Biol Reprod Lalor Postdoctoral Fellowships -- Application Deadline January 15, 2009
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

BOR - Papers in Press, published online ahead of print June 11, 2003.
Biol Reprod 2003, 10.1095/biolreprod.103.017327
This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
69/4/1341    most recent
biolreprod.103.017327v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Wijayagunawardane, M. P.B.
Right arrow Articles by Miyamoto, A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Wijayagunawardane, M. P.B.
Right arrow Articles by Miyamoto, A.
Agricola
Right arrow Articles by Wijayagunawardane, M. P.B.
Right arrow Articles by Miyamoto, A.
BIOLOGY OF REPRODUCTION 69, 1341–1346 (2003)
DOI: 10.1095/biolreprod.103.017327
© 2003 by the Society for the Study of Reproduction, Inc.

Female Reproductive Tract

Tumor Necrosis Factor {alpha} in the Bovine Oviduct During the Estrous Cycle: Messenger RNA Expression and Effect on Secretion of Prostaglandins, Endothelin-1, and Angiotensin II1

Missaka P.B. Wijayagunawardane3,4, Christoph Gabler5, Gary Killian5, and Akio Miyamoto2,6

Department of Animal Science,4 University of Peradeniya, Peradeniya, Sri Lanka John O. Almquist Research Center,5 The Pennsylvania State University, University Park, Pennsylvania, 1680 Department of Agricultural and Life Science,6 Obihiro University of Agriculture and Veterinary Medicine, Obihiro 080-8555, Japan


   ABSTRACT
TOP
 ABSTRACT
INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
Tumor necrosis factor (TNF) {alpha} is an important physiological mediator of cell-to-cell communication. Recent observations suggest that TNF{alpha} is involved in the control of reproductive functions. The present study examined the role of TNF{alpha} in the secretion of factors involved in regulating smooth muscle contraction, such as prostaglandin E2 (PGE2), prostaglandin F2{alpha} (PGF2{alpha}), endothelin-1 (ET-1), and angiotensin II (Ang II), as it was in the original by the cow oviduct at different stages of the estrous cycle using an in vitro microdialysis system. Expression of mRNA for TNF{alpha} and its receptors (TNF{alpha}-R) was also evaluated. For microdialysis, the lumen of a portion (length, 10 cm) of the each oviductal segment was implanted with a dialysis capillary membrane, and TNF{alpha} (100 ng/ml) was infused for 4–8 h during a 16-h incubation period. The microdialysis system maintains cell-to-cell integrity and cell-to-cell communication, and it enables real-time observation of physiological changes in the luminal release of different substances. Concentrations of PG, ET-1, and Ang II in 4-h fractions were measured using second-antibody enzyme immunoassays. Infusion of TNF{alpha} stimulated oviductal secretion of PG, ET-1, and Ang II during the follicular and postovulatory stages, but not during the luteal stage. Expression of TNF{alpha}, TNF{alpha}-R type I, and TNF{alpha}-R type II mRNA was detected in the bovine oviduct by reverse transcription-polymerase chain reaction. High expression of both TNF{alpha}R types and ligands was detected during the follicular and postovulatory stages, whereas low expression was detected during the luteal stage. The results of the present study provide, to our knowledge, the first direct evidence that TNF{alpha} stimulates PG, ET-1, and Ang II secretion and that up-regulation of the TNF{alpha} system occurs in the cow oviduct during the periovulatory period. In conclusion, the TNF{alpha} system may optimize the release of contraction-related substances and modulate local contraction to regulate the oviductal transport of the gametes and embryo.

cytokine, female reproductive tract, oviduct, ovulatory cycle, ovum pick-up/transport


   INTRODUCTION
TOP
 ABSTRACT
 INTRODUCTION
MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
Tumor necrosis factor (TNF) {alpha} is a cytokine that mediates cell-to-cell communication. This cytokine was initially identified as an up- and down-regulator of immunologic, inflammatory, and reparative host responses to injury. The existence of the bioactive form of this proinflammatory cytokine, as well as of its binding sites in the various reproductive tissues, has been reported. It has been postulated that the bioactive form of TNF{alpha} interacts with other substances to control reproductive functions [1–5]. It was reported that specific receptors for TNF{alpha} (TNF{alpha}-R) are present in the bovine endometrium and that TNF{alpha} may trigger the release of luteolytic prostaglandin F2{alpha} (PGF2{alpha}) [3]. In addition, human [6] and rat [7] preimplantation-stage embryos have been shown to express TNF{alpha}-R. Ligands for TNF{alpha} have been shown to be secreted by the supporting tissues of the oviduct and uterus and, in some cases, by the embryo itself [8]. Moreover, the TNF gene is expressed in mouse oviductal epithelial cells [5], and human oviductal fluid contains TNF{alpha} [4].

Local production of prostaglandin E2, (PGE2), PGF2{alpha}, endothelin-1 (ET-1), and angiotensin II (Ang II) as well as their involvement in the cyclic regulation of functions have been studied extensively in the cow oviduct. F-series PGs are involved in oviductal contraction, whereas E-series PGs relax smooth muscle [9]. Moreover, ET-1 is locally involved in muscular oviductal contraction [10]. Local PGs and ET-1 concentrations in the cow oviduct are highest during the periovulatory period of the estrous cycle [11], and PG, ET-1 [12], and Ang II [13] directly increase the contractile amplitude of the cow oviduct in vitro.

