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The Journal of Histochemistry and... Jan 2009Hormone-sensitive lipase (HSL) is a key regulator of cholesterol esters metabolism. The aim of this study was to determine HSL localization in rat female reproductive...
Hormone-sensitive lipase (HSL) is a key regulator of cholesterol esters metabolism. The aim of this study was to determine HSL localization in rat female reproductive organs during the ovarian cycle by IHC methods. HSL was located in the ovarian epithelium. The granulosa cells and oocytes of primordial follicles were immunonegative. In mature follicles, HSL was found in oocytes and theca and granulosa cells. However, HSL expression in theca cells and oocytes decreased during follicular atresia. Luteal cells showed HSL staining in cytoplasm during proestrus and estrus, in the nucleus during metestrus, and in cytoplasm and the nucleus during diestrus. In the tubaric ampulla, HSL was located in the epithelial cells nuclei and in the cilia during proestrus and estrus but mainly in the nucleus during metestrus and diestrus. In the isthmus, cells showed HSL immunolabeling in the nucleus and cilia during proestrus, but only in the cilia during estrus, metestrus, and diestrus. In the uterus, HSL was found in the epithelial cells nuclei. HSL-immunoreactive bands at 84, 67, 54, and 43 kDa were found in rat female reproductive organs. HSL labeling in the nucleus of epithelial and germ cells suggests an as yet unknown function for this protein, probably related to oogenesis and cell proliferation.
Topics: Animals; Estrus; Fallopian Tubes; Female; Immunohistochemistry; Ovary; Rats; Rats, Wistar; Sterol Esterase; Uterus; Vagina
PubMed: 18824635
DOI: 10.1369/jhc.2008.951996 -
Genetics and Molecular Research : GMR Jan 2016Frizzled 3 is an important receptor in the Wnt/β-catenin pathway, a conserved signaling pathway that regulates gene expression and controls diverse developmental...
Frizzled 3 is an important receptor in the Wnt/β-catenin pathway, a conserved signaling pathway that regulates gene expression and controls diverse developmental processes. However, the role of this protein during follicular development in the adult ovary is not known. The present study was designed to investigate the expression and localization of Frizzled 3 mRNA and protein during the estrous cycle in the mouse ovary through in situ hybridization (ISH), real-time quantitative polymerase chain reaction, immunohistochemistry and western blot. ISH results showed that in proestrus, high expression of Frizzled 3 was found in the granulosa and stroma with weak levels in the corpus luteum. In estrus and diestrus, the stroma had high Frizzled 3 expression, but levels were low in granulosa cells and corpus luteum. In the metestrus, moderate expression of Frizzled 3 was found in the stroma but low to no expression was found in luteal cells and follicles. The mRNA and protein levels of Frizzled 3 were found to be the highest in proestrus and diestrus compared to estrus and metestrus (P < 0.05), confirming the ISH results. During estrus and diestrus, high Frizzled 3 expression was observed in the stroma and moderate levels in granulosa cells, and during estrus and proestrus, low expression was seen in the oocyte cell membrane. The western blot results further confirmed this change during the estrous cycle. Together, these results indicate that Frizzled 3 is involved in regulating follicular development and oocyte maturation during the estrous cycle.
Topics: Animals; Corpus Luteum; Diestrus; Estrus; Female; Frizzled Receptors; Gene Expression Regulation, Developmental; In Situ Hybridization; Mice; Mice, Inbred ICR; Oocytes; Ovarian Follicle; Proestrus; Real-Time Polymerase Chain Reaction; Time Factors
PubMed: 26909900
DOI: 10.4238/gmr.15017152 -
Archives of Razi Institute Apr 2022It is known that phoenixin-14 (PNX-14) has a mediatory role in reproduction; however, there is no report on the role of the PNX-14 on epilepsy. Therefore, this study...
