Effects of steroidal hormones on sexual, attack, and search behavior in the isolated male chick

Immature male chickens were treated with testosterone (1 mg/day), Δ₄-androstenedione (1 mg/day), 5α-dihydrotestosterone (5α-DHT; 1 mg/day), 5α-androstanedion (1 mg/day), or estradiol (100 μg/day) in order to assess the effects of these steroids on copulatory behavior, agonistic behavior, and attentional processes. Testosterone, estradiol, and 5α-DHT were most effective in stimulating male copulatory behavior above that of oil-treated controls; whereas Δ₄-androstenedione and 5α-androstanedione had less, but nevertheless significant, effects on this behavior. Testosterone and 5α-DHT facilitated agonistic behavior; however, estradiol, 5α-androstanedione, and Δ₄-androstenedione were ineffective in this capacity. The persistence of response to a given stimulus type was increased by testosterone and decreased by 5α-DHT: 5α-Androstanedione had no discernible effect on this behavior. These findings suggest that in the male chicken the neural structures regulating male copulatory and aggressive behavior as well as attentional processes are differentially sensitive to sex steroids. The effects of all these steroids on somatic structures were assessed.     

Sexual behavior of the female porcupine Hystrix africaeaustralis

Porcupines are sexually active throughout the estrous cycle, and sexual behavior is not affected by the reproductive status and hormonal milieu of females. Increased female-male interactions during estrus are the only indications of behavioral changes during estrus. The lack of aggression shown by females to known males as opposed to aggression shown to strange males, the greater interest shown by males toward certain females, and the superior breeding success of these females suggest a pair-bonding mating system. Limited changes in female receptivity throughout the cycle might be of importance in maintaining the pair-bond.     

Menstrual cycle patterns of hormones and sexual behavior in gorillas

Oppositely sexed pairs of gorillas were tested behaviorally during the menstrual cycle to determine the relationship between hormone concentrations of the female and the frequency of sexual activity by the pair. Five females were tested individually during two cycles with each of two males, but serum samples for hormone assay were obtained from each female only during the first cycle of testing. There was no clear relationship between hormones and behavior for the single cycle in which the serum samples were obtained, with the exception that no copulations occurred after the early luteal phase, when progesterone was greater than 5 ng/ml. Normalized behavioral data from all four test cycles for all pairs suggested that female-solicited copulations were restricted primarily to the periovulatory period. Male sexual initiative (by one of the males) accounted for most copulations temporally dissociated from the periovulatory period. Normalized hormone data for all of the females suggested that (1) attractivity was associated with estradiol concentrations during the follicular phase, (2) proceptivity with estradiol and testosterone at midcycle, whereas (3) receptivity was not associated with hormone patterns or cycle phase. The data suggest that hormones are one of several variables that contribute to the regulation of sexual behavior in gorillas.     

Saccharomyces cerevisiae ARS on a plasmid from Staphylococcus aureus

Staphylococcus aureus plasmid pC194 carries three sequences closely related to a consensus sequence defined previously by analysis of different genetic elements which replicate autonomously in yeast Saccharomyces cerevisiae. Two of these enable the plasmid to replicate in yeast, the third does not. A new consensus sequence A/T T T T A T R T T T, 1 bp shorter than the previous one, can be deduced from our results. Replacement of the T with G at the position 9 of the sequence abolishes its activity. The presence of the two active sequences on pC194 genome can be explained by the A + T-rich base composition of the plasmid.     

Germ-line dependence of the maroon-like maternal effect in Drosophila

We have used pole cell transplantations to construct germ-line mosaics for maroon-like (mal), a maternal effect mutation in Drosophila. Such mosaics allow one to determine the cell type in which a gene is active. We find that the maroon-like maternal effect is (1) autonomous to the germ line and (2) dose sensitive in germ-line mosaics. Aldehyde oxidase activity is used as a histological probe to investigate the tissue and temporal distribution of mal⁺ activity in the developing ovary. The adult ovary shows mal⁺ activity in the germ line at all discernible stages of oogenesis but no activity is observed in the mesodermally derived follicle cells. Differential mal⁺ activity is observed even in the ovary of the third-instar larvae.     

Construction and characterization of plasmid vectors for cloning in Staphylococcus aureus and Staphylococcus carnosus

Several plasmid vectors for cloning in Staphylococcus aureus and S. carnosus have been constructed and characterized. The chimeric plasmids are composed of parts of the following parental plasmids: The chloramphenicol-resistance plasmid, pC194, the tetracycline-resistance plasmid, pMK148, and the erythromycin-resistance plasmid, pE12. All the chimeric plasmids confer two selectable antibiotic-resistance markers on host cells. Insertional inactivation of the various antibiotic-resistance markers occurred at the BclI site of pE12, and the Sau96- or AvaII-site of pMK148; only a slight inactivation of the chloramphenicol-resistance marker occurred at the HaeIII-site of pC194. The chimeric plasmids pCT20 and pCE10 are both stable in S. aureus and S. carnosus. In addition, the hybrid plasmids of pCT20 and pCE10, containing lambda-DNA fragments in various restriction sites between 0.4 and 1.2 kb, are stably maintained. The inserted lambda-DNA fragments appear unchanged.     

