Role of the vomeronasal organ on the estral cycle reduction by pheromones in the rat

The role of he vomeronasal organ on the estral cycle reduction induced by pheromones is studied in adult female wistar rats. The animals were divided in three groups: I, intact rats; II, vomeronasalectomized rats (VNX); and III, sham operated rats (sham). Each group was submitted to another three distinct conditions from the day they were weaned (21 days old): Isolated female rats; with male odors from two adult males of tested sexual potency, and isolated rats again. The isolated intact rats show mainly 5 day length cycles. The groups I and III (intacts and sham) with male odors, show 4 day length cycles. The VNX animals show 5 day cycles in any one experimental conditions. These results support the idea that the vomeronasal organ is the receptor of the male reducing cycle pheromone in the female rat.     

Influence of the oestrous cycle on L-glutamate and L-aspartate transport in rat brain synaptosomes.

Oestrous cycle and sex differences in sodium-dependent transport of L-[3H]glutamate and L-[3H]aspartate were investigated employing well washed synaptosomes prepared from rat brain cortex. Transport was best analysed on the basis of two components, a high and low affinity transport site. Oestrous cycle and sex differences were observed for both substrates. The high affinity transporter displayed highest affinity for glutamate transport in synaptosomes from female rats during proestrous and oestrous. This differed significantly from glutamate transport during dioestrous and in male rats. High affinity aspartate transport displayed highest affinity during oestrous and differed significantly from transport during dioestrous. Maximal velocity of high affinity glutamate transport was higher in synaptosomes from females during dioestrous compared with oestrous and lower in synaptosomes from male rats when compared with female rats in dioestrous and metoestrous. The low affinity sodium-dependent glutamate transporter displayed a 10-fold higher affinity for glutamate during proestrous than during the other three phases of oestrous and in male rats. Exogenously applied oestradiol and progesterone to synaptosomes from male rats showed no effect on glutamate or aspartate transport. No acute effect of oestradiol or progesterone on glutamate currents in oocytes expressing EAAT1 or EAAT2 subtype of glutamate transporter was observed. These results suggest hormonal regulation of high and low affinity sodium-dependent excitatory amino acid transporters over the four day oestrous cycle in synaptosomes from rat cortex. This regulation is unlikely to be due to a direct effect of oestradiol or progesterone on glutamate transporters.     

Hormonal and experiential control of female-male mounting in the female rat

Mounting behavior in the female rat has been studied extensively in same-sex interactions, but not in the heterosexual dyad. The present study examined the display of female mounting of castrated noncopulating male rats (FMM). Ovariectomized (OVX) sexually naive female rats (N = 80) were given either estrogen (E) + progesterone (P), E + oil (O), P + O, or O + O treatment for five tests with castrated male rats. FMMs were observed in both the E + P and E + O females. The influence of the ovarian cycle on FMM was also investigated. Vaginal smears from sexually naive females (N = 16) were taken daily for 12 days immediately after testing with castrated males. FMM frequency was greatest during Proestrus. Finally, OVX females (N = 30) treated with E + P were given either 0, 1, 10 multi-ejaculatory heterosexual experiences with intact, sexually experienced males, prior to tests with intact, copulating or castrated, noncopulating males for five tests. Sexually naive females displayed a greater number of mounts relative to the sexually experienced females when tested with castrated, noncopulating males. In contrast, very few FMMs were observed in females of any group tested with intact, copulating males. These data suggest that FMM occurs naturally in rats as a "super-solicitational" behavior that is modified by hormone treatment and prior heterosexual experience.     

Biological sexual maturation in rats: Interaction among genetic line,sex, age,and body weight

Interaction among genetic line, sex, age and body weight was studied for biological sexual maturation in rats. In female rats vaginal opening, first estrus, and duration of estrus cycle were determined. In male rats first presence of sperm was determined. Age and body weight at vaginal opening, first estrus or first detection of sperm were determined. Differences among five genetic lines were found on all measures of sexual maturation. The random bred control rats differed from the selectively bred lines of rats on majority of the measures, thus suggesting the influence of selective breeding on sexual maturation. The Maudsley lines selected for open-field defecation and Roman lines selected for two-way active avoidance learning showed different patterns of sexual maturation. Age was found to be crucial for the sexual maturation of female rats, whereas body weight was more important for the sexual maturation of male rats. This study is the first to investigate systematically the biological sexual maturation in live male rats and related sex differences. Findings suggest an important genetic component, and effect of selective breeding on the biological sexual maturation of rats.     