Stimulatory effects of TNF{alpha} on the biosynthesis and release of PG [14–19], ET-1 [20–22], and Ang II [23, 24] have been reported in various tissues. However, the role of TNF{alpha} in regulating oviductal secretion of PG, ET-1, and Ang II, which are the main contraction-related substances in the oviduct, is not known. In the present study, we investigated the effect of TNF{alpha} on secretion of PGE2, PGF2{alpha}, ET-1, and Ang II by the cow oviduct using an in vitro microdialysis system. In addition, mRNA expressions for TNF{alpha} and for TNF{alpha}-R were evaluated in oviducts collected from the follicular, postovulatory, and luteal stages.


   MATERIALS AND METHODS
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
RESULTS
 DISCUSSION
 REFERENCES
 
Animals and Sample Collection

Whole reproductive tracts were collected from nonpregnant Holstein cows at a local slaughterhouse within 20 min of slaughter. The stage of the estrous cycle was defined as previously described [11, 25] based on the morphology of the corpus luteum, uterine fluid and cervical mucus characteristics, and luteal progesterone (P4) levels. For microdialysis experiments, oviducts were separated from the uterotubal junction, and the surrounding connective tissues were trimmed. The oviducts were transported to the laboratory in Medium 199 (M199; 25 mmol/L of Hepes, 0.85 g/L of NaHCO3, 60 mg/L of penicillin, 100 mg/L of streptomycin, 56 mg/L of ascorbic acid, and 2 mg/L of amphotericin B; Sigma Chemical Co., St. Louis, MO), maintained at 38°C. The ampullary and isthmus regions of both oviducts were used.

For RNA extraction, oviducts from Holstein cows were collected at the slaughterhouse and transported on ice to the laboratory. Whole oviducts were filled with 1 ml of PBS, and the oviductal contents were squeezed into a 1.5-ml microcentrifuge tube. After centrifugation at 570 x g for 3 min at 4°C, the supernatant was removed, and cell pellets of both oviducts of one cow were combined and stored at -70°C until examined. Cell types (>60% epithelial cells) and oviductal cell viability (>99%) were verified as described previously [26].

In Vitro Microdialysis of Bovine Oviduct

The in vitro microdialysis of the oviduct has been described previously [12, 13]. The lumen of each oviductal segment (length, 10 cm) was implanted with a dialysis capillary membrane (length, 7 cm; diameter, 0.2 mm; cutoff, 1000 kDa; Fresenius SPS 600 Hollow Fiber; Fresenius AG, St. Wendel, Germany) of the microdialysis. The oviductal segment was incubated in M199 with 0.5% BSA (Sigma) in simple organ culture chambers (modified 50-ml Falcon tubes; Becton Dickinson & Co, Franklin Lakes, NJ). Four oviducts were maintained in each organ culture chamber, and the medium was continuously exchanged at a flow rate of 50 ml/h during the whole period of incubation. The chambers were maintained in a water bath at 38°C. Ringer solution was continuously perfused (1.3 ml/h) from one end of the microdialysis using a peristaltic pump; the other end was connected to a fraction collector. After a 2-h preincubation, the perfusate was collected in 4-h fractions for 16 h. Control (Ringer solution only) or TNF{alpha} (100 ng/ml; DAINIPPON Pharmaceutical Co., Ltd., Osaka, Japan) in Ringer solution were infused at 4–8 h of incubation. The 4-h fractions of perfusate were stored at -20°C until extraction for PG and peptides.

Hormone Extraction

Prostaglandin and ET-1 extractions from microdialysis samples were performed as described earlier [27]. The Ang II extraction was done according to the procedure for ET-1. For PG extraction, the 5.2-ml microdialysis fractions were allowed to reach room temperature, adjusted to pH 3.5 with 1 N HCl, and then maintained at room temperature for 1 h. Next, five volumes of diethyl ether were added, and the mixture was shaken for 30 min at a frequency of 250 cycles/min (Shaker SA 31; Yamato Co., Tokyo, Japan). The samples were then allowed to stand for 15 min and placed in a -80°C freezer for 1 h. The upper diethyl ether fraction was decanted and evaporated. The residue was dissolved in 400 µl of assay buffer for enzyme immunoassays (EIAs) for PG (40 mM PBS and 0.1% BSA; pH 7.2). After the diethyl ether extraction, the remaining Ringer solution was used for ET-1 and Ang II extraction. The BSA was added to the samples to a final concentration of 1 mg/ml, and the pH was adjusted to 2.5 with acetic acid. The samples were then applied to a Sep-Pak C18 Cartridge (Waters, Millford, MA), evaporated, and then dissolved in 100 µl of assay buffer for the peptide EIA (42 mM Na2HPO4, 8 mM KH2PO4, 20 mM NaCl, 4.8 mM EDTA, and 0.05% BSA; pH 7.5). As a result of these extractions, PG and peptides (ET-1 and Ang II) were 13- and 52-fold more concentrated, respectively, than in the original samples. Recovery rates were estimated by adding three different concentrations each of PGE2 (1, 0.5, and 0.1 ng/ml), PGF2{alpha} (1, 0.5, and 0.1 ng/ml), ET-1 (10, 5, and 1 pg/ml), and Ang II (1, 10, and 100 pg/ml) to the Ringer solution before extraction. The recovery rates were 78% for PGE2, 75% for PGF2{alpha}, 63% for ET-1, and 90% for Ang II.