Temperature-sensitive periods and autonomy of pleiotropic effects of l(1)Nts1, a conditional notch lethal in Drosophila.

Analysis of temperature-shift experiments using strains homo- and/or hemizygous for a temperature-sensitive (ts) mutation of the Notch locus, l(1)N^ts1, has permitted us to localize temperature-sensitive periods (TSPs) both for lethality and for adult ectodermal morphology defects. Discrete TSPs for lethality are localized to the first half of the embryonic period, to the second larval instar, to the third larval instar, and to a 15 hr period immediately after pupation. TSPs for adult morphology defects are localized to the second and third larval instars for eyeless-headless and duplicated antenna, to the third larval instar for small and rough (spl-like) eye, eye scar, fused leg segments, shortened tarsal leg segments, Notch wings, and extra macrochaetae, and to the early pupal period for extra and missing microchaetae, fa^g-like rough eye and thick wing vein defects. Within the third larval instar, distinct patterns of eye, wing, and leg defects are observed. There is a striking similarity between the adult morphology defects and TSPs of l(1)N^ts1 and those of the larval and adult locomotor mutant, shi^ts1 (C. A. Poodry, L. Hall, and D. T. Suzuki, 1973, Develop. Biol.32, 373–386). Expression of l(1)N^ts1 also has been studied in genetic mosaics, in which we find that the pleiotropic effects of l(1)N^ts1 are autonomously expressed.     

Cell fusion, nuclear fusion, and zygote differentiation during sexual development of Dictyostelium discoideum

The fluorescent nuclear stain Hoechst 33258 was used to study the nuclear events during mating of Dictyostelium discoideum in liquid culture. These studies revealed that cell fusion begins about 11 hr after the sexually compatible cultures are mixed and continues until 26 hr. Approximately 37% of the cells fuse during this 15-hr period. At first the fused cells are relatively small, but by 20 hr the fusion products become evident as morphologically distinct giant cells. Starting at 22 hr these giant cells are transformed into true zygotes as nuclear fusion begins. Both the fusion of amebae and the differentiation of zygote giant cells are Ca²⁺-dependent events as revealed by studies using EGTA. The nuclear events of zygote differentiation involve nuclear swelling, migration, and fusion. The precise timing of these events has been detailed. Of particular interest for genetic analyses via the macrocyst is the presence of a small population of multinucleate cells (maximum level is 1.67% of the cell population) which usually possess 3 or 4 nuclei but may have as many as 10 or more. Although these multinucleate cells contain many nuclei, our evidence suggests that only one is a zygote nucleus. The genetic implications of these data and the potential value of using the mating system for the analysis of cell fusion are discussed.     

ATP concentration in Escherichia coli during oxygen toxicity

Escherichia coli, strain E-26, grown in defined salts medium with glucose as the sole carbon and energy source, contained 1.50 ± 0.16 · 10⁶ molecules of ATP/cell. ATP was extracted with HClO₄ and assayed with a Dupont Luminescence Biometer using the luciferin-luciferase assay. Exposure during exponential growth at 37°C to 4.2 atm of oxygen resulted in complete growth cessation within 5 min, and to cyclic changes in cellular ATP concentration over a 2 h period. However, significant decrease in cellular ATP concentration occurred after growth inhibition in hyperbaric oxygen; hence, lack of ATP was not the cause of growth inhibition from oxygen toxicity.     

13C-N.M.R.-spectral study of the pH behavior of reductively [13C]methylated,glycophorin a glyco-octapeptides and a related glycopentapeptide

The pH dependence of the labeled-carbon resonances of reductively [¹³C] methylated compounds tri-l-Ser, glyco-octapeptide A^M, asialoglyco-octapeptide A^M, glyco-octapeptide A^N, asialoglyco-octapeptide A^N, and a glycopentapeptide was investigated. The results are discussed relative to those previously observed for reductively [¹³C]methylated, intact glycophorins A^M and A^N, and in terms of the mode of display of the MN blood-group specificities by these related glycoproteins. The results indicated that the α-d-NeuAc groups appear to affect the pH-titration results of glyco-octapeptides A^M and A^N. Moreover, comparison of the pH-titration results for reductively [¹³C]methylated glyco-octapeptide A^M and reductively [¹³C]methylated asialoglyco-octapeptide A^M with those of a reductively [¹³C]methylated glycopentapeptide and reductively [¹³C]methylated tri-l-Ser indicated that the other carbohydrate residues present (α-d-GalNAc and β-d-Gal) may also affect the pH-titration results. The reductive-methylation modification appears to affect the chemical shifts of the carbohydrate and peptide carbon atoms of the glycopentapeptide minimally.     

