The postnatal demasculinization of sexual behavior in the Japanese quail (Coturnix coturnix japonica)

Three experiments were performed to analyze the time course of demasculinization in the Japanese quail and to test the activating and organizing effects of estradiol (E2) in adult sexually active birds. In Experiment 1, males and females were castrated at the age of 1 day or 1, 2, 4, and 6 weeks and treated as adults with testosterone (T). The age of castration had no effect on behavior and morphology in males. Plasma gonadotrophins (LH and FSH) were, however, higher in males castrated at or before than in those castrated after 2 weeks of age. This suggests that postnatal testicular secretions have organizing effects on the pituitary activity. Females which were castrated before 1 week of age were less sensitive to the activating effects of T than males, but were not fully demasculinized. The demasculinization of different reproductive characteristics such as male sexual behavior, cloacal gland size, and weight of the syringeal muscles is achieved in females at different times posthatching. In Experiment 2, castration of male and female quail at the ages of 4 days or 4 weeks confirmed that postnatal ovarian secretions contribute to the full behavioral and morphological demasculinization of females. It is easier to elicit mounting in T-treated females when they are tested in their home cage instead of a test arena. This difference was not observed in males. During Experiment 3, it was impossible to demasculinize sexually active adult males or females by treatment with Silastic implants of E2. E2 did not maintain sexual behavior in ovariectomized females showing male sexual behavior when treated with T but maintained the behavior in males.     

Perinatal exposure to 19-Nor-17α-ethynyltestosterone (norethindrone) influences morphology and aggressive behavior of female mice

Pregnant mice were injected with 50 or 100 μg of the synthetic progestin 19-nor-17α-ethynyltestosterone (NET) on Days 14 through 17 of gestation. Others received only the vehicle or were left undisturbed. Exposure to NET during the prenatal period increased anogenital distance of the female offspring as measured on Day 60 but did not influence the duration of adult testosterone (T) exposure required to activate male-like intraspecific fighting behavior. In a second experiment, female mice were exposed to NET either prenatally, postnatally (Day 1), or pre- and postnatally. Exposure during both pre- and postnatal periods increased anogenital distance but only the exposure during the postnatal period enhanced later behavioral sensitivity to the aggression-activating property of T. Thus, NET is similar to testosterone in its ability to virilize morphology and behavior, although, at the dosages administered, behavioral alternation only occurred as a result of treatment during the neonatal period.     

Paranodal dysmyelination and increase in tetrodotoxin binding sites in the sciatic nerve of the motor end-plate disease (med/med) mouse during postnatal development

Motor end-plate disease (med), in the mouse, is a hereditary neuromuscular defect, caused by a single gene mutation and characterized by a progressive muscle weakness. +Med/+med mice die 21-23 days after birth and the neurobiological abnormalities already reported are nerve terminal sprouting and swelling and neurotransmission failures. We studied +med/+med mice at preclinical (9-11 days after birth) as well as at clinically recognized stages of the disease. The nonmyelinated gaps of the nodes of Ranvier in the +med/+med sciatic nerve are found to be significantly widened in +med/+med animals compared to control littermates, even in the preclinical stage, although the nodes of Ranvier are not yet ultrastructurally mature. The maximal binding capacity for [3H]ethylene-diamine tetrodotoxin, expressed in femtomoles per milligram of protein, is significantly increased in +med/+med sciatic nerves. Thus, Na+ channels, which are known to be located mainly at the nodes of Ranvier in normal myelinated axons, are increased in number in +med/+med mice even before the disease becomes clinically established. Both the ultrastructural and biochemical developmental abnormalities of the node of Ranvier rapidly approach their maximal expression as the behavioral signs develop. Such nerve abnormalities may be closely related to the physiological impairment of nerve impulse conduction which leads to the pathophysiological expression of motor end-plate disease.     

