Histological Characteristics of the Equine Oviductal Mucosa at Different Reproductive Stages

The objective of this work was to study cellular changes in the epithelium of the mare’s oviduct. Oviductal samples were taken from mares at different reproductive stages for optical microscopy and Hoechst 33258 staining. Glandular-like structures were observed in 100% of the oviducts. These structures were of the tubular type and were formed by ciliated and nonciliated epithelial cells arranged in a way similar to the epithelial surface. The amount of structures decreased progressively from the ampulla to the isthmus, but did not change through the different reproductive stages. Histological changes in the epithelium of the oviduct were observed associated with the reproductive stages. In the ampulla, the amount of ciliated cells decreased in the anovulatory phase compared with other reproductive stages. Cords of connective tissue lined by epithelium (trabeculae) and dividing completely the lumen of the oviduct were found in 50% of the oviducts. Epithelial cells projected toward the lumen as large vesicles of cytoplasm, sometimes containing a nucleus. The amount of cells presenting nuclear protrusion varied throughout the oviduct, with highest incidence in the ampulla, decreasing progressively toward the isthmus (P < .05). In addition, nuclear protrusions were higher in number during the anovulatory and luteal phases than in the other reproductive stages (P < .05). These nuclear protruding cells appeared to be extruding from the epithelium and showed no signs of apoptosis based on the histological and fluorescent stains used. The existence of these gland-like structures in the oviductal mucosa should be considered when studying the oviductal physiology in mares.     

Freezing of canine semen in an animal-free protein extender

The aims of this study were to study the prevalence of oviduct abnormalities of buffaloes, investigate bacteria accompanying hydrosalpinx, pyosalpinx and salpingitis. In addition, the study was designed to investigate the correlation between bacterial infection of the uterus and oviduct lesions in buffaloes. Bacteriological examinations were performed on hydrosalpinx, pyosalpinx and salpingitis. Hydrosalpinx was found in 28 (6.9%) cases of which 20 (71.4%) were found unilaterally and 8 (28.6%) bilaterally. Pyosalpinx was recorded in 12 (2.9%). Three cases (0.7%) of oviducts filled with blood were recorded. Obstruction of oviducts was recorded in 5 (1.2%). Adhesions between mesosalpinx and perisalpingeal tissues were observed in 7 (1.7) cases. One case of double oviduct was found in the left side of the tract examined. The most prevalent bacteria recovered from hydrosalpinx were Corynebacterium hemolyticum and Actinomyces bovis, 42.8% and 28.6%, respectively. No correlation was noticed between bacteria isolated from the uterus and hydrosalpinx. The most prevalent bacteria recovered from pyosalpinx were Escherichia coli, Archanobacterium pyogenes and Staphylococcus aureus, 33.3%, 26.7% and 16.7%, respectively. Higher rates of leukocyte infiltration (P < 0.01) were observed in the uterine discharge and pyosalpinx than hydrosalpinx. In a conclusion, the current study disclosed that oviductal lesions seem to be an important problem in buffalo cows. In addition, there is no correlation between bacteria isolated from uterus and hydrosalpinx. There is a high correlation between bacteria isolated from uterus and pyosalpinx and salpingitis. The occurrence of pyosalpinx and salpingitis is mainly due to bacterial infection. Bacterial isolates from pyosalpinx and salpingitis might be related to ascending infection from the uterus.     

Gonadotropin releasing hormone receptor gene and protein expression and immunohistochemical localization in bovine uterus and oviducts

Recently GnRH, GnRH-R systems has been demonstrated in various extrahypothalamic and extrapituitary reproductive tissues in different mammalian species, where GnRH acts in an autocrine and or paracrine manner and modulates different biological processes. GnRH-R mRNA has also been demonstrated in bovine ovaries (follicle and corpus luteum) and normal and carcinogenic human endometrium/endometrial cells. This is the first study elucidating presence of GnRH-R mRNA and GnRH-R protein in bovine uterus and oviducts in follicular and luteal phases of the estrous cycle and further localizing the receptors to endometrial and oviductal epithelial cells. To our knowledge this is the first report demonstrating GnRH-R mRNA and protein in mammalian oviducts. We used gene-specific primers and monoclonal GnRH-R antibody to test GnRH-R mRNA and GnRH-R protein through RT-PCR and immunobloting. Immunohistochemistry was employed to localize these receptors to endometrial and oviductal epithelial cells. GnRH-R mRNA and receptor protein were expressed at expected molecular weights of 920bp and 60kD, respectively. Densitometry analysis revealed that expression levels for GnRH-R protein in uterus and oviducts were similar to bovine pituitary. The presence of GnRH receptors in bovine uterus and oviducts is intriguing and it would be imperative to examine the functional role of this system in the regulation of reproductive processes.     

鸡输卵管生物反应器研究进展

鸡输卵管生物反应器是一种很有市场前景的生产药物蛋白的生物反应器。综述了激素对输卵管细胞和卵清蛋白基因的调控、输卵管组织特异性调控元件的研究、转基因鸡的制作途径、新型逆转录病毒载体在转基因鸡制作上的应用和国内研究现状、最新成果及发展前景等。     

Is the Function of the Porcine Sperm Reservoir Restricted to the Ovulatory Period?

