Prenatal exposure to androgen influences morphology and aggressive behavior of male and female mice

To assess the effects of prenatal exposure to androgen on adult aggressiveness in mice, pregnant mice were given injections of 1.5 mg testosterone propionate (TP) or oil from Days 12 to 16 of pregnancy. All offspring were gonadectomized on the day of birth. Neonatal treatment occurred on the day following birth and consisted of one-half of the animals from each prenatal treatment group being injected with 100 μg TP while the other half were injected with oil, yielding four Prenatal/Neonatal treatment groups for each sex. On postnatal Day 60, all offspring were given subcutaneous implants of encapsulated testosterone (T) and tested for 10 min every other day against a male opponent until aggression was observed. Female offspring of TP-treated mothers were indistinguishable from males on external examination at birth. The duration of exposure to T required to induce aggression provides an index of the sensitivity of the neural substrate to T. When arranged from the most sensitive to the least sensitive to the aggression inducing action of T, the four Prenatal/Neonatal treatment groups of females were significantly different from each other: Group TP/TP > Group OIL/TP > Group TP/OIL > Group OIL/OIL. A similar pattern was observed for the male offspring. There were no differences in the proportion of animals per group that exhibited aggression (virtually all animals fought) or the intensity of aggression once exhibited. The results demonstrate that morphological and behavioral masculinization can occur in response to exposure to androgen during prenatal as well as neonatal life in mice.     

Axon growth from limb motorneurons in the locust embryo: the effect of target limb removal on the pattern of axon branching in the periphery

Metathoracic limb buds have been unilaterally ablated from locust embryos at 25 to 30% of embryonic development and the effect of this operation on the axon morphology of the motorneuron fast extensor tibiae (FETi) observed at later embryonic stages. In control embryos this neuron sends a single axon out the main leg nerve, nerve 5, to the extensor tibiae muscle in the femur. In limb ablated embryos the axon of FETi is found in a wide variety of aberrant peripheral nerve pathways and projects to a wide range of foreign muscles. There is a degree of apparent selectivity, but no rigid hierarchy, in the choice of pathway and muscle made by FETi. A high degree of variability is found between one embryo and another in the extent and pattern of axon branching. The axon of FETi is generally found in pathways that correspond to nerves in control embryos but on occasion grows along novel routes. An anteriorly directed dendritic branch, seldom seen in control FETi neurons, is frequently seen in experimental FETis. These findings are discussed in terms of the rules for specific axon growth in normal development.     

Testosterone and photoperiod interact to regulate locomotor activity in male hamsters

Testicular size, plasma testosterone levels, copulatory behavior, and daily locomotor activity are reduced in male hamsters after 10 weeks of exposure to short days. The role of testosterone in the short day-induced decline in locomotor activity was investigated, determining whether or not photoperiod could alter the effect of testosterone on activity. Castrated adult hamsters were allowed to acclimate to running wheels (wired to digital counters) and then were kept on either long (L:D 14:10) or short (L:D 6:18) days for 60 days. On Day 60, half of the animals on each light cycle were implanted with 12-mm-long testosterone-filled Silastic capsules; half received empty capsules. Digital counting of wheel-running activity continued for another 140 days. Blood samples taken on Day 200 confirmed L:D 14:10 and L:D 6:18 testosterone-treated hamsters had equivalent plasma testosterone levels. After an initial decline in activity, L:D 14:10 animals exhibited a progressive rise in mean running activity (from ~2000 to ~5000 wheel revolutions per day) through 100 days after the initiation of testosterone treatment. In contrast, activity levels in testosterone-treated L:D 6:18 animals remained uniform (~2000 wheel revolutions per day) during this time, indicating exposure to short days rendered the hamsters less sensitive to the stimulatory effect of testosterone on activity. Of further interest was a marked increase in activity after 160–200 short days in animals treated with either testosterone-filled or empty capsules. It appears the total amount of daily locomotor activity in the hamster is modulated by circulating testosterone levels in a manner which is dependent upon the environmental photoperiod.     