Biological effects of D2O administration to Coturnix japonica

Levels of 7.8, 18.5 and 26 mole % deuterium oxide were administered sequentially to $\text{Coturnix japonica}$ (Japanese quail) via the drinking water. The primary effect observed was on egg frequency, which decreased from a normal level of 0.89 for 7.8 mole % D₂O to a low of 0.38 during the administration of 26 mole % D₂O. Adverse symptoms, such as hyperexcitability, convulsions, skin ulcerations, comatosity, weight loss, or death, which have been associated with deuterium toxicity in other animals, were not observed in these experiments. The amount of deuterium deposited in the water of the egg was 6.9, 13.98, and 19.83 mole % when 7.8, 18.5 and 26 mole % deuterium respectively was administered. For each period, the deuterium content of egg water rapidly reached a maximum concentration after which the concentration decreased slightly. This dilution effect has not been noted previously in body fluids from other animals.     

Role of vocalizations in the reproductive cycle of ring doves (Streptopelia risoria): effects of hypoglossal nerve section on the reproductive behavior and physiology of the female

The role of vocalizations in the reproductive cycle of female ring doves was investigated. Two-stage bilateral hypoglossal nerve section (HNS) was performed on adult females to alter their cooing and they were observed for courtship behavior with the same stimulus males used in preoperation behavioral tests. The HNS females showed a reduction in both the vocalization and wing flipping components of nest-coos but no changes in other female courtship behaviors. In addition, the HNS females failed to show the typical male courtship-induced follicular growth observed in sham-operated females. The behavior of stimulus males did not differ between groups and therefore could not account for the failure of follicular growth in the HNS females. These data suggest that the female's performance of nest-coos, or factors associated with it, may stimulate her own follicular growth.     

Duration of hormonal deprivation: influences on physiological and behavioral responsiveness to estradiol

A total of 54 ovariectomized female guinea pigs were divided into three groups and tested six times at 2-week intervals for their responsiveness to exogenous ovarian hormones (3 days of 4 micrograms/kg estradiol benzoate plus 1 day of 0.4 mg/kg progesterone) or control injections (0.2 ml oil vehicle). Two weeks after ovariectomy, treatment with estradiol significantly reduced food intake and body weight, and also produced vaginal membrane rupture in 98.1% of the females. When tested for sexual behavior at 4, 6, and 8 hr after the progesterone injection, 29 of the subjects (53.7%) displayed lordosis in response to manual stimulation. Twelve weeks after ovariectomy, the effects of estradiol on food intake, body weight, and vaginal membrane condition had not diminished. However, the overall proportion of females from which lordosis could be elicited declined to 27.8%. Biweekly injections of estradiol benzoate plus progesterone to one of the groups of females did not prevent this decline in the sexual response. Based on these results, it was concluded that the observed reduction in behavioral lordosis does not represent a general decline in the responsiveness of ovariectomized guinea pigs to estrogenic stimulation, but may involve changes in their responsiveness to progesterone or in other mechanisms more specifically associated with sexual behavior.     

Conformational changes following Mn(II) binding to demetalized concanavalin A

A temperature-dependent conformational change occurs following the binding of only one Mn(II) to a concanavalin A monomer. This change is independent of Ca(II) near pH 7 and is characterized by an activation energy of 22.3 kcal mol^?1, a value similar to that attributed to a cis-trans peptide isomerization. Two conformations have been detected in magnetic resonance experiments on solvent water protons where spin lattice relaxation times are influenced by bound Mn(II). Both conformations possess saccharide binding activity and Ca(II) stoichiometrically enhances the rate of conversion to the final, more stable conformation.     

