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Cellular and Molecular Biology... Aug 2018Apelin, which is a new hormone, is secreted especially in the brain by hypothalamus as well as by many other organs like the stomach, fat tissue, and the heart. For...
Apelin, which is a new hormone, is secreted especially in the brain by hypothalamus as well as by many other organs like the stomach, fat tissue, and the heart. For apelin, whose possible effects on many bodily functions like the endocrine system, cardiovascular system and metabolic activities are still under investigation, the reproductive system is also an important target area. The purpose of the present study was to investigate the effects of plasma apelin levels in rats that were in diestrus, pregnancy and lactation periods, and to examine its possible effects on myometrium contractions of pregnant rats and non-pregnant rats that were in diestrus period. The plasma apelin concentrations in female adult Wistar rats were determined with the ELISA method in the diestrus period, and on the 12th, 18th, and 21st days of the pregnancy, and on the 2nd and 10th days of lactation (n=7 for each group). In addition, the effect of apelin at 0.01, 0.1, 1 and 10 μM doses on isometric contractions in the rat uterus on the 21st day of pregnancy and in diestrus period was tested by using isolated organ bath. This protocol was repeated under conditions that were pre-treated with protein kinase C inhibitor in calcium-free medium, and the possible effect of apelin on contractions and the mechanisms that might mediate this effect were investigated. When plasma apelin levels were compared with the rats in diestrus period, the apelin concentrations in the 21-day pregnancy group were high at a significant level (p<0.05); and low at a significant level in the 2-day lactation group (p<0.05). In myometrium contraction trials, it was observed that apelin induced the contractions. Apelin increased the frequency of the myometrium contractions at a significant level when applied at 1 μM and 10 μM concentrations (p<0.05 and p<0.001). Only after the apelin application at 10 μM concentration did the amplitude of the contractions increase at a significant level (p<0.01). In the myometrium of the rats that were on the 21st day of pregnancy, the frequency of the contractions was statistically significant at 0.1 μM, 1 μM and 10 μM doses (p<0.01). In addition, the amplitude of the contractions increased at a statistically significant level at 1 μM and 10 μM dose application (p<0.05 and p<0.01, respectively). The apelin application induced the contractions in calcium-free medium. When apelin was applied after the pre-application with protein kinase C inhibitor, no contractions were observed. The present study showed that apelin levels were increased at the end of pregnancy in rats, and the hormone induced the uterus contractions. This effect does not occur with protein kinase C inhibitor and in calcium-free medium, which shows that protein kinase C pathway might play a role in these mechanism. These findings show that apelin might be an endogenous peptide that plays a role on uterine contractions at birth in rats.
Topics: Animals; Apelin; Benzophenanthridines; Female; Myometrium; Pregnancy; Protein Kinase C; Rats; Rats, Wistar; Uterine Contraction
PubMed: 30213292
DOI: No ID Found -
Animal Reproduction Science Aug 2023Phoenixin-14 (PNX-14) is a regulatory neuropeptide encoded by the SMIM20 gene, which has been implicated in the reproductive cycle by modulating the...
Phoenixin-14 (PNX-14) is a regulatory neuropeptide encoded by the SMIM20 gene, which has been implicated in the reproductive cycle by modulating the hypothalamic-pituitary-gonadal (HPG) axis. Recently, we showed that PNX-14 is downregulated in bitches with cystic endometrial hyperplasia and pyometra. The objective of this study was to determine the expression of Smim20, PNX-14, and its putative receptor GRP173 in the canine ovary (both healthy and those with ovarian cysts), periovarian adipose tissue (PAT) and in the endometrium during the oestrous cycle. The expression was analysed by RT-qPCR and Western blot. In tissue sections, peptides were localised by immunofluorescent assays, and blood plasma concentrations of PNX-14 were detected by EIA. The results demonstrated increased levels of PNX in bitches in the anestrus groups compared to diestrus animals. The expression of GPR173 increased in PAT during the diestrus phase and endometrial tissue in late diestrus bitches. In the ovary, strong signals of PNX-14 and GPR173 were detected in the luteal and follicular cells. Furthermore, bitches with cystic ovaries were characterised by elevated circulating PNX levels and a significantly higher expression of PNX and GPR173 in gonadal tissues, when compared with healthy animals. Moreover, a positive correlation between PNX and progesterone in the blood of healthy bitches was noted, which changed to a negative correlation in females affected by cystic ovaries. These studies expand the knowledge regarding the expression and localization of the PNX/GRP173 system in canine reproductive organs during physiological and pathological conditions.
