Biópsia endometrial na égua: importância no diagnóstico da subfertilidade e principais achados / Endometrial biopsy in the mare: importance in the diagnosis of subfertility and main findings

A biópsia endometrial na égua é uma técnica simples e bastante antiga. É uma importante ferramenta para o diagnóstico de patologia no endométrio nesta espécie utilizada para determinar o estágio do ciclo estral, para detectar a presença de inflamações agudas ou crônicas e para determina a possibilidade do útero levar a gestação a termo. Mais recentemente outras alterações tais como a má-diferenciação e as angiopatias, também estão sendo avaliadas em amostras de biópsia endometrial em éguas, uma vez que também podem interferir no prognóstico reprodutivo destas fêmeas. Esta revisão tem por objetivo descrever a avaliação da biópsia endometrial e as principais alterações que ocorrem no endométrio das éguas.(AU)

Endometrial biopsy in the mare is a simple and quite old technique. It is an important tool for the diagnosis of pathology in the endometrium in this species, used to determine the stage of the estrous cycle, to detect the presence of acute or chronic inflammation and to determine the possibility of the uterus carrying the pregnancy to term. More recently, other alterations, such as poor differentiation and angiopathies, are also being evaluated in endometrial biopsy samples in mares, since they may also interfere with the reproductive prognosis of these females. This review aims to describe the evaluation of the endometrial biopsy and the main changes that occur in the endometrium of mares.(AU)

The equine endometrium and its multiple connections, interrelationships and clinical effects in aged mares

Mares, which make up 90% of animals requiring reproductive assistance in commercial equine production systems, are subjected to strong breeding and reproductive efficiency pressure throughout their lives, but the effects of aging on overall and reproductive systems in particular; it often means that we are forcing the natural homeostasis mechanisms under pressure over time. The objective of this brief communication is to summarize some concepts related to the role and importance of equine endometrium, particularly in old mares, the most challenged category to achieve productive and efficiency goals established by humans. Endometritis is one of the most frequent and recurrent pathologies in equine gynecology and is responsible for enormous economic losses in the industry. Alterations of the microbiota, molecular signals and uterine endocrine microenvironment can lead to infertility, but degenerative phenomena associated with age and parity can lead to endometriosis altering maternal recognition of pregnancy and pregnancy maintenance. Maintaining horses in an adequate nutrition status and environment is essential to achieve pregnancy but also to obtain healthy offspring.(AU)

The in vitro effects of n-3 fatty acids on immune response regulation of bovine ex vivo endometrial explants / Os efeitos in vitro dos ácidos graxos n-3 na regulação da resposta imune de explantes endometriais bovinos ex vivo

Fatty acids are considered metabolic intermediaries, although new facts indicate they also work as signaling molecules with different roles in the immune response. Based on that, in this study, we investigated the anti-inflammatory effects of n-3 polyunsaturated fatty acids (PUFAs) as eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and α-linolenic acid (LNA) in ex vivo bovine endometrial explants. For this, two groups were formed: (1) LPS-challenged and (2) control, both to evaluate the accumulation of proinflammatory cytokines as interleukin 1ß (IL1B) and interleukin 6 (IL6). To develop the study, bovine female reproductive tracts from non-pregnant Angus heifers without evidence of reproductive diseases were selected. Endometrial explants were processed and treated for 24 h with EPA, DHA, and LNA in five different concentrations (0µM, 50µM, 100 µM, 200µM and 400 µM) and then, challenged with LPS for 24 h. Supernatants were collected to evaluate the concentration of IL1B and IL6 by ELISA. Explants treated with EPA from control groups reduced the concentrations of ILB (200µM) and IL6 (400 µM), and IL6 (50 µM; 100 µM) from the LPS-challenged group. DHA decreased the accumulation of IL1B and IL6 at 200 µM on explants from the LPS-challenged group, and 200 µM reduced IL6 from the control group. In contrast, explants treated with LNA only reduced the accumulation of IL1B to 400µM (from both groups). In conclusion, the EPA acid is the best anti-inflammatory option to decrease the concentration of both pro-inflammatory cytokines (IL1B and IL6) from LPS-challenged and control groups in bovine endometrial explants; while LNA evidence to be the last option to promote an anti-inflammatory response.(AU)