It is known that phoenixin-14 (PNX-14) has a mediatory role in reproduction; however, there is no report on the role of the PNX-14 on epilepsy. Therefore, this study aimed to investigate the antiepileptic effects of the PNX-14 on the pentylenetetrazol (PTZ)-induced epilepsy in the stages of the estrous cycle among rats. A total of 168 adult female Wistar rats were randomly divided into seven groups, including control (intracerebroventricular injection was performed with saline), PNX-14 (5 µg), PNX-14 (10 µg), bicuculline (competitive antagonist of GABA receptors; 5 nmol)+PNX-14 (5 µg), bicuculline (BIC) (5 nmol)+PNX-14 (10 µg), saclofen (competitive antagonist of GABA receptors; 2.5 µg)+PNX-14 (5 µg), and saclofen (2.5 µg)+PNX-14 (10 µg) in proestrus, estrus, metestrus, and diestrus. Afterward, the control and treatment groups were followed by intraperitoneal administration of 80 mg/kg PTZ. Initiation time of myoclonic seizures (ITMS), initiation time of tonic-clonic seizures (ITTS), seizure duration (SD), and mortality rate (MR) were monitored and recorded for 30 min. According to the results, PNX-14 alone significantly reduced the SD and seizure mortality in all phases of estrus (<0.05). The injection of PNX-14 with BIC significantly reduced SD and seizure mortality in all estrus phases (<0.05). PNX-14 alone increased both ITMS and ITTS in all phases of estrus (<0.05). Furthermore, the injection of PNX-14 with BIC significantly reduced the effects of the PNX-14 on ITMS and ITTS in all estrus stages (<0.05). These results showed that the antiepileptic activity of PNX-14 was probably mediated by GABA receptors, and this effect was more prominent during the luteal phase than the follicular phase.
Topics: Animals; Female; Rats; Anticonvulsants; Baclofen; Bicuculline; Estrous Cycle; Pentylenetetrazole; Rats, Wistar; Seizures
PubMed: 36284939
DOI: 10.22092/ARI.2022.357297.2014 -
BMC Neuroscience Nov 2023Female sex is a known risk factor of brain disorders with raised intracranial pressure (ICP) and sex hormones have been suggested to alter cerebrospinal fluid (CSF)...
BACKGROUND
Female sex is a known risk factor of brain disorders with raised intracranial pressure (ICP) and sex hormones have been suggested to alter cerebrospinal fluid (CSF) dynamics, thus impairing ICP regulation in CSF disorders such as idiopathic intracranial hypertension (IIH). The choroid plexus (CP) is the tissue producing CSF and it has been hypothesized that altered hormonal composition could affect the activity of transporters involved in CSF secretion, thus affecting ICP. Therefore, we aimed to investigate if expression of various transporters involved in CSF secretion at CP were different between males and females and between females in different estrous cycle states. Steroid levels in serum was also investigated.
METHODS
Female and male rats were used to determine sex-differences in the genes encoding for the transporters Aqp1 and 4, NKCC1, NBCe2, NCBE; carbonic anhydrase enzymes II and III (CA), subunits of the Na/K-ATPase including Atp1a1, Atp1b1 and Fxyd1 at CP. The estrous cycle stage metestrus (MET) and estrous (ES) were determined before euthanasia. Serum and CP were collected and subjected to RT-qPCR analysis and western blots. Serum was used to measure steroid levels using liquid chromatography tandem mass spectrometry (LC-MS/MS).
RESULTS
Significant differences in gene expression and steroid levels between males and ES females were found, while no differences were found between male and MET females. During ES, expression of Aqp1 was lower (p < 0.01) and NKCC1 was higher in females compared to males. CAII was lower while CAIII was higher in ES females (p < 0.0001). Gene expression of Atp1a1 was lower in ES compared to male (p = 0.0008). Several of these choroidal genes were also significantly different in MET compared to females in ES. Differences in gene expression during the estrus cycle were correlated to serum level of steroid hormones. Protein expression of AQP1 (p = 0.008) and CAII (p = 0.035) was reduced in ES females compared to males.