Topics: Female; Animals; Dogs; Neuropeptides; Peptides; Endometrial Hyperplasia; Endometrium; Adipose Tissue; Dog Diseases
PubMed: 37356348
DOI: 10.1016/j.anireprosci.2023.107282 -
Reproduction in Domestic Animals =... Dec 2019In domestic dogs, oocyte maturation rates are low and the percentage of oocytes that remain in the stage of germinal vesicle (GV) regardless of culture conditions is...
In domestic dogs, oocyte maturation rates are low and the percentage of oocytes that remain in the stage of germinal vesicle (GV) regardless of culture conditions is high. The present study was conducted to characterize the proteome of canine oocyte at the germinal vesicle stage using label-free mass spectrometry. Ovaries were collected from 415 adult domestic dogs and oocytes were divided anestrus and diestrus group. Protein lysates were subjected to quantitative proteomic analysis to identify differentially expressed proteins in different status reproductive. All runs for each sample were performed on an Easy nLC1000 nano-LC chromatograph system directly connected to a quadrupole-type Orbitrap mass spectrometer. For identification of peptides and proteins, raw data of the spectra were loaded into MaxQuant software version 1.5.2.8. Proteomic data were analysed according to gene ontology and a protein-protein interaction network. 312 proteins were identified and grouped according to their biological processes, molecular functions and cellular component. Forty-six differentially expressed proteins among diestrus and control group were associated with at least one GO term in the biological process database. Several proteins involved in the cell cycle, fertilization, regulation of transcription and signalling pathways that are essential for the full development of oocytes and fertilization were expressed. This study identified proteins that were absent, and more or less expressed in different status reproductive. These differentially expressed proteins revealed a framework of molecular reorganization within a GV that renders its competency. This knowledge will enable the identification of target competence biomarkers and thus the establishment of more adequate means of cultivation to improve the M-I and II indexes in this species and also to better understand the physiology of the domestic dog, promoting the development of new reproduction biotechniques.
Topics: Anestrus; Animals; Cell Nucleus; Diestrus; Dogs; Female; In Vitro Oocyte Maturation Techniques; Nuclear Proteins; Oocytes; Proteome; Signal Transduction
PubMed: 31484219
DOI: 10.1111/rda.13560 -
Physiology & Behavior Nov 2019Leucaena feed has been reported to cause disruptive effects on livestock reproduction, such as low calving percentages in cows, abortion in female goats and pigs, dead...
Leucaena feed has been reported to cause disruptive effects on livestock reproduction, such as low calving percentages in cows, abortion in female goats and pigs, dead fetuses and fetal resorption in pregnant rats. In this study, the effects of Leucaena on different female reproductive variables were analyzed in two different reproductive conditions: gonadally intact and ovariectomized (OVX) female rats. Leucaena (LEU) was administered to females in both experimental conditions for 30 consecutive days. The effects of the legume extract were compared with those of Daidzein (DAI), a phytoestrogen, and of the female hormone estradiol (E). In intact females, LEU disrupted the estrous cycle and female sexual behavior, decreased the number of follicles and corpora lutea, increased uterine and vaginal epithelium in proestrus and diestrus periods, increased uterine and vaginal relative weights during diestrus, and decreased serum progesterone during proestrus. All these effects were similar to those of DAI but lower than E-induced effects. In OVX females, LEU decreased body weight, induced lordosis, stimulated vaginal epithelium cornification, increased vaginal weight, and augmented vaginal epithelium thickness. Again, these effects were similar to the effects of DAI and lower than the effects observed with E. These results indicate that, in gonadally intact females, LEU can produce antiestrogenic effects in sexual behavior but estrogenic effects on uterine and vaginal weight and epithelia, without modifying serum levels of E. In OVX females, in total absence of endogenous E, LEU induced estrogenic effects on vaginal weight and epithelia, as well as on sexual behavior.