Os ácidos graxos são considerados intermediários metabólicos, embora novos fatos indiquem que eles também atuem como moléculas sinalizadoras com diferentes papéis na resposta imune. Dessa forma, este estudo investigou os efeitos anti-inflamatórios de ácidos graxos poliinsaturados n-3 (PUFAs) como ácido eicosapentaenóico (EPA), ácido docosahexaenóico (DHA) e ácido α-linolênico (LNA) em explantes endometriais ex vivo de bovinos. Para tal, o experimento foi divido em dois grupos: (1) Desafiado-LPS e (2) Controle, para que então pudesse avaliar o acúmulo de citocinas pró-inflamatórias como interleucina 1ß (IL1B) e interleucina 6 (IL6). Foram selecionados tratos reprodutivos de fêmeas bovinas de novilhas Angus não prenhes sem evidência de doenças reprodutivas. Explantes endometriais foram processados e tratados por 24h com EPA, DHA e LNA em cinco concentrações diferentes (0µM, 50µM, 100 µM, 200µM e 400 µM) e, em seguida, desafiados com LPS por mais 24h. Os sobrenadantes foram colhidos para avaliar a concentração de IL1B e IL6 pelo teste de ELISA. Os explantes tratados com EPA dos grupos de controle reduziram as concentrações de ILB (200 µM) e IL6 (400 µM) e no grupo desafiado com LPS houve redução das concentrações de IL6 (50 µM; 100 µM). Nos explantes do grupo desafiado com LPS, o DHA diminuiu o acúmulo de IL1B e IL6 nas concentrações de 200 µM, e no grupo controle reduziu IL6 nas concentrações de 200 µM, enquanto os explantes tratados com LNA reduziram apenas o acúmulo de IL1B a 400 µM (de ambos os grupos). Em conclusão, o ácido EPA provou ser a melhor opção anti-inflamatória para diminuir a concentração de ambas as citocinas pró-inflamatórias (IL1B e IL6) de grupos desafiados com LPS e controle em explantes endometriais bovinos; enquanto o LNA evidencia ser a opção menos viável para promover uma resposta anti-inflamatória.(AU)

Endometrial receptivity in cattle: the mutual reprogramming paradigm

Prior to implantation in cattle, the mucous medium contained in the uterine lumen serves as a working interface for molecular exchange and signaling between the lining endometrium and the embryo. The composition of this luminal fluid changes temporally according to the secretory and reabsorptive activities of the uterus and the embryo, which are under complex regulation. Via this interface, both the embryo and the endometrium reprogram each other's functions to support pregnancy continuation beyond the pre-implantation period. More specifically, the embryo receives elongation signals and the uterus receives anti-luteolytic stimuli. Here, characteristics of the luminal compartment as well as the regulation of its composition to determine the pregnancy outcome will be discussed.(AU)

Altrenogest during early pregnancy modulates uterine glandular epithelium and endometrial growth factor expression at the time implantation in pigs

This study evaluated the effects of supplying altrenogest from day 6-12 of pregnancy on the endometrial glandular epithelium, corpora lutea (CL) morphology, and endometrial and CL gene expression. A total of 12 crossbred females (Landrace × Large White) were used. The females were assigned to 4 treatments according to a random design with a 2 × 2 factorial arrangement, with two categories (sow or gilt) and two treatments (non-treated and treated with altrenogest). On day 6 of pregnancy, animals were allocated to one of the following groups: non-treated (NT, n = 6; 3 sows and 3 gilts), and (T, n = 6; 3 sows and 3 gilts) treated daily with 20 mg of altrenogest, from day 6-12 of pregnancy. All animals were euthanized on day 13 of pregnancy. All CLs were individually weighed, and their volume were determined. The endometrial glandular density (GD), mean glandular area (MGA), and vascular density (VD) were determined by histomorphometric and immunohistochemical analyses. Endometrium samples were collected and analyzed by qRT-PCR to evaluate the abundance of transcripts for VEGF and IGF-I. Females in the T group had higher MGA (P < 0.05) compared to the NT group. There was no effect of treatment on GD or VD for both experimental groups. Sows in the T group had augmented expression of IGF-I (P < 0.05). Progestagen had no detrimental effect on CL morphology. In conclusion, altrenogest improves the uterine environment during the peri-implantation period in pigs without compromising corpora lutea development.(AU)

Alterações degenerativas no útero de éguas: Revisão de literatura / Degenerative alterations in the mare´s uterus: Literature review

As alterações degenerativas do útero das éguas representam um problema importante na criação de equinos. A endometrose é um processo degenerativo crônico caracterizado pelo desenvolvimento de fibrose no endométrio da égua, sendo uma das principais causas de subfertilidade ou infertilidade nestas fêmeas. Algumas outras alterações, tais como a má-diferenciação e as angiopatias, embora não tenham um diagnóstico tão frequente, também podem interferir no prognóstico reprodutivo destes animais. Baseado neste fato, esta revisão tem por objetivo descrever as principais alterações que ocorrem no endométrio das éguas.

Degenerative changes in the mare’s uterus is an important problem in the horse breeding. The endometrosis is a chronic degenerative process characterized by the development of fibrosis in the endometrium of the mare, being one of the main causes of subfertility or infertility in these females. Some alterations, such as maldifferentiation and angiopathies, although they do not have such a frequent diagnosis, may also interfere with the reproductive prognosis of these animals. Based on this fact, this review aims to describe the main changes that occur in the endometrium of mares.