Phylogenetic relationships among Vibrio anguillarum plasmids

Results of restriction endonuclease analysis and Southern blot hybridization suggest that the R-plasmids from Vibrio anguillarum strains isolated in Japan can be divided into at least four groups of homology depending on the time of their isolation and geographical source. Molecular cloning experiments allowed identification of specific restriction endonuclease fragments carrying the genes for either Cm^r or Tc^r as the common sequences between some of these groups of R-plasmids. The Cm^r region from the V. anguillarum R-plasmids was homologous to the Cm^r sequences of an R-plasmid isolated from another fish pathogen, Aeromonas salmonicida. The plasmid pJM1 from V. anguillarum strains isolated in the Pacific Northwest region of the United States, which encodes an iron transport system associated with the high-virulence phenotype of these strains, showed homology with two of the Japanese R-plasmids.     

Isolation of a tetracycline-resistance plasmid excised from a chromosomal DNA sequence in Bacillus subtilis

When Bacillus subtilis GSY908 (recE4-) (H. C. Spatz and T. A. Trautner, 1971, Mol. Gen. Genet. 113, 174-190) protoplasts were infected with Staphylococcus aureus plasmid pNS1 specifying tetracycline resistance (Tcr) (N. Noguchi et al., 1983, Gene 21, 105-112), which was modified such that it either could not replicate or did not carry a functional Tcr gene, a plasmid with a molecular weight of 3.1 X 10(6) (4.9 kb) was generated in Tcr phenotypes. This plasmid, named Tcr pNS1981, exhibited completely different restriction endonuclease cleavage patterns to pNS1 and showed only negligible sequence homology in hybridization experiments. Southern hybridization experiments revealed that pNS1981 arises by excision of a B. subtilis chromosomal DNA sequence. No sequence corresponding to pNS1 was detectable on the chromosome of pNS1981-maintaining B. subtilis. The production of pNS1981 was also observed in B. subtilis RM125 (r-Mm-Mrec+) (T. Uozumi et al., 1977, Mol. Gen. Genet. 152, 65-69.) with almost the same frequency as B. subtilis GSY908. Since the recipient B. subtilis Marburg 168 derivatives stated above are sensitive to Tc, the results indicate that information essential for Tcr is under negative regulatory control in the integrated state on the chromosome. Restriction endonuclease analysis suggested that pNS1981 is essentially the same as pBC16, formerly found in B. cereus (K. Bernhard, H. Schrempf, and W. Goebel, 1978, J. Bacteriol. 133, 897-903).     

The biogenesis of mitochondrial membranes in the yeast Saccharomyces cerevisiae

Membrane lipids of yeast mitochondria have been enriched by growing yeast cells in minimal medium supplemented with specific unsaturated fatty acids as the sole lipid supplement. Using the activity of marker enzymes for the outer (kynurenine hydroxylase) and inner (cytochrome c oxidase and oligomycin-sensitive ATPase) mitochondrial membranes, Arrhenius plots have been constructed using both pro-mitochondria and mitochondria obtained from O₂-adapting cells in the presence of a second unsaturated fatty acid (i.e. linoleate (N₂) to elaidic (O₂)). Transition temperatures which reflect the unsaturated fatty acid enrichment of the new membranes reveal interesting features involved in the mechanism of the assembly of these two mitochondrial membranes. This approach was further enforced with both lipid depletion and mitochondrial protein inhibition studies. Kynurenine hydroxylase which does not require fatty acid for its continued synthesis during aerobiosis seems to be incorporated into the preformed linoleate-anaerobic outer membrane. The newly synthesized activities of inner mitochondrial membrane enzymes on the other hand, appear to integrate their activity into newly formed aerobic-elaidic-rich inner membrane. These latter enzymes show a distinct dependence on fatty acid supplement for their continued synthesis during their aerobic phase. This suggests that O₂-dependent proteo-lipid precursors are formed before these enzymes are integrated into their membrane mosaic. Two separate models are proposed to explain these results, one for the lipid-rich outer mitochondrial membrane and another for the protein-rich inner mitochondrial membrane.     