Measurements of PG, ET-1, and Ang II

The EIAs for the PGE2, PGF2{alpha}, ET-1, and Ang II assays were performed according to the methods of Wijayagunawardane et al. [11], Miyamoto et al. [28], Miyamoto et al. [29], and Hayashi and Miyamoto [30], respectively. Within-assay and between-assay coefficients of variation were 7.3% and 11.4%, respectively, for PGE2; 8.2% and 11.8%, respectively, for PGF2{alpha}; 9.6% and 13.2%, respectively, for ET-1; and 2.67% and 7.99%, respectively, for Ang II. The median effective doses for the PGE2, PGF2{alpha}, ET-1, and Ang II assays were 260, 355, 450, and 150 pg/ml, respectively, and the ranges of the standard curves for these assays were 0.03–14.2, 0.007–7.1, 0.01–20, and 0.024–5 ng/ml, respectively.

RNA Extraction

Total RNA was extracted from flushed oviductal cells following the protocol of Chomczynski and Sacchi [31] using TRIzol reagent (Gibco BRL, Gaithersburg, MD). The yield of extracted total oviductal RNA for each sample was determined by ultraviolet (UV) spectroscopy (optical density, 260 nm). The quantity and integrity of the oviductal RNAs were verified by checking the 28S and 18S ribosomal RNA bands after electrophoresis on a formaldehyde-containing 1% (w/v) agarose gel.

Reverse Transcription-Polymerase Chain Reaction

Genomic DNA contamination was removed from RNA samples by DNA digestion as described previously [32]. Briefly, DNase I treatment was carried out in a 20-µl volume that contained 4 µg of total oviductal RNA, 4 U of DNase I (Ambion, Austin, TX), 50 mM Tris-HCl (pH 8.3), 75 mM KCl, and 3 mM MgCl2. This reaction mixture was incubated at 37°C for 30 min and then heated at 75°C for 5 min to inactivate the DNase. Next, the samples were placed immediately on ice. To each sample was added 140 U of Superscript II reverse transcriptase (Gibco BRL), 2.5 µM random hexamers (Amersham Biosciences, Piscataway, NJ), 0.60 mM dNTPs (Amersham), and 10 mM dithiothreitol for a final volume of 60 µl. Reverse transcription (RT) was performed at 45°C for 30 min, followed by 90°C for 2 min. Commercially synthesized primers (Invitrogen Life Technologies, Carlsbad, CA) (Table 1) were used to amplify specific bovine transcripts.


View this table:
[in this window]
[in a new window]
  		TABLE 1. Primers used for RT-PCR to amplify specific bovine transcripts of TNF{alpha}, TNF{alpha}-RI, and TNF{alpha}-RII

Each polymerase chain reaction (PCR) contained 5 µl of cDNA, 0.2 mM dNTPs, 0.4 µmol of each primer, 0.5 U of Taq DNA Polymerase (Qiagen, Valencia, CA), and a 1:10 dilution of the 10x Qiagen PCR buffer (supplied with the polymerase) in a final volume of 25 µl. The following individual amplification programs (94°C for 30 sec and 60°C for 1 min) were run in a thermocycler (Personal Cycler; Biometra, Göttingen, Germany) for each factor: 34 cycles for TNF{alpha}, 27 cycles for TNF{alpha}-R type I (TNF{alpha}-RI), and 34 cycles for TNF{alpha}-R type II (TNF{alpha}-RII), respectively. All PCR programs started with an initial denaturation step at 94°C for 2 min and ended with a final elongation phase at 72°C for 2 min.

For each sample, 18S rRNA (324 base pairs) was amplified by RT-PCR to check the integrity of the RNA as well as the efficiency of the RT. The relative amount of the ribosomal 18S RNA was similar among the samples for phases of the estrous cycle. Therefore, the 18S rRNA PCR was used as the internal control for the experiments, and 18S primers and the appropriate competitors (both from Ambion) were used in a ratio of 2:3. Twenty-four amplification cycles were performed (94°C, 60°C, and 72°C for 30 sec each). Five microliters from each reaction were run on a 1.5% (w/v) agarose gel containing 1 µg/ml of ethidium bromide. The resultant bands under UV light were documented with the Eagle Eye video system (Stratagene, La Jolla, CA) and analyzed with the Gel-Pro Analyzer software (Media Cybernetics, Silver Spring, MD). All RT-PCR reactions were performed twice for each RNA sample. To ensure that reactions did not reach a plateau in synthesis, preliminary experiments were done with an increasing number of cycles to determine the linear range of cycles for each primer set.

The PCR products were cut from the gel and eluted using a QIA quick gel extraction kit (Qiagen). The purified PCR products were sequenced using their respective forward primers (Penn State Life Sciences Facility, University Park, PA) to confirm specific amplification of PCR products. As negative controls, reactions containing no template (H2O) or non-reverse transcribed RNA were included to verify that the obtained PCR products were not derived from contaminations or genomic DNA. For all oviductal RNA samples, products were obtained for the corresponding cDNA, but not for genomic DNA under the above-described conditions.