Histomorphometric and vascular changes in equine endometrium after the infusion of conceptus fragments

This experiment aimed to verify if the proteins present in a 13th day conceptus induce changes in the equine endometrial ultra-structure, histology, and vascularization, two days after its infusion. Ten healthy cyclic mares were used. Once estrus was confirmed, mares were examined daily to detect ovulation (day 0). After ovulation, mares were examined daily until day seven by transrectal palpation and B-mode and Doppler ultrasonography. In this first cycle, intrauterine biopsies were collected at day seven after ovulation, constituting the Cyclic group (n = 10). In the second cycle, the same mares daily were examined until ovulation was detected. After ovulation, mares were examined daily by transrectal palpation and B-mode and Doppler ultrasonography until day 7. On day 5, after ovulation, fragments from previously collected 13-day-old concepti were infused into the uterus of each mare. Intrauterine biopsies were collected at day 7 in all mares (n = 10), constituting the Fragment group. The percentage of ciliated and flattened cells decreased in the Fragment group. Protruded cells, superficial and intraglandular secretion, glandular lumen and diameter, blood vessel diameter, endometrial vascularization, and immune cells were higher in the Fragment group than in the Cyclic group. In summary, proteins of 13th day equine conceptus fragments infused at day five after ovulation signaled histological and vascular changes in the endometrium at the 7th day after ovulation.

Histomorphometric and vascular changes in equine endometrium after the infusion of conceptus fragments

This experiment aimed to verify if the proteins present in a 13th day conceptus induce changes in the equine endometrial ultra-structure, histology, and vascularization, two days after its infusion. Ten healthy cyclic mares were used. Once estrus was confirmed, mares were examined daily to detect ovulation (day 0). After ovulation, mares were examined daily until day seven by transrectal palpation and B-mode and Doppler ultrasonography. In this first cycle, intrauterine biopsies were collected at day seven after ovulation, constituting the Cyclic group (n = 10). In the second cycle, the same mares daily were examined until ovulation was detected. After ovulation, mares were examined daily by transrectal palpation and B-mode and Doppler ultrasonography until day 7. On day 5, after ovulation, fragments from previously collected 13-day-old concepti were infused into the uterus of each mare. Intrauterine biopsies were collected at day 7 in all mares (n = 10), constituting the Fragment group. The percentage of ciliated and flattened cells decreased in the Fragment group. Protruded cells, superficial and intraglandular secretion, glandular lumen and diameter, blood vessel diameter, endometrial vascularization, and immune cells were higher in the Fragment group than in the Cyclic group. In summary, proteins of 13th day equine conceptus fragments infused at day five after ovulation signaled histological and vascular changes in the endometrium at the 7th day after ovulation.(AU)

Ultrasound monitoring of the uterus and ovaries of dominant and subordinate females of yellow-breasted capuchin (Sapajus xanthosternos) and robust tufted capuchin (Sapajus robustus) in captive colonies during the ovarian cycle and anestrus periods / Acompanhamento ultrassonográfico do útero e ovários de fêmeas dominantes e subordinadas de macacos-prego-do-peito-amarelo (Sapajus xanthosternos) e macacos-prego-de-crista (Sapajus robustus) em colônias cativas, durante o ciclo ovariano e períodos de anestro

The yellow-breasted capuchin (Sapajus xanthosternos) and robust tufted capuchin (Sapajus robustus) are endangered species due to destruction of their natural habitat and predatory chase. However, it is still necessary to elucidate some details of their reproductive physiology in order to obtain better indices in the assisted reproduction of these species. This study aimed to evaluate the ovarian cycle of 13 dominant and subordinate females of S. xanthosternos (n=8) and S. robustus (n=5) using sagittal and transversally scanned ultrasound of their uterus and ovaries. Sonograms were performed every seven days for two months. The ovarian cycle phase and anestrous condition were confirmed by colpocytology. Our results showed different uterine parameters (craniocaudal diameter, dorso-ventral diameter, and transverse diameter) (P<0.05) between anestrous subordinate females and other ovarian cycle phases and social classes. The mean of uterine volume was higher in dominant females than subordinate females in all cycle phases (P<0.05), except in follicular phase. During anestrus, endometrial width was smaller in subordinate females than in dominant females (P<0.05). Subordinate females showed differences in endometrial measures (P<0.05) between anestrous period and follicular and luteal periods. Ovarian measures in dominant females were higher than in subordinate females only during anestrus (P<0.05). In the subordinate females, ovarian parameters were different (P<0.05) between anestrus and follicular and luteal phases. Dominant females showed higher volume of right ovary compared to volume of the left ovary during anestrus and follicular phase (P<0.05). Follicles and corpus luteum were distinguished by ultrasonography in most exams (86.11%). During anestrus, measurable ovarian structures were not observed in both ovaries in dominant and subordinate females. In conclusion, the methodology used in this study allowed to evaluate the ovarian cycle in S. xanthosternos e S. robustus females and that cycle phase/anestrus and social class of the female influenced the size of the uterus and ovaries.(AU)