Effect of estrogen on lysosomal enzyme activities in rat heart

The activities per microgram DNA of five lysosomal enzymes [cathepsin D, cathepsin B, beta-N-acetylglucosaminidase (beta-NAG), beta-glucuronidase, and acid phosphatase] were measured in homogenates of female and male rat (Sprague-Dawley) hearts. Female rats were studied during stages of the estrous cycle and at 3 weeks after ovariectomy. Three-week-postovariectomized female rats and intact male rats were injected subcutaneously with 17 beta-estradiol-3-benzoate. Lysosomal enzyme activities in the male rat heart were more responsive to exogenous estradiol than were activities in the female rat heart. Cathepsin B, beta-NAG, and beta-glucuronidase were increased dramatically in the male rat heart upon short-term administration of estrogen (4 days). In both female and male rat hearts, activities of two lysosomal proteinases, cathepsins B and D, were reduced significantly (approximately 50%) by extended administration of estrogen for 10 days.     

The anorectic effect of fenfluramine is influenced by sex and stage of the estrous cycle in rats

The controls of food intake differ in male and female rats. Daily food intake is typically greater in male rats, relative to female rats, and a decrease in food intake, coincident with the estrous stage of the ovarian reproductive cycle, is well documented in female rats. This estrous-related decrease in food intake has been attributed to a transient increase in the female rat's sensitivity to satiety signals generated during feeding bouts. Here, we investigated whether sex or stage of the estrous cycle modulate the satiety signal generated by fenfluramine, a potent serotonin (5-HT) releasing agent. To examine this hypothesis, food intake was monitored in male, diestrous female, and estrous female rats after intraperitoneal injections of 0, 0.25, and 1.0 mg/kg D-fenfluramine. The lower dose of fenfluramine decreased food intake only in diestrous and estrous females, suggesting that the minimally effective anorectic dose of fenfluramine is lower in female rats, relative to male rats. Although the larger dose of fenfluramine decreased food intake in both sexes, the duration of anorexia was greater in diestrous and estrous female rats, relative to male rats. Moreover, the magnitude of the anorectic effect of the larger dose of fenfluramine was greatest in estrous rats, intermediate in diestrous rats, and least in male rats. Thus our findings indicate that the anorectic effect of fenfluramine is modulated by gonadal hormone status.     

Immunohistochemical evidence of the presence of aromatase P450 in the rat hypophysis

In order to analyze whether aromatase is present in the hypophysis of adult rats, we have performed an immunohistochemical study in young adult male and female rats. Our study has revealed that the hypophysis of adult rats contains aromatase, although marked differences are found between the sexes. The hypophyses of male rats have cells immunoreactive for the enzyme, 34.40% of these hypophyseal cells showing reaction. By contrast, cells from female rats show very little reaction, only 0.84% of them being reactive. No significant differences in the percentage of immunoreactive cells between one phase and another are observed during the estrous cycle. Our results point to the immunohistochemical expression of aromatase in the hypophysis of adult rats and at the same time suggest that its expression is sex-dependent. The enzyme may therefore be involved in the regulation of adenohypophyseal cytology by androgens.     

Repeated light-dark shifts speed up body weight gain in male F344 rats

This study is aimed at verifying the causal relationship of chronic circadian desynchronization and changes in body weight control. Eight male albino F344 rats aged between 12-15 wk were subjected to twice weekly 12-h shifts of the daily light-dark (LD) cycle for 13 wk (3 mo). Continuous circadian phase shifts consisting of intermittent phase delay and advance and reduced circadian amplitudes were consistently displayed in all five experimental rats implanted intraperitoneally with heart rate, body temperature, and activity transponders. The experimental rat maintained a greater body weight during LD shifts and even after 10 days of recovery than that of the age-matched control rat, which was maintained on a regular LD cycle. Body weight gain was greater in the first 2 mo of LD shifts in the experimental rat than in the control rat. Relative to the baseline, food intake and activity percentages were increased and reduced, respectively, for the experimental rats. Features of these results, such as increased body weight gain and food intake, and reduced activity, suggest a causal relationship of chronic circadian desynchronization and changes in body weight control in male albino F344 rats.     

