Mario Cárdenas

Endocrine changes in male stumptailed macaques (Macaca arctoides) as a response to odor stimulation with vaginal secretions.

In mammalian species, social chemosignals are important in modulating endocrine reproductive functions. In nonhuman primates, previous studies have described a high frequency of mounting behavior by females in the follicular and periovulatory phases of the menstrual cycle. Stumptailed macaque females do not signal receptivity by means of sexual swellings, as do others macaques, therefore providing a good model in which to study chemical signaling of reproductive status. We exposed anesthetized stumptailed males to vaginal secretions of either late follicular or menses phase or to saline solution to determine the endocrine changes promoting male sexual behavior. In males exposed to follicular secretions, plasma testosterone concentrations were sustained up to 120 min after exposure. Such an effect was not observed in animals exposed to menses or saline odor sources. A luteinizing hormone surge, occurring 30 minutes after exposure to late follicular phase secretion swabs, preceded this sustained testosterone effect. The fact that late follicular scents induce sustained testosterone concentrations provides support to the idea that stumptailed males draw information concerning female reproductive status from the females vaginal odor.

Unilateral sectioning of the superior ovarian nerve of rats with polycystic ovarian syndrome restores ovulation in the innervated ovary

The present study tested the hypothesis that if polycystic ovary syndrome (PCOS) results from activating the noradrenergic outflow to the ovary, unilaterally sectioning the superior ovarian nerve (SON) will result in ovulation by the denervated ovary, and the restoration of progesterone (P4), testosterone (T) and estradiol (E2) normal serum level. A single 2 mg dose of estradiol valerate (EV) to adult rats results in the development of a syndrome similar to the human PCOS. Ten-day old rats were injected with EV or vehicle solution (Vh) and were submitted to sham surgery, unilateral or bilateral sectioning of the SON at 24-days of age. The animals were sacrificed at 90 to 92 days of age, when they presented vaginal estrus preceded by a pro-estrus smear. In EV-treated animals, unilateral sectioning of the SON restored ovulation by the innervated ovary and unilateral or bilateral sectioning of the SON normalized testosterone and estradiol levels. These results suggest that aside from an increase in ovarian noradrenergic tone in the ovaries, in the pathogenesis of the PCOS participate other neural influences arriving to the ovaries via the SON, regulating spontaneous ovulation. Changes in P4, T and E2 serum levels induced by EV treatment seem to be controlled by neural signals arising from the abdominal wall and other signals arriving to the ovaries through the SON, and presents asymmetry.

Effects on steroid hormones secretion resulting from the acute stimulation of sectioning the superior ovarian nerve to pre-pubertal rats

In the adult rat, neural signals arriving to the ovary via the superior ovarian nerve (SON) modulate progesterone (P4), testosterone (T) and estradiol (E2) secretion. The aims of the present study were to analyze if the SON in the pre-pubertal rat also modulates ovarian hormone secretion and the release of follicle stimulating hormone (FSH) and luteinizing (LH) hormone. P4, T, E2, FSH and LH serum levels were measured 30 or 60 minutes after sectioning the SON of pre-pubertal female rats. Our results indicate that the effects on hormone levels resulting from unilaterally or bilaterally sectioning the SON depends on the analyzed hormone, and the time lapse between surgery and autopsy, and that the treatment yielded asymmetric results. The results also suggest that in the pre-pubertal rat the neural signals arriving to the ovaries via the SON regulate the enzymes participating in P4, T and E2 synthesis in a non-parallel way, indicating that the mechanisms regulating the synthesis of each hormone are not regulated by the same signals. Also, that the changes in the steroids hormones are not explained exclusively by the modifications in gonadotropins secretion. The observed differences in hormone levels between rats sacrificed 30 and 60 min after surgery reflect the onset of the compensatory systems regulating hormones secretion.

Effects of acute unilateral ovariectomy to pre-pubertal rats on steroid hormones secretion and compensatory ovarian responses.