Topics: Animals; Estradiol; Estrogens; Estrous Cycle; Fabaceae; Female; Isoflavones; Organ Size; Ovarian Follicle; Ovary; Phytoestrogens; Plant Extracts; Progesterone; Rats; Rats, Wistar; Reproduction; Sexual Behavior, Animal; Uterus; Vagina
PubMed: 31533020
DOI: 10.1016/j.physbeh.2019.112683 -
Behavioral Neuroscience Oct 2022The ability to remember sequences of events is fundamental to episodic memory. While rodent studies have examined sex and estrous cycle in episodic-like spatial memory...
The ability to remember sequences of events is fundamental to episodic memory. While rodent studies have examined sex and estrous cycle in episodic-like spatial memory tasks, little is known about these biological variables in memory for sequences of events that depend on representations of temporal context. We investigated the role of sex and estrous cycle in rats during training and testing stages of a cross-species validated sequence memory task (Jayachandran et al., 2019). Rats were trained on a two four-odor sequence memory task delivered on opposite ends of a linear track. Training occurred in six successive stages starting with learning to poke in a nose-port for ≥ 1.2 s; eventually demonstrating sequence memory by holding their nose in the port ≥ 1 s for in-sequence odors and < 1 s for out-of-sequence odors. Performance was analyzed across sex and estrous cycle (proestrus, estrus, metestrus, and diestrus), the latter being determined by cellular composition of a daily vaginal lavage. We found no evidence of sex differences in asymptotic sequence memory performance, similar to humans performing an analogous task (Reeders et al., 2021). Likewise, no differences in sequence memory performance were found across the estrous cycle. Some caveats are that males acquired out-of-sequence trials faster during training with a 3-odor sequence, but this apparent advantage did not carry over to the 4-odor sequence. Additionally, males had shorter poke times overall which seem consistent with a decreased overall response inhibition because they occurred regardless of sequence demands. Together, these results suggest sex and estrous cycle are not major factors in sequence memory capacities. (PsycInfo Database Record (c) 2022 APA, all rights reserved).
Topics: Animals; Estrous Cycle; Estrus; Female; Humans; Male; Metestrus; Proestrus; Rats; Spatial Memory
PubMed: 35254840
DOI: 10.1037/bne0000508 -
Heliyon Dec 2022The medial preoptic (MPN) and the ventromedial hypothalamic nuclei (VMN) modulate the estrogen receptor (ER)-dependent female sexual behavior, a response that is...
The medial preoptic (MPN) and the ventromedial hypothalamic nuclei (VMN) modulate the estrogen receptor (ER)-dependent female sexual behavior, a response that is inhibited by tamoxifen (TAM), a modulator of the steroid receptor activation. With the objective to assess TAM action in the brain areas involved in the modulation sexual cues, an animal model on long-term TAM therapy to intact female rats, was used to mimic the 5-year prophylactic TAM therapy offered to women at higher risk of breast cancer. After three months treatment, female sexual behavior with a stud male rat was evaluated. Upon sacrifice, the brains were removed and the MPN and the ventrolateral division of the VMN were screened for the effects of TAM in the expression of ERα, ERβ and progesterone receptor. Results show that TAM inhibited the receptive component of the female sexual behavior. Even though TAM decreased estrogen and progesterone levels to values similar to the ones of estrous and diestrus rats, the biochemical data failed to demonstrate such possible causation for the behavioral response. In fact, TAM administration induced a constant low level of ovarian hormones that changed the pattern of ER and PR expression as well as receptor co-expression in the brain areas regulating the behavioral response, dissimilar to the ones seen in the cycle phases with the same low hormone levels. Nevertheless, present data suggests that by affecting ER- and/or PR-dependent mechanisms, TAM may modulate the hypothalamus, a region known to participate in several social behaviors.
PubMed: 36593822
DOI: 10.1016/j.heliyon.2022.e12362 -
Journal of Psychopharmacology (Oxford,... Dec 2021Acute hypoxia, which is panicogenic in humans, also evokes panic-like behavior in male rats. Panic disorder is more common in women and susceptibility increases during...
Enhanced responsiveness to hypoxic panicogenic challenge in female rats in late diestrus is suppressed by short-term, low-dose fluoxetine: Involvement of the dorsal raphe nucleus and the dorsal periaqueductal gray.