The glycosaminoglycans of estrogen-induced medullary bone in Japanese quail

Glycosaminoglycans were isolated from the femurs of estrogen-treated male Japanese quail. During the 72 h after the injection of estrogen the incorporation of a 1-h pulse of H₂³⁵SO₄ into keratan sulfate increased more than 100-fold in a pattern corresponding to the production of the induced medullary bone. The rate of incorporation into chondroitin 4-sulfate, the only other glycosaminoglycan detected, remained constant throughout the same time period. The rate of incorporation of the 1-h pulse of sulfate into chondroitin 4-sulfate and keratan sulfate was the same at 48 h of estrogen treatment. When birds (48 h estrogen) were allowed to live 6 h after the injection of the isotope, chondroitin 4-sulfate accumulated 5-fold over that found for similar animals labeled for only 1 h. Keratan sulfate, into which the isotope was incorporated at the same rate as the chondroitin sulfate in this experiment, did not accumulate much more in 6 h of labeling than in 1 h of labeling. This suggests that the keratan sulfate turns over more rapidly than the chondroitin 4-sulfate in this tissue. Autoradiography showed that the chondroitin 4-sulfate was associated mainly with the marrow cells near the cortical bone and the keratan sulfate with the newly synthesized medullary bone. These results suggest that keratan sulfate is a specific marker for this secondary bone matrix.     

Fertility of sows injected with exogenous oestradiol and/or gonadotrophins to control post-weaning oestrus

In the first of two experiments 28 multiparous sows were allocated to one of the following treatments 2 days after weaning at approximately 35 days post partum: (1) untreated; (2) i.m. injection 10 μg oestradiol benzoate (OB)/kg body weight (b.wt.); and (3) i.m. injection 20 μg OB/kg b.wt. Sows were bred at first post-weaning oestrus and ovulation rate assessed at slaughter. The mean interval from weaning to oestrus in each group was: (1) 5.6 ± 0.2; (2) 4.7 ± 0.2; and (3) 4.7 ± 0.2 days; the mean ovulation rates in groups 1 and 2 (18.7 ± 0.6 and 17.4 ± 1.8, respectively) were significantly higher (P < 0.01) than that of 12.0 ± 1.7 for treatment 3 sows. Two untreated and one each of the treated sows were not cycling at slaughter.In the second experiment 75 multiparous sows weaned at 28 ± 3 days post partum (day 0) were evenly allocated with respect to parity to one of four treatment groups: (1) untreated; (2) i.m. injection 10 μg OB/kg b.wt. on day 2; (3) PG600 (400 iu PMSG + 200 iu hCG) injection subcutaneous day 0; and (4) combined PG600/OB treatment as in (2) and (3) above. Sows were bred naturally at the first post-weaning oestrus and fertility assessed at farrowing. Control animals had a significantly longer (P < 0.05) weaning to oestrus interval (4.53 ± 0.25 days) compared to treatment 2 (4.03 ± 0.13) treatment 3 (3.97 ± 0.12) and treatment 4 (3.81 ± 0.07) sows. Sows treated with PG600 alone showed a significant increase (P < 0.05) in numbers born live compared to pre-treatment values. A smaller and non-significant increase in numbers born live in control sows (probably related to increasing parity) was not observed in either OB- or PG600/OB-treated animals.These results suggest that with further modification of the treatments, a system may be developed for introducing fixed-time artificial insemination (AI) or mating as a means of controlling the reproductive performance of the weaned sow.     

A new inhibitor of the long-chain fatty acid transfer across the mitochondrial membrane: 2-(3-methylcinnamylhydrazono)-propionate (BM 42.304)

Using two different assay systems to distinguish between overt and inner forms of carnitine palmitoyl transferase (CPT, EC 2.3.1.21) of intact guinea-pig liver mitochondria, we have shown that the hypoglycemic agent 2-(3-methylcinnamylhydrazono)-propionate (BM 42.304) inhibits the activity of carnitine-acylcarnitine translocase of liver mitochondria. The results offer an explanation for the inhibitory effect of the compound on ketogenesis with oleate but not with octanoate in the perfused guinea-pig liver, previously reported by us (Biochem. Pharmacol. 32, 3405-3412, 1983).     

