Embryo‐maternal Communication during the First Days of Embryonic Life

The mechanisms of embryo‐maternal communication during the first days of embryonic life are largely unknown. Using the bovine as a model, the aims of our study were to morphologically characterize the interaction between the pre‐implantation embryo and the epithelium of the maternal ampulla, isthmus and uterotubal junction by light and scanning electron microscopy. For this purpose, oviducts were removed from cows revealing a functional corpus luteum on day 3 after insemination. These were compared to oviducts removed on day 3 (metestrus) of the estrous cycle. Three days after insemination, the majority of the epithelial cells in the ampulla were secretory cells distinctly protruding into the oviductal lumen. Contrary the ampulla of cows on day 3 of the cycle predominantly revealed ciliated cells in the oviductal epithelium. As shown by Periodic Acid Schiff reaction (PAS) with and without amylase digestion, the secretory cells of the ampulla synthesized merely glycoproteins during metestrus, but large amounts of glycogen during pregnancy. In the isthmus no morphological differences were seen between pregnant and cyclic cows. The most conspicuous finding during pregnancy was seen in the uterotubal junction: Vital cumulus cells embedded in between epithelial cells had developed short cytoplasmic processes intensely contacting the epithelial uterine cells. The embryos obtained ex vivo were regularly covered with a thick layer of homogenous extracellular matrix. Contrary embryos produced in vitro– both with and without coculture with oviductal cells –revealed a clearly visible zona pellucida with spongy appearance and numerous pores. Our results imply that already during the first days of life there is intense interaction of the pre‐implantation embryo and the maternal genital tract part of which may be mediated by cumulus cells.     

Laser Micro-dissection and Quantitative RT-PCR as a Tool for Cell Identification

The mechanisms of embryo-maternal communication during the first days of embryonic life are largely unknown. Using the bovine as a model, the aims of our study were to morphologically characterize the interaction between the pre-implantation embryo and the epithelium of the maternal ampulla, isthmus and uterotubal junction by light and scanning electron microscopy. For this purpose, oviducts were removed from cows revealing a functional corpus luteum on day 3 after insemination. These were compared to oviducts removed on day 3 (metestrus) of the estrous cycle. Three days after insemination, the majority of the epithelial cells in the ampulla were secretory cells distinctly protruding into the oviductal lumen. Contrary the ampulla of cows on day 3 of the cycle predominantly revealed ciliated cells in the oviductal epithelium. As shown by Periodic Acid Schiff reaction (PAS) with and without amylase digestion, the secretory cells of the ampulla synthesized merely glycoproteins during metestrus, but large amounts of glycogen during pregnancy. In the isthmus no morphological differences were seen between pregnant and cyclic cows. The most conspicuous finding during pregnancy was seen in the uterotubal junction: Vital cumulus cells embedded in between epithelial cells had developed short cytoplasmic processes intensely contacting the epithelial uterine cells. The embryos obtained ex vivo were regularly covered with a thick layer of homogenous extracellular matrix. Contrary embryos produced in vitro – both with and without coculture with oviductal cells –revealed a clearly visible zona pellucida with spongy appearance and numerous pores. Our results imply that already during the first days of life there is intense interaction of the pre-implantation embryo and the maternal genital tract part of which may be mediated by cumulus cells.     

Light and Scanning Electron Microscopic Studies of Oviductal Epithelium in Thai Swamp Buffalo (Bubalus bubalis) at the Follicular and Luteal Phases

The purpose of this study was to investigate the morphological changes in the epithelium of Thai swamp buffalo oviducts at the follicular and luteal phases by histological technique and scanning electron microscopy. The samples from the infundibulum, ampulla, isthmus and uterotubal junction (UTJ) of the oviduct were taken immediately after slaughter at the local abattoir. Noticeable cyclic changes were observed on the epithelial surface of the infundibulum and ampulla, but few changes were present in the isthmus and UTJ. At the follicular phase, the epithelium of infundibulum and ampulla were densely covered with ciliated cells whose cilia concealed the apical processes of the secretory cells. In contrast, the secretory cells dominated in the epithelium at the luteal phase and most of the ciliated cells were hidden by the bulbous processes of these cells. In the isthmus and UTJ at the follicular and luteal phases, the secretory cells were almost flat or gently rounded and covered with numerous microvilli at their apical surface. In conclusion, the histological and ultrastructural observation of Thai swamp oviduct epithelium revealed marked cyclic changes in the cellular differences associated with the main functions of segmental variations.     