CONCLUSIONS
This study demonstrates for the first time that expression at CP is sex-dependent and markedly affected by the estrous cycle in female rats. Further, expression was related to hormone levels in serum. This opens a completely new avenue for steroid regulation of the expression of CSF transporters and the close link to the understanding of CSF disorders such as IIH.
Topics: Rats; Female; Male; Animals; Choroid Plexus; Membrane Proteins; Sex Characteristics; Chromatography, Liquid; Tandem Mass Spectrometry; Steroids
PubMed: 37946101
DOI: 10.1186/s12868-023-00829-w -
The Journal of Veterinary Medical... Jul 2015We measured the plasma levels of amino acids at various reproductive stages in female rats, including the estrous cycle, pregnancy and lactation, and compared the...
We measured the plasma levels of amino acids at various reproductive stages in female rats, including the estrous cycle, pregnancy and lactation, and compared the resulting amino acid profiles using two- or three-dimensional figures. These figures revealed that the amino acid profiles of pregnant and lactating dams differed considerably from those during the estrous cycle or in male rats. The plasma levels of individual amino acids were almost the same between proestrus, estrus, metestrus and diestrus, and their profiles did not differ significantly. However, the amino acid profiles changed during pregnancy and lactation in dams. The plasma Ser level decreased significantly in mid and late pregnancy, whereas Tyr, Gly and His decreased significantly in the late and end stages of pregnancy, and Trp and Lys significantly decreased and increased at the end of pregnancy, respectively. Much larger changes in amino acid profiles were observed during lactation, when the levels of many amino acids increased significantly, and none showed a significant decrease. Plasma Pro, Ser and Gly levels increased continuously from day 1 until day 15 of lactation, whereas Asn and Met increased significantly from days 1 and 5 respectively until the end of lactation. These results suggest that the profiles of plasma amino acids show characteristic changes according to reproductive stage and that it may be necessary to consider such differences when performing amino acid-based diagnosis.
Topics: Amino Acids; Animals; Estrous Cycle; Estrus; Female; Lactation; Male; Metestrus; Pregnancy; Pregnancy, Animal; Proestrus; Rats; Rats, Wistar
PubMed: 25787929
DOI: 10.1292/jvms.15-0095 -
Infection and Immunity Apr 2022Urinary tract infection (UTI) is one of the most prevalent bacterial infections, particularly in women, children, and the elderly. Uropathogenic Escherichia coli (UPEC)...
Urinary tract infection (UTI) is one of the most prevalent bacterial infections, particularly in women, children, and the elderly. Uropathogenic Escherichia coli (UPEC) is the predominant etiological agent of UTI. Uropathogens are directly instilled in the urinary bladder, bypassing the lower urogenital tract, in the widely used murine model of UTI. We assessed whether vaginal inoculation of UPEC led to UTI and how stages of the estrous cycle would impact bacterial colonization in mice. Mice in proestrus, estrus, metestrus, and diestrus were identified by vaginal cytology and inoculated with UPEC in the vaginal tract. Mice were euthanized 1 day after infection, and bacterial loads in the urogenital tract, liver, and spleen were enumerated. Mice in estrus exhibited the highest and most consistent UPEC burdens in all organs, except the bladder. Vaginal inoculation resulted in bladder colonization in a UPEC strain-specific manner. In contrast, transurethral inoculation of UPEC led to bladder colonization. Importantly, inoculation by both routes led to vaginal and uterine colonization and concomitant systemic dissemination to the spleen and liver. The kinetics of bacterial colonization over 2 weeks following vaginal inoculation was comparable in the urogenital tract. Tissue sections revealed the induction of vaginitis and cystitis upon the vaginal instillation of UPEC. In summary, vaginal inoculation of UPEC in mice during estrus represents a novel approach to investigate infection of the kidneys and genital tract and systemic dissemination from the urogenital tract. Our findings suggest that estrogen primes the urogenital tract to create a conducive milieu for UPEC colonization.
