The induction of pseudopregnancy and pregnancy by mating in albino rats exposed to constant light

Adult female albino rats anovulatory as a result of exposure to constant light (CL rats) were shown to ovulate in response to limited coital stimulation without necessarily becoming either pseudopregnant or pregnant. A higher level of coital stimulation would induce pseudopregnancy provided that intromission was not prevented. The occurrence of an ejaculation was not essential for the induction of pseudopregnancy but the incidence of pseudopregnancy was higher when a series of intromissions included one or more intromissions associated with ejaculation than when it did not. Similar findings regarding the induction of pseudopregnancy were obtained in experiments on female rats maintained under normal light-dark conditions (LD rats). Increasing the interval between intromissions from less than 1 min to as much as 20 min did not reduce the incidence of pseudopregnancy in CL rats. Splitting a set number of intromissions into two groups separated by up to 180 min did not reduce the incidence of pseudopregnancy in CL or LD rats. Conditions for the induction of pregnancy were more critical than for the induction of pseudopregnancy. CL rats showed only a low incidence of pregnancy and, if pregnant, either had small litters or did not deliver living pups even when a high level of coital stimulation from several males, including multiple ejaculations, occurred over a limited period of time. A high success rate was only observed when CL rats were mated with a single male overnight. LD rats showed a higher incidence of pregnancy than CL rats in response to a restricted amount of coital stimulation over a short time period, especially when an ejaculation was the terminal event in the mating sequence. This dependence on the occurrence of an ejaculation as the terminal event was not observed in CL rats, probably because, unlike LD rats, their uteri were not distended with fluid at the time of mating.     

Marking behavior is innate and not learned in the Mongolian gerbil.

We studied whether marking behavior in Mongolian gerbils would be innate or learned behavior. The marking behavior was defined as "animals rubbing their abdominal scent glands on small protruding objects". Between 21 and 90 days of age, Mongolian gerbils, which were kept under such conditions that they would be unable to learn this behavior, were observed at intervals of 5-15 days to find out if there were signs of the behavior or not. Six male and four female Mongolian gerbils were used for observing. Neonate Mongolian gerbils during the age of 3 to 28 days were fostered by ICR mother mice. Weaning Mongolian gerbils were then individually kept away from the others. Marking behavior was observed in 2 out of 6 males at 50 days of age and 2 of 4 females at 60 days and the mean frequency of the marking behavior for 10 min was 3.5 in the males and 5.0 in the females. These results suggest that marking behavior was innate and not learned behavior in Mongolian gerbils.     

Measurement of thymus weight, lumbar node weight and progesterone levels in syngeneically pregnant, allogeneically pregnant, and pseudopregnant mice.

Female CBA mice were mated to fertile CBA males, to vasectomized CBA males, to fertile C57BL males or to vasectomized C57BL males. After allogeneic or syngeneic mating the extent of thymic involution on the 10th day of pregnancy and pseudopregnancy was similar. Lumbar lymph node weight was not affected by pseudopregnancy but increased similarly in allogeneic and syngeneic pregnancies. Serum progesterone levels on the 10th day of pseudopregnancy were similar to those of non-pregnant females, and significantly lower than those of pregnant females. On the 4th to 7th days progesterone levels in pseudopregnant animals were equal to those in pregnant animals. Progesterone levels and thymic involution were similar in syngeneically and allogeneically pregnant females. Progesterone levels were negatively correlated with thymus weight but reached significance only when the mating was allogeneic. It is suggested that there is an interaction between progesterone concentrations and the degree of thymic involution during pregnancy.     

Effect of pelvic endometrial implants on overall reproductive functions of female rats