Statistical Analysis

The absolute amounts of PGE2, PGF2{alpha}, ET-1, and Ang II released into each microdialysis capillary membrane varied among individual oviducts (i.e., among individual cows). Thus, the mean concentration of these substances in the first 4 h of incubation (perfusion with Ringer solution only) was used to calculate the baseline for each hormone (defined as 100%), and all values were expressed as a percentage of the corresponding baseline. This transformation enables an evaluation of relative changes in the hormonal levels between different oviducts. The effect of the TNF{alpha} infusion on the release of PGE2, PGF2{alpha}, ET-1, and Ang II was compared with control values during the same time period using ANOVA followed by the Duncan new multiple-range test. The data were subjected to the arcsine transformation before statistical analysis. Expressions of TNF{alpha}, TNF{alpha}-RI, and TNF{alpha}-RII during different stages of the estrous cycle were analyzed using ANOVA. When ANOVA showed significant differences, the Tukey-Kramer test was used to test the significant differences. A probability level of P < 0.05 was considered to be significant.


   RESULTS
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
DISCUSSION
 REFERENCES
 
During microdialysis, the basal release of PGE2, PGF2{alpha}, ET-1, and Ang II from oviducts of the follicular and postovulatory stages were higher (P < 0.05) than the values obtained from oviducts of the luteal stage (Table 2). This basal release of substances in the control group (Ringer solution only) was constant during the 16-h experimental period. Neither the basal release nor the extent of the stimulation by the infusion of different substances significantly differed between oviducts from the same animal or between the ampullar and isthmic parts of the same oviduct.


View this table:
[in this window]
[in a new window]
  		TABLE 2. Basal release (pg/ml) of PGE2, PGF2{alpha}, ET-1, and Ang II (mean ± SEM in Ringer solution) from microdialyzed oviducts collected during different stages of the estrous cycle (n = 4–5)

Effect of TNF{alpha} Infusion in Microdialyzed Cow Oviduct Collected During Different Stages of the Estrous Cycle

The 4-h infusion of TNF{alpha} in the postovulatory stage resulted in a 2-fold increase in release of PGE2 (P < 0.05 to 0.01), which continued up to 16 h. During the follicular phase, this was only observed during the infusion (P < 0.01). The TNF{alpha} infusion caused an acute and continued, 5- to 10-fold increase in PGF2{alpha} release (P < 0.05 to 0.01) both in follicular- and postovulatory-stage oviducts (Fig. 1). The TNF{alpha} induced an approximately 2-fold increase in the release of ET-1 (P < 0.05) in postovulatory-stage oviducts up to 12 h; however, the stimulation in the follicular-stage oviducts was only observed during infusion (P < 0.001). In postovulatory-stage oviducts, TNF{alpha} induced a 3-fold increase in secretion of Ang II (P < 0.001) between 8 and 12 h of incubation. In follicular-stage oviducts, TNF{alpha} stimulated Ang II secretion (P < 0.01), but only during infusion. During the luteal stage, TNF{alpha} did not stimulate any of the measured substances (Fig. 2).



View larger version (15K):
[in this window]
[in a new window]
  		FIG. 1. Effect of the infusion of TNF{alpha} (100 ng/ml) on the release of PGE2 and PGF2{alpha} in microdialyzed cow oviducts at different stages of the estrous cycle (n = 4–5 cows; mean ± SEM). *P < 0.05, **P < 0.01, and ***P < 0.001 versus the values at the same point in the control group



View larger version (15K):
[in this window]
[in a new window]
  		FIG. 2. Effect of the infusion of TNF{alpha} (100 ng/ml) on the release of ET-1 and Ang II in microdialyzed cow oviducts at different stages of the estrous cycle (n = 4–5 cows; mean ± SEM). *P < 0.05 versus the values at the same point in the control group

Expression of mRNA for TNF{alpha}, TNF{alpha}-RI, and TNF{alpha}-RII

Specific transcripts for TNF{alpha}, TNF{alpha}-RI, and TNF{alpha}-RII were detected in bovine oviductal cells. Each PCR product showed 100% homology to the known bovine genes after sequencing. The same expression pattern for TNF{alpha} and both TNF{alpha}-RI and TNF{alpha}-RII was found during the estrous cycle: higher expression during follicular and postovulatory stages, compared with lower expression during the luteal stage (Fig. 3). This expression pattern was significant (P < 0.05) for both receptor types, as shown by ANOVA. Expression of TNF{alpha} was significantly higher during the postovulatory stage than during the luteal stage (P < 0.05).



View larger version (23K):
[in this window]
[in a new window]
  		FIG. 3. The relative amount of mRNA (integrated absorbance; mean ± SEM) depicted for each PCR result. A) TNF{alpha}. B) TNF{alpha}-RI. C) TNF{alpha}-RII (n = 4 cows). Different letters above the bars denote a significance difference at P < 0.05


   DISCUSSION
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
REFERENCES
 
The present study provides, to our knowledge, the first direct evidence that TNF{alpha} stimulates PG, ET-1, and Ang II secretion in the cow oviduct. Moreover, TNF{alpha} and its receptors are highly expressed in the cow oviduct during the periovulatory period, thus accounting for up-regulation of the TNF{alpha} system compared with the luteal stage. With the microdialysis system, the luminal epithelium of the oviduct collected during different stages of the estrous cycle was gently flushed. This system allows cell-to-cell integrity and cell-to-cell communication to be maintained, and it enables observations of real-time changes in the luminal release of different substances [12, 13].

