult flies. In addition, it was found that the presence of this particular Y chromosome restores wild-type bristle phenotype and development time. Appropriate genetic crosses indicate that the observed effects (increased rRNA synthesis, restoration of wild-type phenotype) are a function of this particular Y chromosome, and are not due to autosomal factors. The results of these experiments suggest that the rate of rRNA accumulation is under genetic control.     

Retention deficit for avoidance training in hypophysectomized rats: Time-dependent enhancement of retention performance with post-training ACTH injections

These experiments examined the effects of hypophysectomy on retention of avoidance training. In addition, the experiments examined the effects, on retention, of post-training ACTH injections administered to hypophysectomized rats. Rats were trained in a visual discriminated avoidance Y maze. Each rat received six training trials followed by six retraining trials the next day. Retention was measured by the number of correct choices during the retraining trials. When trained with a low-footshock intensity (0.8 mA), hypophysectomized rats showed retention performance which was significantly poorer than that of intact animals. There was no significant difference in performance when the animals were trained with a higher footshock intensity (1.4 mA), in part because of poorer retention performance of intact animals under these training conditions. Under both footshock conditions, a single post-training injection of ACTH enhanced later retention performance of hypophysectomized rats. This effect on memory was timedependent; injections delayed 2 or 6 hr after training did not significantly enhance retention. These findings are consistent with the view that hormonal responses to training may modulate later retention of the training experience.     

Initiation of Protein Synthesis Without Formylation in a Mutant of Escherichia coli That Grows in the Absence of Tetrahydrofolate

Starting from a p-aminobenzoate-requiring strain of Escherichia coli (E. coli K-12 AB3292), we have isolated mutants that can grow in the absence of p-aminobenzoate (and thus tetrahydrofolate). The following lines of evidence suggest that at least one of these mutants is capable of initiating protein synthesis without formylation of methionyl-transfer ribonucleic acid (methionyl-tRNA(fMet)). (i) tRNA isolated (and charged in vivo with [(35)S]methionine) from this mutant grown in a p-aminobenzoate-free medium contained less than 0.4% of the total methionine charged to the tRNA as formylmethionine. However, when the mutant was grown in the presence of p-aminobenzoate, 40 to 50% of the total [(35)S]methionine was detected as formylmethionine. (ii) Extracts of the mutant grown in the absence of p-aminobenzoate contained no formyl-tetrahydrofolate, but such extracts did contain formylatable methionyl-tRNA and a functional transformylase. (iii) Tetrahydrofolate-free extracts of the mutant were capable of supporting protein synthesis with viral RNA (from f2) as messenger, but the resulting synthesized proteins contained no formylmethionine, and methionine residues were detected where formylmethionine residues are normally found. In the presence of formyl-tetrahydrofolate, use of a similar extract resulted in the detection of 30 to 40% of the total polypeptide methionine as formylmethionine. (iv) Initiation of protein synthesis in vitro occurred more readily with formyl-tetrahydrofolate-free extracts of the mutant than with similar extracts prepared from the parent strain. However, in the presence of formyl-tetrahydrofolate, initiation of protein synthesis proceeded equally well with both kinds of extracts. tRNA from this mutant and another spontaneously derived mutant was found to be partially deficient in the modified nucleoside ribothymidine (rT). Analysis of extracts showed that the mutants contained decreased levels of the methylase that results in the formation of ribothymidine. In vivo studies with an independently isolated rT(-) strain suggest that the lack of rT in tRNA facilitates the growth of E. coli under conditions where protein synthesis is forced to take place without formylation.     

Effects of 6-thioguanine-containing RNA on in vitro protein synthesis

6-Thioguanine was administered to rats 12 hr after partial hepatectomy at a dose of 40 mg/kg of body weight; 6 hr later, polyadenylic acid-containing RNA was isolated and was used to measure initiation of protein synthesis $\text{in vitro}$ in a wheat germ system. $\text{In vitro}$ initiation was found to be 2.3-fold greater when 6-thioguanine-containing RNA was employed, than when polyadenylic acid-containing RNA isolated from untreated animals was used. The homopolymer, poly(TG), did not promote peptide synthesis in the wheat germ $\text{in vitro}$ system employed.     