The effects of pelvic endometrial implants on the overall reproductive potential of female rats were investigated. After homologous transplantation in the peritoneum, the ectopic endometrium developed into highly vascularized nodes that gradually increased in mass until the 9th week postsurgery and then plateaued. In the presence of these implants, overall reproductive function was adversely affected. The effect was of greatest magnitude during 50-70 days posttransplantation. As compared with values in corresponding controls, ovulation was reduced by 43% (6 of 14) (p < 0.05), mating rate was reduced by 44% (12 of 27) (p < 0.025), and premature termination of pregnancy occurred in 34% (5 of 15) of rats. Wastage of pregnancy, which included complete termination or reduction of fetal number, occurred during the postimplantation course of gestation. Furthermore, 100% of the rats with transplants failed to respond to the copulomimetic stimulation for the induction of pseudopregnancy (p < 0.01, compared with corresponding controls). However, on exposure to vasectomized males, 46% (6 of 13) of these rats exhibited development of pseudopregnancy (p < 0.05, compared with corresponding group receiving copulomimetic stimulation). Increased rate of mating failure and differential pseudopregnancy rates after copulomimetic and natural cervical stimulation suggest that the rats with endometrial explants possibly had an absence or a short appearance of behavioral estrus. Hormonal assessment during the preovulatory phase showed a tendency toward lower mean levels of preovulatory estradiol and significantly lower LH (p < 0.01) and progesterone (p < 0.01) concentrations. The adversely affected reproductive functions may be a secondary consequence of these altered endocrine milieus.     

Non-intromissive mating stimuli are sufficient to enhance sexual behaviors in ovariectomized female rats

When ovariectomized/adrenalectomized female rats, injected with subthreshold doses of estradiol are given copulatory stimulation by a male rat at half hour intervals, the level of lordosis gradually increases over the course of a few hours. We tested the hypothesis that paracopulatory behaviors (behaviors that occur repetitively prior to and between mounts), also generally considered to be heavily dependent on progesterone, are enhanced by this stimulation as well. We have reported previously that the enhancement of copulatory behavior is dependent to a large extent on intromissive stimulation by the male. In the present study, mating stimulation induced high levels of paracopulatory behaviors, as well as lordosis. Surprisingly, though, and in contrast to previous findings, this increase was seen not only in rats receiving intromissive stimulation, but in those receiving non-intromissive stimulation as well. Furthermore, intromissive stimulation induced high levels of rejection behavior. In a subsequent experiment, experimenter-induced, mechanical stimulation increased only rejection behaviors, not copulatory behavior. The results collectively demonstrate that, under the conditions used in these experiments, non-intromissive stimulation is sufficient for inducing both copulatory and paracopulatory behaviors in estradiol-primed rats. However, under the conditions used in these studies, intromissive stimulation increases rejection behaviors.     

Influence of time of mating and paced copulation on induction of pseudopregnancy in cyclic female rats

The present experiment was designed to examine whether changes occur during the course of behavioural oestrus in the sensitivity of the female to copulatory stimulation and in patterns of sexual behaviour which might influence the likelihood of luteal maintenance. Cyclic female rats were mated on the evening of pro-oestrus (21:00 h) or early on the morning of oestrus (05:00 h) and received either 5 or 10 intromissions from males under conditions which allowed or did not allow pacing of contacts with males to occur. During mating, the levels of sexual receptivity, the timing of sexual mounts from males, and pacing behaviours were measured. The occurrence of pseudopregnancy or pregnancy in each animal was determined by examining vaginal smears for 14 days after mating and by examining the uterus for the presence of fetuses at the end of the experiment. At both mating times, pacing of copulation with males increased the likelihood of prolonged luteal activity. However, females were more likely to become pseudopregnant following non-paced mating at 05:00 h than at 21:00 h the previous evening. Of those females receiving an ejaculation during mating, no difference were seen between groups in the incidence of pregnancy. This change in sensitivity to cervical-vaginal stimulation during oestrus was not associated with changes in sexual receptivity or pacing behaviour. The ability of cervical-vaginal stimulation to induce pseudopregnancy therefore increases toward the end of the period of oestrus, but the behavioural mechanisms which regulate receipt of such stimulation remain constant during that time.     

The effects of mating stimulation on c-Fos immunoreactivity in the female hamster medial amygdala are region and context dependent