O macaco-prego-do-peito-amarelo (Sapajus xanthosternos) e o macaco-prego-de-crista (Sapajus robustus) encontram-se em risco de extinção devido a destruição do seu habitat e a caça predatória. Porém, ainda necessita-se elucidar alguns detalhes de sua fisiologia reprodutiva, para obterem-se melhores índices por meio de reprodução assistida. Sendo assim, o objetivo deste trabalho foi acompanhar o ciclo ovariano de 13 fêmeas dominantes e subordinadas de S. xanthosternos (n=8) e S. robustus (n=5) por meio de cortes ultrassonográficos sagitais e transversais do útero e dos ovários. Estas fêmeas foram examinadas uma vez por semana durante dois meses. A fase do ciclo ovariano/anestro foi confirmada pela colpocitologia. Os resultados da pesquisa demonstrou diferença significativa (P<0,05) relacionada ao diâmetro crânio-caudal, diâmetro dorso-ventral e diâmetro transversal entre as fêmeas subordinadas em anestro com todas as outras fases do ciclo ovariano e classes sociais. A média do volume uterino foi maior nas fêmeas dominantes que nas subordinadas em todas as fases do ciclo (P<0,05) a exceção da fase folicular. A largura endometrial, durante o período de anestro, foi menor nas fêmeas subordinadas quando comparada às dominantes (P<0,05). Nas fêmeas subordinadas, houve diferenças (P<0,05) nas mensurações endometriais entre o período de anestro e das fases folicular e lútea. Quanto aos ovários, as médias das medidas observadas nas fêmeas dominantes foram superiores as das subordinadas durante o anestro (P<0,05). As médias das medidas das variáveis ovarianas das fêmeas subordinadas apresentaram diferenças entre o anestro e as fases folicular e lútea (P<0,05). Nas fêmeas dominantes o volume do ovário direito foi maior que o do ovário esquerdo durante o anestro e na fase folicular (P<0,05). Folículos e corpos lúteos foram diferenciados pela ultrassonografia na maioria das coletas (86,11%). Durante o anestro não foram observadas estruturas ovarianas mensuráveis nos ovários em ambas as classes sociais. Concluiu-se que a metodologia empregada neste estudo permitiu o acompanhamento do ciclo ovariano das fêmeas S. xanthosternos e S. robustus e que as fases do ciclo/anestro e a classe social das fêmeas influenciaram as medidas do útero e ovários.(AU)

Métodos de coleta de amostra para exame de citologia endometrial em éguas com endometrite / Sample collection methods for endometrial cytology examination in mares with endometritis

A endometrite é definida como uma inflamação do endométrio, podendo ser aguda ou crônica, infecciosa ou não infecciosa. Essa afecção é de grande importância econômica na medicina veterinária, podendo resultar em morte embrionária, aborto e até tornar a égua infértil, sendo uma das afecções que mais comprometem a sanidade uterina, colocando em risco os índices de prenhez obtidos pós-cobertura, pós-inseminação, e pós-inovulação de embriões. Existem diversos métodos para diagnóstico e o mais utilizado a campo é a citologia endometrial/uterina. Também existem algumas formas de coleta de amostra para análise laboratorial pela citologia, tais como: escova ginecológica, swab uterino guiado por mandril metálico ou plástico descartável e lavado uterino de baixo volume, seguido de sedimentação ou centrifugação podendo ser complementada por citocentrifugação. O diagnóstico pela citologia endometrial por meio da técnica da escova ginecológica é a mais utilizada entre os médicos veterinários, pois trata-se de um método prático para execução e rápido quanto à obtenção dos resultados sobre a presença ou não da endometrite, facilitando a conduta terapêutica quanto à utilização das éguas para reprodução ou descarte. Objetivou-se com esse estudo citaras formas de diagnóstico da endometrite em éguas, enfatizando a citologia endometrial e técnicas para obtenção de amostra, identificando a técnica mais indicada por diversos autores.(AU)

Endometritis is defined as an inflammation of the endometrium, which may be acute or chronic, infectious or noninfectious. This condition is of great economic importance in veterinary medicine, and may result in embryonic death, abortion and even render the mare infertile, being one of the conditions that most compromise uterine health, putting at risk the pregnancy rates obtained post-coverage, postinsemination, and post-embryo transfer. There are several methods for diagnosis and the most used in the field is endometrial/uterine cytology. There are also some forms of sample collection for laboratory analysis by cytology, such as: gynecological brush, uterine swab guided by metallic mandrel or disposable plastic and low volume uterine lavage, followed by sedimentation or centrifugation and may be complemented by cytocentrifugation. Diagnosis by endometrial cytology using the gynecological brush technique is the most used among Veterinary Practitioners, because it is a practical method to execute and fast to obtain the results in the presence of endometritis, facilitating the therapeutic conduct regarding the use of mares for breeding or discarding. The objective of this study was to cite the diagnostic methods of endometritis in mares, emphasizing endometrial cytology and its techniques for obtaining a sample, identifying the most indicated technique by several authors.(AU)

Métodos de coleta de amostra para exame de citologia endometrial em éguas com endometrite / Sample collection methods for endometrial cytology examination in mares with endometritis