Functional consequences of ligand-linked dissociation in hemoglobin from the sea cucumber molpadia arenicola

It has been established that Molpadia hemoglobin tends to dissociate into subunits as oxygen is bound. The kinetics and equilibria of carbon monoxide and ehtylisocyanide binding reported here show a dependence on protein concentration that supports the conclusions that the aggregated hemoglobin has a lower ligand affinity than the dissociated subunits. This is true for the isolated D-chain as well as for the unfractionated hemolysate that contains four distinct polypeptide chains (A-D). This indicates that even homopolymers of Molpadia hemoglobin have lower ligand affinity than the dissociated subunits. At high protein concentration hemolysates of Molpadia hemoglobin show slight cooperativity. The time course of ligand binding to the deoxy D-chain also suggests cooperative interactions, The low affinity of the aggregated state may have a different molecular explanation than in human hemoglobin were tetramers of identical subunits (as in Hb H) show high oxygen affinity. The absence of tyrosine and histidine at the C-tremini of the Molpadia D-chains also suggests a different stabilization of the low affinity deoxy state. An additional functional difference between Molpadia hemoglobin and human hemoglobin is that organic phosphate do not alter the ligand affinity of the sea cucumber hemoglobin.     

Glycogen in the proventriculus of the tsetse fly

Glycogen was detected in the proventriculus of the tsetse fly, Glossina morsitans morsitans, by ultrastructural, histochemical, and biochemical methods. This organ contained ten times or more glycogen on a dry weight basis than was found in the thoracic muscle. Proventriculi of male tsetse contained less glycogen than those of females belonging to the same age group and in teneral flies the amount of glycogen was about 50 per cent lower than in mature, fed flies of the same sex. Although the thoracic muscle of tsetse flies was considerably lower in glycogen than that of blowflies the amounts in the proventriculus of mature females of the two insect species were almost equal. It is suggested that this carbohydrate store may supply the energy required for secretory processes.     

Application of a direct spectrophotometric assay employing a chromogenic substrate for tryptophanase to the determination of pyridoxal and pyridoxamine 5′-phosphates

Improved procedures for the isolation of apotryptophanase and its use in estimation of the vitamin B-6 coenzymes are presented. An excess of the apoenzyme is allowed to react with limiting amounts of pyridoxal-P. Estimation of the holotryptophanase thus formed by use of the chromogenic substrate. S-o-nitrophenyl-l-cysteine, provides a sensitive (1–400 pmol) and conveniently direct spectrophotometric assay for pyridoxal-P. For the specific estimation of pyridoxamine 5′-phosphate, samples are first reduced with NaBH₄ to convert pyridoxal-P to pyridoxine-P (inactive). By nonenzymatic transamination with glyoxylate, pyridoxamine-P is then converted quantitatively to pyridoxal-P and estimated with apotryptophanase. The method gives excellent recoveries of the added coenzymes and indicates that in many tissue extracts pyridoxamine-P surpasses pyridoxal-P in concentration.     

Direct action of ethidium bromide upon mitochondrial oxidative phosphorylation and morphology.

Ethidium bromide (23 nmol/mg of protein) was found to be a potent inhibitor of oxidative phosphorylation, as determined by loss of respiratory control through the inhibition of the ADP-induced state-3 rate of oxygen uptake. A time latency for complete loss of respiratory control was noted, after which 2,4-dinitrophenol (DNP) was ineffective in overcoming this inhibition. In the absence of EDTA, ethidium bromide produced an apparent uncoupling, as evidenced by an increase of state-4 rates of oxygen uptake and loss of respiratory control. As low as 8 nmol of ethidium bromide/mg of protein stimulated mitochondrial adenosine triphosphatase (ATPase) for 5 min. Two to three times this amount of ethidium bromide reduced the amount P_i released. Preincubation of mitochondria with ethidium bromide prevented subsequent release of P_i during incubation with ATP. Likewise, preincubation inhibited the DNP-activated ATPase. The uptake of low levels of [¹⁴C]ADP preincubated with ethidium bromide (14 nmol/mg of protein) and succinate or α-ketoglutarate could apparently be reversed, with loss of radioactivity beginning several minutes after addition of the radioactive nucleotide. Inhibition of oxidative phosphorylation by ethidium bromide may be due to modification of the adenine nucleotide transport system in mitochondria. The production of apparently swollen mitochondria treated in vitro with ethidium bromide and substrates necessary for oxidative phosphorylation, as seen in electron micrographs, further indicates that the compound is capable of acting directly upon mouse liver mitochondrial function and structure.     

Wound healing in the cornea of the chick embryo: III. The influence of pore size of millipore filters on the migration of isolated epithelial sheets in culture

The migratory activity of epithelia isolated from the cornea and the dorsal skin of chick embryos of different ages was examined in vitro. Five types of Millipore filters differing in pore size served as models to represent degrees of unevenness of the substrate instead of the natural wound beds of the corneal stroma and the dorsal dermis. Migration of the epithelium was rapid and extensive when the pore size was below 0.8 μm, but was inhibited or stopped when the pore size reached or exceeded 0.8 μm. The effect of surface properties of the substratum on the motility of the cell membrane and thus on the movement of the cells is discussed.     