Embryonic and larval hemoglobins during the early development of the bullfrog, Rana catesbeiana

The main hemoglobin (Hb) found in Shumway (embryonic) stage 25 bullfrogs is that which we have designated Td-4. The other major tadpole Hbs (Td-1, 2, and 3) predominate during Taylor and Kollros (larval) stages I-XVIII. We propose that Td-4 is an embryonic Hb, whereas Td-1, 2, and 3 are larval (fetal-like) Hbs. Embryonic Hb Td-4 continues to be synthesized during the larval stages. During the larval period, the average peripheral blood Hb profile changes very little with morphological stage or general growth. However, there is great heterogeneity in the embryonic:larval Hb ratio among individual tadpoles of a given stage or weight, apparently due to differential Hb and red cell production by the two active erythropoietic sites, mesonephric kidneys (Td-4), and liver (Td-1, 2, 3).     

Effect of vitamin D deficiency on skeletal development during early growth in the rat

To define the role of vitamin D in early development, female weanling rats were grown to maturity on a vitamin D-deficient diet and mated with normal males. At Day 20 of pregnancy the weight and total body calcium of fetuses were determined. At various times after parturition, pups were sacrificed. Plasma samples were analyzed for calcium and phorphorus, and femurs were characterized as to volume, dry weight, ash weight, and total calcium. The results indicate that vitamin D deficiency with its accompanying hypocalcemia does not impair placental transfer of calcium nor weight gain of the fetus. Vitamin D deficiency does appear to increase calcium accumulation in the fetus. After parturition vitamin D is functional in maintaining a normocalcemia as early as 3 days postpartum and its importance increases with age of the neonate. Bone mineralization is clearly disrupted by Day 14 as judged by calcium content per unit bone volume and the severity of the defect increases with age. Both vitamin D and normal concentrations of calcium and phosphorus appear to be essential for proper skeletal development during early growth postpartum.     

Specific binding and pharmacological interactions of apamin,the neurotoxin from bee venom,with guinea pig colon

This paper describes the interaction of apamin, the bee venom neurotoxin, with its receptor in the guinea pig colon. The pharmacological activity of the toxin was assayed by measuring its contracting effect on guinea pig colon preparations that had been previously relaxed by neurotensin. The IC₅₀ value of apamin in this $\text{in vitro}$ bioassay is 7 nM. These pharmacological data are compared to the binding properties of apamin to smooth muscle membranes prepared from guinea pig colon. The highly radiolabeled monoiododerivative of apamin binds to its colon receptor with a dissociation constant $\text{K}{}_{\mathrm{<mtext>d</mtext>}}{}^1\text{= 36}\text{pM}$. The maximal binding capacity of colonic membranes is 30^dfmol/mg of protein. The dissociation constant of the unmodified toxin is 23 pM. The difference between the toxin concentrations that produce half-maximal effects in the binding and pharmacological studies arises from the different experimental conditions used for the two assays.     

Purification and properties of a lectin from the seeds of Croton tiglium with hemolytic activity toward rabbit red cells

The pH dependence of the chemical shift of the inorganic phosphate (P_i) inside mitochondria observable by ³¹P nmr has been examined and used for the measurement of the internal pH. The pH gradient and the P_i concentration gradient were in the simple relation expected for the neutral exchange process of H₂PO₄^? and OH^?. This P_i distribution across the mitochondrial membrane was not influenced by the cross-membrane electrical potential. Both the P_i, distribution and the pH titration curve of the internal P_i indicate that the activity of the internal P_i can be well represented by the concentration of P_i measured by ³¹P nmr peak intensity. The present results give a sound base for applying ³¹P nmr to study bioenergetics and cell metabolism.     

A Co(III) derivative of concanavalin A

Co(III) has been stoichiometrically incorporated into jack bean concanavalin A. The Co(III) protein still possesses a binding site for an additional divalent transition metal ion which together with Ca(II) can induce the sugar binding ability. No H₂O₂ oxidation of Co(II) occurs with demetallized concanavalin A activated with Ca(II) and Co(II) unless Co(II) is present in a stoichiometric excess. Evidence is presented to indicate that kinetically stable Co(III) is bound to a completely different location than the thermodynamically stable Co(II) protein site.     