Topics: Aged; Animals; Escherichia coli Infections; Escherichia coli Proteins; Estrus; Female; Genitalia; Humans; Kidney; Male; Mice; Urinary Tract Infections; Uropathogenic Escherichia coli
PubMed: 35357220
DOI: 10.1128/iai.00532-21 -
Archivum Immunologiae Et Therapiae... Jun 2013Neutrophils are excreted into the vaginal vault at metestrus during the estrous cycle, and this phenomenon has long been used to determine the phase of the estrous... (Review)
Review
Neutrophils are excreted into the vaginal vault at metestrus during the estrous cycle, and this phenomenon has long been used to determine the phase of the estrous cycle. A much smaller number of neutrophils are also detected in the uterus and the ovary. Recently, we provided several lines of evidence supporting the notion that neutrophils infiltrate into the ovary to regulate the estrous cycle by opioid peptides. Upon inflammation, on the other hand, neutrophils infiltrate into the site of infection to suppress pain by opioid peptides. Thus, opioid peptides are key molecules by which neutrophils play a novel role in regulation of the pain and estrous cycle. In both cases, opioid peptides appear to be secreted by neutrophils stimulated with chemokines, such as MIP-2 and KC in mouse, corticotropin-releasing hormone and IL-1.
Topics: Animals; Chemokine CXCL1; Chemokine CXCL2; Corticotropin-Releasing Hormone; Estrous Cycle; Humans; Interleukin-1; Mice; Neutrophil Infiltration; Neutrophils; Opioid Peptides; Pain; Signal Transduction
PubMed: 23400274
DOI: 10.1007/s00005-013-0218-1 -
Frontiers in Veterinary Science 2023Norwegian Red has been shown to have high levels of estrus behavior under experimental conditions. However, the estrus behaviors of Norwegian Red cows have not been...
INTRODUCTION
Norwegian Red has been shown to have high levels of estrus behavior under experimental conditions. However, the estrus behaviors of Norwegian Red cows have not been studied under commercial conditions.
METHODS
A herd of 89 Norwegian Red cows housed in free stalls on concrete, slatted floors, were continuously video monitored for 21 days. Ovarian cyclicity was confirmed in a final study sample group ( = 18) using milk progesterone concentrations. All mounting and standing activities were recorded, and the duration of mount estrus, standing estrus and the differences between these; prestand and poststand, were determined. The cycle stages metestrus, diestrus and proestrus were estimated based on the starting time and ending time of mount estrus.
RESULTS
All cows in the final study sample group exhibited the primary estrus sign, 'standing to be mounted' during estrus. Two (11%), eleven (61%) and six (33%) cows exhibited the behavior 'standing to be mounted' during metestrus, diestrus and proestrus, respectively. The number of mounts initiated by individual cows was higher during mount and stand estrus than during the rest of the estrous cycle. This study reports a median duration of mount estrus and stand estrus of 21.0 h (interquartile range (IQR) 15.0 to 27.3) and 14.3 h (IQR 12.0 to 18.8), respectively. The median counts per hour of all mount behaviors were 8.6 (IQR 5.6 to 11.3), 1.51 (IQR 0.3 to 3.8) and 1.7 (IQR 0.8 to 6.0) for standing estrus, prestand and poststand, respectively.
DISCUSSION
This study shows that under commercial conditions the Norwegian Red cow displays a high level of mount and stand activity associated with estrus.
PubMed: 37766860
DOI: 10.3389/fvets.2023.1219001 -
Journal of Dairy Science Mar 2022The inverse association between anogenital distance (AGD; the distance from the center of the anus to the base of the clitoris) and fertility, its moderate heritability,...