The effect of urine from castrated male or female guinea pigs and rats on the estrous cycle of guinea pigs and rats, respectively]

A 24-hour reduced cycle duration was observed in 5-day cyclic female rats exposed to the odor of urine from male or female castrated rats. A decrease in the duration of the period of vaginal closure, ranging from 2 to 5 days, was observed in female guinea pigs exposed to the odor of urine from male or female castrated guinea pigs. The pheromonal activity of urine in both species was concluded to be no dependent upon the gonadal function.     

Effects of feminization of male F344 rats on induction of tumors and on nucleic acid alkylation by nitrosobis-(2-oxopropyl)amine

Groups of male and female F344 rats were treated twice weekly by gavage with 2.5 mg of nitrosobis-(2-oxopropyl)amine (BOP) for 35 weeks. Additional groups given the same treatment were male rats castrated at birth, male rats bearing an implant of a pellet containing estradiol and castrated male rats bearing an estradiol pellet. Most rats died with tumors related to the treatment; intact male rats survived the least well of the five groups. Most rats in all groups had alveolar/bronchiolar neoplasms of the lung. Many of the male rats also had follicular cell neoplasms of the thyroid and transitional cell neoplasms of the urinary bladder and kidney pelvis; there were no liver tumors in intact male rats. Almost all female rats and castrated male rats had liver neoplasms, including hepatocellular, cholangiocellular and hemangiosarcomatous neoplasms, but few neoplasms of the thyroid, kidney or bladder. The male rats feminized with estradiol, intact or castrated, had liver neoplasms, mainly cholangiocellular, and also neoplasms of the thyroid. Two rats of each of the five groups were treated at 20 weeks of age with [14C]BOP. As measured by respiration of 14CO2, metabolism of BOP was faster in the two groups of male rats with the estradiol implant than in the other groups. DNA and RNA of the liver were isolated 6 h after treatment. The extent of methylation of liver DNA as 7-methylguanine and O6-methylguanine was higher in the females and in the feminized males than in the intact male rats, but when normalized to the dose of nitrosamine per unit body weight there was little difference among the five groups.     

Effect of bromocriptine and progesterone on the length of the ovarian cycle in 4- and 5-day estrous cyclic rats

To study the effect of prolactin and progesterone on the length of the reproductive cycle in the rat, rats of different estrous cycle length (four and five days, respectively) were injected daily (09.00 h) with either bromocriptine (1 mg/rat) or 70% ethanol vehicle (0.25 ml) from the day of estrus onward, up to the appearance of the next ovulation. Each group of rats was then (16.00, metestrus) also injected with either progesterone (4 mg/rat) or 0.2 ml of olive oil. The effects of these treatments on the length of the estrous cycle was studied by both the recording of vaginal smears daily and by direct visualization of oocyte-cumulus complexes on the ensuing day of estrus (10.00 h-12.00 h). Bromocriptine treatment shortened the length of the cycle by one day in 5-day but not in 4-day cyclic rats, while progesterone treatment lengthened estrous cycles by one day in both groups of rats. Treatment with both bromocriptine and progesterone had no effect on the estrous cycle length of 5-day cyclic rats, but did prolong in one day the cycle of 4-day cyclic rats. These facts suggest that prolactin regulates the length of the ovarian reproductive cycle in the rat through its action on the secretion of progesterone by the corpus luteum.     

Pulmonary antioxidant defences and protein damage during the ageing process of both sexes

The free radical theory holds that the senescence is caused by oxidative damage that results from an imbalance between reactive oxygen and nitrogen species (RONS) and antioxidant defences. Hence, it plays an important role in the field of gerontology. We evaluated, in male and female rats, the activities of the antioxidant enzymes catalase (CAT), glutathione peroxidase (GPx), and total superoxide dismutase (tSOD), as well as oxidative protein damage in pulmonary tissue at 3, 6, 12, and 20 months of age. The results show an increase in the activities of all antioxidant enzymes at 12 months of age in female rats, suggesting an association with the reproductive life cycle. Protein damage in female pulmonary tissues did not change significantly throughout the ageing process. In male rats, the activity of GPx in 20 months of age showed an inter‐gender increase, while the tSOD and GPx showed higher activities in 20 months of age in the intra‐gender analysis. The male lung showed higher protein damage at 6 months of age. These findings suggest that antioxidant enzymatic activity is connected to the reproductive life cycle. Copyright © 2009 John Wiley & Sons, Ltd.     

gamma-Aminobutyric acid activity in the olfactory bulb of the rat during the sexual cycle and response to olfactory stimuli.