Relationships of the Col plasmids E2, E3, E4, E5, E6, and E7: Restriction mapping and colicin gene fusions

Thirteen ColE plasmids representing the E2-E7 types have been compared by restriction mapping. Over 80% of their restriction sites were found to be similarly positioned, indicating that these plasmids share a common structure. Three variants are ColE2-CA42 and ColE7-K317, both of which contain 1.8-kb DNA segments in place of a 2.5-kb segment common to the other plasmids, and ColE6-CT14, which has an additional 5.0-kb DNA segment compared to the other plasmids. The colicin (col), immunity (imm), and colicin release (hic) genes of these plasmids have been localized to regions corresponding to those known for ColE3-CA38 and ColE2-P9, with the imm and hic genes adjacent to the 3' end of the col gene. Active colicin is produced from hybrid col genes containing 5' and 3' ends from different E-type plasmids. The 3'-termini of the fused col genes specify the colicin type.     

A rapid assay for epoxide hydratase activity with benzo(a)pyrene 4,5-(K-region-)oxide as substrate

A rapid radiometric assay for epoxide hydratase activity has been developed using the highly mutagenic [³H]benzo(a)pyrene 4,5-(K-region-)oxide as substrate. By addition of dimethylsulfoxide after the incubation, conditions were found where the unreacted substrate could be separated from the product benzo(a)pyrene-4,5-dihydrodiol(trans) simply by extraction into petroleum ether. The product is then extracted into ethyl acetate and, radioactivity is measured by scintillation spectrometry. This assay allows a rapid measurement of epoxide hydratase activity with an epoxide derived from a carcinogenic polycyclic hydrocarbon as substrate and is at the same time sensitive enough for accurate determination of epoxide hydratase activity in preparations with extremely low enzyme levels such as rat skin homogenate (8–14 pmol of product/mg of protein/min).     

Cell wall assembly in Fucus zygotes. II. Cellulose synthesis and deposition is controlled at the post-translational level

An alkali-insoluble cell wall component of Fucus distichus embryos is shown to be cellulose. Cellulose is not found in eggs, but within 20 min following fertilization is present in cell walls of zygotes. Incorporation of [³H]glucose or NaH¹⁴CO₃ confirms that this cellulose in zygotes of F. distichus and F. vesiculosus is newly synthesized, with the highest rate of synthesis occurring within the first 60 min. Use of specific inhibitors indicates that the increase in cellulose does not require protein or RNA synthesis. The evidence is consistent with the hypothesis that the initiation of cellulose synthesis triggered by fertilization is controlled at the post-translational level.     

Glutathione-related inhibition of prostaglandin metabolism

Placental homogenates contain a heat-stable, dialyzable fraction which specifically inhibits two placental enzymes, both of which possess 15-hydroxyprostaglandin dehydrogenase and 9-ketoprostaglandin reductase activities. The inhibition of the two enzymes is the same. The inhibitor has been resolved into two components by gel filtration on a column of Sephadex LH-20. The component which eluted first has been identified as oxidized glutathione (GSSG), the other as a glutathione-containing material (GSX). Inhibition of the 15-hydroxyprostaglandin dehydrogenase activity is competitive with respect to the prostaglandin substrate (K_i^GSSG = 26 μM, K_i^GSX = 1.4 μM). Inhibition of the 9-ketoprostaglandin reductase activity is also competitive with respect to the prostaglandin substrate (K_i^GSSG = 68 μM). The most effective inhibitor of the 15-hydroxyprostaglandin dehydrogenase is the prostaglandin A₁-glutathione adduct (K_i = 0.27 μM). This compound is not a substrate for oxidation of the 15-hydroxyl group but it is the best substrate found to date for reduction of the 9-keto function.