Endocrine correlates of sexual behavior in the Mohor gazelle (Gazella dama mhorr)

In this study, we quantitatively examined male sexual behavior in relation to fecal estrogen and progesterone concentrations in female Mohor gazelles. We investigated the hypothesis that, during natural mating, males detect cues relating to the potential for successful conception and pregnancy. Time series analysis revealed that males could detect the approach of estrus 2-3 days before female fecal estrogens and estrogen/progestagen (E/P) ratio reached their peak values. Males also paid closer attention to those females excreting higher fecal estrogen concentrations. Mounting and copulation frequencies were positively correlated with both peri-ovulatory fecal estrogen concentrations, and the frequency of pre-copulatory courtship behaviors. These data suggested that males invest their reproductive effort selectively by mating the most fertile females, assuming that estrogen is a valid index of fertility. This assumption was investigated by examining sequential phases of the reproductive cycle for evidence that oocytes and follicles produced in a more estrogenic environment would lead to the formation of the most competent corpora lutea, thereby maximizing the chance of sustaining pregnancy. Associations between sexual behavior and hormone excretion support the hypothesis that males may use this mechanism to assess female fertility.     

Role of growth hormone and prolactin in the control of reproduction: what are we learning from transgenic and knock-out animals?

Growth hormone (GH), insulin-like growth factor (IGF-I), and prolactin (PRL) can influence various aspects of reproductive functions in both females and males. However, the physiological role of PRL and the GH-IGF-I axis in the control of reproduction has been difficult to define, and the recent availability of knock-out (KO) animals allows re-examination of this issue. PRL-receptor (R)-KO and PRL-KO females are sterile because of luteal failure. In addition, these mice have severe deficits in the development of oocytes and early embryos. However, male fertility is not affected in the PRL-KO and in most of the PRL-R-KO animals. IGF-KO animals have an infantile reproductive system and are sterile. GH-R-KO mice can reproduce, but their breeding performance is reduced, particularly in females. These data indicate that IGF-I signaling is required for normal reproductive development and confirm the requirement for PRL for fertility in the female mouse. GH resistance leads to quantitative deficits in reproductive development and functions, but does not preclude fertility in either sex. We suspect that PRL and the GH-IGF-I axis provide partially overlapping (redundant) regulatory inputs to the hypothalamic-pituitary-gonadal axis, and consequently, targeted disruption of either signaling pathway has relatively mild consequences on many functions related to reproduction. Overexpression of heterologous or homologous GH in transgenic animals can lead to severe reproductive deficits, including female sterility in some of the lines. Studies in GH transgenics should allow the identification of mechanisms that mediate the effects of chronic overexposure to GH on reproduction.     

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.     

Background matters: the effects of estrogen receptor alpha gene disruption on male sexual behavior are modified by background strain

One approach to study interactions between behavior and genetics is to use inbred mice with different genetic backgrounds. To examine the effect of background on a specific gene, we conducted a series of experiments with a well-characterized knockout (KO) mouse, the estrogen receptor alpha KO (ERalphaKO). The ERalphaKO mouse has so far been examined in one inbred line, C57BL/6J. Here, we examined the behavior of ERalphaKO mice within three different backgrounds mixed with C57BL/6J; DBA/2J, BALB/c, and A/J. First, we assessed masculine sexual behavior in both intact male and testosterone-treated female offspring. More ERalphaKO males in the DBA/2J (5/12) and BALB/c (5/13) backcrosses displayed intromissions and many ejaculated as compared with males in a C57BL/6J and A/J mixed background. Many fewer ERalphaKO females than males displayed masculine sexual behavior in any of the three hybrid crosses. We assessed fertility in males from the C57BL/6J by DBA/2J cross and found that one of 12 ERalphaKO males sired a litter. Several other characteristics of sexual behavior and physiology were unaffected by genetic background in ERalphaKO mice. Our data suggest that genetic background has dramatic effects on male sexual behavior and its dependence on the ERalpha gene.     

The influence of female age on male mating preference and reproductive success in cabbage beetle,Colaphellus bowringi

The influence of female age on male mating preference and reproductive success has been studied using a promiscuous cabbage beetle, Colaphellus bowringi Baly （Coleoptera： Chrysomelidae）. In a simultaneous choice test, middle-aged females had significantly greater mating success than young and old females. In single pair trials, when paired with middle-aged virgin males, middle-aged females mated faster, copulated longer, and had greater fecundity and fertility than young or old females, while the longevity of males was not significantly affected by female age. This study on C. bowringi suggests that middle-aged females are more receptive to mating, which can result in the highest male reproductive success.     