BACKGROUND
Acute hypoxia, which is panicogenic in humans, also evokes panic-like behavior in male rats. Panic disorder is more common in women and susceptibility increases during the premenstrual phase of the cycle.
AIMS
We here investigated for the first time the impact of hypoxia on the expression of panic-like escape behavior by female rats and its relationship with the estrous cycle. We also evaluated functional activation of the midbrain panic circuitry in response to this panicogenic stimulus and whether short-term, low-dose fluoxetine treatment inhibits the hyper-responsiveness of females in late diestrus.
METHODS
Male and female Sprague Dawley rats were exposed to 7% O. Females in late diestrus were also tested after short-term treatment with fluoxetine (1.75 or 10 mg/kg, i.p.). Brains were harvested and processed for c-Fos and tryptophan hydroxylase immunoreactivity in the periaqueductal gray matter (PAG) and dorsal raphe nucleus (DR).
RESULTS
Acute hypoxia evoked escape in both sexes. Overall, females were more responsive than males and this is clearer in late diestrus phase. In both sexes, hypoxia induced functional activation (c-Fos expression) in non-serotonergic cells in the lateral wings of the DR and dorsomedial PAG, which was greater in late diestrus than proestrus (lowest behavioral response to hypoxia). Increased responding in late diestrus (behavioral and cellular levels) was prevented by 1.75, but not 10 mg/kg fluoxetine.
DISCUSSION
The response of female rats to acute hypoxia models panic behavior in women. Low-dose fluoxetine administered in the premenstrual phase deserves further attention for management of panic disorders in women.
Topics: Animals; Behavior, Animal; Diestrus; Disease Models, Animal; Dorsal Raphe Nucleus; Female; Fluoxetine; Hypoxia; Male; Menstrual Cycle; Panic; Panic Disorder; Periaqueductal Gray; Rats; Rats, Sprague-Dawley; Selective Serotonin Reuptake Inhibitors; Sex Characteristics
PubMed: 34872406
DOI: 10.1177/02698811211058986 -
Biology of Reproduction Jul 2021Zearalenone (ZEN), a nonsteroidal estrogenic mycotoxin, is detrimental to female reproduction. Altered chemical biotransformation, depleted primordial follicles and a...
Zearalenone (ZEN), a nonsteroidal estrogenic mycotoxin, is detrimental to female reproduction. Altered chemical biotransformation, depleted primordial follicles and a blunted genotoxicant response have been discovered in obese female ovaries, thus, this study investigated the hypothesis that obesity would enhance ovarian sensitivity to ZEN exposure. Seven-week-old female wild-type nonagouti KK.Cg-a/a mice (lean) and agouti lethal yellow KK.Cg-Ay/J mice (obese) received food and water ad libitum, and either saline or ZEN (40 μg/kg) per os for 15 days. Body and organ weights, and estrous cyclicity were recorded, and ovaries collected posteuthanasia for protein analysis. Body and liver weights were increased (P < 0.05) in the obese mice, but obesity did not affect (P > 0.05) heart, kidney, spleen, uterus, or ovary weight and there was no impact (P > 0.05) of ZEN exposure on body or organ weight in lean or obese mice. Obese mice had shorter proestrus (P < 0.05) and a tendency (P = 0.055) for longer metestrus/diestrus. ZEN exposure in obese mice increased estrus but shortened metestrus/diestrus length. Neither obesity nor ZEN exposure impacted (P > 0.05) circulating progesterone, or ovarian abundance of EPHX1, GSTP1, CYP2E1, ATM, BRCA1, DNMT1, HDAC1, H4K16ac, or H3K9me3. Lean mice exposed to ZEN had a minor increase in γH2AX abundance (P < 0.05). In lean and obese mice, LC-MS/MS identified alterations to proteins involved in chemical metabolism, DNA repair and reproduction. These data identify ZEN-induced adverse ovarian modes of action and suggest that obesity is additive to ZEN-induced ovotoxicity.
Topics: Animals; Estrogens, Non-Steroidal; Female; Mice; Mycotoxins; Ovary; Proteome; Zearalenone
PubMed: 33855340
DOI: 10.1093/biolre/ioab069 -
F&S Science May 2021To establish if the cessation of testosterone (T) therapy reverses T-induced acyclicity in a transgender mouse model that allows for well-defined T cessation timing.