During mating in hamsters, both tactile and nontactile sensory stimulation experienced by the female affect sexual behavior and progestational neuroendocrine reflexes. To test the interactions of these types of mating stimulation, c-Fos immunohistochemistry measured brain cellular activity during sexual behavior under conditions that included combinations of tactile and nontactile mating stimulation. Test groups received: (1) mating stimulation from a male, females being either fully mated or mated while wearing a vaginal mask, or (2) experimenter applied manual vaginocervical stimulation (VCS)-with or without males present, or (3) handling similar to VCS but without insertions-with or without males present. Numbers of c-Fos immunoreactive cells were counted in specific subdivisions of the posterior medial amygdala (MeP) and ventromedial hypothalamus (VMH). The medial amygdala dorsal and ventral subdivisions responded differentially to components of mating stimulation. The posterodorsal Me (MePD) cellular activation was greatest during mating conditions that included VCS and/or males present. However, the posteroventral Me (MePV) was sensitive to male exposure and not to VCS. Also, MePV and VMH shell responses mirrored each other, both being primarily sensitive to male exposure. In separate tests, manual VCS induced pseudopregnancy, though the procedure was most effective with additional nontactile stimulation from males present. In summary, contextual cues provided by nontactile male stimulation enhance the effect of vaginocervical and other tactile stimulation on reproductive processes. Furthermore, c-Fos expression in the female hamster medial amygdala is region and context dependent.     

Effects of hysterectomy and uterine decidualization on in vivo levels of prostaglandins in the corpus luteum of adult pseudopregnant rats

A possible role of the uterus in regulating content of luteal prostaglandins (PGs) was investigated. Pseudopregnancy was induced in adult virgin female rats by mating them with vasectomized male rats. On Day 5 of pseudopregnancy, decidualization of the uterus was induced or hysterectomy was performed. As controls, intact pseudopregnant animals with a luteal phase of 13 +/- 1 days were used. Measurements of in vivo tissue levels of PGF2 alpha, PGE2, and 6-keto-PGF1 alpha were performed by RIA after homogenization and extraction procedures in CL of pseudopregnancy and remainder of ovaries on Days 5, 13, and 19. Serum levels of progesterone and 20 alpha-dihydroprogesterone were determined by RIA. In hysterectomized animals, PGF2 alpha levels increased 2.5-fold in corpora lutea on Day 13 compared with levels on Day 5 of pseudopregnancy, but were still lower than in control rats undergoing functional luteolysis on Day 13. Decidual-tissue-bearing rats exhibited low levels of PGF2 alpha on Day 13 of pseudopregnancy. On Day 19, when luteolysis had occurred in decidual-tissue-bearing and hysterectomized rats, as judged by plasma levels of progestins, luteal content of PGF2 alpha was elevated to a similar level as that in control animals undergoing functional luteolysis on Day 13. When data pooled from control, decidual-tissue-bearing and hysterectomized rats were analyzed, a highly significant inverse correlation (r = -0.72, n = 46, p less than 0.001) between luteal PGF2 alpha content and ratio of plasma progestins was found.(ABSTRACT TRUNCATED AT 250 WORDS)     

Age-related changes on marking, marking-like behavior and the scent gland in adult Mongolian gerbils (Meriones unguiculatus).

Marking behavior, marking-like behavior [3], and changes of the scent glands were observed in aged Mongolian gerbils. In Experiment 1, changes in the marking and marking-like behavior with aging were evaluated in adult male and female Mongolian gerbils of an inbred strain aged 6 to 36 months. The frequency of marking behavior in males was significantly higher than females throughout the observation period except at 36 months of age. On the other hand, frequency of marking-like behavior in males, but not in females decreased with aging, significantly. In Experiment 2, changes of the scent gland in adult males and females aged 6 to 36 months were morphologically evaluated. Macroscopic examination revealed an increase in the size length and width of the glands of males aged 12 months and females aged 6 months. Histologically the glands of all the males and females aged 6 months developed moderately or well. Some of the 12-month-old males and females showed acinar atrophy of the glands, and all the females aged 18 months or more had highly atrophied scent glands. From these results, we concluded that there is no relationship between the changes of marking behavior and those of the scent glands in aged male Mongolian gerbils, and assume that marking behavior in aged animals does not have an important meaning as marking. In Experiment 3, marking and marking-like behavior in castrated adult Mongolian gerbils aged 16 weeks were observed. The result showed that marking behavior, not marking-like behavior was inhibited after castration. From these findings, we consider that generally marking behavior in Mongolian gerbils consists of androgen-dependent marking behavior and androgen-independent marking behavior (marking-like behavior).     