Group mating among Norway rats I. Sex differences in the pattern and neuroendocrine consequences of copulation

The copulatory pattern of groups of rats (Rattus norvegicus) was studied in the laboratory in a seminatural environment. In a given mating session, every oestrous female copulated with each male; likewise, every male copulated with each oestrous female. While individual males and females experienced similar amounts of copulation, there were dramatic sex differences in sequence and temporal pattern. Males mated in a multiple intromission pattern and had more ejaculatory series when several females were in oestrus. In contrast, females received intromissions and ejaculations in a random order, not in the sequence of a male ejaculatory series. Males copulated at shorter intervals than females did, a temporal sex difference that was determined by the pattern of female solicitations and male approaches. These sex differences are used to discuss the different units of analysis that are appropriate for male and female sexual behaviour in this species. Furthermore, the sex differences in the temporal pattern of copulation which emerged during group mating parallel the known sex differences in the temporal parameters of the neuroendocrine reflexes which mediate successful reproduction in the domestic strain.     

The ontogeny of social relationships with group companions among free-ranging infant rhesus monkeys II. Differentiation and attractiveness

Infants are frequently with their mothers when troop members initiate interaction with them. To what extent are the infants' companions attracted specifically to them rather than to the mother? A simple method is presented for approaching this question and for assessing the extent to which the relationships of 11 free-ranging rhesus (Macaca mulatta) infants on Cayo Santiago have acquired qualities independent of (differentiated from) those of the mother. Companions of all ages tend to be attracted specifically to infants between 1 and 14 weeks of age and not simply to their mothers. However, this tendency is less marked for sibling companions than for more distantly related companions. No differences are found between infants of high- and low-ranking mothers. Infantile attractiveness is not necessarily sustained when infants are 17 to 30 weeks of age. The degree to which relationships with infant companions and with older female companions have, differentiated by 17 to 30 weeks is negatively related to the companion's age. Infants show early and increasingly marked tendencies to develop differentiated relationships with one another; however, they show no signs of doing so with adult female companions by 30 weeks of age.     

Frequency and intensity of chick distress calls: Effects on maternal foodcalling in chickens

The food call of broody domestic hens was used to measure maternal response to four frequency components found in chick distress calls (2,3,4 and 5 kHz) and to variations in distress call intensity (0—86 dB). Foodcalling increased significantly with frequency of the pure-tone test pulse; response to a taped distress call occurred between 40 dB and 86 dB intensity with a maximum at 60–65 dB. The results suggest that the mother uses the higher frequency components in recognizing the distress call, but responds maximally within a specific intensity range. The selective advantage of such behaviour is discussed.     

The ontogeny of social relationships with group companions among free-ranging infant rhesus monkeys I. Social networks and differentiation

Social networks of infant rhesus monkeys (Macaca mulatta) in a free-ranging, lineage-based group on Cayo Santiago are described by assessing the extent to which four measures of positive interaction between infants and finely-divided categories of companions are associated with (1) degree of relatedness through maternal lines; (2) sex of the companion; (3) age of the companion; and (4) dominance rank of the infant's lineage. The results suggest that the infant's social network mirrors that of its mother both in the first weeks of life and as late as 30 weeks of age. Infants have more positive social interaction with close kin than with distant kin or with unrelated individuals, and thus function as members of their lineage from the beginning. They associate more with female companions than with male companions, and more with younger immatures than with older immatures. Finally, infants in the top-ranking lineage spend more time with their own relatives than do infants in other lineages. The fact that these patterns change little as the infant gains independence from the mother supports suggestions that early maternal influence serves to pass on aspects of the mother's social network. It is suggested that the ontogeny of early social relationships resembles a process of differentiation.     

A favourable assessment of the O.P.T.--histamine reaction in the method of von Redlich and Glick

During the course of an investigation of the properties of triglyceride lipase in nerve endings of the central nervous system (1) there arose a need for rapid determinations of a lipase in a large number of membrane preparations. This communication reports a procedure for the determination of glyceride lipase in which fatty acid, formed from a radioactive substrate by the lipase, is separated from glycerides on DEAE-cellulose paper.     