OBJECTIVE
To establish if the cessation of testosterone (T) therapy reverses T-induced acyclicity in a transgender mouse model that allows for well-defined T cessation timing.
DESIGN
Experimental laboratory study using a mouse model.
SETTING
University-based basic science research laboratory.
ANIMALS
A total of 10 C57BL/6NHsd female mice were used in this study.
INTERVENTION(S)
Postpubertal C57BL/6NHsd female mice were subcutaneously implanted with T enanthate (n = 5 mice) or placebo (n = 5 mice) pellets. Pellets were surgically removed after 6 weeks to ensure T cessation, after which the mice were followed for four estrous cycles after the resumption of cyclicity.
MAIN OUTCOME MEASURE(S)
Primary outcomes included daily vaginal cytology and weekly T levels before, during, and after T enanthate or placebo pellet implantation and removal. Secondary outcomes included ovarian follicle distribution and corpora lutea numbers, body metrics, and terminal diestrus hormone levels.
RESULT(S)
T-treated mice (100%) resumed cycling within one week of T pellet removal after six weeks of T therapy. T levels were significantly elevated during T therapy and decreased to control levels after surgical pellet removal. No detectable differences were observed in the follicle count, corpora lutea formation, diestrus hormone levels, or body metrics after four estrous cycles, with the exception of persistent increased clitoral area between T-treated mice and controls. One T-treated mouse was sacrificed early due to vaginal prolapse and not included in subsequent analyses.
CONCLUSION(S)
Our results demonstrated a close temporal relationship between estrous cycle return and T levels dropping to control levels following T pellet removal. The return of regular cyclic ovulatory function is also supported by the formation of corpora lutea and the lack of detectable differences in key reproductive parameters as compared to controls four cycles after T cessation. These results may be relevant to understanding the reversibility of T-induced amenorrhea and possible anovulation in transgender men interested in pausing T to pursue pregnancy or oocyte donation. Results may be limited by the duration of T treatment, lack of functional testing, and physiological differences between mice and humans.
Topics: Animals; Disease Models, Animal; Female; Heptanoates; Mice; Mice, Inbred C57BL; Ovarian Follicle; Pregnancy; Testosterone; Transgender Persons
PubMed: 35559746
DOI: 10.1016/j.xfss.2021.01.008 -
Theriogenology Jan 2020Hemodynamics of uterine vascularization is modified throughout pregnancy to meet the increasing demand of the growing fetuses and triplex doppler ultrasonography is...
Hemodynamics of uterine vascularization is modified throughout pregnancy to meet the increasing demand of the growing fetuses and triplex doppler ultrasonography is widely used in human medicine to study the uterine arteries and assess the fetal and placental conditions. The aim of our study was to confirm this observation in the bitch, to evaluate differences between bitches of different sizes and to study abnormal pregnancies. Forty-four bitches were monitored during the estrous period to determine ovulation and every 10 days from ovulation to 50 days post-ovulation: the resistivity (RI) and pulsatility (PI) indexes of the right uterine artery were measured as well as usual assessment of fetal development and follow up of the luteal function. Thirty-three out of forty-four bitches were pregnant, including 6 abnormal pregnancies (resorption of more than 10% of the embryos). We also divided them in four weight categories: 8 were small (<10 kg), 13 medium (10-25 kg), 13 large (>25-40 kg) and 10 were giant breeds (>40 kg). We observed that RI and PI decreased over time and were significantly lower for pregnant bitches compared to non-pregnant ones from 30 days post-ovulation. In contrast, RI and PI did not significantly vary with the size of the bitches and we could not determine a significant impact of abnormal pregnancies either. In conclusion, we found no significant difference related to the size of bitches in the RI and PI. The only significant difference between pregnant and non-pregnant bitches was observed from 30 days post-ovulation.
Topics: Animals; Biological Evolution; Body Size; Diestrus; Dogs; Female; Pregnancy; Progesterone; Ultrasonography, Doppler; Uterus
PubMed: 31541785
DOI: 10.1016/j.theriogenology.2019.08.035