Characterization of the DNA of a Nonoccluded Baculovirus, Hz-1V

The DNA of the nonoccluded baculovirus (Hz-1V) obtained from the IMC-Hz-1 cell line was characterized by physicochemical and restriction endonuclease techniques. Hz-1V DNA isolated from purified virus had buoyant densities of 1.58 and 1.54 g/ml in CsCl-ethidium bromide density gradients, which corresponded to supercoiled and to relaxed circular and linear DNA, respectively. Neutral CsCl equilibrium centrifugation indicated that the Hz-1V DNA had a buoyant density of 1.7024 g/ml, which corresponded to a guanine-plus-cytosine (G+C) content of 43%. Thermal denaturation indicated a high G+C domain(s) in the Hz-1V genomic DNA. The domain(s), which included about 11% of the total genomic DNA, exhibited a T(m) of 97 degrees C. The remaining portion (89%) of the DNA had a T(m) of 86.5 degrees C. The T(m)s corresponded to G+C contents of 42 and 67%, respectively. The mean genetic complexity of Hz-1V DNA determined by DNA reassociation kinetic analysis was found to be 152 x 10(6). A possible rapidly reassociating component comprising approximately 13% of the genome was observed. The mean molecular weights from restriction endonuclease digests were 159 x 10(6) for both HindIII and EcoRI. Genomic heterogeneity was found in both the wild-type Hz-1V stock and in two plaque isolates. Of 12 single-plaque isolates, 3 basic restriction endonuclease DNA fragment patterns were observed. The molecular size estimates from electron microscopic contour lengths of uncloned viral DNA ranged from 70 to 158 megadaltons, and the mode was the 130- to 140-megadalton class.     

Control of protein synthesis in Escherichia coli: Lack of correlation with changes in intracellular pools of ATP,GTP, and ppGpp

Intracellular pools of ATP, GTP, and ppGpp have been measured in Escherichia coli after an energy source shift-down from glucose minimal to succinate-minimal medium. In a Tic⁺ strain (ATCC 10798), which reduces translational initiation after the down-shift, the rate of protein labeling falls to about 30% of its preshift rate within the first minute after shift and reaches a minimum of 17% by 6 min after shift. The ATP pool in this strain remains constant for about 10 min after shift, then declines gradually to about 60% of its initial level. The temporal discrepancy between protein synthesis and the decline in the ATP pool indicates that a decrease in intracellular ATP is not necessary for the control of protein synthesis. In a Tic^? strain (W1), which cannot control translational initiation under these conditions, the decline in the ATP pool is somewhat more rapid and more pronounced (to 40%) than in the Tic⁺ strain, indicating that the decline in the ATP pool is not sufficient to trigger control of translational initiation. The intracellular GTP pool in the Tic⁺ strain remains constant for 2 min after shift, then declines gradually to reach a minimum of 45% of its initial level at 20 min after shift. The pattern is in general similar in the the Tic^? strain, although the ultimate decline in GTP is more pronounced (to 29%). These data indicate that the decline in GTP is not sufficient and probably unnecessary to elicit control of translational initiation. Intracellular levels of ppGpp increase with very similar kinetics in relA⁺Tic⁺ (ATCC 10798) and relA⁺Tic^? (W1) strains, indicating that elevated ppGpp levels are not sufficient to elicit control of translation. In a relA^?Tic⁺ strain (NF162), or in a relA⁺Tic⁺ strain treated with rifampin, the ppGpp pool does not increase significantly after shift-down although translational initiation is reduced. Thus, an increase in the ppGpp pool is not necessary to control of translational initiation.     

Sexual maturation among members of a transported troop of Japanese macaques (Macaca fuscata)

Data on the sexual maturation of a transported natural troop of Japanese macaques were collected during the 1973–74 and 1974–75 breeding seasons. Analysis of the data revealed that the sexual maturation of many monkeys was delayed one to two years. It is suggested that the delay of sexual maturation is related to a failure of the pubescent-aged monkeys to attain appropriate weight levels after transportation. This study was partially supported by a University of Oregon, Department of Anthropology PHS Biomedical Science Grant 50-262-1112.     

Maintenance of adult hamster pancreas cells on fibroblastic cells

Summary The maintenance of primary cultures of adult hamster pancreatic cells on layers of irradiated C3H/10T1/2 cells was studied. Various types of pancreatic cells, acinar, islet and ductular cells could be identified in the cultures by light and electron microscopy. Morphologically the various pancreatic cells retained many differentiated characteristics of their respective in vivo cell types. Insulin production was maintained at near Day 1 levels for the 16 d in culture for which it was measured. Colonies of epithelial cells continued to grow during a 20 d culture period. It is believed that this procedure for maintaining functional and growing pancreas cells in culture may be a useful in vitro model for studying the initiation of pancreatic carcinogenesis. Supported by Grant R01 CA 20022 and Contract N01 CP33278 from the National Cancer Institute, National Institutes of Health, Bethesda, Maryland.