The uterotubal junction (UTJ) and caudal isthmus are recognized as a functional pre-ovulatory sperm reservoir (SR). Spermatozoa are released from the SR in a complex and concerted action. However, whether this functionality is restricted only to the ovulatory period is still open to debate. Our study was aimed to analyze the presence of spermatozoa within the UTJ (SR), isthmus (ISTH) and ampulla (AMP) after laparoscopic intrauterine insemination (LIUI) either in the peri- (PERI) or post-ovulatory (POST) period or at mid cycle (MID). Each uterine horn of estrus synchronized gilts (n=12) was inseminated with 20 ml sperm (29.5×10⁶ cells/ml). Oviducts were recovered 7 h after LIUI and separated into the UTJ, ISTH and AMP, and sections were flushed with 10 ml PBS+EDTA solution. After centrifugation, the sperm pellet was evaluated by Čeřovský staining. The median sperm numbers in the PERI, POST and MID groups were 578, 171 and 789 in the UTJ; 545, 233 and 713 in the ISTH; and 496, 280 and 926 in the AMP, respectively, and there were differences between the POST and MID groups (P<0.05) but not between the oviductal sections of each group (P>0.05). Compared with the MID group, the percent of intact sperm cells was higher (P<0.01) in the PERI and POST groups (32.8 vs. 66.4 and 76.8%). Also, the percentages of aberrations in the acrosome and tail were higher (P<0.05) in the MID group. Based on this, it can be assumed that the sperm reservoir is active during different phases of the estrus cycle. However, the mid-cycle oviduct environment considerably impairs sperm cell quality.     

Detection of Coxiella burnetii, the agent of Q fever, in oviducts and uterine flushing media and in genital tract tissues of the non pregnant goat

The aim of the present study was the detection and quantification of Coxiella burnetii DNA in the flushing media (oviducts and uterine horns) and genital tract tissues of non pregnant goats from 20 goats chosen at random from 86 goats originating from 56 different breeding herds in south-west France. The serological prevalence rate of C. burnetii in the study population was 70.3%.The DNA of C. burnetii was identified using conventional PCR in the flushing media from the oviducts and uterus in 8/20 goats (40%) and in genital tract tissues (oviduct, uterus and ovary) in 5/20 goats (25%). This study clearly shows for the first time that the media used to flush the oviducts or uterine horns, collected using the standard embryo harvesting technique in goats, are susceptible to infection with C. burnetii. The 16 conventional PCR-positive samples were also analyzed using real-time PCR. The bacterial load of the oviduct and uterine flushing media varied from 2.9 × 10⁴ to 7.5 × 10⁶ bacteria per flushing medium, while the bacterial load of the tissue samples varied from 1.0 × 10² to 1.5 × 10⁵ bacteria per mg of tissue. The infection of genital tract flushing media and tissues is a risk factor for the transmission of C. burnetii from donor to recipient during embryo transfer or to the embryo and fetus when gestation is pursued to term.     

Ovarian steroids modulate tumor necrosis factor-α and nitric oxide-regulated prostaglandin secretion by cultured bovine oviductal epithelial cells

Ovarian steroids assure an optimum environment for the final maturation of oocytes, gamete transport, fertilization, and early embryonic development. The aim of experiment 1 was to examine the influence of ovarian steroids on tumor necrosis factor-α (TNF-α)- or nitric oxide (NO)-regulated prostaglandin (PG), and nitrite/nitrate (NO₂/NO₃) secretion by cultured bovine oviductal epithelial cells (BOECs). BOECs were pretreated with 17β-estradiol (E₂; 10⁻⁹ M) and/or progesterone (P₄; 10⁻⁷ M) for 24 h. For the next 24 h, BOECs were treated with TNF-α (10 ng/mL) or spermine nitric oxide complex (NONOate; 10⁻⁵ M). Prostaglandin F_2α and PGE₂ secretion was measured in medium by ELISA. The pretreatment of cells with P₄ (progesterone), E₂ (17 β-estradiol), or E₂/P₄ augmented TNF-α-induced PGF_2α and PGE₂ secretion (P < 0.01). The pretreatment of cells with E₂ or E₂/P₄ increased NONOate-induced PGF_2α and PGE₂ secretion (P < 0.01). TNF-α induced NO₂/NO₃ production by BOECs. The pretreatment of cells with E₂ augmented only TNF-α-induced NO₂/NO₃ production (P < 0.05). The aim of experiment 2 was to examine the influence of TNF-α, NO, and ovarian steroids on the protein content of enzymes specifically involved in PG and NO production, PG synthases, and NO synthases (NOSs). BOECs were treated with TNF-α (10 ng/mL) or NONOate (10⁻⁵ M). TNF-α increased the protein content of PGG/H synthase, PGF synthase, and PGE synthase (P < 0.05) and endothelial and inducible NOSs (P < 0.05). Nitric oxide increased the protein content of PGF synthase, PGE synthase, endothelial NOS, and inducible NOS (P < 0.05). These results show possible linkage between TNF-α and NO, modulated by ovarian steroids, in the regulation of PG synthesis by BOECs that may be important for triggering the process of oviductal contractions.     

不同胚胎移植方式的移植效率比较

在人源化抗体转基因猪的研究工作中，将所设计的转基因胚胎成功植入能繁发情受体母猪的输卵管内是推进后续研究工作的基础。所以，本研究对比了两种胚胎移植方法对62例受体母猪的手术效果，结果表明，采用输卵管伞部移植方式的移植效率（60%）显著高于刺破输卵管移植方式（31.8%）。