Involvement of matrix metalloproteinases (MMP-2, -7, -9) and their tissue inhibitors (TIMP-2, -3) in the chicken oviduct regression and recrudescence.

This study was undertaken to examine mRNA and protein expression, localization of selected MMPs and their tissue inhibitors (TIMPs), and the activity of MMPs in all segments of the chicken oviduct (infundibulum, magnum, isthmus, shell gland, vagina) during a pause in egg laying induced by food deprivation. Control chickens (n = 18) were fed ad libitum, while experimental birds (n = 18) were fasted for 5 days, followed by being fed every other day, and then fed daily from day 10 onwards. Chickens were decapitated on day 6 (when the oviduct was regressed), day 13 (during oviduct recrudescence), and days 17-20 (oviduct rejuvenation) of the experiment. A pause in laying occurred between days 6 and 13 and resumption of laying before days 17-20. Real-time polymerase chain reaction and western blot revealed different expression of MMP-2, -7, -9 and TIMP-2 and -3 on mRNA and protein levels, respectively, as well as activity of MMP-2 and -9 by activity assay in the oviduct parts. Immunohistochemistry showed cell- and tissue-dependent localization of the examined members of the MMP system. Regression of the oviduct was accompanied predominantly by an increase in the relative expression of MMP-2, -7 and -9, and TIMP-2 and -3 mRNAs. The protein expression levels and localization of MMPs and TIMPs did not show pronounced changes during the examined period. During oviduct regression and recrudescence, elevated activity of MMP-2 and -9 was found. In summary, the results showing the stage of reproductive cycle-dependent changes as well as cell- and tissue-dependent differences in the expression of selected MMPs and TIMPs, and activity of MMP-2 and MMP-9 point to the significance that these molecules might be the local regulators of remodeling and functions of the hen oviduct.

Histochemical structure and immunolocalisation of the hyaluronan system in the dromedary oviduct.

We investigated the local modulation of some histochemical properties of oviducts of the dromedary (Camelus dromedarius), focusing on the immnolocalisation of hyaluronic acid (HA) synthases (HAS2 and HAS3), hyaluronidases (HYAL2 and HYAL1) and the HA receptor CD44 in the ampulla and isthmus. Abundant acidic mucopolysaccharides (glycosaminoglycans) were detected by Alcian blue staining along the luminal surface of both ciliated and non-ciliated epithelial cells (LE). Staining for HAS2 was higher in the primary epithelial folds of the ampulla compared with the isthmus, especially in secretory cells, adluminal epithelial surface and supranuclear cell domain. HAS3 staining was stronger in the LE of the isthmus than ampulla. HYAL2 was detected in the LE in the ampulla and isthmus and was more intense in the adluminal projections of secretory cells. HYAL1 was weakly detected in the LE with no difference between the ampulla and isthmus. Strong CD44 immunostaining was present in the LE of the ampulla and isthmus. CD44 staining was higher in secretory cells than in ciliated epithelial cells and was higher in the supranuclear region than the basal region of the cytoplasm. In conclusion, we provide evidence that HA synthesis and turnover occur in the camel oviduct. Differences in HAS2 and HAS3 expression suggest regional differences in the molecular size of HA secreted in oviductal fluid that may influence oviduct-gamete interaction in the camel.

Estrogen regulation of phosphoserine phosphatase during regression and recrudescence of female reproductive organs.

Phosphoserine phosphatase (PSPH) is a well-known mediator of l-serine biosynthesis in a variety of tissues and its dysregulation causes various diseases, specifically most cancers. However, little is known about the expression and hormonal regulation of PSPH gene in the female reproductive tract. Therefore, in the current study, we focused on relationships between PSPH expression and estrogen during growth, development, differentiation, remodeling and recrudescence of the chicken oviduct and in the progression of epithelial-derived ovarian carcinogenesis in laying hens. The results revealed that PSPH mRNA and protein levels increased in the glandular (GE) and luminal epithelial (LE) cells in the developing oviduct of chicks treated with exogenous estrogen. Additionally, PSPH mRNA and protein expression was up-regulated in GE and LE of the oviduct in response to endogenous estrogen during the recrudescence phase after induced molting. Furthermore, PSPH mRNA and protein were predominantly detected in GE of cancerous, but not normal ovaries. In conclusion, PSPH is a novel estrogen-responsive gene involved in development of the oviduct of chicks and recrudescence of the oviduct of laying hens after molting. PSPH is also a potential target molecule that may help elucidate mechanism responsible for the progression of epithelial cell-derived ovarian carcinogenesis and be of use in therapeutic applications as a biomarker for early diagnosis of epithelial cell-derived ovarian cancer in laying hen as well as women.