Studies of spontaneous oviductal contractions in the cow revealed low amplitude and frequency during the luteal phase [33] and a gradual increase in amplitude and frequency concomitant with a rapid decrease in P4 levels [34]. Both amplitude and frequency reach their maximal value during estrus, and values quickly diminish over the next 3 days [33, 34]. During the active periovulatory period, elevated local distributions of PG and ET-1 were observed in the cow oviduct [11], as was the stimulation of local oviductal PG and ET-1 [12, 13, 35, 36] secretion. In the present study, oviducts collected around the time of ovulation had TNF{alpha}-stimulated release of all the investigated substances.

Infusion of TNF{alpha} into microdialyzed cow oviducts collected during the follicular and postovulatory stages clearly stimulated release of PG in the present study. The TNF{alpha} is known to stimulate both PGE2 and PGF2{alpha} secretion by the bovine endometrium through activation of phospholipase (PL) A2 and arachidonic acid conversion [14, 15]. The TNF{alpha}-induced PG release by cultured human amnion cells [16], gingival fibroblasts [17], and brain microvessel endothelial cells [18] through activation of cyclooxygenase (COX)-2 has also been recorded. Moreover, TNF{alpha}-stimulated PGE2 biosynthesis by amnion-derived AV3 cells was accompanied by increased prostaglandin H synthase 2 (PGHS) mRNA expression [19]. Therefore, the increased PG secretion by the cow oviduct following infusion of TNF{alpha} that we observed may be mediated through the activation of PLA2, COX-2, or PGHS.

In the present study, TNF{alpha} stimulated ET-1 release in the oviduct. In human mesangial cells [20] and bovine pulmonary arterial smooth muscle cells [37], TNF{alpha}-stimulated synthesis and release of ET-1 was the result of up-regulation of prepro-ET-1 mRNA. Moreover, TNF{alpha} stimulated ET-converting enzyme-1 mRNA expression in human normal bronchial epithelial cells, with a significant increase in ET-1 expression at both the mRNA and peptide levels [21]. In addition, TNF{alpha} induced higher levels of ET-1 in human ciliary nonpigmented epithelial cells, and this was accompanied by protein kinase C activation [22]. Elevated ET-1 levels in the cow oviduct induced by TNF{alpha} may facilitate oviductal contraction during the periovulatory period. Our recent study showed that the addition of ET-1 to organ culture medium rapidly increases the contractile amplitude of periovulatory bovine oviducts [12]. On the other hand, TNF{alpha} has been reported to increase ET-B receptor mRNA expression in human endothelial cells [38] and ET-B receptor-mediated contraction in rat superior mesenteric artery [39]. Thus, TNF{alpha} may further intensify the ET-1 action on the oviduct through up-regulation of ET-B receptors.

The results of the present study clearly demonstrate that TNF{alpha} stimulates the release of Ang II into the microdialysis system. In cultured fibroblasts, TNF{alpha} has been reported to up-regulate type 1 Ang II-receptor mRNA and protein fibroblasts [23, 24]. In pregnant rat [40] and sheep [41], Ang II increased PG production, and Ang II stimulated release of ET-1 [42] and induction of prepro-ET-1 mRNA by the endothelial cells [43]. We recently observed that Ang II stimulates the secretion of PG and ET-1 in the microdialyzed cow oviduct [13]. Therefore, Ang II may be a local intermediator of TNF{alpha} that stimulates PG and ET-1 secretion during the periovulatory period, in addition to directly acting on oviductal smooth muscle.

We observed a clear, stage-specific mRNA expression for TNF{alpha} and for TNF{alpha}-R during the estrous cycle. Progesterone is a potent inhibitor of TNF{alpha} mRNA expression and TNF{alpha} protein production in the endometrium of ruminants [1] and in activated mouse macrophages [44]. Thus, the low expression of mRNA for TNF{alpha} and for TNF{alpha}-R observed in the present study during the luteal stage may result from the inhibitory action of P4. High levels of TNF{alpha} and TNF{alpha}-R expression in the cow oviduct during the follicular and postovulatory stages are consistent with a previous report that the level of TNF{alpha} was highest in the mouse uterus during proestrus and/or estrus [45], a time when the estradiol 17ß:P4 ratio is increasing.

In the reproductive tract, a variety of immune cells are present, and their number and distribution vary in a tissue-specific manner with the stage of the estrous cycle [2]. Thus, a potential source for the TNF{alpha} in the oviduct may be immune cells present in the oviduct. Cyclic changes in the number of immune cells have been reported that may account for the stage differences in TNF{alpha} mRNA expression observed in the present study. However, because the embryo produces and secretes TNF{alpha} [46, 47] during oviductal passage, the ability of the embryo to act as a source of TNF{alpha} in the oviduct cannot be excluded. Human embryos at the 6- to 8-cell stage secrete TNF{alpha} [46], and a dramatic enhancement in TNF{alpha} secretion occurs in bovine oocytes 48 h after fertilization [47]. The minute quantities of TNF{alpha} secreted by the embryo may act locally to enhance the production of PG, ET-1, and Ang II in the oviduct, which may result in an active oviductal contraction in the microenvironment around the embryo. This may ensure that the embryo migrates into the uterus at the optimal time.