Ultrasonic vocalizations induced by sex and amphetamine in M2, M4, M5 muscarinic and D2 dopamine receptor knockout mice

Adult mice communicate by emitting ultrasonic vocalizations (USVs) during the appetitive phases of sexual behavior. However, little is known about the genes important in controlling call production. Here, we study the induction and regulation of USVs in muscarinic and dopaminergic receptor knockout (KO) mice as well as wild-type controls during sexual behavior. Female mouse urine, but not female rat or human urine, induced USVs in male mice, whereas male urine did not induce USVs in females. Direct contact of males with females is required for eliciting high level of USVs in males. USVs (25 to120 kHz) were emitted only by males, suggesting positive state; however human-audible squeaks were produced only by females, implying negative state during male-female pairing. USVs were divided into flat and frequency-modulated calls. Male USVs often changed from continuous to broken frequency-modulated calls after initiation of mounting. In M2 KO mice, USVs were lost in about 70-80% of the mice, correlating with a loss of sexual interaction. In M5 KO mice, mean USVs were reduced by almost 80% even though sexual interaction was vigorous. In D2 KOs, the duration of USVs was extended by 20%. In M4 KOs, no significant differences were observed. Amphetamine dose-dependently induced USVs in wild-type males (most at 0.5 mg/kg i.p.), but did not elicit USVs in M5 KO or female mice. These studies suggest that M2 and M5 muscarinic receptors are needed for male USV production during male-female interactions, likely via their roles in dopamine activation. These findings are important for the understanding of the neural substrates for positive affect.     

Prostaglandin-F2 alpha stimulates reproductive behavior of female paradise fish (Macropodus opercularis)

The function of prostaglandin-F2 alpha (PGF2 alpha) in reproductive behavior of adult female paradise fish (Macropodus opercularis) was evaluated. In experiment 1, females were allowed to spawn normally with a male and were left with that male until the following day when testing occurred. Subjects were then randomly assigned to one of three groups: PGF2 alpha (N = 8), vehicle control (N = 3), or a handling control (N = 3); for testing, females were removed, injected, then returned to the same tank. Seven of the eight subjects injected with 500 ng PGF2 alpha initiated complete reproductive behavior following injection. Vehicle and handling control subjects did not show any female reproductive behaviors. Prostaglandin-induced increases in sexual behaviors were seen from 15 to 75 min after the injection and were maximal approximately 45 min postinjection. In experiment 2, mature but unspawned (and presumably sexually naive) females were placed for 1 hr with males which were actively nestbuilding and parenting, following which subjects were either injected with PGF2 alpha (N = 6) or given a vehicle injection (N = 6). Experimental females did not behave differently from control females. Neither groups was observed to approach the male or initiate spawning behaviors following treatment; all subjects in both groups spawned on the following day. In experiment 3, sexually mature females were given extended familiarity with the male and the test tank. Pairs which had not spawned after 6 days (unspawned) were randomly assigned to experimental (N = 6) or control (N = 5) groups. Pairs which had spawned 1 day previously (N = 6) formed a "prespawned" comparison group. Females from the experimental and prespawned groups were injected with PGF2 alpha (500 ng), while the control females received a vehicle injection. Half of the "prespawned" females performed spawning acts while none of the unspawned or control females did so. Detailed behavioral analyses showed little or no effect of PG treatment, despite greater familiarity with the male. In experiment 4, females were placed with males and observed until the early signs of spawning, at which time they were injected with indomethacin (a PG synthesis inhibitor) or vehicle. The behavior of females treated with 35 micrograms (N = 3) or 70 micrograms (N = 6) of indomethacin was unaffected by indomethacin and largely indistinguishable from controls (N = 6). In summary, exogenous prostaglandins reinstate sexual behavior in female paradise fish. However, responsiveness to prostaglandins is influenced by prior sexual experience.(ABSTRACT TRUNCATED AT 400 WORDS)     

Reproductive disruption in adult female and male rats prenatally exposed to mesquite pod extract or daidzein