Glutamic acid decarboxylase activity in the main and accessory olfactory bulbs throughout the sexual cycle of the rat was studied. The effect of male pheromonal secretion on enzyme activity during proestrus and estrus day was also tested. The enzyme activity showed circadian rhythm during the estrous cycle. This rhythm was disrupted during diestrus-2 afternoon in the main bulb and came back during proestrus afternoon. A different pattern of enzyme activity was present in the accessory bulb, since the circadian rhythm was altered during proestrus morning, returning during estrus afternoon. Male odor exposition did not change enzyme profile activity during proestrus day and during estrus morning in the main bulb. In contrast, in the accessory bulb the olfactory stimuli induced opposite changes to that found in rats from the vivarium during proestrus. Comparison of enzyme activity in olfactory stimuli-deprived rats with that of pheromone-stimulated rats during proestrus showed that male odor exposure specifically affects accessory bulb enzyme activity. It is concluded that the changes of the olfactory bulb GABAergic system during proestrus and estrus day, or that evoked by odor stimuli, demonstrate the discriminative response of this system between the accessory olfactory bulb and the main olfactory bulb.     

Influence of the estrous cycle on heterosexual aggression in two strains of rats (S3 and WEzob)

Two strains of rats (S₃ and WEzob), which show different levels of aggression in the laboratory, were tested in repeated heterosexual confrontations. Daily 15-min observations were made of the interactions between a female throughout a complete estrous cycle and the same male partner. In both strains the topography of aggression was similar in males and females, but the frequency of specific parameters varied. Males showed more offensive and females more defensive patterns. The overall level of aggression was very low on the day of estrous, when the female was sexually receptive. There were no differences in any elements of female or male behavior between the other 3 days of the cycle. The results support previous conclusions from single-sex encounters that in rats there is no sexual dimorphism in the ability to show aggression.     

Hormonal regulation of rat renal cytochrome P450s by androgen and the pituitary.

The hormonal regulation of rat renal cytochrome P450s, P450 4A2 (K-5) and K-2, was investigated. The level of P450 4A2 in male rats was five times that in female rats and accounted for some 90% of total cytochrome P450, measured photometrically. Lauric acid omega- and (omega-1)-hydroxylation activities of renal microsomes of male rats were also higher than those of female rats. The sex differences in lauric acid hydroxylation activity seemed to arise from the differences in P450 4A2 concentrations, according to an immunochemical study. P450 K-2 was a female-dominant form in rat kidneys. The level of P450 K-2 in renal microsomes of male rats was one-tenth that of P450 4A2. Castration of male rats decreased the levels of P450 4A2 and treatment of castrated male rats with testosterone reversed the decrease. The castration of male rats decreased the lauric acid hydroxylation of the renal microsomes to the level of female rats. The administration of testosterone to castrated male rats reversed the decrease. Hypophysectomy of male rats decreased the level of P450 4A2 and the administration of growth hormone reversed the decrease when intermittent injections mimicking the male secretory pattern were given, although continuous administration mimicking the female secretory pattern did not. Castration of male rats did not affect the level of P450 K-2, but testosterone decreased its level. Hypophysectomy of male rats increased the level of P450 K-2 and growth hormone decreased its level in hypophysectomized rats. These results suggested that the expression of P450 4A2 was regulated by androgen or growth hormone and regulation of P450 4A2 was different from that of P450 K-2. To explore the regulation of renal cytochrome P450 further, testosterone was given to control (intact) or hypophysectomized adult female rats. P450 4A2 was induced in the kidneys of both control and hypophysectomized female rats to close to the level of male rats. Thus, P450 4A2 was directly regulated by testosterone as well as growth hormone, and the regulation of the male-dominant form in rat kidneys was different from that of the male-specific form in the rat liver, which is regulated mostly by growth hormone.     