Analyzing the joint effects of two antibodies and the design of molecularly engineered vaccines

A new analytical approach to determine how antibodies relate to each other in producing immunity is presented. One purpose of this analysis is to decide on the antigenic composition of vaccines. Regression analyses and single discrete analyses are shown to be inappropriate to this task. The proposed analyses makes multiple cut points on two pre-exposure antibody levels. At each resulting discrete classification, additive and multiplicative interaction parameters are calculated. Various models of joint antibody action are shown to produce two general patterns of interaction terms with this type of analysis. One pattern characterizes situations where each antibody, or something substituting for the action of each antibody, is always needed to eliminate all infections. This may be because the antigens to which the antibodies are directed are on separate microbes or because microbes can only be neutralized by the joint action of both antibodies. A second pattern characterizes situations where both antibodies act cooperatively but given high enough levels only one may be needed. This may be because the antibodies have the same molecular effect or because they act by separate means against the same organisms in an innoculum.     

Mobilization of iron from specifically labeled reticulocyte ghosts.

Specifically labeled 59Fe ghosts have been prepared by incubation of whole reticulocytes with 59Fe3+-transferrin-CO3(2)-- followed by washing and ghost isolation. The binding of 59Fe by the membrane fraction is quite stable over a wide range of conditions, but iron mobilization occurs on incubation with chelating agents or cell lysate. The time course of 59Fe mobilization by unlabeled reticulocyte lysate exhibits five apparently zero-order phases. The rate of iron mobilization is linearly dependent on the concentration of 59Fe ghosts present in the incubation mixture. In contrast, the relative concentration of lysate appears to exhibit a saturation dependence with regard to membrane iron mobilization. Bathophenanthroline sulfonate follows a multiphasic time course of iron mobilization similar to that found with the lysate. Lysate from mature erythrocytes was found to mobilize iron with kinetics that are identical to reticulocyte lysate. The number and duration of the phases is independent of the mobilizing agent. The role of the membrane fraction in regulating the rate of iron release to cytosol was also investigated by the repetitive incubation of 59Fe ghosts with fresh lysate. The rate of 59Fe mobilization depended on the condition of the ghost with regard to prior 59Fe depletion. This publication emphasizes the active role of the membrane fraction in determining the rate at which iron will become available to the cytosol and the possibility that cytosol factors modulate the action of membrane bound components.     

Conditions for the various types of co-operativity allowed by the adair,equation and their relation with the algebraic character of binding polynomials

A convenient way to obtain for any number, n, of sites, the functions of the constants of the Adair equation that decide the type of co-operativity of ligand binding to a non-dissociating protein is given and is illustrated by the examples n = 4 and n = 5. These functions are invariants of the binding polynomial and various of its derivatives.Although there are some simple sufficient conditions (inequalities relating successive Adair constants) for some co-operativity types, the full necessary and sufficient conditions even for uniform positive and negative co-operativity depend on very complicated functions of the constants for n > 4.However there are alternative ways of writing binding polynomials known as canonical forms. Up to at least n = 5, and probably beyond, the conditions that are complicated in terms of Adair constants are very simple in terms of the constants of canonical forms. For instance any fourth-degree polynomial can be written in the form p(x - α)⁴ + q(x - β)⁴ + 6μ (x - α)²(x - β)² although in three different ways. For one of these ways, the sign of μ distinguishes between mixed and uniform co-operativity. For any kind of mixed co-operativity μ > 0, while μ < 0 corresponds to uniform co-operativity. Advantages of the use of canonical forms are briefly commented on.     

The role of the anterior hypothalamus-preoptic area in the regulation of male reproductive behavior in the lizard, Anolis carolinensis: lesion studies.

Bilateral radiofrequency lesions of the anterior hypothalamic-preoptic area (AH-POA) abolish male Courtship and Agonistic behaviors in both intact and castrated/androgen-treated Anolis carolinensis. Control lesions caudal and dorsal to the AH-POA have no effect on these behaviors. Lesions rostral to the AH-POA also cause a significant decline in the behavior of intact animals. Histological examination of testes showed that lesions including and rostral to the AH-POA cause testicular collapse and regression to initial stages of spermatogenesis, indicating interruption of gonadotropin secretion. Castration and subcutaneous implantation of androgen restores Courtship and Agonistic behaviors in Rostral and Caudal Control groups to prelesion levels, while AH-POA lesioned animals remain at their low behavioral levels. Neither AH-POA or control lesions cause significant changes in animals' body weight.