不同生活史阶段长爪沙鼠尿标记行为的遗传和空间距离效应

气味标记普遍存在于哺乳动物类群中,它能向同类传达领地占有、社会等级及繁殖状态等多种信息。2014年4月至10月,我们在内蒙古锡林郭勒正蓝旗对长爪沙鼠 (Meriones unguiculatus) 野外种群进行了标志重捕,采用中立场的方式测定了不同社群间同性别长爪沙鼠的尿标记行为。通过建立洞群样地坐标后用勾股弦方法计算群间距离,并用微卫星DNA分子标记估算个体间的亲缘系数,以探讨野生动物标记行为是否受个体间空间和遗传距离的影响。广义线性混合效应分析显示,雄鼠间的尿标记强度在繁殖期和贮食期受空间距离或遗传距离的影响均不显著。繁殖期雌鼠间的尿标记强度受空间或遗传距离的影响亦不明显,但在贮食期,遗传距离对雌鼠间标记的强度有显著影响,且遗传距离与空间距离对标记强度有明显的交互作用。本研究结果暗示亲缘关系和空间距离对长爪沙鼠的尿标记行为具有一定影响,这与长爪沙鼠的繁殖或贮食行为相适应。     

Blockage of induced pseudopregnancy by electrochemical stimulation of the limbic system.

Pseudopregnancy induced by cervical stimulation was inhibited by acute electrochemical stimulation of the corticomedial amygdala or dorsal hippocampus under sodium pentobarbital anesthesia (40 mg/kg) in adult, cyclic female Sprague-Dawley rats. The degree to which pseudopregnancy was blocked depended on temporal conditions of brain stimulation and sodium pentobarbital administration. Pentobarbital alone had a suppressing effect on the incidence of pseudopregnancy, especially when it preceded cervical stimulation. Limbic stimulation before cervical stimulation had a tendency to potentiate the suppression of pseudopregnancy by pentobarbital. After cervical stimulation, hippocampal stimulation tended to inhibit the development of pseudopregnancy, potentiating the pentobarbital suppression, while amygdala stimulation tended to override the pentobarbital blockage of pseudopregnancy. These findings suggest a negative influence of these two limbic structures and pentobarbital on the secretion of prolactin.     

Conditions for establishment of reflex ovulation in light estrous rats.

Continual anovulatory state associating with persistent vaginal cornification (light estrus) was induced by placing 4-day cycling rats under continuous lighting (LL). Uterine cervical stimulation was applied at arbitrary solar hours to light estrous rats showing continual vaginal estrus for more than 2 weeks. The ovulation was induced between 14 and 16 hr after the stimulation dissociating entirely with solar hours. Injection of anti-LHRH serum 5 min after the stimulation but not later than 20 min blocked this ovulation. Ovulation thus induced was always followed by pseudopregnancy with continual leucocytic vaginal smear lasting 10.70 days. The change in concentrations of peripheral serum progesterone during this period was almost similar to that of normal pseudopregnancy except extremely low levels observed at the start and end. Effectiveness of the cervical stimulation for induction of ovulation in light estrous rats was related to not only the duration of light estrus but also the time after transfer to LL, suggesting that the neural mechanism of ovulation in light estrous rats shifted from that of the spontaneous to reflex ovulators due to the extinction of environmental photic cue.     

Sex difference and response to testosterone in gabaergic cells of the medial preoptic nucleus and ventral bed nuclei of the stria terminalis in gerbils

The medial preoptic nucleus (MPN) and ventral bed nuclei of the stria terminalis (BST) are needed to maintain mating in sexually experienced male gerbils and rats. The gerbil ventral BST is also activated with mating, as assessed by Fos expression, as is the medial MPN (MPNm) of both species. In gerbils, many of those mating-activated cells contain glutamic acid decarboxylase (GAD), the enzyme that synthesizes γ-aminobutyric acid (GABA). Some of those cells are projection neurons, but others may release GABA locally. Through actions in the medial preoptic area, GABA inhibits and testosterone (T) promotes male sex behavior. Thus, T may promote mating, in part, by decreasing GAD in MPNm or ventral BST cells. In rats, T increases GAD mRNA in the central MPN (MPNc), where MPN GABAergic cells are densest, but mating behavior does not change in sexually experienced males when the MPNc is ablated. Therefore, this study focused on the MPNm and ventral BST to ask whether their GABAergic cells respond to T or are sexually dimorphic. This was done by visualizing cells immunoreactive (IR) for GAD₆₇, an isoform found primarily in cell bodies, in male and female gerbils and in castrated males with and without T. At both sites, males had more GAD₆₇-IR cells than females, and T decreased GAD₆₇-IR cell numbers in males. Thus, the MPNm and ventral BST have GABAergic cells that are sexually dimorphic and in which T decreases GAD, consistent with local effects of T and GABA on mating.     