A endometrite é definida como uma inflamação do endométrio, podendo ser aguda ou crônica, infecciosa ou não infecciosa. Essa afecção é de grande importância econômica na medicina veterinária, podendo resultar em morte embrionária, aborto e até tornar a égua infértil, sendo uma das afecções que mais comprometem a sanidade uterina, colocando em risco os índices de prenhez obtidos pós-cobertura, pós-inseminação, e pós-inovulação de embriões. Existem diversos métodos para diagnóstico e o mais utilizado a campo é a citologia endometrial/uterina. Também existem algumas formas de coleta de amostra para análise laboratorial pela citologia, tais como: escova ginecológica, swab uterino guiado por mandril metálico ou plástico descartável e lavado uterino de baixo volume, seguido de sedimentação ou centrifugação podendo ser complementada por citocentrifugação. O diagnóstico pela citologia endometrial por meio da técnica da escova ginecológica é a mais utilizada entre os médicos veterinários, pois trata-se de um método prático para execução e rápido quanto à obtenção dos resultados sobre a presença ou não da endometrite, facilitando a conduta terapêutica quanto à utilização das éguas para reprodução ou descarte. Objetivou-se com esse estudo citaras formas de diagnóstico da endometrite em éguas, enfatizando a citologia endometrial e técnicas para obtenção de amostra, identificando a técnica mais indicada por diversos autores.

Endometritis is defined as an inflammation of the endometrium, which may be acute or chronic, infectious or noninfectious. This condition is of great economic importance in veterinary medicine, and may result in embryonic death, abortion and even render the mare infertile, being one of the conditions that most compromise uterine health, putting at risk the pregnancy rates obtained post-coverage, postinsemination, and post-embryo transfer. There are several methods for diagnosis and the most used in the field is endometrial/uterine cytology. There are also some forms of sample collection for laboratory analysis by cytology, such as: gynecological brush, uterine swab guided by metallic mandrel or disposable plastic and low volume uterine lavage, followed by sedimentation or centrifugation and may be complemented by cytocentrifugation. Diagnosis by endometrial cytology using the gynecological brush technique is the most used among Veterinary Practitioners, because it is a practical method to execute and fast to obtain the results in the presence of endometritis, facilitating the therapeutic conduct regarding the use of mares for breeding or discarding. The objective of this study was to cite the diagnostic methods of endometritis in mares, emphasizing endometrial cytology and its techniques for obtaining a sample, identifying the most indicated technique by several authors.

Aspects of embryo-maternal communication in establishment of pregnancy in cattle

Establishment of pregnancy in mammals requires reciprocal molecular communication between the conceptus and endometrium that modifies the endometrial transcriptome and uterine luminal milieu to support pregnancy. Due to the small size of the early embryo and elongating conceptus relative to the volume of the uterine lumen, collection of endometrium adjacent to the developing conceptus is difficult following conventional uterine flushing methods in cattle. Use of endometrial explants in culture can overcome this challenge and reveal information about the dialogue between the developing embryo and the uterus. The aim of this short review is to summarize some of our recent findings in relation to embryo maternal interaction during bovine pregnancy establishment and to put them in the wider context of fertility in cattle.

Aspects of embryo-maternal communication in establishment of pregnancy in cattle

Establishment of pregnancy in mammals requires reciprocal molecular communication between the conceptus and endometrium that modifies the endometrial transcriptome and uterine luminal milieu to support pregnancy. Due to the small size of the early embryo and elongating conceptus relative to the volume of the uterine lumen, collection of endometrium adjacent to the developing conceptus is difficult following conventional uterine flushing methods in cattle. Use of endometrial explants in culture can overcome this challenge and reveal information about the dialogue between the developing embryo and the uterus. The aim of this short review is to summarize some of our recent findings in relation to embryo maternal interaction during bovine pregnancy establishment and to put them in the wider context of fertility in cattle.(AU)

Modeling early embryo-maternal interactions in vitro

Environmental conditions experienced during early embryonic development influence growth, metabolism, and gene expression of the embryo as well as the epigenetic profile of the offspring. The environment of the early embryo consists of the luminal fluid within the oviduct and uterus and the epithelial cells composing this fluid. Whether the embryo is able to shape its own microenvironment by interacting with the epithelial lining of the oviduct/uterus and which factors potentially interfere with or regulate these interactions remains to be elucidated. As early embryonic signals and the respective maternal responses are subtle and local events, it is challenging to study them in vivo. Therefore, adequate in vitro-models optimally mimicking the contact zone between the maternal reproductive tract and the early embryo are needed to a) elucidate basic mechanisms involved in early embryonic development and b) reduce the number of experimental animals used for such studies. Functional epithelial cells are generally defined by a polarized distribution of organelles and proteins. Proper polarization is tightly connected with physiological cell behavior and in vivo-like reactivity of the epithelium. Therefore, this review summarizes current strategies for in vitro preservation of epithelial cell polarity. It presents recent advances in 3D culture of female reproductive tract epithelia and embryo-epithelial cocultures. A special emphasis is set on compartmentalized culture systems, powerful tools for studying early embryo-maternal interactions in vitro. In such systems, cultured epithelial cells are manipulable from their basolateral as well as their apical cell pole, allowing concomitant application of embryonic as well as maternal effectors from the appropriate cellular compartment.

Modeling early embryo-maternal interactions in vitro

Environmental conditions experienced during early embryonic development influence growth, metabolism, and gene expression of the embryo as well as the epigenetic profile of the offspring. The environment of the early embryo consists of the luminal fluid within the oviduct and uterus and the epithelial cells composing this fluid. Whether the embryo is able to shape its own microenvironment by interacting with the epithelial lining of the oviduct/uterus and which factors potentially interfere with or regulate these interactions remains to be elucidated. As early embryonic signals and the respective maternal responses are subtle and local events, it is challenging to study them in vivo. Therefore, adequate in vitro-models optimally mimicking the contact zone between the maternal reproductive tract and the early embryo are needed to a) elucidate basic mechanisms involved in early embryonic development and b) reduce the number of experimental animals used for such studies. Functional epithelial cells are generally defined by a polarized distribution of organelles and proteins. Proper polarization is tightly connected with physiological cell behavior and in vivo-like reactivity of the epithelium. Therefore, this review summarizes current strategies for in vitro preservation of epithelial cell polarity. It presents recent advances in 3D culture of female reproductive tract epithelia and embryo-epithelial cocultures. A special emphasis is set on compartmentalized culture systems, powerful tools for studying early embryo-maternal interactions in vitro. In such systems, cultured epithelial cells are manipulable from their basolateral as well as their apical cell pole, allowing concomitant application of embryonic as well as maternal effectors from the appropriate cellular compartment.(AU)

Endometrial prostaglandin F2α in vitro production and its modulation regarding dominant follicle position in cattle / Produção in vitro de PGF F2α endometrial e sua modulação referente ao folículo dominante em bovinos

Prostaglandin F2α (PGF2α) determines luteolysis in cattle, and the ability to manipulate its endogenous synthesis is indispensible for large-scale animal breeding. Estradiol (E2) and progesterone (P4) modulate several molecular pathways in endometrial cells, including the synthesis of PGF2α; however, its specific mechanisms are still not totally known. This study investigated the production in vitro and possible modulation of endometrial PGF2α due to a local effect of endogenous E2 in the ipsilateral uterine horn (UH) containing the dominant follicle (DF) or from P4 in ipsilateral horn containing the corpus luteum (CL). The PGF2α stimulators oxytocin (OT) and phorbol 12,13-dibutyrate (PDBu) were incubated with endometrial explants, and PGF2α content was measured. For that, cycling cows were synchronized, the development of DF and CL was examined by ultrasonography and on the seventh day of the estrous cycle, endometrial explants were collected and cultured in medium supplemented with 10-6 M PDBu or 10-6 M OT or non-supplemented. Media samples were collected immediately after treatment and 60 min later. Radioimmunoassay showed that the PGF2α content of the UH ipsilateral to the DF was 49% less than that of the contralateral UH (8.22 ± 0.95 vs. 12.24 ± 0.95 pg/mL/mg tissue, respectively; P < 0.01). However, the PGF2α levels did not differ between the UHs as a function of the CL position (9.46 ± 0.95 vs. 11 ± 0.95 pg/mL/mg; P > 0.05). The cellular stimulators promoted an increase in PGF2α synthesis (P < 0.02), and the effects differed among the animals (P < 0.04). The PGF2a production was higher in the explants treated with PDBu rather than OT (13.68 ± 1.16 vs. 10.01 ± 1.16 pg/mL/mg tissue, respectively; P < 0.05). In conclusion, PGF2α synthesis is modulated by the presence of the DF (local E2) but not the CL (local P4), and both PDBu and OT stimulated PGF2a synthesis.(AU)

A prostaglandina F2α (PGF2α) determina a luteólise em bovinos. A capacidade de manipular sua síntese endógena é indispensável para a produção animal em grande escala. O estradiol (E2) e a progesterona (P4) modulam diversas vias moleculares das células endometriais, incluindo a síntese de PGF2α; no entanto, pouco se sabe sobre seus mecanismos específicos. Este trabalho investigou a produção in vitro e a possível modulação da PGF2α endometrial devido a um efeito local do E2 endógeno no corno uterino ipsilateral ao folículo dominante (FD) ou da P4 no corno ipsilateral ao corpo lúteo (CL). Os estimuladores de PGF2α oxitocina (OT) e 12,23-dibutirato de forbol (PDBu) foram incubados com explantes endometriais, e o conteúdo de PGF2α foi mensurado. Para tal, vacas cíclicas foram sincronizadas, o desenvolvimento de FD e CL foi examinado por ultrassonografia, e no 17º dia do ciclo estral os explantes endometriais foram coletados e cultivados em meio ou suplementados com PDBu 10-6M ou 10-6M OT. As amostras de meio foram coletadas imediatamente após o tratamento e sessenta minutos depois. O radioimunoensaio mostrou que o conteúdo de PGF2α do corno ipsilateral ao FD foi 49% menor que o do corno contralateral (8,22 ± 0,95 vs. 12,24 ± 0,95 pg/mL/mg de tecido, respectivamente, P < 0,01). No entanto, os níveis de PGF2α não diferiram entre os cornos em função da posição do CL (9,46 ± 0,95 versus 11 ± 0,95 pg/mL/mg; P > 0,05). Os estimuladores celulares promoveram um aumento na síntese de PGF2α (P < 0,02), e os efeitos diferiram entre os animais (P < 0,04). A produção de PGF2α foi maior nos explantes tratados com PDBu em comparação à OT (13,68 ± 1,16 versus 10,01 ± 1,16 pg/mL/mg de tecido, respectivamente, P < 0,05). A conclusão obtida foi que a síntese de PGF2α é: modulada pela presença do FD (E2 local), mas não do CL (P4 local); e estimulada por PDBu e OT.(AU)

Endometrial prostaglandin F2α in vitro production and its modulation regarding dominant follicle position in cattle / Produção in vitro de PGF F2α endometrial e sua modulação referente ao folículo dominante em bovinos

Prostaglandin F2α (PGF2α) determines luteolysis in cattle, and the ability to manipulate its endogenous synthesis is indispensible for large-scale animal breeding. Estradiol (E2) and progesterone (P4) modulate several molecular pathways in endometrial cells, including the synthesis of PGF2α; however, its specific mechanisms are still not totally known. This study investigated the production in vitro and possible modulation of endometrial PGF2α due to a local effect of endogenous E2 in the ipsilateral uterine horn (UH) containing the dominant follicle (DF) or from P4 in ipsilateral horn containing the corpus luteum (CL). The PGF2α stimulators oxytocin (OT) and phorbol 12,13-dibutyrate (PDBu) were incubated with endometrial explants, and PGF2α content was measured. For that, cycling cows were synchronized, the development of DF and CL was examined by ultrasonography and on the seventh day of the estrous cycle, endometrial explants were collected and cultured in medium supplemented with 10-6 M PDBu or 10-6 M OT or non-supplemented. Media samples were collected immediately after treatment and 60 min later. Radioimmunoassay showed that the PGF2α content of the UH ipsilateral to the DF was 49% less than that of the contralateral UH (8.22 ± 0.95 vs. 12.24 ± 0.95 pg/mL/mg tissue, respectively; P < 0.01). However, the PGF2α levels did not differ between the UHs as a function of the CL position (9.46 ± 0.95 vs. 11 ± 0.95 pg/mL/mg; P > 0.05). The cellular stimulators promoted an increase in PGF2α synthesis (P < 0.02), and the effects differed among the animals (P < 0.04). The PGF2a production was higher in the explants treated with PDBu rather than OT (13.68 ± 1.16 vs. 10.01 ± 1.16 pg/mL/mg tissue, respectively; P < 0.05). In conclusion, PGF2α synthesis is modulated by the presence of the DF (local E2) but not the CL (local P4), and both PDBu and OT stimulated PGF2a synthesis.(AU)

A prostaglandina F2α (PGF2α) determina a luteólise em bovinos. A capacidade de manipular sua síntese endógena é indispensável para a produção animal em grande escala. O estradiol (E2) e a progesterona (P4) modulam diversas vias moleculares das células endometriais, incluindo a síntese de PGF2α; no entanto, pouco se sabe sobre seus mecanismos específicos. Este trabalho investigou a produção in vitro e a possível modulação da PGF2α endometrial devido a um efeito local do E2 endógeno no corno uterino ipsilateral ao folículo dominante (FD) ou da P4 no corno ipsilateral ao corpo lúteo (CL). Os estimuladores de PGF2α oxitocina (OT) e 12,23-dibutirato de forbol (PDBu) foram incubados com explantes endometriais, e o conteúdo de PGF2α foi mensurado. Para tal, vacas cíclicas foram sincronizadas, o desenvolvimento de FD e CL foi examinado por ultrassonografia, e no 17º dia do ciclo estral os explantes endometriais foram coletados e cultivados em meio ou suplementados com PDBu 10-6M ou 10-6M OT. As amostras de meio foram coletadas imediatamente após o tratamento e sessenta minutos depois. O radioimunoensaio mostrou que o conteúdo de PGF2α do corno ipsilateral ao FD foi 49% menor que o do corno contralateral (8,22 ± 0,95 vs. 12,24 ± 0,95 pg/mL/mg de tecido, respectivamente, P < 0,01). No entanto, os níveis de PGF2α não diferiram entre os cornos em função da posição do CL (9,46 ± 0,95 versus 11 ± 0,95 pg/mL/mg; P > 0,05). Os estimuladores celulares promoveram um aumento na síntese de PGF2α (P < 0,02), e os efeitos diferiram entre os animais (P < 0,04). A produção de PGF2α foi maior nos explantes tratados com PDBu em comparação à OT (13,68 ± 1,16 versus 10,01 ± 1,16 pg/mL/mg de tecido, respectivamente, P < 0,05). A conclusão obtida foi que a síntese de PGF2α é: modulada pela presença do FD (E2 local), mas não do CL (P4 local); e estimulada por PDBu e OT.(AU)