In the present study we analyzed the existence of asymmetry in the secretion of steroid hormones in pre-pubertal female rats treated with unilateral ovariectomy (ULO) or unilateral perforation of the abdominal wall (sham-surgery). Treated rats were sacrificed at different times after surgery. Since sham-surgery had an apparent effect on the age of first vaginal estrous (FVE) and serum levels hormone, the results of the sham surgery groups were used to assess the effects of their respective surgery treatment groups. On the day of FVE, compensatory ovulation (CO) and compensatory ovarian hypertrophy (COH) were similar in animals with ULO, regardless of the ovary remaining in situ. In ULO treated animals, progesterone (P4) levels were higher than in animals with sham-surgery one hour after treatment but lower in rats sacrificed at FEV. Left-ULO resulted in lower testosterone (T) concentration 48 and 72 hours after surgery. In rats with Right-ULO lower T concentrations were observed in rats sacrificed one or 72 hours after surgery, and at FVE. ULO (left or right) resulted in lower estradiol (E2) concentrations one or 72 hours after treatment. In rats with Left-ULO, E2 levels were higher 48 hours after surgery and at FVE. Left-ULO resulted in higher levels of follicle stimulating hormone (FSH) five hours after surgery and at FVE. FSH levels were higher in rats with Right-ULO sacrificed on FVE. The present results suggest that in the pre-pubertal rat both ovaries have similar capacities to secrete P4, and that the right ovary has a higher capacity to secrete E2. Taken together, the present results support the idea that the effects of ULO result from the decrease in glandular tissue and changes in the neural information arising from the ovary.

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.

Seasonality of LH, testosterone and sperm parameters in spider monkey males (Ateles geoffroyi)

There are no reported data on hormonal fluctuations in black‐handed spider monkey males. On previous research about the reproductive physiology of this monkey we have found that during the dry season females show ovulatory estrogen peaks and males present the best quality semen. As part of an ongoing research, in this study we assessed seasonal variations in the concentration of serum luteinizing hormone (LH) and testosterone (T) in three adult spider monkey males to corroborate the seasonal reproductive synchrony. At the same time sperm count and motility were evaluated to search for any correlation between those sperm parameters and hormonal concentrations. We took blood and semen samples (by electroejaculation) of anesthetized males throughout the rainy (June–September) and dry (October–May) months. Our results revealed that T and LH were higher throughout the dry season and there was a significant correlation between T concentration and sperm count. Although higher during the dry season, sperm motility tended to correlate with testosterone and LH levels. These results demonstrated that black‐handed spider monkeys have a tendency to show a seasonal pattern of reproduction being the dry season the most likely time to achieve fertilization. Am. J. Primatol. 71:427–431, 2009.

Effects of unilaterally microinjecting ethanol in the preoptic-anterior hypothalamic areas of rats on ovulation.

BACKGROUND Intragastric or intraperitoneal ethanol (EtOH) treatment inhibits reproductive functions in females and male rats. The area of the hypothalamus where these effects take place is unknown. As the participations of the preoptic-anterior hypothalamic area (POA-AHA) in regulating ovulation is asymmetric, this study aims to analyze the effects on 17β-estradiol(E2 ), progesterone, luteinizing hormone (LH), follicle-stimulating hormone (FSH) serum levels, the messenger ribonucleic acid (mRNA) expression of estrogen receptor alpha (ERα) and beta (ERβ), and ovulation resulting from unilaterally microinjecting water or an EtOH solution into either side of the POA-AHA. METHODS The treatment consisted of microinjecting a 8.6 μM EtOH solution into either side of the POA-AHA. The study was performed on groups of adult cyclic rats at 09.00 hours on diestrus-1, and sacrificed on diestrus-2 at 13.00, on proestrus at 09.00 or 17.00 or on estrus at 09.00 hours. Ovulation rates were assessed in rats sacrificed on estrus. Hormonal serum levels were measured using radioimmunoassay, and as a function of ERα and ERβ mRNA expression in each side of the POA-AHA by reverse transcriptase polymerase chain reaction. RESULTS EtOH treatment blocked ovulation and the preovulatory release of LH, and lowered E2 levels. Irrespective of the treated POA-AHA side, ERα mRNA expression was consistently lower in the left POA-AHA and higher on the right. EtOH treatment in the left POA-AHA decreased FSH serum levels and lowered ERβ mRNA expression. In turn, EtOH treatment on the right POA-AHA resulted in higher FSH levels and ERβ mRNA expression. CONCLUSIONS The present results show that EtOH blocks the preovulatory surge of LH on the POA-AHA. The effects of EtOH treatment of preovulatory FSH surge on the POA-AHA are asymmetric (stimulative on the right and inhibiting in the left). The effects of EtOH treatment on preovulatory LH and FSH surge could be explained by the inhibition of ERα and ERβ mRNA expression, respectively.

