DigitalUM :: Browsing by Subject "Oviduct"

Browsing by Subject "Oviduct"

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Open Access
Bovine oviduct epithelial cells suppress the phagocytic activity of neutrophils towards sperm but not for bacteria in vitro: Immunofluorescence and electron microscopic observations
(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2020) Marey, Mohamed Ali; Matsukawa, Haruhisa; Sasaki, Motoki; Ezz, Mohamed Aboul; Yousef, Mohamed Samy; Takahashi, Ken-ichi; Miyamoto, Akio
Previously, we reported that polymorpho- nuclear neutrophils (PMNs) are constantly existent in the bovine oviduct fluid during the pre-ovulatory stage under physiological conditions. Moreover, incubation of PMNs with bovine oviduct epithelial cells-conditioned medium (BOEC-CM) resulted in suppression of their phagocytic activity for sperm. During pathophysiological conditions, cows may be inseminated by infected semen which exposes oviductal PMNs to allogenic sperm simultaneously with pathogens. This study aimed to visually investigate the role of oviduct epithelium in regulating the phagocytic behavior of PMNs toward sperm as a physiological stimulus, with Escherichia coli (E. coli) as a pathological stimulus. In our experiment, PMNs were incubated for 2 h in BOEC-CM. Phagocytosis was then assayed by co-incubation of these PMNs either with sperm, E. coli, or latex beads. BOEC- CM significantly suppressed the direct phagocytosis of PMNs for sperm, but did not affect their phagocytic activity for E. coli or latex beads. Additionally, an investigation with scanning electron microscopy revealed that BOEC-CM suppressed the formation of DNA-based neutrophil extracellular traps (NETs) for sperm entanglement. BOEC-CM did not alter NETs formation towards E. coli. A quantification of NETs formation using an immunofluorescence microscopy showed that the areas of NETs formation for E. coli were significantly larger than those formed for sperm. Our data clearly show that the bovine oviduct, through secretions, protects sperm from phagocytosis by PMNs and eliminates bacterial dissemination through maintaining the phagocytic activity of PMNs towards bacteria
Open Access
Caracterización de exosomas producidos por células oviductales in vivo e in vitro, en la especie bovina
(Facultad de Veterinaria y el Servicio de Publicaciones de la Universidad de Murcia, 2023) Toledo Guardiola, Santa María; Matás Parra, Carmen; Rueda Gomariz, Almudena
Las vesículas extracelulares (VEs), exosomas y micro vesículas son un tipo de estructuras heterogéneas pre-sentes en la mayoría de los fluidos orgánicos incluyendo el fluido oviductal. Las VEs contienen varios compuestos derivados de la célula original, como proteínas, lípidos, ARNm, miARN y ADN. Las VEs en el oviducto son producidas por las células epiteliales y entre sus funciones se encuentran: la interacción con los espermatozoides, mantener la viabilidad de éstos, participar en la maduración de los ovocitos y en el proceso de fecundación. Durante la fecundación in vitro y con el fin de mejorarla imitando las condiciones in vivo, numerosos investigadores han utilizado cultivos de células del epitelio oviductal bovino (CEOB) con notables mejoras. Estas células producen, entre otros componentes VEs, por ello, en este trabajo hemos planteado un estudio com-parativo de VEs presentes en el fluido oviductal (FO) bovino recogido en momentos próximos a la ovulación (in vivo) y de aquellas VEs producidas en cultivos de CEOB a los 7 días de cultivo (in vitro) comparando el tamaño, la distribución de la población y la concentración de proteína en ambos tipos. Las VEs se identificaron mediante microscopía electrónica, su tamaño mediante dispersión de luz láser y la concentración de proteínas mediante el método Bradford. Los resultados mostraron que el tamaño de las VEs fue similar entre ambos grupos experimentales. Por otro lado, sí que se observaron diferencias en cuanto a la concentración de proteínas. Las VEs obtenidas in vivocontenían mayor cantidad de proteína en su cargo que en las VEs obtenidas in vitro.En cuanto a identificación de las VEs mediante microscopía electrónica de transmisión, solo pudieron ser observadas aquellas obtenidas in vivo. Este hecho podría deberse al lugar de dónde han sido recogidas, al mét-odo de cultivo de células epiteliales oviductales bovinas o la escasez en su producción.
Open Access
Comparative View on the Oviductal Environment during the Periconception Period.
(2020) González-Brusi, Leopoldo; Algarra, Blanca; Izquierdo Rico, María José; Avilés Sánchez, Manuel; Jíménez Movilla, María; Moros Nicolás, Carla; Biología Celular e Histología
The oviduct plays important roles in reproductive events: sperm reservoir formation, final gamete maturation, fertilization and early embryo development. It is well known that the oviductal environment affects gametes and embryos and, ultimately, the health of offspring, so that in vivo embryos are better in terms of morphology, cryotolerance, pregnancy rates or epigenetic profile than those obtained in vitro. The deciphering of embryo–maternal interaction in the oviduct may provide a better understanding of the embryo needs during the periconception period to improve reproductive efficiency. Here, we perform a comparative analysis among species of oviductal gene expression related to embryonic development during its journey through the oviduct, as described to date. Cross-talk communication between the oviduct environment and embryo will be studied by analyses of the secreted or exosomal proteins of the oviduct and the presence of receptors in the membrane of the embryo blastomeres. Finally, we review the data that are available to date on the expression and characterization of the most abundant protein in the oviduct, oviductin (OVGP1), highlighting its fundamental role in fertilization and embryonic development.
Open Access
Cytomorphological changes in the rabbit oviductal epithelium after human chorionic gonadotropin treatment
(Murcia : F. Hernández, 1997) Bondi, A.M.; Gabrielli, M.G.; Marchetti, L.; Materazzi, G.; Menghi, Giovanna
An electron microscopic investigation was performed to examine the ultrastructural changes occurring in the rabbit oviductal epithelium after human chorionic gonadotropin (HCG) administration. Mainly, the non-ciliated secretory cells proved to be affected by the hormonal treatment which resulted in qualitative and quantitative modifications of the secretory patterns differently expressed in the ampulla and isthmus. Thus, morphological evidence of intense secretion was observed in both the oviduct regions at preovulatory stages. Following ovulation, timing of expression of active secretory patterns in the ampulla and isthmus correlated well with the rate of gamete transport and relative functional roles of the oviductal regions in the reproductive process. At present, HCG-induced changes concerning the ciliated cells seem to consist of the occurrence of secretory granules responsible for the appearance of "mixed cells".
Open Access
Effect of cumulus cell removal and sperm pre‐incubation with progesterone on in vitro fertilization of equine gametes in the presence of oviductal fluid or cells
(2019-05-30) Douet, C; Reigner, F; Goudet, G; Moros Nicolás, Carla; Biología Celular e Histología
In spite of many attempts to establish an in vitro fertilization (IVF) technique in the equine, no efficient conventional IVF technique is available. The presence of oviductal fluid or oviductal cells during IVF help to improve embryo production in vitro but is not sufficient to reach high fertilization rates. Thus, our aim was to perform equine IVF either after sperm preincubation with oviductal fluid or in the presence of oviductal cells, and to evaluate the effect of cumulus removal from the oocyte or sperm preincubation with progesterone. In experiment 1 and 2, IVF was performed in the presence of porcine oviduct epithelial cells. The removal of cumulus cells from equine oocytes after in vitro maturation tended to increase the percentage of fertilization when fresh sperm was used (1/33 vs 4/31, p > 0.05) but had no effect when frozen sperm was used (1/32 vs 1/32). Equine sperm preincubation with progesterone did not significantly influence the fertilization rate when fresh or frozen sperm was used (2/14 vs 2/18 for fresh, 1/29 vs 1/25 for frozen). In experiment 3 and 4, IVF was performed after preincubation of sperm with porcine oviductal fluid. The removal of cumulus cells tented to increase the percentage of fertilization when fresh sperm was used (1/24 vs 3/26, p>0.05). Sperm preincubation with progesterone did not significantly influence the fertilization rate when fresh or frozen sperm was used (2/39 vs 2/36 for fresh, 2/37 vs 1/46 for frozen), but two 3-4 cells stage zygotes were obtained with fresh sperm preincubated with progesterone. This is an encouraging result for the setting up of an efficient IVF procedure in equine.
Open Access
Genes Encoding Mammalian Oviductal Proteins Involved in Fertilization are Subjected to Gene Death and Positive Selection
(2018) Fouchécourt, Sophie; Goudet, Ghylène; Monget, Philippe; Moros Nicolás, Carla; Biología Celular e Histología
Oviductal proteins play an important role in mammalian fertilization, as proteins from seminal fluid. However, in contrast with the latter, their phylogenetic evolution has been poorly studied. Our objective was to study in 16 mammals the evolution of 16 genes that encode oviductal proteins involved in at least one of the following steps: (1) sperm–oviduct interaction, (2) acrosome reaction, and/or (3) sperm–zona pellucida interaction. Most genes were present in all studied mammals. However, some genes were lost along the evolution of mammals and found as pseudogenes: annexin A5 (ANXA5) and deleted in malignant brain tumor 1 (DMBT1) in tarsier; oviductin (OVGP1) in megabat; and probably progestagen-associated endometrial protein (PAEP) in tarsier, mouse, rat, rabbit, dolphin, and megabat; prostaglandin D2 synthase (PTGDS) in microbat; and plasminogen (PLG) in megabat. Four genes [ANXA1, ANXA4, ANXA5, and heat shock 70 kDa protein 5 (HSPA5)] showed branch-site positive selection, whereas for seven genes [ANXA2, lactotransferrin (LTF), OVGP1, PLG, S100 calcium-binding protein A11 (S100A11), Sperm adhesion molecule 1 (SPAM1), and osteopontin (SPP1)] branch-site model and model-site positive selection were observed. These results strongly suggest that genes encoding oviductal proteins that are known to be important for gamete fertilization are subjected to positive selection during evolution, as numerous genes encoding proteins from mammalian seminal fluid. This suggests that such a rapid evolution may have as a consequence that two isolated populations become separate species more rapidly
Open Access
Glycoconjugates within the oviduct and their functional significance with special reference to marsupials
(Murcia : F. Hernández, 2010) Chapman, Jamie A.; Chuah, M.I.; Breed, William G.
In placental (eutherian) mammals, a number of important events take place within the oviduct including the pre-fertilisation maturation of gametes (including sperm storage), sperm-egg interactions, egg activation and early embryonic development. Many of these events involve interactions of glycoconjugates; both on the surface of the gametes and with the secretions of the oviductal epithelium and these have best been studied in eutherian mammals. In marsupials, however, while the oviduct is known to produce the extracellular egg coat, the mucoid layer, that comes to surround the zona pellucida, its role in the maturation of gametes is only now being elucidated, particularly in the oocyte. This review emphasises what is known of the structure and function of the oviduct and its secretions in marsupials and briefly compares it with data from eutherians. In particular, knowledge of oviductal glycoconjugates in the structure of the post-ovulatory oocyte and its vestments around the time of fertilisation in Australian marsupials is outlined.
Open Access
Mimicking the temperature gradient between the sow’s oviduct and uterus improves in vitro embryo culture output
(Oxford University Press, 2020-07-09) García Martínez, Soledad; Latorre Reviriego, Rafael Manuel; Sánchez-Hurtado, M. A.; Sánchez-Margallo, F. M.; Bernabò, N.; Romar Andrés, Raquel; López Albors, Octavio Miguel; Coy Fuster, Pilar; Anatomía y Anatomía Patológica Comparada
This work was designed to determine temperature conditions within the reproductive tract of the female pig and study their impact on ARTs. Temperatures were recorded using a laparo-endoscopic single-site surgery assisted approach and a miniaturized probe. Sows and gilts were used to address natural cycle and ovarian stimulation treatments, respectively. According to in vivo values, IVF was performed at three temperature conditions (37.0°C, 38.5°C and 39.5°C) and presumptive zygotes were cultured in these conditions for 20 h, while further embryo culture (EC) (21–168 h post-insemination) was maintained at 38.5°C. After 20 h, different fertility parameters were assessed. During EC, cleavage and blastocyst stages were evaluated. Sperm membrane fluidity at the experimental temperatures was studied by using differential scanning calorimetry and fluorescence recovery after photobleaching techniques. An increasing temperature gradient of 1.5°C was found between the oviduct and uterus of sows (P < 0.05) and when this gradient was transferred to pig in vitro culture, the number of poly-nuclear zygotes after IVF was reduced and the percentage of blastocysts was increased. Moreover, the temperature transition phase for the boar sperm membrane (37.0°C) coincided with the temperature registered in the sow oviduct, and sperm membranes were more fluid at 37.0°C compared with those of sperm incubated at higher temperatures (38.5°C and 39.5°C). These data suggest that there may be an impact of physiological temperature gradients on human embryo development.
Open Access
Progesterone induces the release of bull spermatozoa from oviductal epithelial cells
(CSIRO Publishing, 2019-04-29) Romero Aguirregomezcorta, Jon; Cronin, S.; Donnellan, E.; Fair, S.; Anatomía y Anatomía Patológica Comparada; Facultad de Veterinaria
The mechanism that causes the detachment of spermatozoa from the oviductal reservoir around the time of ovulation remains to be elucidated. Because the cumulus cells of the bovine oocyte are known to secrete progesterone (P4), and P4 has been shown to act upon cation channels of spermatozoa (CatSper) in human spermatozoa, it was hypothesised that P4 could induce hyperactivation due to an influx of extracellular calcium, and this would facilitate detachment of spermatozoa from oviductal epithelial cells. Therefore, this study aimed to investigate the role and mechanism of action of P4 in the release of spermatozoa from bovine oviduct epithelial cells (BOEC). Initial dose–response assessments on sperm hyperactivation determined the optimum concentration of P4 (10 nM), mibefradil (a non-specific Ca2+ channel antagonist; 5 µM), NNC 55-0396 dihydrochloride (NNC; a CatSper antagonist; 2 µM), mifepristone (a classical and membrane P4 receptor antagonist; 400 nM) and AG205 (a membrane P4 receptor antagonist; 10 µM). BOEC explants were incubated with frozen–thawed bovine spermatozoa for 30 min, following which loosely bound spermatozoa were removed. Two experiments were completed. In Experiment 1, BOECs were treated for 30 min with either no treatment, P4, NNC, mibefradil, P4 + mibefradil, P4 + NNC, P4 + mibefradil + NNC or P4 + EGTA. In Experiment 2, BOECs were treated for 30 min with either no treatment, P4, mifepristone, AG205, mifepristone + AG205, P4 + mifepristone, P4 + AG205 or P4 + mifepristone + AG205. The number of spermatozoa remaining bound per millimetre squared of BOEC explant was determined. Progesterone stimulated the release of bound spermatozoa from BOEC explants, whereas NNC, mibefradil and EGTA inhibited this release. The release of spermatozoa by P4 was inhibited in the presence of both mifepristone and AG205, whereas the combination of both had the greatest inhibitory action on P4 release of spermatozoa. These findings suggest the presence of a P4 membrane receptor on bovine spermatozoa and that P4-induced release of spermatozoa from BOECs is likely mediated by extracellular Ca2+.
Open Access
The immunolocalization of HIF-2α, GLUT1 and CAIX in porcine oviduct during the estrous cycle
(Wiley, 2022-06-09) Párraga Ros, Ester; Latorre Reviriego, Rafael Manuel; Aparicio Gozález, Mónica; Boronat Belda, Talía; López Albors, Octavio Miguel; Anatomía y Anatomía Patológica Comparada
Oxygen (O2) rates in the oviduct are essential to human and animal reproduction. These rates are regulated by the activity of hypoxia markers such as the hypoxia-inducible factors (HIFs), the glucose transporters (GLUT), and the carbonic anhydrase (CA). In the porcine model, scarce studies have been reported regarding these markers and their effects in reproduction are unknown. The objective was to characterize the immunolocalization of HIF-2α, GLUT1, and CAIX in porcine oviducts throughout the estrous cycle. Oviducts (ampulla and isthmus) of adult sows (n = 45) were collected for histological and immunohistochemical analysis with HIF-2α, GLUT1, and CAIX markers. The percentage of immunopositive area was quantified, and the differences among phases of the estrous cycle were analyzed (folicular, early luteal, and late luteal). The three markers showed epithelial presence mainly. Significantly lower expression of HIF-2α was found in the luteal phases, especially in the isthmus. GLUT1 expression did not change throughout the estrous cycle, but differences were found between the ampulla and isthmus. CAIX expression showed the highest, with a significant downward trend throughout estrous cycle. The ubiquitous expression of hypoxia markers shows the porcine oviduct physiology in relation to O2. The differential expression of HIF-2α, GLUT1, and CAIX in different subcompartments of the oviduct throughout the estrous cycle contributes to improve the knowledge of the cell physiology of the oviduct, which can be useful in fertilization studies.
Open Access
The presence of the u-opioid receptor in the isthmus of mare oviduct
(Murcia : F. Hernández, 2008) Desantis, S.; Albrizio, M.; Ventriglia, G.; Deflorio, M.; Guaricci, A.C.; Minoia, R.; De Metrio, G.
The presence of the μ-opioid receptor and the type of glycosylation in the third extra-cellular loop of this receptor was investigated in the isthmus of mare oviduct during oestrus by means of immunoblotting and immunohistochemistry combined with enzymatic (Nglycosidase F and O-glycosidase) and chemical (ßelimination) treatments. Immunoblotting analysis showed that the μ -opioid receptor consists of two peptides with molecular weights of around 65 and 50 kDa. After N-deglycosylation with N-glycosidase F an additional immunoreactive peptide was observed at around 30 KDa. The cleavage of O-glycans by Oglycosidase failed in immunoblotting as well as in immunohistochemistry investigations, revealing that the third extra-cellular loop of the μ -opioid receptor expressed in mare isthmus oviduct contains some modifications of the Galß(1-3)GalNAc core binding to serine or threonine. Immunohistochemistry revealed the μ-opioid receptor in the mucosal epithelium, some stromal cells, muscle cells and blood vessels. In ciliated cells the μ-opioid receptor showed N-linked glycans, since the immunoreactivity was abolished after Nglycosidase F treatment, whereas it was preserved in the apical region after ß-elimination. Most non-ciliated cells expressed the μ-opioid receptor with both N- and Olinked oligosaccharides, as revealed by the abolition of immunostaining after N-glycosidase F and ßelimination. Stromal cells, endothelial and muscle cells of blood vessels expressed the μ -opioid receptor containing both N- and O-linked oligosaccharides. Myosalpinx myocytes expressed the μ-opioid receptor with O-linked oligosaccharides. The immunopositive myocytes formed a circular coat in the intrinsic musculature, whereas they were arranged in some isolated, oblique bundles in the extrinsic musculature. In conclusion, the μ-opioid receptor could have a role in the production and the movement of isthmus lumen content that contributes to ensuring the effective condition of the sperm in the mare oviduct.
Open Access
Timing of oviductal fluid collection, steroid concentrations and sperm preservation method affect in vitro fertilization efficiency.
(2014) Ballester, L.; Soriano Ubeda, C. M.; Matas Parra, Carmen; Romar Andrés, Raquel; Coy Fuster, Pilar; Romero Aguirregomezcorta, Jon; Fisiología

Browsing by Subject "Oviduct"

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