In recent studies, we showed that LH in combination with a basal level of P4 and a high concentration of E2 has a maximal stimulatory effect on the oviductal secretion of PG and ET-1 during the periovulatory period [12, 35, 36]. The findings of the present study that TNF{alpha} stimulates maximal secretion of PGE2, PGF2{alpha}, ET-1, and Ang II in the cow oviduct during the periovulatory period suggest that TNF{alpha} may act synergistically with LH, E2, and P4. In conclusion, the TNF{alpha} system in the oviduct may act to optimize the release of contraction-related substances and, thereby, modulate local contraction to regulate oviductal transport of the gametes and embryo.


   ACKNOWLEDGMENTS
 
The authors thank Dr. D. Schams (Technical University of Munich, Germany) for ET-1 antiserum, Dr. S. Ito (Kansai University of Medicine, Moriguchi, Japan) for PG antiserum, Dr. K. Wakabayashi (Gunma University, Maebashi, Japan) for Ang II antiserum, Fresenius AG for the microdialysis capillary membranes, and DAINIPPON Pharmaceutical Co., Ltd., for human recombinant TNF{alpha}.


   FOOTNOTES
 
1 Supported by National Research Council of Sri Lanka (Grant 99-28) and the 21st Century COE Program (A-1), Ministry of Education, Culture, Science and Technology, Japan. M.P.B.W. is a postdoctoral fellow supported by Japan Society for the Promotion of Science (JSPS). Back

2 Correspondence. FAX: 81 155 49 5593; akiomiya{at}obihiro.ac.jp Back

3 Current address: Department of Agricultural and Life Science, Obihiro University of Agriculture and Veterinary Medicine, Obihiro 080-8555, Japan Back

Received: 19 March 2003.

First decision: 16 April 2003.

Accepted: 4 June 2003.