The inverse association between anogenital distance (AGD; the distance from the center of the anus to the base of the clitoris) and fertility, its moderate heritability, and high variability reported in dairy cattle make AGD a promising candidate for further exploration as a reproductive phenotype. In addition to heritability, repeatability (i.e., consistency in measurements taken at different time points) is important for a reproductive phenotype to be considered useful in genetic selection. Therefore, our primary objective was to determine the repeatability of AGD from birth to breeding age (≈16 mo) in Holstein heifer calves, and during different stages of the estrous cycle, gestation, and lactation in Holstein cows. We also determined the associations among AGD, height (at the hip), and body weight (BW) at birth. In calves (n = 48), we recorded BW (kg) and height (cm) at birth and measured AGD (mm) at approximately 0, 2, 6, 9, 12, and 16 mo of age. In cows, AGD was measured at different stages of the estrous cycle (proestrus, estrus, metestrus and diestrus; n = 20), gestation (30, 90, 180, and 270 d; n = 78), and lactation (30-300 d in milk in 30-d increments; n = 30). Calf height and BW at birth had a weak positive association with AGD at birth. The AGD increased linearly from birth to breeding age, but there was no association between the AGD at birth and at breeding age in heifers. Although any 2 consecutive AGD measurements were correlated, 6 mo was the earliest age at which AGD was moderately correlated (r = 0.41) with that of breeding-age heifers. The AGD was neither influenced by the different stages of estrous cycle nor lactation and remained highly repeatable (r ≥ 0.95). Although AGD measurements at 30, 90, and 180 d of gestation (126.9, 126.7, and 127.7 mm, respectively) were strongly correlated (r ≥ 0.97) with each other, AGD at 270 d of gestation (142.8 mm) differed from AGD at all earlier stages of gestation. In summary, AGD measured at birth did not reflect AGD at breeding age in heifers, but AGD measurements in cows had high repeatability at all stages of the estrous cycle, gestation, and lactation, except at 270 d of gestation. Therefore, AGD could be measured reliably at any of the aforesaid physiological states in cows due to its high repeatability, except during late gestation. The earliest gestational stage when pregnancy-associated increase in AGD occurred, however, could not be definitively established in the present study.
Topics: Animals; Body Weight; Cattle; Female; Fertility; Lactation; Milk; Pregnancy; Reproduction
PubMed: 35033344
DOI: 10.3168/jds.2021-21419 -
International Journal of Molecular... Aug 2021Cluster of differentiation 73 (CD73, also known as ecto-5'-nucleotidase) is an enzyme that converts AMP into adenosine. CD73 is a surface enzyme bound to the outside of...
Cluster of differentiation 73 (CD73, also known as ecto-5'-nucleotidase) is an enzyme that converts AMP into adenosine. CD73 is a surface enzyme bound to the outside of the plasma membrane expressed in several cells and regulates immunity and inflammation. In particular, it is known to inhibit T cell-mediated immune responses. However, the regulation of CD73 expression by hormones in the uterus is not yet clearly known. In this study, we investigated the expression of CD73 in ovariectomized mice treated with estrogen or progesterone and its regulation in the mouse uterus during the estrous cycle. The level of CD73 expression was dynamically regulated in the uterus during the estrous cycle. CD73 protein expression was high in proestrus, estrus, and diestrus, whereas it was relatively low in the metestrus stage. Immunofluorescence revealed that CD73 was predominantly expressed in the cytoplasm of the luminal and glandular epithelium and the stroma of the endometrium. The expression of CD73 in ovariectomized mice was gradually increased by progesterone treatment. However, estrogen injection did not affect its expression. Moreover, CD73 expression was increased when estrogen and progesterone were co-administered and was inhibited by the pretreatment of the progesterone receptor antagonist RU486. These findings suggest that the expression of CD73 is dynamically regulated by estrogen and progesterone in the uterine environment, and that there may be a synergistic effect of estrogen and progesterone.
Topics: 5'-Nucleotidase; Animals; Estrogens; Estrous Cycle; Female; Gene Expression Regulation; Mice; Mice, Inbred ICR; Progesterone; Progestins; Uterus
PubMed: 34502315
DOI: 10.3390/ijms22179403