Announcement

Abstract

The Gastrin content has been estimated in the antral mucosae of male and female rats in different phases of the estrous cycle. Gastrin content appears to be highest in the rats at di‐estrous cycle end lowest in the rats at pro‐estrous. There is not much difference between the gastrin content at estrous and pro‐estrous. The gastrin content in the antral mucosa of male rats is higher than in rats a both estrous and pro‐estrous but it is about the same amount in rats at di‐estrous. The possible role of the circulating sex hormones on the storage of gastrin in the antral mucosa of rats has been discussed.     

Circadian variations in plasma growth hormone and prolactin in the infant rat: Comparison with the adult pattern

Radioimmunoassayable (RIA) plasma growth hormone (GH) and prolactin (PRL) levels were determined at 3 hr intervals during a controlled 24-hr light-dark cycle in 10-day-old male and female rats; parallel measurements were made of brain monoamines (MA's), dopamine (DA), norepinephrine (NE) and serotonin (5-HT) concentration. Plasma GH and PRL and brain MA levels found in infant rats were compared to the same determinations made during the 24-hr cycle in 50-day-old male rats. GH levels were rather uniform and did not show circadian periodicity in the plasma of infant rats, while PRL levels showed a diurnal surge in the late afternoon hr (1800). In adult rats, GH levels exhibited wide fluctuations during the 24-hr cycle and no circadian periodicity, while PRL levels showed one diurnal (1500–1800) and one nocturnal (2400) surge. A pulsatile GH secretion was found in adult rats sampled at 15 min intervals over a period of 2 hr, which seemed to be lacking in infant rats. In the brain of infant rats, DA and NE levels exhibited circadian patterns which resembled the ones present in the brain of adult rats, whereas no circadian variations were present in 5-HT levels.     

Serial copulatory behavior of male hamsters and rats

The copulatory behavior of mammalian males is generally characterized by the male's repeated approaches to and mounting of the female. The mounting behavior can lead to intromission, and after several intromissions, an ejaculation occurs. Following ejaculation, the male refrains from sexual activity for a period of time, the so-called "post-ejaculatory interval (PEI)". Most mammals will return to copulate again. Both male hamsters and rats were used and each animal performed five series of copulations with a proestrous female. From the 1st to 5th series of copulations the hamsters showed a shorter PEI than the rats. In addition, the PEI of the hamsters showed no change after each ejaculation, while the rats gradually showed a significantly increased PEI during the five series of copulations.     

Effects of estrogen and testosterone on the metabolism of mevalonate by the shunt pathway

Mevalonate is metabolized by a sterol-forming and a non-sterol-forming, also called the "shunt", pathway. Effects of estrogen and testosterone administration on the shunt activity were examined in male and female Wistar and Sprague-Dawley rats. Shunt activity was determined in vivo from the yield of expired 14CO2 following [5-14C]mevalonate injection. Total mevalonate utilized was determined from the yield of expired 14CO2 following [1-14C]mevalonate injection. In the female, about 45% of mevalonate appears to be metabolized via the shunt, and in the male about 20%. This difference between male and female rats is dependent on both testosterone and estrogen, and apparently on testosterone to a greater extent. Thus estrogen treatment produced a 20-35% increase in shunt activity over castrated controls, while castration of males without hormonal treatment resulted in about a 50% increase in shunt activity, and testosterone administration returned castrated male and female shunt activity to that of intact males, or nearly so. Light/dark cycle had no effect in vivo on shunt activity, but may be critical in demonstrating sex differences in shunt activity in kidney slices.     

Effects of prolonged administration of morphine on the estrus cycle of the rat

Ten female rats, of the age of three months, were treated for 24 days with increasing doses (10 mg/kg daily in the first week, 20 mg/kg in the second week, 30 mg/kg in the third week and 40 mg/kg in the last three days of treatment) of morphine acetate injected intra peritoneum, in order to cause a chronic intoxication (13). Five rats were utilized as controls. In all cases the vaginal smears was daily examined in order to evaluate the possible changes induced by the drug administration. The results obtained show that morphine produces important alterations in the estrous cycle of all the treated animals. In fact from the second day of treatment all animals had the cycle blocked in metaestrous. Afterwords a great variability occurred with a complete lack of estrous for the whole experimental period in five rats and very irregular succession and length of the phases into other ones. After the interruption of the treatment a resumption of the normal cycle was observed.