Plasma gonadotropins and prolactin in pseudopregnancy in the rat

Radioimmunoassay presented a method of measuring plasma levels of FSH,LH and prolactin in pseudopregnant rats. Plasma prolactin levels doubled 15 minutes following cervical probing (p .01) on the day of estrus. Plasma LH levels were not significantly elevated. Due to the use of ether anesthesia at blood collecting 3 hr before and 15 minutes after stimulation, only 1 of 16 rats developed pseudopregnancy. On Day 4 of pseudopregnancy in rats mated with vasectomized males; plasma LH was lower (p .05) than in normal rats on the first day of diestrus, perhaps due to the suppressive action of ovarian progesterone and some estrogen. FSH was higher than in normal rats (p .05) perhaps due to the lesser sensitivity of FSH to the inhibitory effect of progesterone. Large decidoumata developed by Day 9 in uterine horns traumatized on Day 4 (153 plus or minus 8 mg uterus weight compared to 1510 plus or minus 204 mg). Thus, the corpora remain functional after LH and prolactin are at normal and subnormal levels. On Day 9 plasma prolactin was lower than at Day 1 of diestrus (p .001). Plasma FSH was elevated (p .01). Plasma LH was unchanged. The degree of rise of LH levels 5 days following ovariectomy on Day 4 of psuedopregnancy or on the first day of diestrus was greater in the former group (p .02), perhaps due to rebound of LH from suppression by ovarian steroids. FSH rose equally in both groups. Prolactin remain about the same. Increased prolactin release by the adenohypophysis was briefer than expected.     

Induction of luteal activity and progesterone secretion in the nonpregnant one-humped camel (Camelus dromedarius)

Corpora lutea (n = 20) were detected in 5 one-humped female camels studied during a period of 4 months. Complete mating by a vasectomized male, male introduction into the pen of females without mating, or a progesterone decrease from a previous corpus luteum were followed by a similar progesterone pattern. A maximal plasma concentration of 4.5 +/- 1.5 ng progesterone/ml (2.7-8.8 ng/ml) occurred 8.55 +/- 1.32 days (6-11 days) after the inducing stimulus. Luteal regression, beginning 8.65 +/- 1.18 days after the stimulus, was completed at Day 11.55 +/- 1.05. Morphological development of ovarian structures, detected by rectal palpation, was in synchrony with the progesterone increase, but there was a prolonged period of regression. Females accepted mating up to 7 days after the ovulatory stimulus, when progesterone levels were as high as 3.5 ng/ml. This study establishes the absence of pseudopregnancy in the one-humped female camel, and offers opportunities for improving the management of reproduction. It also shows that ovulatory stimuli other than mating can be effective in these animals.     

Genetic consequences of group living in Mongolian gerbils

Social behavior can shape the local population genetic structure of mammals. Group living can increase pairwise genetic relatedness of mammals at a local level but differentiate the genetic structure at a population level through offspring philopatry and nonrandom mating. Our study aimed to test the hypothesis that social groups of Mongolian gerbils (Meriones unguiculatus) would consist of genetically related individuals due to offspring philopatry and would have distinct genetic structures because of restricted gene flow among social groups and nonrandom mating. We genotyped 327 wild gerbils, live captured from 28 social groups in Inner Mongolia, China, using nine microsatellite loci. The within-group pairwise genetic relatedness coefficient averaged 0.28 ± 0.14 (standard deviation), whereas the average pairwise genetic relatedness coefficient of the whole gerbil population was 0.0 ± 0.2. Additionally, the value of the global F statistic (F(st)) was 0.21, suggesting a substantial genetic differentiation among social groups of Mongolian gerbils. The Bayesian clustering divided the 327 gerbils into 23 distinct genetic clusters. Therefore, our results show that high within-group genetic relatedness and among-group genetic differentiation are the genetic consequences of group living in social mammals because of restricted gene flow, female philopatry, and nonrandom mating within social groups.     