An increase in estradiol facilitates the onset of paternal behavior in the dwarf hamster (Phodopus campbelli)

ABSTRACT In the dwarf hamster (Phodopus campbelli), activational effects of testosterone (T) and estradiol (E2) in the regulation of paternal behavior have been repeatedly rejected because peripheral concentrations of E2 do not change across the reproductive cycle of males. Further, castration no affected paternal behavior despite that both T and E2 concentrations decreased significantly. However, the role of these hormones has not been evaluated in models of castration and hormonal replacement in virgin males. Here, we analysed the effects of E2 and T in paternal behavior in virgin male dwarf hamster (Phodopus campbelli). Thirty paternal (PAT) males were bilaterally castrated; of them, 10 were implanted with T, 10 with E2 and 10 males received no treatment. Other 10 PAT males underwent sham‐castration. Seventeen aggressive (AGG) males were also bilaterally castrated; of these, 10 AGG received E2 replacement, 7 were not treated. Other 7 AGG males were submitted to sham‐castration. Following treatments, paternal behavior tests were conducted again. T and E2 levels in plasma were quantified by radioimmunoassay (RIA). The results showed that the treatments did not affect the paternal behavior of males that were initially paternal. Neither castration nor sham‐castration surgery affected the behavior of AGG males. However, when these males were treated with E2 and the concentrations of this hormone increase significantly they became paternal. Our data suggest that an increase in E2 levels shifted infanticidal behavior to paternal behavior in dwarf hamster. HighlightsAn increase in E2 facilitates transitions from aggressive males to paternal.Changes in T and E2 had no effect in paternal activities in paternal males from the beginning.Low concentrations of E2 may be the cause of infanticidal behavior.

Neuronal activation associated with paternal and aversive interactions toward pups in the Mongolian gerbils ( Meriones unguiculatus )

ABSTRACT Approach/avoid model is used to analyze the neural regulation of maternal behavior in the laboratory rat. This model proposes that the medial preoptic area (mPOA) and bed nucleus of stria terminalis (BNST) are brain regions involved in facilitating mechanisms. By contrast, anterior hypothalamic nucleus (AHN), ventromedial hypothalamic nucleus (VMH), and periaqueductal gray participate in the inhibiting mechanisms of neural regulation of maternal behavior. We hypothesized that there are also facilitating and inhibiting mechanisms in the neural regulation of paternal behavior. Here, we determined which neural areas are activated during paternal and aversive interactions with pups in the Mongolian gerbils (Meriones unguiculatus). By testing paternal behavior, we selected 40 males aggressive toward pups and 20 paternal males. These males were organized into six groups of 10 animals in each group: aggressive males that interacted with pups (AGG‐pups) or candy (AGG‐candy), paternal males that interacted with pups (PAT‐pups) or candy (PAT‐candy), and males with testosterone (T)‐induced paternal behavior that interacted with pups (IPAT‐pups) or candy (IPAT‐candy). After interacting with pups or candy, the brains were extracted and analyzed for immunoreactivity (ir) with c‐fos. Males that interacted with pups had significantly higher c‐fos‐ir in the mPOA/BNST than males that interacted with candy. Males that displayed aggression had significantly higher c‐fos‐ir in the AHN, VMH, and periaqueductal gray than aggressive males that interacted with candy. These results suggest that in the neural regulation of paternal behavior in the Mongolian gerbil underlie positive and negative mechanisms as occurs in maternal behavior. HIGHLIGHTSAnalyzing the neural regulation of paternal behavior with the approach/avoid model is novel.mPOA/BNST are part of the positive mechanism of neural regulation of paternal behavior.AHN/VMH are part of the negative mechanism of neural regulation of paternal behavior.A decrease in the testosterone level was observed in aggressive males that interacted with the pups.Neural regulation of paternal behavior is according to the approach/avoid model.

Inhibition of serotonin reuptake in the prepubertal rat ovary by fluoxetine and effects on ovarian functions.

Fluoxetine (FLX), a selective serotonin reuptake inhibitor is an antidepressant in the treatment of mood disorders. Its impact on reproductive processes is incompletely known. The present study analyzed the reproductive effects of FLX in prepubertal female rats. Two experiments were conducted. First (acute administration), 30-day-old female rats were injected intraperitoneally with 5mg/kg of fluoxetine-hydrochloride, and were terminated 24, 48 or 72h after the treatment. Second (subchronic administration), FLX was injected on days 30-33 of age, and the animals were terminated the day of first estrus. In acute treatment estradiol concentration increased to 72h. In subchronic treatment increased serotonin concentration in ovaries and decreased the number of ova shed. An increase in number of atretic follicles and oocyte fragmentation was observed in these animals. The results suggest that FLX acts on the ovary or hypothalamus-pituitary axis resulting in modifications of the follicular development and ovulation.