Phytoestrogens are considered to be endocrine disruptors, since they can alter the endocrine system, thus disturbing many reproductive events. The intake of diets containing a high content of phytoestrogens has increased worldwide in human populations and in domestic animals. Phytoestrogens in maternal blood can pass through the placenta to the fetus in high amounts and can have long-term organizational effects. Mesquite (Prosopis sp) is a leguminous plant widely used to feed several livestock species, and is also used in the human diet. In this study we assessed the effects of exposure to mesquite pod extract during the periconception and pregnancy periods on the reproduction of male and female descendants. The females of three experimental groups received one of the following treatments: 1) vehicle injection; 2) mesquite pod extract or 3) the isoflavone daidzein during the periconception and pregnancy periods. Estrous cyclicity, sexual behavior and hormones, as well as uterine and vaginal epithelia were evaluated in the female descendants. In the males, sexual behavior and hormones, apoptosis in testicular cells and sperm quality were evaluated. In females the following was observed: alterations in estrous cycles, decreased sexual behavior, estradiol and progesterone levels, increased uterine and vaginal epithelia. In males, we observed a decrease in sexual behavior, testosterone and sperm quality, and apoptosis increased in testicular cells. All these effects were similar to those caused by daidzein. These results indicate that prenatal exposure to mesquite pod extract or daidzein, administered to females before and during pregnancy, can disrupt normal organizational-activational programming of reproductive physiology in female and male descendants.     

Fmr1基因敲除对小鼠生理发育和繁殖性能的影响

目的对清洁级FVB.KO和FVB的生理发育指标和繁殖性能进行分析比较,探讨Fmr1基因敲除对小鼠生理发育和繁殖性能的影响。方法挑选10周龄清洁级FVB.KO和FVB各20只（雌雄各半）,采取1∶1同居,全部同胞兄妹近交繁殖,测定新生仔鼠生理发育指标和品系繁殖性能。结果 FVB.KO在耳廓分离、体毛长出、门齿萌发、眼睑开裂等生理发育指标上和FVB比较接近,在平均每窝产仔数和离乳率等方面偏低,在雌雄比例上有显著差异。结论 Fmr1基因敲除对小鼠生理发育和繁殖性能影响较小,对后代雌雄比例可能有一定的影响。     

Cellular and morphological traits of oocytes retrieved from aging mice after exogenous ovarian stimulation.

The present study aims to shed light on the origin of abnormal oocytes ovulated by aged females. In order to reach this goal, cellular and morphological traits of ovulated oocytes from hybrid (C57Bl/6JIco female x CBA/JIco male) female mice retrieved after exogenous ovarian stimulation at the age of 12, 40-42, 50-52, or 57-62 wk were analyzed. Aging of female mice was associated with 1) decreased number of ovulated oocytes; 2) increased percentage of cumulus-free oocytes; 3) raised percentage of oocytes with intracellular mitochondrial aggregates; 4) reduced percentage of oocytes displaying a normal distribution of chromosomes in the metaphase-II plate; 5) increased percentage of normal oocytes exhibiting a DNA-containing polar body (PB); 6) higher percentage of oocytes with chromosome scattering; 7) increased percentage of chromosome-scattered oocytes without a DNA-containing PB and with intracytoplasmic mitochondrial aggregates; 8) raised percentage of oocytes exhibiting chromosome decondensation; 9) lower percentage of chromosome-decondensed oocytes lacking both a DNA-containing PB and intracytoplasmic mitochondrial aggregates; 10) increased percentage of abnormal/degenerated oocytes; 11) reduced percentage of abnormal/degenerated oocytes displaying cellular fragmentation; and 12) higher percentage of abnormal/degenerated oocytes with mitochondrial aggregates exhibiting no nuclear/chromosomal DNA fluorescence, cellular fragmentation, milky or dark cytoplasm, or cellular remains enclosed by the zona pellucida. Although several studies suggest aging females may ovulate aged or overripened oocytes, these data support the hypothesis that old females ovulate an increased percentage of atretic/apoptotic oocytes coming from rescued follicles that would have become atretic earlier in life.     

Effects of reproductive aging and postovulatory aging on the maintenance of biological competence after oocyte vitrification: insights from the mouse model

Cryopreservation of female reproductive cells allows preservation of fertility and provides materials for research. Although freezing protocols have been optimized, and there is a high survival rate after thawing, the in vitro fertilization (IVF) pregnancy rate is still lower in cycles with cryopreserved oocytes, thus highlighting the importance of identifying intrinsic limiting factors characterizing the cells at time of freezing. The aim of the present study is to investigate in the mouse model the impact of reproductive aging and postovulatory aging on oocyte biological competence after vitrification. Metaphase II oocytes were vitrified soon after retrieval from young and reproductively old mice. Part of the oocytes from young animals was vitrified after 6 h incubation (in vitro aged oocytes). All classes of oocytes showed similar survival rate after vitrification. Moreover, vitrification did not alter chromosomal organization in young cells, whereas in vitro aged and old oocytes presented an increase of slightly aberrant metaphase configurations. Compared to fresh young oocytes, in vitro aged and old oocytes showed increased ROS levels which remained unchanged after vitrification. By contrast, cryopreservation significantly increased ROS production in young oocytes. Both the aging processes negatively impacted oocyte ability to undergo pronucleus formation and first cleavage after vitrification by stimulating cellular fragmentation. These results could be helpful for establishing the correct time table for cryopreservation in the laboratory routine and improving its application in reproductively old females. Moreover, our observations highlight the importance of oxidative stress protection during vitrification procedures.     