Avian prostatic acid phosphatase: estrogen regulation in the oviduct and epithelial cell-derived ovarian carcinomas.

Prostatic acid phosphatase (ACPP) is a glycoprotein that is mainly synthesized and secreted by glandular epithelial cells (GE) of the prostate, and it is well known as a biomarker for prostate cancer. Although ACPP was used as prognostic/diagnostic indicator and studied to elucidate regulatory mechanism(s) during several decades in humans, its role is not clearly understood. Gene profiling data using a chicken DNA microarray revealed that ACPP increased significantly during remodeling and recrudescence of the oviduct in response to estrogen. Thus, in this study, we investigated the expression and hormonal regulation of ACPP gene in the reproductive tracts of chickens. ACPP was specifically detected in the luminal cells (LE) and GE of chicken oviduct, and diethylstilbestrol (a synthetic nonsteroidal estrogen) stimulated its expression during development of the oviduct. In addition, ACPP mRNA and protein were localized to LE and GE during the regeneration phase of the oviduct of laying hens during induced molting. Furthermore, ACPP mRNA and protein were abundant in GE of ovarian carcinoma, but not in normal ovaries. Moreover, strong expression of ACPP protein was detected in epithelial cells of cancerous ovaries from women. Collectively, results of the present study are the first to show that ACPP is a novel estrogen-stimulated gene in the oviductal epithelial cells of the chicken and that its expression increases significantly in epithelial cells of ovarian carcinoma, which indicates that it may be a candidate biomarker for diagnosis of epithelia-derived ovarian cancer in women.

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.

Are glucocorticoids auto- and/or paracrine factors in early bovine embryo development and implantation?

We determined the transcript content of three genes involved in the metabolism of glucocorticoids (GC) in bovine in vitro fertilized embryos (2-blastomere stage until hatched blastocyst), trophoblast as well as the oviduct (Day 2-4 of the estrous cycle) and endometrium (Day 16 of the cycle and pregnancy). Since mRNA expression of the glucocorticoid receptor and two enzymes responsible for GC production (11ß-HSD1 and 2) was demonstrated in the embryos in all pre-implantation stages as well as in the endometrium and oviduct, it is suggested that GC may serve as auto-/paracrine factors in the development of bovine pre-implantation embryos.

Localization of lysyl oxidase in hen oviduct: implications in egg shell membrane formation and composition.

Lysyl oxidase activity was found in the isthmus (the membrane-forming region) of the hen's oviduct in a copper-rich region proximal to the shell gland. Desmosine and isodesmosine, cross-linking compounds associated with mature elastin, were found in hydrolysates of the shell membrane, confirming the necessity for lysyl oxidase in its biosynthesis. Shell membranes from hens fed a copper-deficient diet or a diet supplemented with beta-aminopropionitrile had a reduced content of desmosine and isodesmosine.

Estrogenic induction of ornithine decarboxylase in vivo and in vitro.

Injection of estrogens (17beta-estradiol or diethylstilbestrol) into immature chicks results in a marked (30- to 50-fold) increase in the ornithine decarboxylase activity of oviductal homogenates within a 4-hour period. Similar stimulations were obtained when estrogen was injected into hypophysectomized or castrated rats and the uterus was examined for decarboxylase activity. An elevation of decarboxylase activity was obtained in vitro when oviducts from immature chicks were incubated in the presence of estrogen. These data indicate a direct action of estrogen on oviduct tissue to promote a rapid increase in the activity of a specific enzyme and represent the first example of a completely in vitro enzyme response to estrogen.

Steroid hormones: effects on adenyl cyclase activity and adenosine 3',5'-momophosphate in target tissues.

The adenyl cyclases of chick oviduct and rat prostate were not stimulated by estrogen and testosterone, respectively, suggesting that growth and differentiation of these target tissues are not mediated by adenosine 3',5'-monophosphate. Estrogen acutely activated adenyl cyclase in the castrate rat uterus, but this was prevented by administration of DL-propranolol, suggesting that the effect was mediated by catecholamines. Progesterone produced a delayed stimulation of oviduct adenyl cyclase preceding and concomitant with the induction of synthesis of avidin.

Transgenic cyclooxygenase-2 expression and high salt enhanced susceptibility to chemical-induced gastric cancer development in mice.

Cyclooxoygenase (COX)-2 overexpression is involved in gastric carcinogenesis. While high-salt intake is a known risk factor for gastric cancer development, we determined the effects of high salt on gastric chemical carcinogenesis in COX-2 transgenic (TG) mice. COX-2 TG mice were developed in C57/BL6 strain using the full-length human cox-2 complementary DNA construct. Six-week-old COX-2 TG and wild-type (WT) littermates were randomly allocated to receive alternate week of N-methyl-N-nitrosourea (MNU, 240 p.p.m.) in drinking water or control for 10 weeks. Two groups of mice were further treated with 10% NaCl during the initial 10 weeks. All mice were killed at the end of week 50. Both forced COX-2 overexpression and high-salt intake significantly increased the frequency of gastric cancer development in mice as compared with WT littermates treated with MNU alone. However, no additive effect was observed on the combination of high salt and COX-2 expression. We further showed that MNU and high-salt treatment increased chronic inflammatory infiltrates and induced prostaglandin E(2) (PGE(2)) production in the non-cancerous stomach. Whereas high-salt treatment markedly increased the expression of inflammatory cytokines (tumor necrosis factor-alpha, interferon-gamma, interleukin (IL)-1 beta and IL-6) in the gastric mucosa, COX-2 overexpression significantly altered the cell kinetics in the MNU-induced gastric cancer model. In conclusion, both high salt and COX-2 overexpression promote chemical-induced gastric carcinogenesis, possibly related to chronic inflammation, induction of PGE(2), disruption of cell kinetics and induction of inflammatory cytokines.

Glycosidase determination in bovine oviducal fluid at the follicular and luteal phases of the oestrous cycle.

Gamete recognition and binding of spermatozoa to the oviduct are carbohydrate-mediated processes in which several glycosidases are thought to have a role, although this has not been demonstrated unequivocally. Oviducal fluid is the biological milieu in which fertilisation and early embryo development take place, but the enzyme composition of oviducal fluid is largely unknown. The aim of the present study was to determine glycosidase activity and protein content in bovine oviducal fluid (bOF) and the volume of fluid collected per oviduct. Oviducts obtained from a slaughterhouse were classified as either in the follicular or luteal phase on the basis of ovarian luteal morphology. Oviducal fluid was aspirated, centrifuged and the volume determined. Samples were then frozen until assay. Substrates conjugated to 4-methylumbelliferyl were used to screen for the activity of seven glycosidases at pH 7.2. The results indicate that bOF has alpha-l-fucosidase, beta-N-acetyl-glucosaminidase, beta-d-galactosidase, alpha-D-mannosidase and beta-N-acetyl-galactosaminidase activity during both phases of the cycle, with the specific activity of the latter two enzymes being higher during the follicular phase. There was no N-acetyl-neuraminidase or alpha-d-galactosidase activity detected in bOF at either phase of the oestrous cycle at pH 7.2, but activity for both glycosidases was detected at pH 4.4. There were no differences in protein concentration or the volume of bOF collected between the two phases of the cycle. These findings indicate that oviducal fluid exhibits glycosidase activity, with specific variations throughout the oestrous cycle, suggesting that these enzymes play a role in carbohydrate-mediated events.

Expression of superoxide dismutases in the bovine oviduct during the estrous cycle.

The superoxide dismutases (SODs) are first-line enzymatic antioxidants that dismute superoxide anion (O(2)(-)*) to produce hydrogen peroxide (H(2)O(2)). The primary objective was to characterize, by western blot analysis, the expression of two SODs, the cytosolic (Cu,ZnSOD or SOD1) and the mitochondrial (MnSOD or SOD2) forms in three sections of the oviduct, i.e. isthmus (I), ishtmic-ampullary junction (IA), and ampulla (A), during the estrous cycle. The Cu,ZnSOD and MnSOD proteins were mostly expressed in the ampulla (I

Metabolic barrier against bisphenol A in rat uterine endometrium.

Exposure to environmental chemicals with estrogenic activity during the early stage of pregnancy can seriously affect embryonic development and the maintenance of pregnancy. To estimate the metabolism and pharmacodynamics of a xenoestrogen, bisphenol A, in a reproductive organ, the metabolite of bisphenol A was analyzed after incubating a rat uterine sac in buffer solutions containing the chemical. When the inner or the outer side of the uterine sac was exposed to bisphenol A, the concentration of the parent chemical was decreased in buffer solution and then, only one metabolite, bisphenol A-glucuronide, was observed only in the outer, that is, the maternal, side. A small amount of the parent chemical could pass through the uterine sac without being modified. Uridine diphosphate (UDP)-glucuronosyltransferase (UGT) was shown by immunohistochemical staining analysis to be distributed in epithelial cells of the endometrium, oviduct, and uterine glands. Based on measurements of enzyme activity and on Western blot analysis, UGT activity toward bisphenol A and UGT protein were identified in the microsomal fractions prepared from rat uterus. UGT isoforms, such as UGT1A1, 1A2, 1A5, 1A6, and 1A7, were expressed, and MRP-1 (multidrug resistance-associated protein) and MRP-3, which are well-known to be transporters of various drug-glucuronides, were detected in the rat uterus by reverse transcription-PCR. These results elucidate the rat uterine barrier system by showing that most bisphenol A perfused into the uterus was glucuronidated in the epithelium, resulting in transport of glucuronides to the maternal side.

The -6.1-kilobase chicken lysozyme enhancer is a multifactorial complex containing several cell-type-specific elements.

In the chromatin domain of the chicken lysozyme gene of myeloid and oviduct cells, which both have the potential to activate the gene, a developmentally stable DNase I-hypersensitive site is formed around 6.1 kb upstream of the gene. This implies that this DNA region, which has previously been demonstrated to function as a transcriptional enhancer element in myeloid cells, is intimately involved in the cell-type-specific activation of the lysozyme gene locus. Deletion analysis identifies a 157-bp minimal fragment that confers the same promacrophage-specific enhancer activity as the originally described 562-bp -6.1-kb enhancer fragment. By introducing specific point mutations, we demonstrate in transient gene transfer experiments that the minimal fragment consists of at least six adjacent elements, each substantially contributing to enhancer function. The compact multifactorial enhancer complex includes a nuclear factor I (NF-I)/TGGCA binding site, homologies to AP1, and octanucleotide or enhancer core consensus motifs. Point mutation of the NF-I binding site results in the loss of NF-I binding in vitro and enhancer activity in vivo after gene transfer. Surprisingly, four overlapping oligonucleotides, each consisting of at least two elements of the -6.1-kb enhancer, confer myeloid-cell-specific enhancer activity. We found several myeloid-cell-specific DNA-binding proteins interacting with the -6.1-kb enhancer, a result consistent with that described above. Therefore, we suggest that more than a single trans-acting factor mediates the cell type specificity of the -6.1-kb enhancer.

Spatial distribution of cannabinoid receptor 1 and fatty acid amide hydrolase in the cat ovary and oviduct.

Involvement of the endocannabinoid system in female reproduction has been extensively described in humans with the cognate receptors and ligands being found in the ovaries and genital tract. In human, an imbalance of the endocannabinoid system is linked with both ectopic pregnancy and infertility. In bovine species anandamide levels regulate aspects of sperm-oviduct interaction. Here we report the immunohistochemical distribution of cannabinoid receptor 1 (CB1R) and fatty acid amide hydrolase (FAAH) in cat ovary and oviduct, using paraffin-embedded tissue samples and commercially available antibodies. We found a differential expression of both CB1R and FAAH during different stages of ovarian function and in the oviduct. CB1R was detected only in tertiary follicle granulosa cells while more immature follicles were negative. FAAH was instead found in ovarian pre-antral follicles, the oocyte cytoplasm, and in granulosa cells of primary, secondary and tertiary follicles. Secondary and tertiary follicles were also FAAH immunoreactive. Luteal cells were immunopositive for both CB1R and FAAH. Because CBR1 in oviduct was found only in ciliated cells, it might represent a specific marker at least in cats. In contrast, FAAH immunoreactivity was observed in both ciliated and non-ciliated cells. The present study may thus serve as the starting point for further investigations on the role of the endocannabinoid system in cat reproduction. Additional work will be needed to assess whether the morphological distribution of CB1R and FAAH changes in different conditions such as pre-pubertal age, follicular phase of the sexual cycle and pregnancy.

Expression and localization of cyclooxygenases in the oviduct of laying hens during the ovulatory cycle.

Prostaglandins (PGs) play important roles in regulation of the functions of the hen oviduct. However, little is known about the expression and localization of the rate-limiting cyclooxygenases (COX-1 and COX-2) in the oviduct. The aim of this study was to determine the COXs expression and localization in the different segments of the oviduct and to investigate changes in their expression levels during the ovulatory cycle of laying hens. White Leghorn laying hens were killed at 0, 4, 7, 16 and 24 h after oviposition, and samples from the infundibulum, magnum, isthmus, uterus, and vagina were collected. Gene and protein expressions were examined by real-time PCR and western blot, respectively, for both COX-1 and COX-2. Localization of COX-1 and COX-2 in the hen oviduct was determined by immunohistochemistry and PCR analysis of samples collected by laser capture microdissection (LCM). The expression level of COX-1 was highest in the infundibulum, while that of COX-2 was significantly higher in the uterus than in the other segments. The expression levels of COX-1 in the infundibulum and COX-2 in the uterus were higher at 0 and 24 h after oviposition, just prior to subsequent ovulation and oviposition. Western blot analysis confirmed the presence of COX-1 and COX-2 in all oviductal segments. The density of COX-2 was the highest in the uterus, and did not change during the ovulatory cycle. COX-1 and COX-2 were localized in the surface epithelium of all oviductal segments besides the uterine tubular glands. We conclude that both COXs are differentially expressed in the different oviductal segments with a temporal association to ovulation and oviposition. COX-1 and COX-2 may play an important role in the infundibulum and uterus, respectively, and COX-2 may be one of the factors regulating the induction of oviposition.

Expression and localization of matrix metalloproteinases (MMP-2, -7, -9) and their tissue inhibitors (TIMP-2, -3) in the chicken oviduct during pause in laying induced by tamoxifen.

Induced pause in egg laying simulates natural molting events in which the hen's reproductive organs regress and rejuvenate. Such processes require extracellular matrix remodeling that is maintained, at least in part, by the action of proteolytic enzymes known as matrix metalloproteinases (MMPs). Nevertheless, information concerning the expression and hormonal regulation of MMP system members in chickens is scarce. Therefore, MMP-2, -7, and -9 and their tissue inhibitors (TIMP-2, -3) expression and localization were investigated in all segments of the domestic hen oviduct (infundibulum, magnum, isthmus, shell gland, vagina) during a pause in egg laying induced by tamoxifen (TMX)-an estrogen receptor modulator. Hy-Line Brown hens were treated daily with TMX (n = 6) at a dose of 6 mg/kg of body weight or a vehicle (n = 6) until complete cessation of egg laying (for 7 days). Chickens were decapitated on Day 7 of the experiment. Real-time polymerase chain reaction and Western blotting revealed section-dependent expression of MMP-2, -7, -9 and TIMP-2 and -3. Immunohistochemistry found tissue and cell-dependent localization of examined proteins in the wall of the oviduct. The MMP-2, TIMP-2, and TIMP-3 were localized mainly in the luminal epithelium, MMP-7 in the luminal and glandular epithelium, whereas MMP-9 was detected only in the connective tissue. Treatment of chickens with TMX markedly elevated the relative expression of MMP-7 and MMP-9 mRNA in the oviduct, but did not affect MMP-2, TIMP-2, and TIMP-3 mRNA levels. However, TMX increased the MMP-2 protein level in the infundibulum, shell gland, and vagina as well as activity of MMP-2 evaluated by gelatin zymography. The results obtained indicate that MMP-2, MMP-7, and MMP-9 are involved in chicken oviduct regression. Moreover, changes in the expression and activity of chosen MMPs after TMX treatment may indicate a contribution of estrogen in the regulation of transcription, translation, and/or the activity of selected elements of the MMP system.

Differential expression of cyclooxygenase 1 and cyclooxygenase 2 in the bovine oviduct.

The aim of the present study was to investigate the enzymes for the local prostaglandin (PG) biosynthesis present in the bovine oviduct during the estrous cycle to influence early reproductive events. Bovine oviducts were classified into four phases: pre-ovulatory, post-ovulatory, early-to-mid luteal, and late luteal phase, subdivided further into ipsi- or contralateral site and separated into ampulla or isthmus. Oviductal cells were gained by flushing the oviductal regions. Quantitative real-time reverse transcriptase-PCR was performed for the secretory and cytosolic phospholipases A(2) (sPLA(2)IB, cPLA(2)alpha, and cPLA(2)beta) and cyclooxygenases (COX-1 and COX-2) as the first step enzymes of PG synthesis. COX-1 and cPLA(2)beta showed significant highest mRNA expression around and before ovulation compared with the luteal phase respectively. sPLA(2)IB and cPLA(2)alpha mRNA expression was unregulated during the estrous cycle. Regional differences in mRNA content were found for sPLA(2)IB with higher mRNA expression in the ampulla than in the isthmus. Western blot analysis revealed the highest COX-1 protein content in the early-to-mid luteal phase. Immunohistochemistry demonstrated that COX-1 was localized in epithelial and smooth muscle cells, whereas COX-2 was only localized in epithelial cells. COX-2 showed a differential distribution within the epithelial cell layer suggesting a regulation on a cellular level, although the COX-2 mRNA and protein amounts did not vary throughout the estrous cycle. A COX activity assay of oviductal cells revealed that COX activity originated predominantly from COX-1 than from COX-2. Treatment of primary oviductal cells with 10 pg/ml 17beta-estradiol or 10 ng/ml progesterone resulted in a higher expression of COX-2 and cPLA(2)alpha, but not of the other enzymes. The expression pattern of these enzymes suggests that an estrous-cycle dependent and region-specific PG synthesis in the bovine oviduct may be required for a successful reproduction.

17-beta-Estradiol upregulates COX-2 in the rat oviduct.

We investigated the regulation of cyclooxygenase-2 (COX-2) by 17-beta-estradiol (E2) in the rat oviduct. We observed that COX-2 is expressed mainly in proestrous and estrous stages, periods under estrogenic influence. While exogenous administration of E2 (1 microg/rat) significantly increased COX-2 protein levels, progesterone did not modify it. COX-2 was mainly localized on oviductal epithelial cells from estrogenized rat. Induction of COX-2 expression by E2 was partially reverted by tamoxifen (1 mg/rat), an E2 receptor antagonist. Estradiol treatment also increased prostaglandins (PGs) synthesis: 6-keto-PGF(1alpha) (40%), a stable metabolite of prostacyclin (PGI2), PGF(2alpha) (40%) and PGE2 (50%). Tamoxifen completely suppressed this enhancement. In order to discriminate which isoform of COX was implicated in the stimulatory effect of E2 on PGs synthesis, oviducts were preincubated with meloxicam (Melo: 10(-9)M) or NS-398 (10(-7)M), two selective COX-2 inhibitors. Both Melo and NS-398 abolished the increase of PGs synthesis stimulated by E2. All together, these data indicate that E2 could upregulate COX-2 expression and activity in the rat oviduct and that the stimulatory effect of E2 may be receptor-mediated.

Acid glycosidases in the isthmus of the hen oviduct and egg shell membranes.

Specific activities of seven acid glycosidases: beta-hexosaminidase, alpha- and beta-galactosidase, alpha- and beta-mannosidase, alpha-glucosidase and alpha-fucosidase were determined in various parts of the domestic hen oviduct (infundibulum, isthmus, shell gland and vagina). The activity of most enzymes was the highest in the isthmus. Multiple forms of all acid glycosidases from the isthmus were separated by strong anion exchange chromatography at pH 6.0. The isoelectric points of the isthmus forms of beta-hexosaminidase, beta-galactosidase and alpha- and beta-mannosidase were determined by chromatofocusing. For the first time the high beta-galactosidase activity was found in hen egg shell membranes.