   REFERENCES
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 

  1. Hansen PJ. Interactions between the immune system and the ruminant conceptus. J Reprod Fertil Suppl 1995 49:69-82[Medline]
  2. Kaushic C, Frauendorf E, Rossoll RM, Richardson JM, Wira CR. Influence of the estrous cycle on the presence and distribution of immune cells in the rat reproductive tract. Am J Reprod Immunol 1998 39:209-216
  3. Miyamoto Y, Skarzynski DJ, Okuda K. Is tumor necrosis factor {alpha} a trigger for the initiation of endometrial prostaglandin F2{alpha} release at luteolysis in cattle?. Biol Reprod 2000 62:1109-1115[Abstract/Free Full Text]
  4. Srivastava MD, Lippes J, Srivastava BI. Cytokines of the human reproductive tract. Am J Reprod Immunol 1996 36:157-166
  5. Hunt JS, Chen HL, Hu XL, Pollard JW. Normal distribution of tumor necrosis factor-{alpha} messenger ribonucleic acid and protein in the uteri, placentas, and embryos of osteopetrotic (op/op) mice lacking colony-stimulating factor-1. Biol Reprod 1993 49:441-452[Abstract]
  6. Sharkey A, Dellow K, Blayney M, Macnamee M, Charnock-Jones S, Smith S. Stage-specific expression of cytokine and receptor messenger ribonucleic acids in human preimplantation embryos. Biol Reprod 1995 53:974-981[Abstract]
  7. Pampfer S, Moulaert B, Vanderheyden I, Wuu Y, De Hertogh R. Effect of tumour necrosis factor {alpha} on rat blastocyst growth and glucose metabolism. J Reprod Fertil 1994 101:199-206[Abstract/Free Full Text]
  8. Kaye P, Harvey M. The role of growth factors in preimplantation development. Prog Growth Factor Res 1995 6:1-24[CrossRef][Medline]
  9. Spilman C, Harper M. Effects of prostaglandins on oviductal motility and egg transport. Gynecol Invest 1975 6:186-205[Medline]
  10. Rosselli M, Imthurn B, Macas E, Keller P. Endothelin production by bovine oviduct epithelial cells. J Reprod Fertil 1994 101:27-30[Abstract/Free Full Text]
  11. Wijayagunawardane MPB, Miyamoto A, Cerbito WA, Acosta TJ, Takagi M, Sato K. Local distributions of oviductal estradiol, progesterone, prostaglandins, oxytocin and endothelin-1 in the cyclic cow. Theriogenology 1998 49:607-618[CrossRef][Medline]
  12. Wijayagunawardane MPB, Miyamoto A, Taquahashi Y, Gabler C, Acosta TJ, Nishimura M, Killian G, Sato K. In vitro regulation of local secretion and contraction of the bovine oviduct: stimulation by luteinizing hormone, endothelin-1 and prostaglandins, and inhibition by oxytocin. J Endocrinol 2001 168:117-130[Abstract]
  13. Wijayagunawardane MPB, Miyamoto A, Taquahashi Y, Acosta TJ, Nishimura M, Sato K. Angiotensin II and atrial natriuretic peptide in the cow oviductal contraction in vitro: direct effect and local secretion of prostaglandins, endothelin-1, and angiotensin II. Biol Reprod 2001 65:799-804[Abstract/Free Full Text]
  14. Skarzynski DJ, Miyamoto Y, Okuda K. Production of prostaglandin F2{alpha} by cultured bovine endometrial cells in response to tumor necrosis factor {alpha}: cell type specificity and intracellular mechanisms. Biol Reprod 2000 62:1116-1120[Abstract/Free Full Text]
  15. Murakami S, Miyamoto Y, Skarzynski DJ, Okuda K. Effects of tumor necrosis factor-{alpha} on secretion of prostaglandins E2 and F2{alpha} in bovine endometrium throughout the estrous cycle. Theriogenology 2001 55:1667-1678[CrossRef][Medline]
  16. Spaziani EP, Benoit RR, Tsibris JC, Gould SF, O'Brien WF. Tumor necrosis factor-{alpha} upregulates the prostaglandin E2 EP1 receptor subtype and the cyclooxygenase-2 isoform in cultured amnion WISH cells. J Interferon Cytokine Res 1998 18:1039-1044[Medline]
  17. Nakao S, Ogtata Y, Shimizu E, Yamazaki M, Furuyama S, Sugiya H. Tumor necrosis factor {alpha} (TNF-{alpha})-induced prostaglandin E2 release is mediated by the activation of cyclooxygenase-2 (COX-2) transcription via NF{kappa}B in human gingival fibroblasts. Mol Cell Biochem 2002 238:11-18[CrossRef][Medline]
  18. Mark KS, Trickler WJ, Miller DW. Tumor necrosis factor-{alpha} induces cyclooxygenase-2 expression and prostaglandin release in brain microvessel endothelial cells. J Pharmacol Exp Ther 2001 297:1051-1058[Abstract/Free Full Text]
  19. Hansen WR, Marvin KW, Potter S, Mitchell MD. Tumour necrosis factor-{alpha} regulation of prostaglandin H synthase-2 transcription is not through nuclear factor-{kappa}B in amnion-derived AV-3 cells. Placenta 2000 21:789-798[CrossRef][Medline]
  20. Foschi M, Sorokin A, Pratt P, McGinty A, La Villa G, Franchi F, Dunn MJ. Preproendothelin-1 expression in human mesangial cells: evidence for a p38 mitogen-activated protein kinase/protein kinase-C-dependent mechanism. J Am Soc Nephrol 2001 12:1137-1150[Abstract/Free Full Text]
  21. Saleh D, Furukawa K, Tsao MS, Maghazachi A, Corrin B, Yanagisawa M, Barnes PJ, Giaid A. Elevated expression of endothelin-1 and endothelin-converting enzyme-1 in idiopathic pulmonary fibrosis: possible involvement of proinflammatory cytokines. Am J Respir Cell Mol Biol 1997 16:187-193[Abstract]
  22. Prasanna G, Dibas A, Brown K, Yorio T. Activation of protein kinase C by tumor necrosis factor-{alpha} in human nonpigmented ciliary epithelium. J Ocul Pharmacol Ther 1998 14:401-412[Medline]
  23. Gurantz D, Cowling RT, Villarreal FJ, Greenberg BH. Tumor necrosis factor-{alpha} upregulates angiotensin II type 1 receptors on cardiac fibroblasts. Circ Res 1999 85:272-279[Abstract/Free Full Text]
  24. Cowling RT, Gurantz D, Peng J, Dillmann WH, Greenberg BH. Transcription factor NF-{kappa}B is necessary for up-regulation of type 1 angiotensin II receptor mRNA in rat cardiac fibroblasts treated with tumor necrosis factor-{alpha} or interleukin-1ß. J Biol Chem 2002 277:5719-5724[Abstract/Free Full Text]
  25. Wijayagunawardane MPB, Cerbito WA, Miyamoto A, Acosta TJ, Takagi M, Miyazawa K, Sato K. Oviductal progesterone concentration and its spatial distribution in cyclic and early pregnant cows. Theriogenology 1996 46:1149-1158