Defects in reproductive functions in PACAP-deficient female mice

Pituitary adenylate cyclase-activating polypeptide (PACAP) is a highly conserved neuropeptide and widely expressed in both brain and peripheral tissues, including several reproductive organs (e.g., testis and ovary). PACAP stimulates syntheses of several sexual hormones and steroids, suggesting it has possible roles in reproductive function. In this study, the role of PACAP in female reproductive functions such as fertility, mating behavior and maternal behaviors were investigated by using mice lacking PACAP (PACAP(-/-)). PACAP(-/-) females showed reduced fertility (the number of parturitions relative to the number of pairings). Mating experiments using vasectomized males revealed that mating frequency and its intervals in some PACAP(-/-) females were quite different (zero to eight times/4 weeks), whereas the frequency was relatively constant (two to three times/4 weeks) in wild-type females. In PACAP(-/-) females, maternal crouching behavior tended to decrease compared to wild-type females, although the influence of litter size on maternal behavior needs to be considered. These data suggest a role for endogenous PACAP in female reproductive processes.     

Paternal behavior in the Mongolian gerbil (Meriones unguiculatus): Estrogenic and androgenic regulation

Here, we analyzed the effects of testosterone (T) and its metabolites, estradiol (E₂) and dihydrotestosterone (DHT), on the onset of paternal behavior in virgin male Mongolian gerbils (Meriones unguiculatus). We hypothesized that T and E₂, but not DHT, would facilitate the onset of paternal behavior. Seventy males displaying aggression toward pups were selected through a paternal behavior screening test. Forty males were bilaterally castrated. Of them, 10 were implanted with T, 10 with E₂, and 10 with DHT, and 10 received no treatment. Another 30 males underwent a sham procedure. In these gerbils, T, E₂ and DHT were measured to obtain the basal levels of these hormones. After treatment, the paternal behavior test was conducted again. Blood samples were obtained immediately after the administration of the test for the quantification of T, E₂ and DHT by radioimmunoassay. Surprisingly, 100% of the males that received T, E₂ and DHT implants stopped being aggressive and became paternal. Castrated and sham-operated males displayed no changes in their aggressive behaviors. This is the first report that T and its metabolites are involved in neuroendocrine mechanisms that inhibit aggression toward pups and facilitate paternal behavior in virgin male Mongolian gerbils. In addition, this is the first report of regulation of paternal behavior in a rodent by estrogenic and androgenic pathways.     

长爪沙鼠血清雌性激素的比较研究

目的测定长爪沙鼠、NIH小鼠和SD大鼠的血清雌二醇(E2)和孕酮(P)水平,为研究长爪沙鼠的生殖和胚胎工程提供基础资料。方法用放射免疫分析法测定上述动物生产前后血清雌二醇(E2)和孕酮(P)水平,进行统计学处理和分析。结果不同阶段雌性沙鼠E2值差异无显著性(P>0.05),P值差异有显著性(P<0.01);E2值比较,处女期沙鼠与同期NIH和SD差异有显著性(P<0.01),而在经产期的动物间水平接近(P>0.05);P值比较,处女期沙鼠与NIH接近,与SD差异有显著性(P<0.01);而经产期沙鼠在三种动物中是最高的(P<0.01)。结论长爪沙鼠血清E2、P的含量具种属特异性,并随动物的生理发育时期而变化。     

食物诱导长爪沙鼠集群贮食的野外实验设计及其在社群研究中的应用

长爪沙鼠为典型的群居性鼠类。在自然条件下该鼠的集群贮食活动随时可为人工投放的种子食物所导引。本文依据组群个体合作贮食的习性,并结合剪趾和染毛双重标记的重捕跟踪方法设计了着重观测社群结构、群内个体序位、领域边界及相关社群行为的野外实验。据2(X)2年5月对4个相邻洞群的观测结果分析表明,应用此法便于在复杂的社群环境中直接确认组群成员及其属性,亦便于分辨同一组群或不同组群成员之间发生的行为事件及相关过程,且能在重复实验中得以验证。为在野外条件下定量研究长爪沙鼠社群行为提供了一种便捷可靠的观测方法。