Cellular events during ovine implantation and impact for gestation

The establishment of pregnancy in sheep includes elongation of the blastocyst into a filamentous conceptus, pregnancy recognition, production of histotroph, attachment of the conceptus to the endometrium for implantation, and development of synepitheliochorial placentation. These processes are complex, and this review describes some of the molecular events that underlie and support successful pregnancy. The free-floating sheep blastocyst elongates into a filamentous conceptus and metabolizes, or is responsive to, molecules supplied by the endometrium as histotroph. Amongst these molecules are SPP1, glucose and fructose, and arginine that stimulate the MTOR nutrient sensing system. The placental trophectoderm of elongating conceptuses initiate pregnancy recognition and implantation. The mononucleate cells of the trophectoderm secrete IFNT, which acts on the endometrial LE to block increases in estrogen receptor α to preclude oxytocin receptor expression, thereby preventing oxytocin from inducing luteolytic pulses of PGF2α. In addition, IFNT increases expression of IFN stimulated genes in the endometrial stroma, including ISG15, a functional ubiquitin homologue. Implantation is the initial step in placentation, and includes sequential pre-contact, apposition, and adhesion phases. Implantation in sheep includes downregulation of Muc1 and interaction of GLYCAM1, galectin 15 (LGALS15) and SPP1 with lectins and integrins (αvβ3). Sheep have synepitheliochorial placentation in which mononucleate trophectoderm cells fuse to form binucleate cells (BNCs). BNCs migrate and fuse with endometrial LE cells to form trinucleate syncytial cells, and these syncytia enlarge through continued BNC fusion to form syncytial plaques that form the interface between endometrial and placental tissues within the placentome. The placentae of sheep organize into placentomal and interplacentomal regions. In placentomes there is extensive interdigitation of endometrial and placental tissues to provide hemotrophic nutrition to the fetus. In interplacentomal regions there is epitheliochorial attachment of endometrial LE to trophectoderm, mediated through focal adhesion assembly, and areolae that take up histotroph secreted by endometrial GE.

Cellular events during ovine implantation and impact for gestation

The establishment of pregnancy in sheep includes elongation of the blastocyst into a filamentous conceptus, pregnancy recognition, production of histotroph, attachment of the conceptus to the endometrium for implantation, and development of synepitheliochorial placentation. These processes are complex, and this review describes some of the molecular events that underlie and support successful pregnancy. The free-floating sheep blastocyst elongates into a filamentous conceptus and metabolizes, or is responsive to, molecules supplied by the endometrium as histotroph. Amongst these molecules are SPP1, glucose and fructose, and arginine that stimulate the MTOR nutrient sensing system. The placental trophectoderm of elongating conceptuses initiate pregnancy recognition and implantation. The mononucleate cells of the trophectoderm secrete IFNT, which acts on the endometrial LE to block increases in estrogen receptor α to preclude oxytocin receptor expression, thereby preventing oxytocin from inducing luteolytic pulses of PGF2α. In addition, IFNT increases expression of IFN stimulated genes in the endometrial stroma, including ISG15, a functional ubiquitin homologue. Implantation is the initial step in placentation, and includes sequential pre-contact, apposition, and adhesion phases. Implantation in sheep includes downregulation of Muc1 and interaction of GLYCAM1, galectin 15 (LGALS15) and SPP1 with lectins and integrins (αvβ3). Sheep have synepitheliochorial placentation in which mononucleate trophectoderm cells fuse to form binucleate cells (BNCs). BNCs migrate and fuse with endometrial LE cells to form trinucleate syncytial cells, and these syncytia enlarge through continued BNC fusion to form syncytial plaques that form the interface between endometrial and placental tissues within the placentome. The placentae of sheep organize into placentomal and interplacentomal regions. In placentomes there is extensive interdigitation of endometrial and placental tissues to provide hemotrophic nutrition to the fetus. In interplacentomal regions there is epitheliochorial attachment of endometrial LE to trophectoderm, mediated through focal adhesion assembly, and areolae that take up histotroph secreted by endometrial GE.(AU)

Partial ablation of endometrial glands in dogs after exposure to progestin during the neonatal period

Bitches with uteri devoid of endometrial glands should be sterile, and consequently could contribute to the population control of dogs. Considering that an inadequate exposure of the female reproductive system to steroids can lead to the formation of the uterine gland knock-out (UGKO) phenotype in some species, the aim of this study was to evaluate the effect of serial applications of medroxyprogesterone acetate (MPA) from birth until the age of six months on the development of endometrial glands in bitches. For this purpose, 16 female mongrel dogs from different litters were distributed into either an MPA group (n = 8), animals treated with 10 mg kg sc (Promone-E®, Pfizer, Brasil) at 3-week intervals, from day one after birth until the age of six months, or a control group (n = 8), composed of animals that only received a 0.9% NaCl solution in place of MPA. At six months of age, ovariohysterectomy was performed and uterine horn samples were collected for histological and immunohistochemical examinations. The bitches from the MPA-treated group presented a 35% decrease in the number of endometrial glands, a larger diameter of the endometrial glands, a greater epithelial height, as well as a greater thickness of the uterine wall, endometrium, and myometrium. However, no significant differences were observed between the two groups in the expression of ER-α, ER-β, and PR on the surface epithelium and endometrial stroma. Therefore, the serial application of MPA from birth until the age of 6 months do not completely ablate the development of the endometrial glands in bitches, but impair it by 35%.