  26. Gabler C, Lauer B, Einspanier A, Schams D, Einspanier R. Detection of mRNA and immunoreactive proteins for acidic and basic fibroblast growth factor and expression of the fibroblast growth factor receptors in the bovine oviduct. J Reprod Fertil 1997 109:213-221[Abstract/Free Full Text]
  27. Acosta TJ, Miyamoto A, Ozawa T, Wijayagunawardane MPB, Sato K. Local release of steroid hormones, prostaglandin E2 and endothelin-1 from bovine mature follicles in vitro: effects of luteinizing hormone, endothelin-1 and cytokines. Biol Reprod 1998 59:437-443[Abstract/Free Full Text]
  28. Miyamoto A, Tashiro Y, Nakatsuka T, Meyer H, Taguchi K, Abe N, Fukui Y. Effect of tumor necrosis factor-{alpha} and interleukin-1 on local release of progesterone, prostaglandin F2{alpha} and oxytocin in microdialyzed ovine corpus luteum in vitro. Assist Reprod Technol Androl 1995 8:21-32
  29. Miyamoto A, Kobayashi S, Arata S, Ohtani M, Fukui Y, Schams D. Prostaglandin F2{alpha} promotes the inhibitory action of endothelin-1 on the bovine luteal function in vitro. J Endocrinol 1997 152:R7-R11[Abstract/Free Full Text]
  30. Hayashi K, Miyamoto A. Angiotensin II interacts with prostaglandin F2{alpha} and endothelin-1 as a local luteolytic factor in the bovine corpus luteum in vitro. Biol Reprod 1999 60:1104-1109[Abstract/Free Full Text]
  31. Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 1987 162:156-159[Medline]
  32. Huang Z, Fasco MJ, Kaminsky LS. Optimization of DNase I removal of contaminating DNA from RNA for use in quantitative RNA-PCR. Biotechniques 1996 20:1012-1020[Medline]
  33. Bennett W, Watts T, Blair W, Waldhalm S, Fuquay J. Patterns of oviductal motility in the cow during the oestrous cycle. J Reprod Fertil 1988 83:537-543[Abstract/Free Full Text]
  34. Ruckebusch Y, Bayard F. Motility of the oviduct and uterus of the cow during the oestrous cycle. J Reprod Fertil 1975 43:23-32[Abstract/Free Full Text]
  35. Wijayagunawardane MPB, Choi YH, Miyamoto A, Kamishita H, Fujimoto S, Takagi M, Sato K. Effect of ovarian steroids and oxytocin on the production of prostaglandin E2, prostaglandin F2{alpha} and endothelin-1 from cow oviductal epithelial cell monolayers in vitro. Anim Reprod Sci 1999 56:11-17[CrossRef][Medline]
  36. Wijayagunawardane MPB, Miyamoto A, Sato K. Prostaglandin E2, prostaglandin F2{alpha} and endothelin-1 production by cow oviductal epithelial cell monolayers: effect of progesterone, estradiol-17ß, oxytocin and luteinizing hormone. Theriogenology 1999 52:791-801[CrossRef][Medline]
  37. Barker S, Khan NQ, Wood EG, Corder R. Effect of an antisense oligodeoxynucleotide to endothelin-converting enzyme-1c (ECE-1c) on ECE-1c mRNA, ECE-1 protein and endothelin-1 synthesis in bovine pulmonary artery smooth muscle cells. Mol Pharmacol 2001 59:163-169[Abstract/Free Full Text]
  38. Smith PJ, Teichert-Kuliszewska K, Monge JC, Stewart DJ. Regulation of endothelin-B receptor mRNA expression in human endothelial cells by cytokines and growth factors. J Cardiovasc Pharmacol 1998 31:suppl 1S158-S160
  39. Uddman E, Moller S, Adner M, Edvinsson L. Cytokines induce increased endothelin ET(B) receptor-mediated contraction. Eur J Pharmacol 1999 376:223-232[CrossRef][Medline]
  40. Dubin N, Ghodgaonkar R. Angiotensin II and [Sar1, Ile5, Ala8]angiotensin II effect on contractile activity and prostaglandin production of in vitro pregnant rat uteri. Endocrinology 1980 107:1855-1860.[Abstract]
  41. Magness R, Rosenfeld C, Faucher D, Mitchell M. Uterine prostaglandin production in ovine pregnancy: effects of angiotensin II and indomethacin. Am J Physiol 1992 263:H188-H197
  42. Emori T, Hirata Y, Ohta K, Kanno K, Eguchi S, Imai T, Shichiri M, Marumo F. Cellular mechanism of endothelin-1 release by angiotensin and vasopressin. Hypertension 1991 18:165-170[Abstract/Free Full Text]
  43. Imai T, Hirata Y, Emori T, Yanagisawa M, Masaki T, Marumo F. Induction of endothelin-1 gene by angiotensin and vasopressin in endothelial cells. Hypertension 1992 19:753-757[Abstract/Free Full Text]
  44. Miller L, Hunt JS. Regulation of TNF-{alpha} production in activated mouse macrophages by progesterone. J Immunol 1998 160:5098-5104[Abstract/Free Full Text]
  45. De M, Sanford T, Wood G. Interleukin-1, interleukin-6, and tumor necrosis factor {alpha} are produced in the mouse uterus during the estrous cycle and are induced by estrogen and progesterone. Dev Biol 1992 151:297-305[CrossRef][Medline]
  46. Zolti M, Ben-Rafael Z, Meirom R, Shemesh M, Bider D, Mashiach S, Apte R. Cytokine involvement in oocytes and early embryos. Fertil Steril 1991 56:265-272[Medline]
  47. Gurevich M, Shemesh M. Induction of cyclooxygenase and prostaglandin E2 production by the bovine pre-embryo. Reprod Fertil Dev 1994 6:687-691[CrossRef][Medline]



This article has been cited by other articles:


Home page
 ReproductionHome page
S. P Kodithuwakku, A. Miyamoto, and M. P B Wijayagunawardane
Spermatozoa stimulate prostaglandin synthesis and secretion in bovine oviductal epithelial cells
Reproduction, June 1, 2007; 133(6): 1087 - 1094.
[Abstract] [Full Text] [PDF]

Home page
 J EndocrinolHome page
M Zerani, C Boiti, C Dall'Aglio, L Pascucci, M Maranesi, G Brecchia, C Mariottini, G Guelfi, D Zampini, and A Gobbetti
Leptin receptor expression and in vitro leptin actions on prostaglandin release and nitric oxide synthase activity in the rabbit oviduct
J. Endocrinol., May 1, 2005; 185(2): 319 - 325.
[Abstract] [Full Text] [PDF]

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
69/4/1341    most recent
biolreprod.103.017327v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Wijayagunawardane, M. P.B.
Right arrow Articles by Miyamoto, A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Wijayagunawardane, M. P.B.
Right arrow Articles by Miyamoto, A.
Agricola
Right arrow Articles by Wijayagunawardane, M. P.B.
Right arrow Articles by Miyamoto, A.

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS