Bovine Oviduct Research Articles

Summer heat stress affects prostaglandin synthesis in the bovine oviduct

Summer heat stress (HS) negatively affects reproductive functions, including prostaglandin (PG) F2α secretion in the endometrium, and decreases fertility in cattle. In the present study, we examined the effects of elevated temperatures on PG synthesis in oviductal epithelial cells. The epithelial cells obtained from the ampulla and isthmus of the oviduct were incubated at various temperatures (38.5, 39.5, 40.0, and 40.5 °C) for 24 h. In the ampulla, PGE₂ concentration was higher at 40.5 °C than at 38.5 °C, while PGF2α production was not affected by the temperatures in this range. The expressions of microsomal PGE synthase 1 (PTGES (mPGES1)), cytosolic PGES (PTGES3 (cPGES)), and heat shock protein 90 (HSP90AA1 (HSP90)) mRNAs and proteins were higher at 40.5 °C than at 38.5 °C in the ampullary epithelial cells. Seasonal changes in the expressions of PGES and HSP90AA1 mRNAs in oviductal tissues were also investigated. The expressions of PTGES3 and HSP90AA1 mRNAs were higher in the ampullary tissues in summer than in winter. In summary, elevated temperatures stimulated PGE₂ production in the ampullary oviduct by increasing the expressions of PGESs and HSP90AA1, which can activate cPGES. The overall results suggest that HS upsets PG secretions and reduces oviductal smooth muscle motility, which in turn could decrease gamete/embryo transport through the oviduct.

Progesterone receptors mRNA expression in bovine oviduct during estrous cycle

Progesterone receptors mRNA expression in bovine oviduct during estrous cycle

IL-10 release by bovine epithelial cells cultured with Trichomonas vaginalis and Tritrichomonas foetus

Trichomonas vaginalis and Tritrichomonas foetus are parasitic protists of the human and bovine urogenital tracts, respectively. Several studies have described the cytotoxic effects of trichomonads on urogenital tract epithelial cells. However, little is known about the host cell response against trichomonads. The aim of this study was to determine whether T. foetus and T. vaginalis stimulated the release of the cytokine interleukin (IL)-10 from cultured bovine epithelial cells. To characterise the inflammatory response induced by these parasites, primary cultures of bovine oviduct epithelial cells were exposed to either T. vaginalis or T. foetus. Within 12 h after parasite challenge, supernatants were collected and cytokine production was analysed. Large amounts of IL-10 were detected in the supernatants of cultures that had been stimulated with T. foetus. Interestingly, T. vaginalis induced only a small increase in the release of IL-10 upon exposure to the same bovine cells. Thus, the inflammatory response of the host cell is species-specific. Only T. foetus and not T. vaginalis induced the release of IL-10 by bovine oviduct epithelial cells.

Long-term culture of primary bovine oviduct epithelial cells: Impact of insulin and EGF supplementation

Long-term culture of primary bovine oviduct epithelial cells: Impact of insulin and EGF supplementation

Regulation of Innate Immune Function in Bovine Oviduct Epithelial Cells in Culture: The Homeostatic Role of Epithelial Cells in Balancing Th1/Th2 Response

This study aimed to investigate the role of epithelial cells in regulating innate immunity in bovine oviduct epithelial cell (BOEC) culture. We studied the effect of Escherichia coli lipopolysaccharide (LPS) and its interaction with ovarian steroids, estradiol (E2) and progesterone (P4), and luteinizing hormone (LH) at concentrations observed during the preovulatory period on immune responses in BOEC culture. Immunohistochemistry of oviduct tissue showed intensive expression of Toll-like receptor-4 (TLR-4) and TLR-2 in epithelial cells. A dose of 10 ng/ml LPS stimulated TLR-4, cyclooxygenase-2 (COX-2), nuclear factor kappa B inhibitor A (NFKBIA), interleukin 1β (IL-1β) and tumor necrosis factor α (TNF-α) expression, indicating an early pro-inflammatory response. A dose of 100 ng/ml LPS did not induce expression of these genes but stimulated TLR-2, IL-10,IL-4 and microsomal prostaglandin E synthase-1 (mPGES-1) expression and PGE2 secretion, indicating an anti-inflammatory response. Ovarian steroids and LH completely block LPS (10 ng/ml)-induced TLR-4, IL-1β and TNF-α expression as well as LPS (100 ng/ml)-induced TLR-2 expression. Taken together, this study suggests the existence of an early signaling system to respond to infection in the BOEC. In addition, ovarian steroids and LH may play a critical role in inducing homeostasis and in controlling hyperactive pro-inflammatory responses detrimental to epithelial cells, sperm and the embryo.

96 EXPRESSION OF THE LONG FORM OF THE LEPTIN RECEPTOR IN BOVINE OVIDUCT EPITHELIAL, UTERINE EPITHELIAL, AND BLASTOCYST-STAGE EMBRYOS

Critical reproductive events of animals are affected via metabolic cues that directly modulate the hypothalamic-pituitary-gonadal axis. The adipose-derived 146-amino-acid protein-hormone leptin has been associated with puberty and reproduction in many mammals and may be of critical importance in the regulation of reproduction. Leptin receptors are expressed in the brain, pituitary, and gonads, and there is growing evidence that leptin may be required for normal embryonic implantation and development in some animals. To date 6 isoforms of the leptin receptor have been identified; the long form (Ob-Rb) is the only isoform to be associated with signal transduction. The objective of this experiment was to test the hypothesis that the Ob-Rb isoform is expressed in cultured bovine oviduct and uterine epithelial cells, in vivo-derived mid-luteal uterine biopsies, and expanded bovine blastocyst-stage embryos. For cells, cell lines from mid-luteal abattoir-derived reproductive tracts (n = 5) were used; for uterine biopsies (n = 20), biopsies were collected from both uterine horns at the greater curvature from mature beef cows (n = 4) in mid-luteal phase; expanded IVP blastocyst-stage embryos (n = 40) were produced from abattoir-derived ovaries. For fluorescence microscopy samples were incubated in rabbit anti-human leptin receptor (#4781-L, Linco Research, St. Charles, MO) followed by Alexa Fluor 488 anti-rabbit IgG (#A-11008, Molecular Probes, Carlsbad, CA). We confirmed that the Ob-Rb isoform is expressed in both oviductal and uterine in vitro-cultured epithelial cells and uterine biopsies. All oviduct but not all uterine-cultured epithelial cells were immunopositive for the Ob-Rb isoform. All uterine biopsies were positive for the Ob-Rb isoform along the luminal side of the uterine endometrium with an apparent lack of staining occurring in myometrial tissue. Moreover, we demonstrated that all bovine blastocysts were immunopositive with staining occurring within the trophoblast cell type but not the inner cell mass. These results show for the first time that bovine oviduct and uterine epithelial cells express the long form of the leptin receptor and support earlier findings that blastocyst-stage embryos contain Ob-Rb isoforms. This suggests that the Ob-Rb isoform is a component of the bovine maternal-embryonic complex; however, its function in pre-implantation embryo development remains to be clarified.

123 GENISTEIN TRANSPORT ACROSS THE BOVINE OVIDUCT EPITHELIUM

The oviduct plays a vital role in regulating the environment surrounding the gametes and early mammalian embryo. However, the permeability of the oviduct to circulating dietary-derived compounds remains relatively unknown. The present study has investigated the barrier properties of the oviduct epithelium in vitro to the movement of genistein, a soya isoflavone and analogue of 17β-estradiol that naturally occurs in the diet and has been reported to exert teratogenic effects. Bovine oviduct epithelial cells (BOECs) were isolated from abattoir-derived reproductive tracts at the mid-luetal phase. The purity of the cell isolate was confirmed using flow cytometry to determine the proportion of cells that expressed CK18 (epithelia), and Vimentin (fibroblasts). Cells were seeded at a density of 1 × 106 mL–1 on polyethylene terephthalate transwell porous supports (Corning) and maintained between two media-filled chambers for 10 to 12 days, until they formed a polarised confluent monolayer, as confirmed by transepithelial resistance (TEER) greater than 700 Ωcm2. To assess the rate of transport, genistein was added to the basal compartment at physiologically relevant levels (100 µM) and the apical compartment was sampled at regular time points for 120 mins. The concentration of genistein in the apical and basal media was measured by HPLC. Furthermore, we compared the rate of genistein transport at physiological (39°C) and room temperature to indicate whether transport was temperature dependent. Rates of transport are expressed as mean apparent permeability coefficient and were compared between groups by a Student’s t-test. Genistein crossed the bovine oviduct epithelium at a linear rate that was higher than spontaneous diffusion across a blank membrane support (12.7 × 10–3 cm–2 µM–1 v. 7.32 × 10–3 cm–2 µM–1, n = 4; P = 0.0075). The rate of genistein transport by epithelial cells was unchanged when cells were assayed at room temperature (12.7 × 10–3 cm–2 µM–1 v. 13.11 × 10–3 cm–2 µM–1, n = 3; P = 0.76), respectively. No significant difference in the directionality of transport was found. Furthermore, TEER was maintained at approximately 700 Ωcm2, indicating that the cells remained confluent for the duration of the experiment. These data suggest that the bovine oviduct epithelial cell monolayer facilitates genistein movement from the basal to the apical compartment in vitro. Furthermore, the observation that the rate of transport is unchanged by temperature suggests a passive, trans-cellular, physicochemical mechanism, rather than an active biological process. Regardless of the mechanism, the oviduct epithelium is permeable to genistein, and may even facilitate its transport into the lumen, suggesting that gametes and early embryos could be exposed to this compound in vivo. This is relevant given the previously reported finding indicating that this naturally occurring dietary isoflavone has detrimental effects on early development (Newbold et al. 2001 Cancer Res. 61, 4325–4328). Furthermore, the results demonstrate the potential use of this epithelial model in characterising the transport or barrier properties of the oviduct epithelium towards a range of circulating xenobiotics.

Immunolocalization of Progesterone Receptors in the Canine Oviduct around Ovulation

In the bitch, oocyte maturation, sperm storage, fertilization and early embryo development take place within the oviducts under high and increasing circulating progesterone concentrations. To investigate the potential effects of progesterone on the canine oviduct, nuclear progesterone receptors (PR) were localized. Oviducts were collected by ovariectomy from adult Beagle bitches during anestrus, after the onset of proestrus but prior to the Luteinizing Hormone (LH) peak (Pre-LH), after the LH peak but before ovulation (Pre-ov) and on Days 1, 4 and 7 after ovulation (n = 3 bitches per stage). The cellular distribution of PR was studied by immunohistochemistry (IHC) in the ampulla, isthmus and tubal part of the utero-tubal junction (UTJ). Plasma progesterone and 17β-oestradiol were assayed on the day of surgery. PR were specifically expressed in the nuclei of epithelial, stromal and muscular cells in the ampulla, isthmus and UTJ. The IHC scores did not vary from one oviductal region to another. However, the epithelium displayed higher scores than the stroma at anestrus, Pre-ov, Days 4 and 7, and also higher scores than muscle at Days 4 and 7 (p < 0.05). Immunohistochemistry scores in the stroma and muscle decreased at Days 4 and 7 compared with previous stages (p < 0.05). Furthermore, muscular IHC scores were positively correlated with circulating 17β-oestradiol concentrations and negatively correlated with circulating progesterone concentrations (p < 0.05). In conclusion, PR were identified in the canine oviduct, with differences in expression between tissues and times around ovulation, suggesting that progesterone may regulate tubal functions and reproductive events in this species.

Efficiency of in vitro embryo production of yak (Bos grunniens) cultured in different maturation and culture conditions

A study was conducted to evaluate the potential of immature oocytes from the ovaries of yak. The effects of maturation times (23–30 h), hormones (FSH, LH, E2), sera (fetal calf serum, FCS, or oestrous cow serum, ECS) and culture system (co-culture with granulosa cell, GC, or with bovine oviduct epithelia cells, BOEC, tissue culture medium, TCM, 199 absence co-culture cell, and synthetic oviduct fluid with amino acids, SOFaa) on the in vitro development of in vitro matured and fertilised yak oocytes were examined. Immature oocytes surrounded with compacted cumulus cell were cultured for 23–30 h in TCM 199 supplemented with 10% FCS and hormones. In vitro fertilisation (IVF) was performed with frozen-thawed, caffeine and heparin treated spermatozoa from Datong yak. Oocytes were incubated with 1×106/ml spermatozoa for 16–18 h and then cultured in co-culture system with GC or BOEC and/or SOFaa for 7–8 days, respectively. Cleavage and development to blastocysts were recorded on days 2 and 8, respectively, after the start of culture. 27–28 h was superior to other culture times and addition hormones (FSH, LH, E2) were superior to absence either of them for oocyte maturation. The best maturation of oocyte in TCM 199 with 5.0 mg/L LH, 0.5 mg/L FSH, 1 mg/L E2. FCS was superior to ECS for development to blastocysts. Co-culture with GC was superior to SOFaa and TCM 199 absence co-culture cell for cleavage, development to blastocysts. The results show that choice of culture conditions has marked effect on the development of in vitro matured and fertilised yak oocytes. The present results indicate that the co-culture with GCs is the most important factor for IVF to development into blastocysts of yak oocytes matured in vitro.

Expression and localization of nodal in bovine oviduct and uterus during different functional stages of oestrus cycle and pregnancy

Members of TGF-β superfamily play a major role in the endometrial changes involved in the establishment and maintenance of pregnancy. Their deregulated expression and action could lead to absolute or partial failure of embryo implantation. Nonetheless, the precise function and mechanism of many of these cytokines remain unclear. Nodal, a transforming growth factor beta (TGF-β) superfamily member, was characterized in the human and rodent uterus and implicated in the tissue remodeling events during menstruation and embryo implantation. In order to study its possible role in the cattle reproductive process, we have analyzed Nodal expression pattern and localization in the oviduct and uterine horn during the oestrus cycle and early pregnancy (day 20). Nodal was detected both in oviduct and uterus during either the oestrus cycle or pregnancy; however, it shows a differential expression profile in the uterine horn at dioestrus and pregnancy, decreasing 1.5 and 1.4 folds in comparison with oestrus. Nodal immunostaining intensity was observed in stromal and in epithelial cells of the surface and the glandular epithelium. The staining pattern correlates with the RT-qPCR expression profile. This work is the first to evidence the presence of Nodal in the bovine reproductive tract; our data suggest that Nodal is a novel cytokine that would be involved in the remodelling occurring in the endometrium of cattle during the oestrus cycle and in the embryo implantation. The identification of new molecules that participate in endometrium cycling and/or pregnancy may be useful for predicting the ability of the uterine tissue to establish and maintain pregnancy or for detecting the infertility processes. These results highlight Nodal as a possible novel marker of the fertility process, nevertheless further studies should be done to determine its role in the reproductive system.

Local immune tolerance via Toll-like receptor 2 and 4 in the bovine oviduct epithelial cells

Local immune tolerance via Toll-like receptor 2 and 4 in the bovine oviduct epithelial cells

The polyol pathway in the bovine oviduct

The oviducts likely provide optimized micro-environments for the final maturation of gametes, fertilization, and early embryo development. Hexoses, including glucose, fructose, and sorbitol, are involved in these critical reproductive events. Monosaccharide production is controlled, in part, by the polyol pathway and requires two enzymes: an aldose reductase (AR) that reduces glucose into sorbitol, followed by its oxidation into fructose by sorbitol dehydrogenase (SDH). We analyzed the expression of AR and SDH in the isthmus and ampulla of the bovine oviduct at the proliferative, mid-luteal, and late-luteal phases of the estrous cycle by quantitative PCR and immunoblots. Immunochemistry and an assay of SDH activity were also performed. The quantity of hexoses in whole sections of isthmus and ampulla were determined by liquid chromatography coupled to mass spectrometry. In sum, AR expression was restricted to the isthmus, while SDH was mostly expressed in the isthmic-ampullary junction and the ampulla, specifically concentrated in the luminal epithelium of the oviduct. The estrous cycle had no impact on protein expression of AR and SDH. Instead, the levels of AR and SDH expression were associated with higher ratios of sorbitol to fructose in the isthmus (1.6) than in the ampulla (4.1; P = 0.005). These results are discussed in light of physiological events occurring in the oviduct.

MRNA Expression of Selected Candidate Genes in the Bovine Oviduct Is Influenced by the Energy Status as a Long-Term Effect.

mRNA Expression of Selected Candidate Genes in the Bovine Oviduct Is Influenced by the Energy Status as a Long-Term Effect.

Role of the Oviduct in Early Embryo Development

This review highlights the role of the oviduct in early embryo development, which has to fulfil many aligned and well-tuned tasks during early embryogenesis. The oviductal lining is subjected to dynamic changes to timely accomplish gamete transport, fertilization and embryo development and to deliver a competent and healthy conceptus to the endometrium which can implant and develop to term. Although knowledge about the role of the oviduct is limited, we know that embryos are very sensitive to the environment in which they develop. The success of in vitro embryo production techniques demonstrates that it is possible to bypass the oviduct during early development and, to a certain extent, replicate the conditions in vitro. However, comparative studies show that embryos developed in vivo are superior to their in vitro produced counterparts, underlining our relatively poor knowledge of the biology of the oviduct. Oviduct activity is orchestrated by various factors, depending on cyclic dynamics, which crucially affect the success of tubal transfer and/or (re-)collection of embryos in embryo transfer studies. This paper reviews data which demonstrate that in vivo culture of embryos in the bovine oviduct is a useful tool for the assessment of embryos developed under various conditions (e.g. superovulation vs single ovulation, lactating dairy cows vs non-lactating cows). It is concluded that more work in the field of early embryo development within the oviduct would contribute to improved ART protocols leading to healthy pregnancies and offspring.

Interaction between neurotrophin 4 and gonadotrophin in bovine oviducts

The expression and localization of neurotrophin 4 (NT4) and its receptor, tyrosine kinase B (TRKB), in the bovine oviduct, and their interaction with gonadotrophins in bovine oviduct epithelial cells (BOECs), were examined. Transcripts for NT4 and TRKB were detected by reverse transcription polymerase chain reaction (RT-PCR) in bovine oviducts in the follicular and luteal phases, and their proteins were immunolocalized in BOECs. Based on real time PCR, NT4 mRNA did not differ significantly between the two phases of the cycle, although TRKB mRNA expression was higher (P < 0.05) in the luteal phase than that in follicular phase. The BOECs were treated with various concentrations of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) in vitro; for NT4, mRNA and protein were higher (P < 0.05) than those in the control (based on real time PCR and enzyme-linked immunosorbent assay (ELISA) assays). The effects of NT4 and the TRKB inhibitor (K252a) on the expression of LH receptor (LHR) and FSH receptor (FSHR) in the oviduct epithelial cells were also studied using a monolayer culture model. Expression levels of LHR and FSHR mRNA in BOECs treated with various concentrations of NT4 were higher (P < 0.05) than those in the control. However, these expressions were blocked by treatment with K252α. We concluded that neurotrophin 4 may have a role in regulating the function of bovine oviducts by interacting with gonadotrophins.

Nitric oxide in the bovine oviduct: Influence on contractile activity and nitric oxide synthase isoforms localization

The oviducts of 64 Holstein cows in luteal (early I, early II and late) and follicular phases were evaluated to determine the protein expression and mRNA transcription of different nitric oxide synthase isoforms (eNOS, iNOS, nNOS) as well as the effect of nitric oxide (NO) on spontaneous contractility in vitro. The expression patterns of nitric oxide synthase (NOS) isoforms in isthmus and ampulla (n = 6 for each phase) were determined by immunohistochemistry, reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot analysis. In the contractility studies, longitudinal and circular isolated strips of isthmus and ampulla (n = 10 for each phase) of oviducts located ipsilateral to the luteal structure or preovulatory follicle were treated as follows: a) L-arginine, an endogenous NO donor (10−8 to 10−3m), b) Nω-nitro-L-arginine methyl ester (L-NAME), a NOS inhibitor (10−5m) and L-arginine (10−3m), c) methylene blue (MB), an inhibitor of soluble guanylate (10−5m) and L-arginine (10−3m) and d) sodium nitroprusside (SNP), an exogenous NO donor (10−8 to 10−4m). Immunohistochemical evaluation revealed that endothelial NOS (eNOS) expression detected in epithelial layer of isthmus and ampulla was strong in early I luteal phase, moderate in follicular phase and weak in other phases. Neuronal NOS (nNOS) immunoreactivity was strong in isthmus and moderate in ampulla, and staining of nerve fibers was observed mostly in early I luteal and follicular phases. All eNOS, nNOS and inducible NOS (iNOS) isoforms were detected by RT-PCR. eNOS and iNOS proteins were evident, whereas nNOS was undetectable by Western blot analysis in the tissue examined. L-arginine applied alone or after L-NAME did not alter or increase the contractile tension of the strips in most tissues examined. However, L-arginine applied after MB increased contractile tension in the strips of ampulla and longitudinal isthmus from early I luteal phase and circular isthmus from follicular phase but decreased it in isthmus from early II luteal phase. SNP differentially modulated oviductal contraction depending on the type of muscular strips and period examined. These results showed the estrous phase-dependent changes related to endogenous NO system which might be of physiological importance to the oviduct for secretory and ciliary functions involved in gametes and embryo(s) transportation.

Expression of nuclear progesterone receptor and progesterone receptor membrane components 1 and 2 in the oviduct of cyclic and pregnant cows during the post-ovulation period

BackgroundProgesterone (P4) may modulate oviductal functions to promote early embryo development in cattle. In addition to its nuclear receptor (PR), P4 may mediate its actions through P4 receptor membrane component 1 (PGRMC1) and its relative, PGRMC2. Two successive experiments were undertaken to characterise the expression of PR, PGRMC1 and PGRMC2 in the bovine oviduct during the post-ovulation period, and to relate their expression to the presence of an embryo, the proximity of the CL and to the region of the oviduct.MethodsIn the first experiment (Exp. I), whole oviduct sections were collected from Holstein cows at Day 1.5, Day 4 and Day 5 post-ovulation (n = 2 cows per stage). The expression of PR, PGRMC1 and PGRMC2 was studied in the ampulla and isthmus by RT-PCR, western-blot and immunohistochemistry. In Exp. II, oviduct epithelial cells were collected from cyclic and pregnant Charolais cows (n = 4 cows per status) at Day 3.5 post-ovulation and mRNA expression of PR, PGRMC1 and PGRMC2 was examined in the ampulla and isthmus by real-time quantitative PCR.ResultsIn Exp. I, PR, PGRMC1 and PGRMC2 were expressed in all oviduct samples. PGRMC1 was mainly localised in the luminal epithelium whereas PR and PGRMC2 were localised in the epithelium as well as in the muscle and stroma layers of the oviduct. The expression was primarily nuclear for PR, primarily cytoplasmic for PGRMC1 and both nuclear and cytoplasmic for PGRMC2. In Exp. II, mRNA levels for PR, PGRMC1 and PGRMC2 were not affected by either the pregnancy status or the side relative to the CL. However, the expression of PR and PGRMC2 varied significantly with the region of the oviduct: PR was more highly expressed in the isthmus whereas PGRMC2 was more highly expressed in the ampulla.ConclusionsThis is the first evidence of PGRMC2 expression in the bovine oviduct. Our findings suggest that P4 regulates the functions of the bovine oviduct in a region-specific manner and through both classical and non classical pathways during the post-ovulation period.

The effect of DDT and its metabolite (DDE) on prostaglandin secretion from epithelial cells and on contractions of the smooth muscle of the bovine oviduct in vitro

The insecticide DDT and its metabolite (DDE), due to their lipolytic nature and resistance to biodegradation, are accumulated in the living tissues. In cows, DDT and DDE were found to affect prostaglandin (PG) secretion from the endometrium and contractions of the myometrium.In this study, the impact of both xenobiotics (0.1, 1, 10 or 100ng/ml) on the function of epithelial cells and muscle strips of bovine oviducts from 1 to 5day of the oestrous cycle was examined. Therefore the concentration of PGE2 and PGFM (a metabolite of PGF2α) in culture media, mRNA expression of genes involved in PGs synthesis in epithelial cells and the force and amplitude of strips contractions were measured after 2 and 24 or 48h of incubation.Neither DDT nor DDE affected the viability of cells after 48h (P>0.05). Both DDT and DDE increased the concentrations of PGFM in culture medium and secretion of PGE2 after only 2h of cell culture (P<0.05). Similar effects were seen for the influence of DDE on amount of PGFM after 48h, while DDT decreased secretion of PGE2 (P<0.05). DDT after 2h increased (P<0.05) mRNA expression of PGF2α synthase (PGFS), while both xenobiotics decreased (P<0.05) mRNA expression of cyclooxygenase-2 (COX-2) after 24h. DTT also increased the force of isthmus contractions after 2h, as did both xenobiotics after 48h (P<0.05). Moreover, after 2 and 48h, DDE stimulated the amplitude of contractions of the isthmus as well as the ampulla, (P<0.05). The effect of both compounds on oviduct contractions was diminished by indomethacin, which blocks PG synthesis.We conclude that oviductal secretion of prostaglandins is affected, by DDT and DDE. The influence of these xenobiotics on PGF2α and PGE2 secretion and ratio may be part of the mechanism by which both DDT and its metabolite disturb the contractions of oviductal muscle.

Effect of aracnotoxin from Latrodectus mactans on bovine sperm function: modulatory action of bovine oviduct cells and their secretions

Latrodectus mactans' aracnotoxin (Atx) induces changes in sperm function that could be used as a co-adjuvant in male contraceptive barrier methods. This effect includes the suppression of intracellular reactive oxygen species (ROS), an event necessary for capacitation, chemotaxis and acrosome reaction (AR). The sperm that are not trapped by the barrier method can reach the oviduct before fertilisation and be exposed to the secretions of the oviducts. This study evaluated the effect of bovine tubal explants (TU) and conditioned media (CM) from the ampullar and isthmal regions on spermatozoa exposed to Atx. Thawed bovine sperm were incubated with Atx, TU and CM from the ampullar and isthmal regions for 4 h and then DNA integrity, intracellular ROS and lysophosphatidylcholine-induced AR were determined. Spermatozoa exposed to Atx and co-incubated with TU and CM for 4 h produced an increase in sperm DNA damage, a decrease in ROS production and a decrease in %AR, compared with the control. A similar result was obtained from the co-incubation of spermatozoa with Atx. In conclusion, the effect of Atx is not modified by tubal cells or their secretions and this opens the door to future studies to evaluate the application of synthetic peptides obtained from Atx as a co-adjuvant of contraceptive barrier methods.

56 COMPARISON OF THE MORPHOLOGY OF VARIOUS REGIONS OF THE CATTLE OVIDUCT IN FOUR PHASES OF THE OVARIAN CYCLE

The oviduct provides the environment necessary for the gamete transport, completion of spermatozoa capacitation, oocyte fertilization and the early development of embryos. In cattle, all of these processes take place between Day 0 to 4 of the ovarian cycle (Day 0 is the day of ovulation). In previous studies, temporal changes in the bovine oviduct morphology were evaluated by dividing the ovarian cycle into luteal and follicular phases. In order to understand the relation between the bovine oviduct morphology and processes occurring there, the ovarian cycle has been further divided into four phases: I (Day 0–4), II (Day 5–10), III (Day 11–17) and IV (Day 18–20), with the day of ovulation considered Day 0 (1980 J. Dairy Sci. 63, 155–160). The aim of the study was to evaluate the oviduct morphology of the infundibulum, ampulla and isthmus in 4 phases of the ovarian cycle. Research material comprised cattle oviducts (classified into 1 of the 4 phases of the cycle based on ovarian morphology), dissected into infundibulum, ampulla and isthmus and subsequently sectioned and processed for histological preparations (hematoxylin and eosin, H&amp;E, staining). Diameters of transverse cross-sections of oviduct and its lumen and thickness of tunicas: mucosa, muscularis and serosa were evaluated in relation to the region of oviduct and the phase of ovarian cycle. Values are given in μm. Statistical analysis was carried out by 1-way analysis of variance and comparisons of mean values were made with the Tukey honestly significant difference test (Statgraphics Plus 5); P &lt; 0.05 was considered to reflect the presence of statistical significance. The comparison of the diameters of transverse cross-sections (A) of oviduct and its lumen (B) shows significant statistical differences between ampulla and isthmus within the phases: A-I (4507.26 vs 2524.47), II (4510.53 vs 2540.67), III (4503.28 vs 2534.07), IV (4500.73 vs 2533.90); B-I (4191.10 vs 1950.88), II (4173.63 vs 1986.33), III (4198.53 vs 1966.88) and IV (4192.50 vs 1959.33). There are no differences among 4 phases of the ovarian cycle. The thickness of tunicas muscularis and serosa of infundibulum (I: 26.81 vs 196.85; II: 27.03 vs 201.80; III: 26.22 vs 199.45; IV: 23.97 vs 198.01), ampulla (I: 91.51 vs 214.50; II: 90.72 vs 212.55; III: 88.61 vs 213.30; IV: 89.65 vs 206.28) and isthmus (I: 202.29 vs 216.52; II: 199.24 vs 207.74; III: 200.90 vs 212.38; IV: 200.38 vs 210.86) show only statistically significant differences within the phases, whereas the tunica mucosa shows only statistically significant differences between phases and the term of the height of epithelium at the base of mucosal folds (I: 26.49; II: 25.20; III: 24.14; IV: 29.96) and their apical parts (I: 28.09; II: 26.01; III: 25.45; IV: 30.96). In conclusion, differences in oviduct morphology are mainly region specific, whereas the epithelium morphologically infundibulum, ampulla and isthmus show variation in the 4 phases of the ovarian cycle. Supported by Grant MNiSW N311236137.