Photoreversible inhibition by ultraviolet light of germ line development in Smittia sp. (Chironomidae, Diptera)

Pole cell formation in embryos of the parthenogenetic midge, Smittia sp., can be delayed or inhibited by irradiation of the posterior egg pole with ultraviolet light (uv). This leaves the schedule of nuclear divisions and chromosome eliminations virtually unaffected. However, uv irradition delays the precocious migration to the posterior pole of one nucleus, which normally becomes included in the first pole cell. This effect is photoreversible, i.e., mitigated by application of blue light after uv. Photoreversibility indicates that a nucleic acid component is involved as an effective target. During normal development of Smittia a number of chromosomes are eliminated during mitosis V, not only from somatic nuclei but also in the germ line. In the latter, this mitosis takes place during the first gonial division in the larva. After uv irradiation, the first pole cell nucleus has undergone supernumerary mitoses before pole cell formation and, as a result, is driven into mitosis V precociously as the pole cell divides. This is frequently associated with chromosome elimination from pole cells, which in turn is correlated with subsequent disappearance of already formed pole cells. Adults derived from embryos without pole cells do not form ovaries. Pole cell formation, pole cell preservation, and ovary development are separately inhibited by uv, and inhibition of each step is photoreversible. The results are discussed in the context of germ cell determination, protection against chromosome elimination, and the role of chromosomes limited to the germ line.     

Regulation of cytosolic HMG-CoA reductase activity in pea seedlings: contrasting responses to different hormones, and hormone-product interaction, suggest hormonal modulation of activity

Isoprenoid products added to reaction mixtures have no effect on HMG-CoA reductase activity. However, in vivo treatment with stigmasterol, cholesterol, ubiquinone or 4-hydroxybenzoic acid inhibits activity in etiolated tissues. The hormones abscisic acid (ABA) and gibberellic acid (GA) have opposite effects: ABA inhibits, and GA has little effect alone but in combined treatments completely overcomes inhibition by ABA or sterol. The results indicate that hormonal modulation contributes to the regulation of cytosolic HMG-CoA reductase activity.     

Effect of catecholamines and ovarian hormones on cyclic AMP accumulation in rat hypothalamus

Effect of different monoamines and estradiol were studied on cyclic AMP (cAMP) accumulation in hypothalami from 21 day old female rats. Incubation for 5 min with 10^?4M epinephrine, norepinephrine or dopamine resulted in an increase in cAMP accumulation in the hypothalamus. Incubation of hypothalamic tissue with estradiol (4 × 10^?7M to 2 × 10^?5M) also resulted in an increase in cyclic AMP levels. The increase caused by estradiol was observed only after 50 min of incubation period. The estradiol induced increase in cyclic AMP accumulation was abolished by both α and β blockers. These results suggest that the estradiol-induced increase in cyclic AMP may be mediated by a prior increase in catecholamines in the hypothalamic tissue.     

Reactions of the ferri-ferrocytochrome-c system with superoxide/oxygen and CO2/CO2 studied by fast pulse radiolysis

The reduction of ferricytochrome c by O₂⁻ and CO₂⁻ was studied in the pH range 6.6–9.2 and Arrhenius as well as Eyring parameters were derived from the rate constants and their temperature dependence. Ionic effects on the rate indicate that the redox process proceeds through a multiply-positively charged interaction site on cytochrome c. It is shown that the reaction with O₂⁻ and correspondingly with O₂ of ferrocytochrome c) is by a factor of approx. 10³ slower than warranted by factors such as redox potential. Evidence is adduced to support the view that this slowness is connected with the role of water in the interaction between O₂⁻/O₂ and ferri-ferrocytochrome c in the positively charged interaction site on cytochrome c in which water molecules are specifically involved in maintaining the local structure of cytochrome c and participate in the process of electron equivalent transfer.     

Isolation of germ line-dependent female-sterile mutation that affects yolk specific sequestration and chorion formation in Drosophila

Two loci on the X chromosome have been implicated in choriogenesis by in situ hybridization of poly A-containing RNA from choriogenic eggchambers to Drosophila polytene chromosomes (A. C. Spradling and A. P. Mahowald (1979): 7E and 12E. At least two genes coding for major eggshell proteins map to region 7E (A. C. Spradling, M. E. Digan, A. P. Mahowald, M. Scott, and E. A. Craig (1980). In an effort to elucidate the functional role of the 12E gene product, 3600 EMS-treated X chromosomes were screened for recessive female-sterile mutations that mapped within the region 11F10-12F1. Four independent female-sterile mutations were recovered, three of which fell into one complementation group (fs29, fs117, and fs445). Mapping by analysis of recombinant progeny as well as of trans heterozygotes utilizing other deficiency chromosomes showed that the three noncomplementing mutations all mapped to region 12E1-12F1. Studies comparing chorion morphology and protein synthesis indicate localized perturbations in the extracellular assembly of eggshell components in mutant eggchambers. The germ line dependence of the mutations was established using germ line mosaics constructed by pole cell transplantation. Analysis of eggchamber protein accumulation patterns showed reduced amounts of yolk polypeptides (YPs) in the mutants. The elevated concentrations of YPs found in mutant hemolymph coupled with the normal YP biosynthetic patterns and active uptake of trypan blue by mutant oocytes suggest that 12E sequences play a role in yolk-specific sequestration.