Male mice ultrasonic vocalizations enhance female sexual approach and hypothalamic kisspeptin neuron activity

Vocal communication in animals is important for ensuring reproductive success. Male mice emit song-like “ultrasonic vocalizations (USVs)” when they encounter female mice, and females show approach to the USVs. However, it is unclear whether USVs of male mice trigger female behavioral and endocrine responses in reproduction. In this study, we first investigated the relationship between the number of deliveries in breeding pairs for 4 months and USVs syllables emitted from those paired males during 3 min of sexual encounter with unfamiliar female mice. There was a positive correlation between these two indices, which suggests that breeding pairs in which males could emit USVs more frequently had more offspring. Further, we examined the effect of USVs of male mice on female sexual behavior. Female mice showed more approach behavior towards vocalizing males than devocalized males. Finally, to determine whether USVs of male mice could activate the neural system governing reproductive function in female mice, the activation of kisspeptin neurons, key neurons to drive gonadotropin-releasing hormone neurons in the hypothalamus, was examined using dual-label immunocytochemistry with cAMP response element-binding protein phosphorylation (pCREB). In the arcuate nucleus (Arc), the number of kisspeptin neurons expressing pCREB significantly increased after exposure to USVs of male as compared with noise exposure group. In conclusion, our results suggest that USVs of male mice promote fertility in female mice by activating both their approaching behavior and central kisspeptin neurons.     

Reproductive potential of male silver foxes Vulpes vulpes after long selection for the domesticated behavior type

Age peculiarities of reproductive activity and fertility during the reproductive season as well as the hormonal and the testis spermatogenic function were studied in silver foxes Vulpes vulpes from the population selected for a long time for domesticated behavior. It has been established that during the first reproductive season the male sexually activity (the coupled females’ number) was reduced as compared with subsequent seasons, and about one third of the males were not coupled at all with females; however, the young (one-year old) males did not differ from adults by the testis weight, ejaculate spermatozoid number, and the fraction of abnormal spermatozoids. Comparison of indexes of spermatogenesis and the testicular hormonal function in the young males with a decreased and “normal” sexual activity did not reveal any statistically significant differences. It is suggested that the decreased (or absent) sexual activity in young silver fox males can be a response of the population to the human-created environment.     

幼小鼠对种内几种化学信息的识别和反应

50年代以来,国际上相继开展了一些关于化学信息在小家鼠繁殖中作用的研究,并在实验室中揭示了一些反应:如集群雌鼠气味使雌鼠性周期延长的Lee-Boot效应,雌鼠气味能解除集群雌鼠对性周期的抑制、并促使它们同期发情的Whitten效应,以及陌生雄鼠气味中断妊娠的Bruce效应等等。但是,在成鼠间起作用的几种化学信息,是否能被幼鼠感受,对幼鼠的发育和性成熟是否产生影响,还是一个未知的问题。 影响小鼠发育的因素很多,除环境因子外,近来还出现了有关社群因素中外激素对小鼠性成熟影响的报道,发现雄鼠或给予雄激素的雌鼠有促进幼雌鼠性成熟的作用,范志勤发现父本气味和陌生雄鼠气味对幼雌鼠性成熟均有促进作用。但是,目前关于种内雌鼠气味、集群雌鼠气味、幼鼠气味影响性成熟的研究尚不多见。有关各种气味对性成熟效应的比较的报道亦少。本文的目的在于通过实验方法,阐明种内的几种化学信息,诸如父本、陌生雄性、雌性、集群雌性、幼雌等几种气味,对幼雌小鼠性成熟的影响,并据此了解幼鼠识别这几种信息的能力。     

Female sexual behavior in hypogonadal mice with GnRH-containing brain grafts

Hypogonadal female mice respond to GnRH-containing fetal preoptic area (POA) implants in the third ventricle with vaginal opening and persistent vaginal estrus, ovarian, and uterine development and increased gonadotropin secretion. When these females are mated with normal males, reflex ovulation results in pregnancy. In the present study, POA implants derived from neonatal pups, whether male or female, were also capable of supporting reproductive development in the hypogonadal female mice. Evaluation of female sexual behavior in the mice with grafts showed that these mice responded to normal males with comparable levels of lordosis as are seen in normal female mice on the proestrous days of their cycles.