Scanning electron microscopic study of the functional anatomy of the porcine oviductal mucosa

A three-dimensional model has not been clearly established for the porcine oviductal mucosa. The oviducts of 12 cyclic sows were examined by Scanning Electron Microscopy to study the structure and nature of the mucosal arrangement of the oviduct. Epithelial cyclic changes were also studied. The oviductal infundibulum is an asymmetric funnel-shaped structure surrounding the ostium, in which a wide and a narrow side can be distinguished. The mucosa is more complex in the narrow side, showing numerous and tortuous longitudinal primary folds, while the mucosa becomes simpler in the wide side. Around the ostium abdominale wide secondary folds form cul-de-sacs, with their opening pointing in ovarian direction. Areas between folds throughout the lumen of the oviduct show a high-degree of complexity. Inter-fold spaces are occupied by a system of irregular grooves and pockets, with the presence of basal crypts in the caudal oviduct. Marked variations were observed in the oviductal epithelium depending on the oviductal segment, basal or apical areas of the folds, and phase of the oestrous cycle. Cyclical changes were observed in the infundibulum and ampulla, so that prominent and numerous ciliated cells lined apical areas of the folds in the follicular phase; whereas secretary cells were predominant throughout all areas of epithelial surface in the luteal phase. The oestrous cycle phase appeared do not affect the epithelial population cells of the caudal segments of the oviduct: ciliated and secretory cells uniformly lined apical and basal areas of the folds. The topography of the oviduct provides a complex system of regulation, which may influence not only the passage of cells, but also movement of fluid within the oviductal canal.     

Morphometric and ultrastructural changes in ciliated cells of the oviductal epithelium in prolific Chinese Meishan and Large White pigs during the oestrous cycle.

There are some differences in reproductive features between the Chinese Meishan (CM) pig and Large White (LW) pig. The aim of the present study was to investigate the quantitative changes and ultrastructural features of ciliated cells in the various regions of the CM and LW pig oviduct during the follicular and luteal phases of the oestrous cycle. In the fimbrial and ampullar epithelia at the follicular phase, the ciliated cells were more plentiful than in the isthmic region in both pigs. In the CM pigs, there was a striking decrease in the percentage and cell height of ciliated cells in the luteal phase compared with the follicular phase. Although similar quantitative changes were observed in the LW pig oviduct, these changes were less dramatic than that in the CM pig oviduct. In both pigs, the percentage and cell height of ciliated cells in the isthmus were unchanged between the follicular and luteal phases. The ultrastructure of ciliogenic and ciliated cells was observed. In the fimbrial and ampullar epithelia during the follicular phase, most of the ciliated cells showed normal morphology, having many elongated cilia and mitochondria, but in the CM pig oviduct during the luteal phase many of the ciliated cells possessed immature cilia and swollen mitochondria. Cells undergoing ciliogenesis were frequently observed in the fimbriae and ampulla, and occasionally in the isthmus. Cytoplasmic protrusions containing variable numbers of ciliary axonemal complexes occurred in the fimbrial and ampullar epithelium in the CM pigs at the luteal phase, suggesting that deciliation of cells occurs by membrane-bound cilia packets forming at the apices of cells and pinching off. These results demonstrate that there are regional variations in the cyclic changes associated with the oviductal ciliated cells of the pigs, while there are marked morphometrical and ultrastructural changes in oviductal ciliated cells of the CM pigs compared with that of the LW pigs.     

Three‐Dimensional Architecture of the Ovine Oviductal Mucosa

The objective of this work was to establish for the first time a complete three‐dimensional model of the ovine oviductal mucosa. The oviducts of 15 cyclic ewes were examined combining the direct examination of the mucosa, by scanning electron microscopy (SEM) and histology, with the SEM observation of resin moulds of the oviductal lumen. Around the ostium abdominale, all longitudinal primary folds and wide secondary are seen to form cul‐de‐sacs, with their opening pointing in the ovarian direction were observed. At the connection of the ampulla to the isthmus, there is a sharp change in the morphology, from a high folded structure to a smoother one. In the utero‐tubal junction, the primary folds broaden and become more voluminous, the lumen has a slit‐like appearance, and secondary folds form cul‐de‐sacs with their opening oriented towards the uterus. The areas between the folds throughout the lumen of the oviduct show a high degree of complexity. The presence of crypts was observed in all the regions studied, branched in the ampulla and spiniform in the isthmus. Marked variations were observed in the oviductal epithelium depending on the oviductal segment and the basal or apical areas of the folds. The variations found regarding the phase of the oestrous cycle were similar to those described in studies of other species. The anatomy of the oviductal mucosa provides a complex system that seems to be designed to regulate the movement of fluids and the passage of cells within the oviductal canal.     

Interaction of Tritrichomonas foetus and the bovine oviduct in an organ culture model

Tritrichomonas foetus is an extracellular parasite of the reproductive tract in cattle. The mechanism by which T. foetus causes abortion in cattle is largely unknown. There are no studies of infection in the cow oviducts, almost all published papers are related to vagina infection and few articles focusing on the uterus. The aim of the present study was to establish a working model of bovine oviduct epithelial cells and submit these cells to Tritrichomonas foetus interaction. Twenty bovine oviducts were obtained from cows at a commercial abattoir and T. foetus was injected through the isthmus into the oviduct lumen. The whole oviduct was analyzed by scanning and transmission electron microscopy. The results reported here demonstrate that: (1) fresh whole oviducts can be used as a good model to study parasite-host cell interaction; (2) cow oviduct epithelium has been shown to consist of two cell types: ciliated and nonciliated secretory cells, and T. foetus displayed great specificity for the nonciliated cells localized in the deeper oviduct folds; (3) T. foetus adheres as single separate cells, and maintains the flagella externalized; (4) differently from T. vaginalis, T. foetus does not change its shape during the adhesion process; and (5) oviduct cells exhibited morphological characteristics of apoptosis after trichomonadal interaction.     

Meiosis Resumption of Canine Oocytes Cultured in the Isolated Oviduct

The aim of this study was to investigate the effects of culture in isolated oviducts relative to meiotic maturation, the time required to resume meiosis and the viability of the canine oocytes. For this purpose, cumulus–oocyte complexes and isthmus–ampullar tracts of the oviducts were collected from bitches undergoing ovariohysterectomies and destined to two experiments of culture. In experiment 1, the oocytes were cultured for 24 or 30 h: (1) in 100 μl drops under oil; (2) on the mucosal epithelium of the open oviducts; (3) in the ligated oviducts. In experiment 2, oocytes were cultured in the ligated oviduct for 24, 30 and 48 h. A group of control oocytes was not cultured (0 h). The results showed that within 30 h of culture, a higher proportion of oocytes (p < 0.001) resumed meiosis in the ligated oviduct (63.8%) than in drop (20.4%) or in the open oviduct (27.1%). Moreover, 24 and 30 h of culture assured higher proportions of meiosis resumption than 48 h (69.2 and 59.1% vs 35.8%, p < 0.005). Oocyte resumption of meiosis was mainly determined by oocytes at meiotic stages preceding metaphase I, while stages between metaphase I and II in the ligated oviduct ranged between 12.5 and 31.9%. The extension of the culture time up to 48 h in the oviduct increased oocyte degeneration significantly (59.3%, p < 0.0001) compared with 24 and 30 h (18.7 and 27.3%, respectively) and the oviductal epithelium showed nuclear picnosis and degeneration following culture. The present study suggests that the close physical interaction between the canine oocytes and the oviductal tract positively affects oocyte maturation, and meiosis is resumed within 30 h of culture. Moreover, the oocyte survival is better preserved within 30 h in the ligated oviduct compared with the conventional culture in drop or to the culture in the open oviduct, but the ligated oviduct does not assure viability of the oocytes up to 48 h of culture.     

Morphological Changes and Proliferative Activity in the Oviductal Epithelium during Hormonally Defined Stages of the Oestrous Cycle in the Bitch

The aim of the present study was to investigate morphological changes and proliferative activities in the epithelium of the canine oviduct with regard to the part of the oviduct – possibly indicating the existence of a locally restricted sperm reservoir – and the stage of the oestrous cycle. Nine healthy adult nulliparous bitches were submitted to ovariohysterectomy at three stages of the cycle: anoestrus (n = 3), late follicular phase (n = 3) and mid‐luteal phase (n = 3). The whole oviduct ranging from the utero‐tubal junction (UTJ) to the infundibulum (IN) was collected, divided into UTJ, IN plus six segments of equal length, i.e. eight oviductal specimens per animal were studied by light microscopy. Morphological characteristics of ovaries and endometrium were recorded macroscopically and verified histologically. The height of oviduct epithelial cells and percentage of ciliated cells (CC) were assessed and the respective data analysed statistically. Proliferative activity was immunohistochemically visualized by means of Ki‐67 antigen detection. Blood was collected and concentrations of oestradiol‐17β and progesterone (P₄) were measured. Within the IN and five of the six tissue samples collected from the ampulla and isthmus in anoestrous bitches, the oviductal surface epithelium consisted of low cuboidal cells demonstrating a uniform dark staining intensity. Only a very few scattered lighter staining CC could be detected. Under the influence of oestrogens during late follicular phase, the oviductal epithelium was highly differentiated. Lighter stained CC with apically located nuclei were easily distinguishable from basophilic secretory cells with apical cytoplasmic protrusions. Cell height and percentage of CC were significantly higher than in anoestrus (p ≤ 0.05). During mid‐luteal phase, high levels of P₄ were associated with differentiated and dedifferentiated cells as well as cells in regression seen in the mucosal folds of all samples. The percentage of CC and cell height were significantly lower than during late follicular phase (p ≤ 0.05). Further signs of dedifferentiation consisted of a loss of cilia, a pinching off of the apical cytoplasm as well as the presence of debris and macrophages within the oviductal lumen. In the oviductal part of UTJ and the caudal isthmus hormone‐dependent variations in cellular morphology were less distinct. Changes in cell height were minimal and did not differ significantly throughout the oestrous cycle. Hypertrophic cells with large nuclei were predominantly present at these sites, but did not consistently demonstrate signs of ciliation or secretion. Sporadic proliferating activity, visualized by means of Ki‐67 antigen, was mainly seen in some cells of the late follicular phase samples. Thus, overall proliferative activity is generally very low or may occur within a relatively short period of time. It therefore cannot be excluded, that periods exhibiting higher mitotic rates are not included in the present study. It should, however, be mentioned that cells demonstrating morphological signs of apoptosis can only be seen very sporadically within a few specimens during mid‐luteal phase, thus, reflecting low proliferative capacities and minimal cellular turnover found during this study. The results of the present study strongly indicate that oestrogens cause hypertrophy and differentiation, whereas P₄ induces gradual dedifferentiation or regression of the oviductal epithelium. Furthermore, they reveal clearly visible changes in the morphology of the tubal epithelium during the oestrous cycle. Depending on the tubal segment, these are, however, variably expressed. Whether the low degree of cellular variation of the UTJ and caudal isthmus is caused by specific hormone concentrations at these sites or specific regulatory mechanisms and may be associated with specific functional properties such as the formation of a locally restricted sperm reservoir needs further investigations.     

In Vivo and In Vitro Sperm Interaction with Oviductal Epithelial Cells of Llama

Sperm reservoirs in South American Camelids would be crucial for successful fertilization. Since ovulation occurs approximately 36 h after mating, the maintenance of the sperm viability in the oviduct waiting for the ovum is a critical reproductive event. Our study aimed at determining whether the isthmus or the utero tubal junction (UTJ) could function as a sperm reservoir in llama by means of in vivo and in vitro experiments. For the in vivo experiments, the oviducts of adult females with a dominant follicle bigger than 7 mm were examined for the presence of sperm at 6, 18, 24, 28 and 35 h after mating. The results using scanning and transmission electron microscopy showed ultrastructural differences between isthmus and UTJ with respect to (1) predominance of secretory cells in the UTJ and ciliated cells in the isthmus epithelium and (2) cytoplasmic bulbous projection of the secretory cells in the UTJ. Sperm adhered by a mucus‐like substance were seen only in the UTJ at 6, 18, 24 and 28 h post‐mating. Lack of sperm adhered to oviductal mucosa was observed around ovulation (35 h). In vitro experiments demonstrated higher ability of UTJ epithelial cell explants with respect to isthmus explants to bind sperm in a co‐cultured system. The anatomical features and the presence of a sperm bonding agent in the UTJ together with the in vitro differential binding of sperm to UTJ explants strongly suggest that both may be feasible mechanisms that facilitate sperm storage in this oviductal region in llama.     

Regional and Cyclic Variations in the Ultrastructural Features of Secretory Cells in the Oviductal Epithelium of the Chinese Meishan Pig

The Chinese Meishan pig is prolific breed and it is considered that this pig has a capacity with higher rates of embryonic survival. The oviductal secretory cells may affect the embryonic development and survival. The aim of the present study was to investigate the ultrastructural features of secretory cells in the various regions of the Chinese Meishan pig oviduct during the follicular and luteal phases of the oestrous cycle. In the ampullar secretory cells, numerous secretory granules with moderately electron-dense matrices were present in the supranuclear cytoplasm and exocytosis of secretory granules was observed. The number of secretory granules was dramatically reduced in the ampullar secretory cells in the luteal phase. During the follicular phase in the fimbrial epithelium, the secretory cells contained rough endoplasmic reticulum and Golgi apparatus, but most cells had few small granules. In the luteal phase, the secretory cells in the ampullar and fimbrial epithelia extended beyond the luminal border of the ciliated cells. In the isthmus, many granules were present in the cytoplasm of secretory cells throughout the oestrous cycle, but the number of secretory granules was reduced in the luteal phase. The cytomorphometric data revealed that the height of ciliated cells decreased substantially in the fimbriae and ampulla at the luteal phase, while that of non-ciliated cells was less affected. These results suggest that the drastic reduction of cell height of ciliated cells cause the extrusion of most secretory cells beyond the ciliated cells in the fimbriae and ampulla during the luteal phase. In summary, our ultrastructural observations of Chinese Meishan pig oviduct revealed marked cyclic changes in the ultrastructural features of secretory cells. In particular, the ultrastructural features and the numbers of secretory granules were distinctive for each particular segment. These findings should provide insight into the regional and cellular differences in functions of secretory cells of the Chinese Meishan pig oviduct.     

Detection of the Matrix Metalloproteinases MMP‐2 and MMP‐9 and Tissue Inhibitors of Metalloproteinases TIMP‐1 and TIMP‐2 in Llama (Lama glama) Oviduct

Matrix metalloproteinases (MMPs) and their inhibitors (TIMPs) are involved in several reproductive events like oocyte–spermatozoa interaction and semen liquefaction. In order to study their role in the llama oviductal reproductive process, MMP activity in oviductal fluid (OF) was assayed. Considering that llama genome sequences are partially known, a strategy to procure cDNA sequences of MMP‐2, MMP‐9, TIMP‐1 and TIMP‐2 was designed. Afterwards, their expression patterns in the different llama oviductal segments were assayed. Gelatine zymograms detected 62 and 94 kDa protease activities that matched MMP‐2 and pro‐MMP‐9, respectively. Expression pattern analysis showed that MMP and TIMP mRNAs were present in ampulla, isthmus, utero‐tubal junction (UTJ) and papilla. Altogether, these findings support the argument that MMPs/TIMPs are produced in the oviduct and secreted into the oviductal lumen. Our results encourage further studies to elucidate the role of these proteins in reproductive oviductal events.     

The Sow Endosalpinx at Different Stages of the Oestrous Cycle and at Anoestrus: Studies on Morphological Changes and Infiltration by Cells of the Immune System

The aim of this study was to investigate the morphological changes of the sow endosalpinx and the distribution of leukocytes throughout the oestrous cycle and at anoestrus. Nineteen crossbred sows (Swedish Landrace x Swedish Yorkshire) at late dioestrus (three), prooestrus (three), oestrus (three), early dioestrus (three), dioestrus (three) and anoestrus (four) were used. Oviductal samples from three different parts (isthmus, ampulla and infundibulum), taken immediately after slaughter, were fixed, embedded in plastic resin and stained with toluidine blue or stored in a freezer at -70 degrees C until analysed by immunohistochemistry (prooestrus and anoestrus) with an avidin-biotin peroxidase method. Quantitative and qualitative examinations of oviductal epithelium and subepithelial connective tissue were performed by light microscopy. During all stages, a lower degree of morphological changes (pseudostratification, mitosis and secretory granules) was found in the isthmus compared with ampulla and infundibulum. In ampulla and infundibulum, pseudostratification, mitotic activity and secretory granules of the epithelium were high at prooestrus/oestrus. Cytoplasmic protrusions of epithelial cells with some extruded nuclei were prominent in ampulla and infundibulum at all stages except for oestrus and early dioestrus. Lymphocytes as well as CD2- and CD3-positive cells were the predominant immune cells in the epithelial layer. The numbers of lymphocytes and CD3-positive cells did not differ among segments and stages. Numbers of CD2-positive cells did not differ between prooestrus and anoestrus while the numbers were significantly higher in the infundibulum than in ampulla and isthmus. Neutrophils were only occasionally found and mainly in the infundibulum. In the subepithelial connective tissue layer, the two most commonly observed immune cell types were lymphocytes and plasma cells. The numbers of lymphocytes as well as CD2- and CD3-positive cells was lower in isthmus than in the other segments (p < or = 0.001). Higher numbers of plasma cells (p < or = 0.001) were found in infundibulum than in ampulla and isthmus. The numbers of lymphocytes and plasma cells were not significantly different between stages of the oestrous cycle. However, the number of neutrophils differed and were highest at prooestrus in ampulla and infundibulum. The numbers of CD2-, CD3- and CD79-positive cells did not differ between prooestrus and anoestrus whereas for CD14- and SWC3-positive cells, the numbers were higher at prooestrus (p < or = 0.05) than at anoestrus. In the oviduct, the morphology differed in ampulla and infundibulum with oestrous cycle stages, which indicates an effect by ovarian steroid hormones. The immune cell infiltration was less influenced by cyclic changes. However, the immune cell infiltration (in the connective tissue) in the upper part, especially infundibulum, differed significantly from the one in the lower part, isthmus, indicating different immune functions within various parts of the oviduct.     

Hypothyroidism Affects Differentially the Cell Size of Epithelial Cells Among Oviductal Regions of Rabbits

Oviductal regions show particular histological characteristics and functions. Tubal pathologies and hypothyroidism are related to primary and secondary infertility. The impact of hypothyroidism on the histological characteristics of oviductal regions has been scarcely studied. Our aim was to analyse the histological characteristics of oviductal regions in control and hypothyroid rabbits. Hypothyroidism was induced by oral administration of methimazole (MMI) for 30 days. For both groups, serum concentrations of thyroid and gonadal hormones were determined. Sections of oviductal regions were stained with the Masson's trichrome technique to analyse both epithelial and smooth muscle layers. The percentage of proliferative epithelial cells (anti‐Ki67) in diverse oviductal regions was also quantified. Data were compared with Student t‐test, Mann–Whitney U‐test, or Fischer's test. In comparison with the control group, the hypothyroid group showed: (i) a low concentration of T3 and T4, but a high level of TSH; (ii) similar values of serum estradiol, progesterone and testosterone; (iii) a large size of ciliated cells in the ampulla (AMP), isthmus (IST) and utero‐tubal junction (UTJ); (iv) a large size of secretory cells in the IST region; (v) a low percentage of proliferative secretory cells in the fimbria‐infundibulum (FIM‐INF) region; and (vi) a similar thickness of the smooth muscle layer and the cross‐sectional area in the AMP and IST regions. Modifications in the size of the oviductal epithelium in hypothyroid rabbits could be related to changes in the cell metabolism that may impact on the reproductive functions achieved by oviduct.     

The localization and connections of suboviductal lymphatic vessels in the bovine uterine broad ligament

The distribution of oviductal lymphatic vessels in the bovine mesosalpinx and bursa ovarica, and their communications with other lymphatics in the uterine broad ligament were examined after exposition of all lymphatic pathways with varicoloured microfil and/or ink-gelatin mixture. However, filling of the lumen of the oviductal infundibulum lymphatics was difficult or impossible because of its slender walls. Lymphatics of the isthmus and ampulla with short precollectors formed branches 1.5-2.0 cm long on both sides: dorsal and ventral of the uterine broad ligament. Collectors with slender and long lymphangions, visible after filling their lumen, encircled the alymphatic area in the parainfundibullar mesosalpinx. The greatest number of lymphatic branches, which originated from the paraisthmal part of the oviductal ampulla were observed in the subovarian area. The characteristic feature was their immediate proximity with uterine and oviductal arterial vessels. Subsequent studies have provided convincing evidence that there are direct connections of lymphatics leaving the oviduct and ovarian sac with lymphatics emerging from the uterus and ovary (after bilateral filling of lymphatics pathways in the whole broad liagment of the uterus). The performed investigations showed particularly characteristic feature in the cow--mixing lymph leaving various reproductive organs in the area of the right and left ligament--before two long collector branches reach the nearest lymphatic node/-s.     

奶牛输卵管平滑肌前列腺素类受体的分析研究

本研究选用了前列腺素类化合物(Prostaglandin E2（PGE2）、Prostaglandin F2α（PGF2α）、Prostaglandin D2（PGD2)）和受体选择性激动剂（Butaprost、U-46619）,用多道生理信号采集系统测定其对体外分离的奶牛输卵管峡部及壶腹部平滑肌收缩的影响,以此初步揭示受体的种类。结果发现,PGE2和Butaprost均抑制了奶牛输卵管峡部、壶腹部的自发性收缩运动,呈现明显的浓度依存性舒张反应;PGF2α在高浓度(1～10 μmol/L)下,使峡部和壶腹部的自发性收缩运动有了显著增强;PGD2在低浓度时轻微抑制了壶腹部平滑肌的收缩,但在高浓度(1～10 μmol/L)时加强了峡部和壶腹部平滑肌的收缩;U-46619从低浓度便引起了输卵管峡部和壶腹部平滑肌的强烈收缩。结果表明,EP4、EP2、FP、DP和TP受体同时存在于奶牛输卵管平滑肌上,能与内源性前列腺素类结合并参与调节输卵管平滑肌的自发性收缩。     

The distribution and immunolocalization of fibroblast growth factors (FGFs) in the rat oviduct during early pregnancy and the post-partum period

The mammalian oviduct provides a favourable environment for several reproductive processes, including ovum transport, sperm capacitation, fertilization and pre-implantation embryonic development. This environment is regulated by cyclic ovarian steroids, that is oestrogen, and growth factors. Fibroblast growth factors (FGFs) regulate the differentiation and growth of various cell types in the female genital tract. This study aimed to determine the localization of FGF1, FGF2, FGF receptor 1 (FGFR1) and 2 (FGFR2) in the rat oviduct, by immunohistochemistry, on day 5 of pregnancy and post-partum days 1, 3 and 5, and to demonstrate the possible functions of these proteins during early pregnancy and the post-partum period. On all examination days, cytoplasmic and nuclear FGF1 immunoreactivity was detected in the epithelium lining the infundibulum, ampulla and isthmus of the oviduct. Immunoreactivity was much stronger in the basal bodies of the cilia on the epithelium lining the infundibulum and ampulla. FGF1 immunoreactivity was also detected in stromal cells, myocytes and endothelial cells. Cytoplasmic FGF2 immunoreactivity was observed in the tunica muscularis, vascular myocytes and endothelial cells. While strong cytoplasmic FGF2 immunoreactions were observed in the stromal cells of the lamina propria, the luminal epithelium, some stromal cells and smooth muscle cells displayed a rather weak FGFR1 and FGFR2 immunoreactivity. Immunoreaction intensity did not differ between the periods examined. This study shows that FGF1, FGF2, FGFR1 and FGFR2 are produced by rat oviduct cells during pregnancy and the post-partum period, and reproductive physiology is regulated not only by hormonal mechanisms, but also by growth factors.     

Immunolocalization of nerve growth factor (NGF) and its receptors (TrkA and p75LNGFR) in the reproductive organs of Shiba goats

The objective of this study was to determine the immunolocalization of NGF and its receptors (TrkA and p75LNGFR) in the reproductive tract of the Japanese Shiba goats. Five adult goats were used in this study and sections of ovaries, uteri and oviducts were immunostained by the avidin-biotin-peroxidase complex method (ABC). The results showed that NGF and its receptors (TrkA and p75LNGFR) were expressed in granulosa cells, theca cells, interstitial cells and lutein cells in ovaries. Immunoreactions for NGF, TrkA and p75LNGFR were also detectable in epithelial cells and muscle cells of the ampulla and isthmus of the oviduct, and in epithelial cells and uterine glands of the uterus. These results strongly suggest autocrine and paracrine regulation of reproductive function by NGF in the reproductive tract of female Shiba goats.     

Increased ectopic fat cells in the longitudinal muscularis layer of the oviduct isthmus in obese Japanese Black cows

In obese humans, mesenchymal stem cells differentiate to become ectopic fat cells in muscles. These ectopic fat cells inhibit the contraction of vascular smooth muscles. Stem cells have been recently identified in the human oviduct, a structure important in reproduction. We therefore investigated the number of Oil Red O (ORO)‐positive cells in the oviducts of control Japanese Black cows (n = 6; body condition score [BCS], 3.0 on a 5‐point scale) compared to those with diet‐induced obesity (n = 5; BCS, 4.0). We stained the ampulla and isthmus collected on the second day after ovulation with ORO and then counted the positive cells in each layer in 10 cross‐sections of the ampulla or isthmus. The obese group (23.4 ± 3.4 in the 10 sections) had larger numbers of ORO‐positive cells in the longitudinal muscularis of the isthmus (P < 0.05) than did the control group (15.0 ± 2.4). ORO‐positive cells were also observed in all other layers of the isthmus and ampulla; however, the number of cells in these layers did not differ significantly between obese cows and controls. Whether this observed increase in ORO‐positive cells in the oviducts of obese cows affects their reproduction warrants further study.