help button home button Endocrine Society Endocrinology
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Purchase Article
Right arrow View Shopping Cart
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 Request Copyright Permission
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Lazarus, M.
Right arrow Articles by Urade, Y.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Lazarus, M.
Right arrow Articles by Urade, Y.
Endocrinology Vol. 143, No. 6 2410-2419
Copyright © 2002 by The Endocrine Society

REPRODUCTION-DEVELOPMENT

Immunohistochemical Localization of Microsomal PGE Synthase-1 and Cyclooxygenases in Male Mouse Reproductive Organs

Michael Lazarus, Craig J. Munday, Naomi Eguchi, Shigeko Matsumoto, Gary J. Killian, Bruno Kilunga Kubata and Yoshihiro Urade

Department of Molecular Behavioral Biology (M.L., C.J.M., N.E., S.M., B.K.K., Y.U.) and Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Corporation (Y.U.), Osaka Bioscience Institute, Osaka 565-0874, Japan; and Department of Dairy and Animal Science (G.J.K.), J. O. Almquist Research Center, Pennsylvania State University, University Park, Pennsylvania 16802

Address all correspondence and requests for reprints to: Dr. Yoshihiro Urade, Department of Molecular Behavioral Biology, Osaka Bioscience Institute, Furuedai 6-2-4, Suita, Osaka 565-0874, Japan. E-mail: . uradey{at}obi.or.jp


   Abstract
Top
 Abstract
Introduction
 Materials and Methods
 Results
 Discussion
 References
 
We investigated the tissue distribution and cellular localization of microsomal PGE synthase-1 (mPGES-1) and cyclooxygenase (COX)-1 and -2 in the male mouse reproductive organs. Northern blotting revealed that the mPGES-1 mRNA was expressed intensely in the epididymis and weakly in the lung, spleen, skin, kidney, colon, and brain. In the male reproductive tract, the expression of mPGES-1 increased from the testis to the cauda epididymis and was highest in the vas deferens when examined by Northern blotting, RT-PCR, and Western blotting. By immunohistochemistry, mPGES-1 was detected in Leydig cells of the testis and in epithelial cells of the epididymis, vas deferens, and seminal vesicles. In addition, the caput and cauda regions of the epididymis and the vas deferens in this order showed a progressive increase in the expression of COX-1 mRNA and immunoreactivity, whereas COX-2 was dominantly expressed in the vas deferens. COX-1 was localized in epithelial cells of the caput, corpus and cauda epididymis and of the vas deferens, and COX-2 was evident in epithelial cells of the distal cauda epididymis and vas deferens. These results show that mPGES-1 is expressed coordinately with COX-1 and COX-2 and is involved in PGE2 production in male genital organs.


   Introduction
Top
 Abstract
 Introduction
Materials and Methods
 Results
 Discussion
 References
 
PROSTAGLANDINS WERE originally discovered in 1935 by von Euler (1) as a vasodilatory substance in human seminal fluid. The E-series of PGs were isolated from the seminal fluid of humans and sheep, and their structures were determined in 1962 (2, 3). PGE2 exhibits a wide variety of biological activities in vertebrate and invertebrate reproduction (4, 5). During the past decades, several attempts have been made to elucidate the enzymes involved in PGE2 synthesis. However, little is known about the cellular localization of the PGE2-producing enzymes in mammalian genital organs.

In the presence or absence of the reduced form of glutathione, PGE synthase (PGES, EC. 5.3.99.3) catalyzes the isomerization of the 9,11-endoperoxide group of PGH2, a common precursor of various prostanoids, to PGE2 having 9-keto and 11-hydroxy groups (6). PGH2 is produced from arachidonic acid by cyclooxygenase (COX)-1 or COX-2 (7). A variety of enzymes have already been characterized as PGES (8, 9, 10, 11, 12, 13). The glutathione-dependent, membrane-bound PGES-1 (mPGES-1) was partially purified from the microsomal fraction of bovine and sheep vesicular glands (14, 15). Recently, cDNA for human mPGES-1 was identified (16) and its gene cloned (17). Coordinate up-regulation of mPGES-1 and COX-2 was previously reported to occur in a human lung cancer cell line, A549, after treatment with IL-1ß (16, 18), in rat osteoblasts and peritoneal macrophages after stimulation with IL-1ß, TNF-{alpha}, and lipopolysaccharide (LPS) (19), in mouse peritoneal macrophages after stimulation with LPS (20), and in human rheumatoid synovial cells after treatment with IL-1ß and TNF-{alpha} (21). Moreover, the induction of mPGES-1 was demonstrated to take place in rat astrocytes after treatment with ß-amyloid (22) in human dermal fibroblasts and vascular smooth muscle cells after stimulation with IL-1ß, TNF-{alpha}, phorbol 12-myristate 13-acetate, and LPS (23) and in granulosa cells of the bovine ovarian follicles after administration of gonadotropins (24).

In the present study, we determined the tissue distribution and cellular localization of mPGES-1 and COXs in male mouse genital organs to gain insight into the nature of PGE2-producing cells and the site of PGE2 action. We showed immunohistochemically that mPGES-1 and COX-1 were localized in the caput, corpus, and cauda epididymis as well as in the vas deferens, whereas COX-2 was dominantly expressed in the vas deferens. These results indicate that mPGES-1 is an efficient downstream enzyme for the production of PGE2 by coordinate expression with COX-1 and COX-2 in the mouse male reproductive tract.


   Materials and Methods
Top
 Abstract
 Introduction
 Materials and Methods
Results
 Discussion
 References
 
Animals and materials
Inbred C57BL/6 strain mice were purchased from Shizuoka Laboratory Animal Center (Shizuoka, Japan). Mice deficient in COX-2 (COX-2-/-) were kindly provided by Prof. Makoto Taketo, Kyoto University (25). All procedures used in handling animals were approved by the Animal Research Committee of the Osaka Bioscience Institute. All commercially available chemicals were of analytical grade. Oligonucleotides were purchased from Amersham Pharmacia Biotech (Buckinghamshire, UK).

Isolation of Leydig cells and seminiferous tubules from mouse testis
Leydig cells and seminiferous tubules were isolated from mouse testis by a method previously described (26). The enrichment of the Leydig cells was confirmed by determining the expression of mRNA for 3-ß-hydroxysteroid dehydrogenase (3ßHSD), an enzyme essential for the production of steroid hormones in the Leydig cells (27).

Northern blot analysis
Mouse tissues were homogenized in Isogen solution (1 ml/mg tissue; Nippon Gene, Toyama, Japan). Total RNA was extracted from the homogenates as described previously (28), separated on 1.2% (wt/vol) agarose-2% formaldehyde gels, and blotted onto Hybond-XL membranes (Amersham Pharmacia Biotech) for 15 h. The RNA was cross-linked to the membrane by UV irradiation at a wavelength of 254 nm in a CL-1000 UV cross-linker (Funakoshi, Tokyo, Japan). The membrane was prehybridized for 2 h at 42 C with a mixture of 5x sodium citrate-buffered saline (SSC), 0.2% (wt/vol) SDS, 5x Denhardt’s reagent, and 100 µg heat-denatured salmon sperm DNA. Under the same conditions, the membranes were hybridized with cDNAs for either the coding region of mouse mPGES-1 (518 bp), COX-1 (285 bp), COX-2 (300 bp), or glyceraldehyde 3-phosphate dehydrogenase (GAPDH; 452 bp), with appropriate probes labeled with [32P]-dCTP by use of a Rediprime labeling kit (Amersham Pharmacia Biotech). After hybridization, the membranes were rinsed with 2x SSC and 0.1% (wt/vol) SDS at 25 C and washed with 0.2x SSC and 0.1% (wt/vol) SDS at 50 C. The membranes were then exposed to BAS imaging plates (Fujiix, Tokyo, Japan).

RT-PCR analysis
First-strand cDNA was transcribed from 1 µg total RNA of mouse tissues or Leydig cells with random primers by avian myeloblastosis virus reverse transcriptase (Takara Shuzo, Kyoto, Japan). PCR was performed using an ExTaq PCR kit (Takara Shuzo) with primers specific for mouse GAPDH (5'-TGAACGGGAAGCTCACTGG-3' and 5'-TACAGCAACAGGGTGGTGGA-3'; expected PCR product size, 307 bp); mouse 3ßHSD (5'-CCCATACAGCAAAAAGATGGCTGAG-3' and 5'-GTGTCATCTGAGATGTAGTAGAACT-3'; expected PCR product size, 315 bp); mouse mPGES-1 (5'-CTGCTGGTCATCAAGATGTACG-3' and 5'-CCCAGGTAGGCCACGGTGTGT-3'; expected PCR product size, 293 bp); mouse COX-1 (5'-CTTTGCACAACACTTCACCCACC-3' and 5'-AGCAACCCAAACACCTCCTGG-3'; expected PCR product size, 285 bp); and COX-2 (5'-GCATTCTTTGCCCAGCACTT-3' and 5'-AGACCAGGCACCAGACCAAAGA-3'; expected PCR product size, 277 bp) in a GeneAmp PCR system (PE Biosystems, Foster City, CA). After denaturation at 95 C for 5 min, the reactions were cycled 30 times with denaturation at 95 C for 15 sec, annealing at 55 C for 15 sec, and elongation at 74 C for 30 sec.

For semiquantitative PCR, we amplified the cDNA by using a LightCycler and a LightCycler-DNA Master SYBR Green I kit (Roche Diagnostics, Mannheim, Germany). The reactions were cycled 40 times with denaturation at 95 C for 3 sec, annealing at 57 C for 5 sec, and elongation at 72 C for 10 sec. Temperature gradients for denaturation, annealing, and elongation were 20 C/sec, 2 C/sec, and 20 C/sec, respectively. Fluorescence was acquired after heating at 20 C/sec to a temperature of 2 C below the product melting temperature and holding this temperature for 1 sec. Quantification data were analyzed with LightCycler analysis software. All PCR products were visualized with UV light after electrophoresis in a 2% (wt/vol) agarose gel containing ethidium bromide to verify that only specific amplification had occurred and subsequently sequenced to confirm their origin from the intended mRNAs.

Western blot detection
Mouse tissues or Leydig cells were homogenized with an Ultra Turrax blender (Janke & Kunkel, Staufen, Germany) in PBS (1 ml/100 mg material) in the presence of protease inhibitor Complete (Roche Diagnostics). Cellular debris was removed by centrifugation at 7,000 x g and 4 C for 20 min. The microsomal fractions were then obtained by centrifugation at 100,000 x g and 4 C for 1 h. Microsomal proteins were resolved by 14% or 6% SDS-PAGE (Tefco, Tokyo, Japan) and transferred onto Immobilon polyvinyl difluoride (PVDF) membranes (Millipore Corp., Bedford, MA) at 200 mA for 1 h. After blockage of nonspecific binding sites with BlockAce (Dainippon Seiyaku, Osaka, Japan) for 1 h at 25 C, the membranes were incubated at 4 C overnight with guinea pig antimouse-mPGES-1 IgG (2 µg/ml; 20), rabbit antimouse-COX-1 or antimouse-COX-2 polyclonal antibody (1 µg/ml; Cayman, Ann Arbor, MI), or goat antihuman COX-2 polyclonal antibody (1 µg/ml; Santa Cruz Biotechnology, Inc., Santa Cruz, CA) as appropriate, applied in 10% BlockAce and PBS containing 0.2% (vol/vol) Tween-20. As secondary antibody, horseradish peroxidase-coupled donkey antiguinea pig, antigoat, or antirabbit IgG (Jackson ImmunoResearch Laboratories, West Grove, PA) was used at a concentration of 10 µg/ml for 1 h at room temperature. Detection was performed with an enhanced chemoluminescence kit (Amersham Pharmacia Biotech) according to the manufacturer’s instructions.

Immunoperoxidase staining
After deep anesthesia with ether, mice were perfused via the left ventricle of the heart with 20 mM sodium phosphate (pH 7.4) containing 4% paraformaldehyde and 4% sucrose and then with Bouin’s solution (29). Genital organs were removed and soaked overnight at 4 C in Bouin’s fixative. After embedment in paraffin, the tissues were sectioned (10 µm) and mounted on slides. The sections were then deparaffinized in xylene and rehydrated in ethanol with increasing concentrations of water. Endogenous peroxidase activity was quenched with 0.3% hydrogen peroxide in PBS for 30 min at 25 C. The tissues were then treated with 0.3% pepsin in 0.01 M hydrochloric acid for 5 and 30 min for COXs and mPGES-1 immunostaining, respectively, at room temperature. Thereafter, nonspecific binding sites were blocked for 1 h with 10% sheep serum (Life Technologies, Inc., Tokyo, Japan) in PBS that contained 0.1% Triton X-100 and 0.1% sodium azide. Primary and secondary antibodies were diluted in PBS containing 1% goat serum and 0.1% Triton X-100. Tissue sections were exposed to anti-COX antibodies (5 µg/ml) and antimouse-mPGES-1 antibody (4 µg/ml) for 1 and 2 d, respectively, at 4 C. After 1 h of incubation at 4 C with biotinylated antiguinea pig or antirabbit IgG (7.5 µg/ml; Vector Laboratories, Inc., Burlingame, CA), tissues were rinsed three times in PBS containing 0.1% Triton X-100. A Vectastain Elite avidin-biotin-peroxidase kit (Vector Laboratories, Inc.) with diaminobenzidine substrate was used according to the manufacturer’s protocol. Tissues were counterstained with hematoxylin. Finally, sections were dehydrated, rinsed in xylene, and mounted with Mount-Quick (Daido Sangyo, Tokyo, Japan).

Immunofluorescence staining
The tissue sections were prepared and incubated with antibodies against mPGES-1 or COXs as described above. For visualization of mPGES-1 or COXs immunoreactivity, tissues were treated with either anti-guinea pig or antirabbit IgG (Chemicon, Temecula, CA) conjugated with fluorescein isothiocyanate or rhodamine, respectively, for 1 h at 25 C. The immunoreactivity of cells was analyzed by confocal microscopy using a laser-scanning microscope (Carl Zeiss, Oberkochen, Germany).


   Results
Top
 Abstract
 Introduction
 Materials and Methods
 Results
Discussion
 References
 
Northern blotting of mPGES-1 in various mouse tissues
The expression level of mRNA for mPGES-1 in various mouse tissues was investigated by probing Northern blots with the cDNA for mouse mPGES-1 (Fig. 1Go). The mRNA for mPGES-1 was highly expressed in the epididymis, weakly in the lung, spleen, skin, kidney, colon, and brain but was barely detectable in many other tissues. Three different sizes of the mRNA for mPGES-1 were found, at positions of 1.8, 2.4, and 4.4 kb. The 1.8-kb mRNA was dominantly expressed in the epididymis, kidney, and brain, whereas the 1.8- and 4.4-kb mRNAs predominated in the spleen and lung and the 1.8- and 2.4-kb mRNAs were expressed in the colon. All three transcripts were expressed with similar intensity in the skin.



View larger version (38K):
[in this window]
[in a new window]
  		Figure 1. Northern blot of mPGES-1 mRNA in various mouse tissues. Total RNA (6 µg each) from various mouse tissues was used. Hybridization signals of random-primed cDNA probes specific for mPGES-1 and GAPDH were detected by using a digital imaging system.

 
Expression of mPGES-1, COX-1, and COX-2 in mouse male genital organs
The mRNA expression for mPGES-1, COX-1, and COX-2 in mouse genital organs (Fig. 2AGo) was investigated in detail by Northern blotting (Fig. 2BGo). The mPGES-1 mRNA was highly expressed in the cauda epididymis and vas deferens and weakly in the caput epididymis but was barely detectable in the testis. Two different sizes of the mRNA for mPGES-1 were found, at positions of 1.8 and 4.4 kb, with the former being dominantly expressed. The COX-1 mRNA was expressed strongly in the cauda epididymis and vas deferens and moderately in the caput epididymis but was not detectable in the testis and seminal vesicles, whereas the mRNA for COX-2 was observed to be dominantly expressed in the vas deferens.



View larger version (69K):
[in this window]
[in a new window]
  		Figure 2. Expression of mRNAs and immunoreactive proteins for mPGES-1, COX-1, and COX-2 in mouse male genital organs. A, Photo of the male mouse genital organs showing the locations of the five segments analyzed by Northern blotting, RT-PCR, and Western blotting. The five regions are the testis (1), caput (2) and cauda (3) epididymis, vas deferens (4), and seminal vesicles (5). Northern blotting (B), semiquantitative RT-PCR (C), and Western blotting (D) of mPGES-1, COX-1, and COX-2 in mouse testis (lane 1), caput (lane 2) and cauda (lane 3) epididymis, vas deferens (lane 4), and seminal vesicles (lane 5). B, Total RNA (20 µg each) from various mouse tissues was used for Northern blot analysis. C, The relative amounts of mRNAs for mPGES-1, COX-1, and COX-2 to GAPDH mRNA were determined by RT-PCR. The chart inset shows the low expression of mRNA for mPGES-1 in the testis and seminal vesicles on a different scale. The means ± SEM of three independent experiments are shown. D, Microsomes of mouse genital organs (15 µg protein) were electrophoresed and subsequently transferred onto a PVDF membrane. The membranes were immunostained with anti-mPGES-1 (a), antimouse COX-1 (b) and COX-2 (c), and antihuman COX (d) antibodies. Molecular weights of mPGES-1, COX-1, and COX-2 (x 10-3) are indicated on the left.

 
We also compared the mRNA expression for mPGES-1, COX-1, and COX-2 in male mouse genital organs by semiquantitative PCR (Fig. 2CGo). The mRNA expression level of mPGES-1 relative to that of GAPDH increased from the testis (0.008) to the caput and cauda epididymis (0.084 and 0.560, respectively) and to the vas deferens (0.970) in a step-wise manner but was low in the seminal vesicles (0.060). Thus, the mPGES-1 expression was the highest in the vas deferens among the various mouse tissues. Similar to that of mPGES-1, the mRNA expression level of COX-1 relative to that of GAPDH increased step-wisely from the caput and cauda epididymis (0.56 and 1.16, respectively) to the vas deferens (2.25). The COX-2 mRNA was observed to be low to moderate in the caput and cauda epididymis (0.06 and 0.84, respectively) but was highly expressed in the vas deferens (26.61). Thereby, the mRNA expression level of COX-2 relative to that of GAPDH in the vas deferens was 12-fold higher than that of COX-1 mRNA. The mRNAs for COX-1 and COX-2 were not detected in the testis or seminal vesicles.

We also investigated the expression of mPGES-1 mRNA in the female mouse reproductive organs, i.e. uterus, ovary, and oviduct. No mRNA for mPGES-1 was detectable in the uterus, ovary, or oviduct by Northern blotting analysis (data not shown). Semiquantitative RT-PCR revealed that the mPGES-1 mRNA expression level relative to that of GAPDH was very low in the uterus (0.003), ovary (0.006), and oviduct (0.005).

Previously, we generated a polyclonal antimouse mPGES-1 antibody highly specific for mPGES-1 (20). Thus, we used this antibody to investigate the tissue distribution of mPGES-1 in the mouse male genital organs by Western blotting (Fig. 2DGo, a). The mPGES-1-immunoreactive protein was detected strongly in the microsomal fraction of mouse vas deferens (lane 4) but weakly to moderately in those of the caput and cauda epididymis (lanes 2 and 3). No immunoreactivity was detectable in the membrane fraction of the testis or seminal vesicles (lanes 1 and 5). When the cytosolic fractions of those tissues were used, no immunoreactive protein was detected (data not shown).

By Western blotting analysis of microsomal fractions of mouse genital organs with the antimouse COX-1 antibody from Cayman (Fig. 2DGo, b), the immunoreactive signal for COX-1 was weak to moderate in the caput (lane 2) and cauda epididymis (lane 3), intense in the vas deferens (lane 4) but was not detected in the testis or seminal vesicles (lanes 1 and 5). We also applied antibodies against COX-2, i.e. antimouse COX-2 (Cayman) and antihuman COX-2 (Santa Cruz Biotechnology), for Western blot analysis of the microsomal fractions of mouse genital organs (Fig. 2DGo, c and d). An immunoreactive band for COX-2 (Mr = 72,000) was detectable exclusively in the microsomal fraction of the mouse vas deferens with the antimouse COX-2 antibody (Cayman; Fig. 2DGo, c, lane 4). The antihuman COX-2 antibody (Santa Cruz Biotechnology) cross-reacted with the mouse COX-2 but also with mouse COX-1 (Fig. 2DGo, d, lanes 2–4). Immunoreactivity for COX-2, which appeared as a double band of higher molecular weight, compared with that for COX-1, was observed only in the vas deferens, whereas a COX-1-immunoreactive protein was present in the microsomal fractions of the caput and cauda epididymis and vas deferens (lanes 2, 3, and 4, respectively).

Localization of mPGES-1 in Leydig cells of the mouse testis
We determined the cellular localization of mPGES-1 in the mouse testis by immunoperoxidase staining with the polyclonal antibody selective for mPGES-1 (Fig. 3Go). Positive immunostaining for mPGES-1 was observed in the Leydig cells of the interstitium of the testis but not in the seminiferous tubules (Fig. 3Go, A and B). In adjacent sections of the testis stained with the anti-mPGES-1 antibody preabsorbed with recombinant mPGES-1, no positive signals were detected (Fig. 3CGo). No positive signals were detected in the testis immunostained with the antimouse COX-1 or antimouse COX-2 antibody (data not shown).



View larger version (71K):
[in this window]
[in a new window]
  		Figure 3. Immunolocalization of mPGES-1 in mouse testis. Light micrographs of sections of the testis reacted with antimouse-mPGES-1 antibody (A and B, bars, 40 and 10 µm, respectively) or with the antibody preabsorbed with excess amounts of recombinant mouse mPGES-1 (C, bar, 40 µm). B, A higher-magnification micrograph shows positive staining for mPGES-1 in the Leydig cells.

 
We compared the expression of mPGES-1, COX-1, and COX-2 in enriched fractions of Leydig cells and seminiferous tubules of the mouse testis by RT-PCR and Western blot analyses (Fig. 4Go). The mRNAs for mPGES-1 and COX-1 were detected in Leydig cells (Fig. 4AGo, lane 1) but barely in the seminiferous tubules (lane 2). No mRNA for COX-2 was detected in the Leydig cells or seminiferous tubules (lanes 1 and 2). Western blotting with the antimouse mPGES-1 and COX-1 antibodies (Fig. 4BGo) showed immunoreactive proteins corresponding to mPGES-1 and COX-1 at positions Mr = 17,000 and 70,000, respectively, in the microsomal fraction of the Leydig cells (lane 1) but not in that of the seminiferous tubules (lane 2). Immunoreactivity for COX-2 was not detectable in the microsomal fraction of the Leydig cells or seminiferous tubules (data not shown).



View larger version (41K):
[in this window]
[in a new window]
  		Figure 4. Expression of mRNAs and immunoreactive proteins for mPGES-1 and COX-1 in Leydig cells and seminiferous tubules of mouse testis. A, RT-PCR for mPGES-1, COX-1, COX-2, 3ßHSD, and GAPDH was performed using cDNAs prepared from Leydig cells (lane 1) or seminiferous tubules (lane 2) of the mouse testis and from the mouse vas deferens (lane 3). B, Microsomes (45 µg protein) of Leydig cells (lane 1) and seminiferous tubules (lane 2) of the mouse testis were electrophoresed and subsequently transferred onto a PVDF membrane. The membranes were immunostained with anti-mPGES-1 and antimouse COX-1 antibodies. Molecular weights of mPGES-1 and COX-1 (x 10-3) are indicated on the left.

 
Immunohistochemical localization of mPGES-1, COX-1, and COX-2 in the epididymis
In the epididymis, immunoreactivity for mPGES-1 was localized in the tubular epithelial cells of the caput, corpus, and cauda portions. Positive staining for mPGES-1 increased from the caput via the corpus to the cauda region of the epididymis (Fig. 5AGo). The caput epididymis had an occasional epithelial cell stained positively for mPGES-1, whereas the corpus region showed a progressive increase in the number of mPGES-1-positive cells. The epithelium of the cauda epididymis was intensely and uniformly positive for mPGES-1. In adjacent sections of the cauda epididymis stained with the anti-mPGES-1 antibody preabsorbed with recombinant mPGES-1, no positive signals were detected (data not shown).



View larger version (133K):
[in this window]
[in a new window]
  		Figure 5. Immunolocalization of mPGES-1, COX-1, and COX-2 in mouse epididymis. A, A composite low-magnification micrograph illustrates the epididymis immunostained with mouse-mPGES-1 polyclonal antibody, including the initial segment (IS), caput (Cpt), corpus (Cps), and cauda (Cd) regions (bar, 1 mm). Higher-magnification micrographs surrounding the composite tissue represent that the staining intensity is increased from the caput of the epididymis via its corpus to the cauda epididymis (bars, 40 µm). B and C, Micrographs of the immunolocalization of COX-1 (B) and COX-2 (C) in the caput (left photos), corpus (middle photos), and cauda (right photos) epididymis. The arrows in the right photo of C indicate positive immunostaining for COX-2 in epithelial cells of the cauda epididymis (bars, 40 µm).

 
A weak to moderate positive signal for COX-1 was detected in the epithelium of the caput and corpus epididymis, whereas an intense and uniform positive signal was detected in the epithelium of the cauda epididymis (Fig. 5BGo). In contrast, only the distal cauda epididymis had cells in the epithelium occasionally stained for COX-2 (Fig. 5CGo, right photo). The weak immunostaining in the smooth muscle cells encapsulating the epithelium of the cauda epididymis was also evident in COX-2-/- mice (data not shown).

The different cell types of the epididymis, stained with anti-mPGES-1 and COX-1 antibodies, are shown in Fig. 6Go. No mPGES-1 immunoreactivity was observed in the columnar epithelial cells of the efferent ducts (Fig. 6AGo). In the epididymis head, the only cells that stained positively for mPGES-1 were the narrow (Fig. 6BGo) and apical (Fig. 6CGo) cells of the initial segment and the apical cells of the caput epididymis (Fig. 6DGo). In contrast, the immunoreactivity for mPGES-1 was detected in all principal cells, but not clear cells, of the epithelium of the cauda epididymis (Figs. 5AGo and 6EGo). Similar to mPGES-1, COX-1 was expressed in all principal cells, but not clear cells, of the cauda epididymis (Figs. 5BGo and 6FGo). No signal for COX-2 was detectable in the clear cells of the cauda epididymis (data not shown).



View larger version (108K):
[in this window]
[in a new window]
  		Figure 6. Immunolocalization of mPGES-1 and COX-1 in various cell types of the mouse epididymis. Light micrographs of sections of the epididymis reacted with antimouse-mPGES-1 (A-E) or anti-COX-1 antibody (F). A, The immunostaining for mPGES-1 was not detected in the epithelial cells of the efferent ducts (bar, 35 µm). B–F, High-magnification micrographs illustrate positive immunostaining for mPGES-1 in the narrow (B, bar, 10 µm) and apical cells (C, bar, 5 µm) of the initial segment, in the apical cells of the caput epididymis (D, bar, 8 µm), and in the principal cells of the cauda epididymis (E, bar, 6 µm) and for COX-1 in the principal cells of the cauda epididymis (F, bar, 6 µm). Arrows in E and F indicate the absence of immunostaining for mPGES-1 or COX-1 in the clear cells of the cauda epididymis.

 
Immunohistochemical localization of mPGES-1, COX-1, and COX-2 in the vas deferens
In the vas deferens, strong immunoreactivity for mPGES-1, COX-1, and COX-2 (Fig. 7Go, A–C) was evident in the epithelium throughout the vas deferens. No positive signals for mPGES-1, COX-1, and COX-2 were detected in smooth muscle tissues surrounding the epithelium of the vas deferens (Fig. 7Go, A–C).



View larger version (95K):
[in this window]
[in a new window]
  		Figure 7. Immunolocalization of mPGES-1, COX-1, and COX-2 in mouse vas deferens. A–C, Light micrographs of the vas deferens immunostained with anti-mPGES-1 (A), anti-COX-1 (B), and anti-COX-2 (C) antibodies (bars, 40 µm). D–I, Confocal micrographs (bars, 10 µm) of the vas deferens after staining for mPGES-1 (D and G), COX-1 (E), and COX-2 (H). The overlay of mPGES-1 with COX-1 (F) and COX-2 (I).

 
Confocal microscopic analysis revealed the colocalization of mPGES-1 and COX-1 or COX-2 (Fig. 7Go, D–I). In good agreement with the results of the light microscopic analysis, mPGES-1, COX-1, and COX-2 were colocalized in the cytoplasmic region but not in the nucleus of epithelial cells of the vas deferens.

Cellular localization of mPGES-1 in the seminal vesicles
When we immunostained the seminal vesicles of mice with the anti-mPGES-1 antibody, mPGES-1 was detected in the epithelium of the seminal vesicular gland (Fig. 8AGo). In adjacent sections of the seminal vesicles, stained with the anti-mPGES-1 antibody preabsorbed with recombinant mPGES-1, no positive signals were detected (Fig. 8BGo). We also used antimouse COX-1 and COX-2 antibodies to immunostain the seminal vesicles of mice. COX-1 was weakly detected in the epithelium of the seminal vesicles (Fig. 8CGo), but no positive staining for COX-2 was found in the seminal vesicles (Fig. 8DGo).



View larger version (140K):
[in this window]
[in a new window]
  		Figure 8. Immunolocalization of mPGES-1, COX-1, and COX-2 in mouse seminal vesicles. Light micrographs of the seminal vesicles reacted with antimouse-mPGES-1 antibody (A, bar, 40 µm), with antimouse-mPGES-1 antibody preabsorbed with excess amounts of recombinant mouse mPGES-1 (B, bar, 40 µm) or with antimouse-COX-1 (C, bar, 40 µm) or COX-2 (D, bar, 40 µm) antibody. Each inset in A and C shows a high-magnification micrograph of positive immunostaining (bar, 5 µm).

 

   Discussion
Top
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
References
 
In this study, we demonstrated that mPGES-1 is intensely expressed in the male genital organs of mice with mPGES-1 expression levels increasing from the testis to the cauda epididymis and was the highest in the vas deferens (Fig. 2Go). These results are in agreement with previous studies on PGE2 production in the genital organs of rodents, in which low, intermediate, and high concentrations of PGE2 were detected in the testis, different segments of the epididymis, and vas deferens, respectively (30, 31, 32).

RT-PCR, Western, and immunohistochemical analyses revealed that mouse mPGES-1 was present in the Leydig cells of the testis (Figs. 3Go and 4Go). It was previously demonstrated that PGE2 suppressed the gonadotropin-induced formation of T in Leydig cells of rats (33). Therefore, PGE2 produced by mPGES-1 may be involved in the regulation of T production in rodents. It has been also shown that PGs stimulate seminiferous tubule contractility through direct action on the peritubular myoid cells (34), suggesting that these cells may be a target for PGE2 secreted from the Leydig cells to control the contractility of the seminiferous tubule.

Because of its smooth muscle relaxant activity, PGE2 is thought to play an important role in the transport of sperm through the epididymis and its subsequent expulsion at coitus in the vas deferens and for the release of a seminal vesicle gland fluid (31, 35, 36, 37). We hypothesized that PGE2 produced by mPGES-1 in epithelial cells of the epididymis, vas deferens, and seminal vesicles is released toward the basal surface of the epithelium and acts on the surrounding smooth muscle layer (Figs. 5–8GoGoGoGo). In addition, it has been established that PGE2 is constitutively released into the semen of rodents from the vas deferens rather than the prostate or seminal vesicles (38), although the concentration of PGE2 in the semen of rodents is low, compared with other species, e.g. ram or human (39). This suggests that mPGES-1-derived PGE2 is also released apically from epithelial cells of the epididymis, vas deferens, and seminal vesicles into the lumen of these organs to regulate the sperm fertility, as previously described (40).

A remarkable similarity was found in the tissue distribution profiles of mPGES-1 and COX-1 in the Leydig cells of the mouse testis, different segments of the epididymis, vas deferens, and seminal vesicles (Figs. 2Go and 4–8GoGoGoGoGo). Conversely, COX-2 was dominantly expressed in the distal portion of the cauda epididymis and in the vas deferens. The mPGES-1 immunoreactivity was colocalized with that of COX-1 and COX-2 in tubular epithelial cells (Fig. 7Go). The functional coupling of mPGES-1 to COX, preferentially to COX-2, was demonstrated in human embryo kidney 293 cells cotransfected with plasmids for mPGES-1 and COX-1 or COX-2 (19). The histological colocalization of mPGES-1 and COX-2 was also recently reported in vascular endothelial cells of rat brain after stimulation with IL-1ß (41) or LPS (42) and in mouse peritoneal macrophages after LPS treatment (20). However, in male mouse genital organs, mPGES-1 is expressed coordinately with COX-1 and COX-2 as judged by the tissue distribution profiles.

Although COX-2 is generally referred to as the inducible isozyme (43), it was constitutively expressed in epithelial cells of the cauda epididymis and the vas deferens (Figs. 5Go and 7Go). The constitutive expression of COX-2 has also previously been observed in the rat (44) and human (45) vas deferens, neurons (46) and leptomeningeal cells (47) in the rat brain, and the macula densa of the rat kidney (48). Therefore, COX-2 in the vas deferens may act along with the COX-1 isozyme to form a larger amount of PGH2 than in other locations for the production of large quantities of PGE2 by mPGES-1. The concentration of PGF2{alpha} was also considerably higher in the vas deferens, compared with that in other male mouse genital organs (30). Therefore, COX-2 in the vas deferens may also contribute to the active production of PGF2{alpha}.

The physiological roles of COX-1 and COX-2 were investigated by using COX-1- or COX-2-deficient mice. The COX-1-deficient mice were fertile with limited parturition defects of the mutant female (49), whereas the COX-2-deficient mice showed multiple female reproductive failures (50). The coexpression of both COX-1 and COX-2 in the mouse vas deferens may explain the apparently normal fertility of male COX-1- or COX-2-deficient mice by providing a rescue pathway for prostaglandin production through the remaining COX isozyme.

In conclusion, the immunohistochemical localization of mPGES-1 and COXs in mouse reproductive organs implicates that mPGES-1 was colocalized with COX-1 and COX-2 in the male genital organs to produce PGE2 involved in the male reproductive system. We also found that epithelial cells of the mouse vas deferens represent an interesting model to investigate the diverse functional coupling between COXs and terminal prostanoid synthases, i.e. COX-1 or COX-2 and mPGES-1 or PGF synthase.


   Acknowledgments
 
We thank Dr. Osamu Hayaishi for encouragement of this study. We also acknowledge Drs. T. Okada and P. Kabututu for valuable discussions as well as Ms. S. Sakae, Ms. M. Yamaguchi, and Ms. T. Nishimoto for their secretarial assistance.


   Footnotes
 
This work was supported in part by grants-in-aid for scientific research from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (Grant 12558078 to Y.U. and Grant 13557016 to N.E.) and by grants from the Takeda Science Foundation (to M.L., C.J.M., and Y.U.), Science and Technology Agency (Grant 199041 to M.L. and Grant 298141 to B.K.K.), the U.S. Department of Agriculture (Grant 97-35203-9806 to G.J.K.), and Osaka City.

Abbreviations: COX, Cyclooxygenase; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; 3ßHSD, 3-ß-hydroxysteroid dehydrogenase; LPS, lipopolysaccharide; mPGES-1, microsomal PGES-1; PGES, prostaglandin E synthase; PVDF, polyvinyl difluoride; SSC, sodium citrate-buffered saline.

Received November 20, 2001.

Accepted for publication February 28, 2002.


   References
Top
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
 References
 

  1. von Euler US 1935 Über die spezifische blutdrucksenkende Substanz des menschlichen Prostata- und Samenblasensekretes. Klin Wochenschrift 14:1182–1183
  2. Bergström S, Rhyhage R, Samuelsson B, Sjövall J 1962 The structure of prostaglandin E1, F1 and F2. Acta Chem Scand 16:501–502
  3. Samuelsson B 1963 Isolation and identification of prostaglandins from human seminal fluid. J Biol Chem 238:3229–3234[Free Full Text]
  4. Tyler MJ, Shearman DJ, Franco R, O’Brien P, Seamark RF, Kelly R 1983 Inhibition of gastric acid secretion in the gastric brooding frog, Rheobatrachus silus. Science 220:609–610[Abstract/Free Full Text]
  5. Narumiya S, Sugimoto Y, Ushikubi F 1999 Prostanoid receptors: structures, properties, and functions. Physiol Rev 79:1193–1226[Abstract/Free Full Text]
  6. Miyamoto T, Yamamoto S, Hayaishi O 1974 Prostaglandin synthetase system-resolution into oxygenase and isomerase components. Proc Natl Acad Sci USA 71:3645–3648[Abstract/Free Full Text]
  7. Smith WL, Dewitt DL 1996 Prostaglandin endoperoxide H synthases-1 and -2. Adv Immunol 62:167–215[Medline]
  8. Ujihara M, Tsuchida S, Satoh K, Sato K, Urade Y 1988 Biochemical and immunological demonstration of prostaglandin D2, E2, and F2{alpha} formation from prostaglandin H2 by various rat glutathione S-transferase isozymes. Arch Biochem Biophys 264:428–437[CrossRef][Medline]
  9. Chang M, Hong Y, Burgess JR, Tu CP, Reddy CC 1987 Isozyme specificity of rat liver glutathione S-transferases in the formation of PGF2{alpha} and PGE2 from PGH2. Arch Biochem Biophys 259:548–557[CrossRef][Medline]
  10. Ogorochi T, Ujihara M, Narumiya S 1987 Purification and properties of prostaglandin H-E isomerase from the cytosol of human brain: identification as anionic forms of glutathione S-transferase. J Neurochem 48:900–909[CrossRef][Medline]
  11. Beuckmann CT, Fujimori K, Urade Y, Hayaishi O 2000 Identification of mu-class glutathione transferases M2–2 and M3–3 as cytosolic prostaglandin E synthases in the human brain. Neurochem Res 25:733–738[CrossRef][Medline]
  12. Tanioka T, Nakatani Y, Semmyo N, Murakami M, Kudo I 2000 Molecular identification of cytosolic prostaglandin E2 synthase that is functionally coupled with cyclooxygenase-1 in immediate prostaglandin E2 biosynthesis. J Biol Chem 275:32775–32782[Abstract/Free Full Text]
  13. Watanabe K, Kurihara K, Suzuki T 1999 Purification and characterization of membrane-bound prostaglandin E synthase from bovine heart. Biochim Biophys Acta 1439:406–414[Medline]
  14. Ogino N, Miyamoto T, Yamamoto S, Hayaishi O 1977 Prostaglandin endoperoxide E isomerase from bovine vesicular gland microsomes, a glutathione-requiring enzyme. J Biol Chem 252:890–895[Abstract/Free Full Text]
  15. Moonen P, Buytenhek M, Nugteren DH 1982 Purification of PGH-PGE isomerase from sheep vesicular glands. Methods Enzymol 86:84–91[Medline]
  16. Jakobsson PJ, Thoren S, Morgenstern R, Samuelsson B 1999 Identification of human prostaglandin E synthase: a microsomal, glutathione-dependent, inducible enzyme, constituting a potential novel drug target. Proc Natl Acad Sci USA 96:7220–7225[Abstract/Free Full Text]
  17. Forsberg L, Leeb L, Thoren S, Morgenstern R, Jakobsson P 2000 Human glutathione dependent prostaglandin E synthase: gene structure and regulation. FEBS Lett 471:78–82[CrossRef][Medline]
  18. Thoren S, Jakobsson PJ 2000 Coordinate up- and down-regulation of glutathione-dependent prostaglandin E synthase and cyclooxygenase-2 in A549 cells. Inhibition by NS-398 and leukotriene C4. Eur J Biochem 267:6428–6434[Medline]
  19. Murakami M, Naraba H, Tanioka T, Semmyo N, Nakatani Y, Kojima F, Ikeda T, Fueki M, Ueno A, Oh-ishi S, Kudo I 2000 Regulation of prostaglandin E2 biosynthesis by inducible membrane-associated prostaglandin E2 synthase that acts in concert with cyclooxygenase-2. J Biol Chem 275:32783–32792[Abstract/Free Full Text]
  20. Lazarus M, Kubata BK, Eguchi N, Fujitani Y, Urade Y, Hayaishi O 2002 Biochemical characterization of mouse microsomal prostaglandin E synthase-1 and its colocalization with cyclooxygenase-2 in peritoneal macrophages. Arch Biochem Biophys 397:336–341[CrossRef][Medline]
  21. Stichtenoth DO, Thoren S, Bian H, Peters-Golden M, Jakobsson P-J, Crofford LJ 2001 Microsomal prostaglandin E synthase is regulated by proinflammatory cytokines and glucocorticoids in primary rheumatoid synovial cells. J Immunol 167:469–474[Abstract/Free Full Text]
  22. Satoh K, Nagano Y, Shimomura C, Suzuki N, Saeki Y, Yokota H 2000 Expression of prostaglandin E synthase mRNA is induced in ß-amyloid treated rat astrocytes. Neurosci Lett 283:221–223[CrossRef][Medline]
  23. Soler M, Camacho M, Escudero JR, Iniguez MA, Vila L 2000 Human vascular smooth muscle cells but not endothelial cells express prostaglandin E synthase. Circ Res 87:504–507[Abstract/Free Full Text]
  24. Filion F, Bouchard N, Goff AK, Lussier JG, Sirois J 2001 Molecular cloning and induction of bovine prostaglandin E synthase by gonadotropins in ovarian follicles prior to ovulation in vivo. J Biol Chem 276:34323–34330[Abstract/Free Full Text]
  25. Dinchuk JE, Car BD, Focht RJ, Johnston JJ, Jaffee BD, Covington MB, Contel NR, Eng VM, Collins RJ, Czerniak PM, Gorry SA, Trzaskos JM 1995 Renal abnormalities and an altered inflammatory response in mice lacking cyclooxygenase II. Nature 378:406–409[CrossRef][Medline]
  26. Ojeifo JO, Byers SW, Papadopoulos V, Dym M 1990 Sertoli cell-secreted protein(s) stimulates DNA synthesis in purified rat Leydig cells in vitro. J Reprod Fertil 90:93–108[Abstract]
  27. Bain P, Yoo M, Clarke T, Hammond S, Payne A 1991 Multiple forms of mouse 3ß-hydroxysteroid dehydrogenase/delta5delta4 isomerase and differential expression in gonads, adrenal glands, liver, and kidneys of both sexes. Proc Natl Acad Sci USA 88:8870–8874[Abstract/Free Full Text]
  28. Chomczynski P, Sacchi N 1987 Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162:156–159[Medline]
  29. Gerena RL, Eguchi N, Urade Y, Killian GJ 2000 Stage and region-specific localization of lipocalin-type prostaglandin D synthase in the adult murine testis and epididymis. J Androl 21:848–854[Abstract]
  30. Badr FM, Barcikowski B, Bartke A 1975 Effect of castration, testosterone treatment and hereditary sterility on prostaglandin concentration in the male reproductive system of mice. Prostaglandins 9:289–297[CrossRef][Medline]
  31. Gerozissis K, Dray F 1977 Selective and age-dependent changes of prostaglandin E2 in the epididymis and vas deferens of the rat. J Reprod Fertil 50:113–115[CrossRef][Medline]
  32. Gerozissis K, Dray F 1983 In vitro prostanoid production by the rat vas deferens. J Reprod Fertil 67:389–394[Abstract]
  33. Calkins JH, Sigel MM, Nankin HR, Lin T 1988 Interleukin-1 inhibits Leydig cell steroidogenesis in primary culture. Endocrinology 123:1605–1610[Abstract]
  34. Tripiciano A, Filippini A, Ballarini F, Palombi F 1998 Contractile response of peritubular myoid cells to prostaglandin F2{alpha}. Mol Cell Endocrinol 138:143–150[CrossRef][Medline]
  35. Cosentino MJ, Takihara H, Burhop JW, Cockett AT 1984 Regulation of rat caput epididymidis contractility by prostaglandins. J Androl 5:216–222[Abstract/Free Full Text]
  36. Free MJ, Jaffe RA, Morford DE 1980 Sperm transport through the rete testis in anesthetized rats: role of the testicular capsule and effect of gonadotropins and prostaglandins. Biol Reprod 22:1073–1078[Abstract]
  37. Hedqvist P, von Euler US 1972 Prostaglandin-induced neurotransmission failure in the field-stimulated, isolated vas deferens. Neuropharmacology 11:177–187[CrossRef][Medline]
  38. Klein LA, Stoff JS 1987 Prostaglandin synthesis is independent of androgen levels in rat male genitalia. J Lab Clin Med 109:402–408[Medline]
  39. Ventura WP, Freund M 1973 Evidence for a new class of uterine stimulants in rat semen and male accessory gland secretions. J Reprod Fertil 33:507–511[Medline]
  40. Joyce CL, Nuzzo NA, Wilson Jr L, Zaneveld LJ 1987 Evidence for a role of cyclooxygenase (prostaglandin synthetase) and prostaglandins in the sperm acrosome reaction and fertilization. J Androl 8:74–82[Abstract/Free Full Text]
  41. Ek M, Engblom D, Saha S, Blomqvist A, Jakobsson PJ, Ericsson-Dahlstrand A 2001 Inflammatory response: pathway across the blood-brain barrier. Nature 410:430–431[Medline]
  42. Yamagata K, Matsumura K, Inoue W, Shiraki T, Suzuki K, Yasuda S, Sugiura H, Cao C, Watanabe Y, Kobayashi S 2001 Coexpression of microsomal-type prostaglandin E synthase with cyclooxygenase-2 in brain endothelial cells of rats during endotoxin-induced fever. J Neurosci 21:2669–2677[Abstract/Free Full Text]
  43. O’Banion MK 1999 Cyclooxygenase-2: molecular biology, pharmacology, and neurobiology. Crit Rev Neurobiol 13:45–82[Medline]
  44. McKanna JA, Zhang MZ, Wang JL, Cheng H, Harris RC 1998 Constitutive expression of cyclooxygenase-2 in rat vas deferens. Am J Physiol 275:R227–R233
  45. Kirschenbaum A, Liotta DR, Yao S, Liu XH, Klausner AP, Unger P, Shapiro E, Leav I, Levine AC 2000 Immunohistochemical localization of cyclooxygenase-1 and cyclooxygenase-2 in the human fetal and adult male reproductive tracts. J Clin Endocrinol Metab 85:3436–3441[Abstract/Free Full Text]
  46. Breder CD, Dewitt D, Kraig RP 1995 Characterization of inducible cyclooxygenase in rat brain. J Comp Neurol 355:296–315[CrossRef][Medline]
  47. Beuckmann CT, Lazarus M, Gerashchenko DR, Mizoguchi A, Nomura S, Mohri I, Uesugi A, Kaneko T, Mizuno N, Hayaishi O, Urade Y 2000 Cellular localization of lipocalin-type prostaglandin D synthase (ß-trace) in the central nervous system of the adult rat. J Comp Neurol 428:62–78[CrossRef][Medline]
  48. Harris RC, McKanna JA, Akai Y, Jacobson HR, Dubois RN, Breyer MD 1994 Cyclooxygenase-2 is associated with the macula densa of rat kidney and increases with salt restriction. J Clin Invest 94:2504–2510
  49. Langenbach R, Morham SG, Tiano HF, Loftin CD, Ghanayem BI, Chulada PC, Mahler JF, Lee CA, Goulding EH, Kluckman KD, Kim HS, Smithies O 1995 Prostaglandin synthase 1 gene disruption in mice reduces arachidonic acid-induced inflammation and indomethacin-induced gastric ulceration. Cell 83:483–492[CrossRef][Medline]
  50. Lim H, Paria BC, Das SK, Dinchuk JE, Langenbach R, Trzaskos JM, Dey SK 1997 Multiple female reproductive failures in cyclooxygenase 2-deficient mice. Cell 91:197–208[CrossRef][Medline]



This article has been cited by other articles:


Home page
 ReproductionHome page
M. Mewe, I. Wulfsen, R. Middendorff, and C. K Bauer
Differential modulation of bovine epididymal activity by oxytocin and noradrenaline
Reproduction, September 1, 2007; 134(3): 493 - 501.
[Abstract] [Full Text] [PDF]

Home page
 Pharmacol. Rev.Home page
B. Samuelsson, R. Morgenstern, and P.-J. Jakobsson
Membrane Prostaglandin E Synthase-1: A Novel Therapeutic Target
Pharmacol. Rev., September 1, 2007; 59(3): 207 - 224.
[Abstract] [Full Text] [PDF]

Home page
 Biol. Reprod.Home page
J. L. Tomsig, S. Usanovic, and T. T. Turner
Growth Factor-Stimulated Mitogen-Activated Kinase (MAPK) Phosphorylation in the Rat Epididymis Is Limited by Segmental Boundaries
Biol Reprod, October 1, 2006; 75(4): 598 - 604.
[Abstract] [Full Text] [PDF]

Home page
 EndocrinologyHome page
T. Ishikawa and P. L. Morris
A Multistep Kinase-Based Sertoli Cell Autocrine-Amplifying Loop Regulates Prostaglandins, Their Receptors, and Cytokines
Endocrinology, April 1, 2006; 147(4): 1706 - 1716.
[Abstract] [Full Text] [PDF]

Home page
 EndocrinologyHome page
M. Mewe, C. K. Bauer, D. Muller, and R. Middendorff
Regulation of Spontaneous Contractile Activity in the Bovine Epididymal Duct by Cyclic Guanosine 5'-Monophosphate-Dependent Pathways
Endocrinology, April 1, 2006; 147(4): 2051 - 2062.
[Abstract] [Full Text] [PDF]

Home page
 GutHome page
D Wang and R N DuBois
PROSTAGLANDINS AND CANCER
Gut, January 1, 2006; 55(1): 115 - 122.
[Full Text] [PDF]

Home page
 Hum ReprodHome page
D. M. Duffy, C. L. Seachord, and B. L. Dozier
Microsomal prostaglandin E synthase-1 (mPGES-1) is the primary form of PGES expressed by the primate periovulatory follicle
Hum. Reprod., June 1, 2005; 20(6): 1485 - 1492.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Gastrointest. Liver Physiol.Home page
Y. Shinji, T. Tsukui, A. Tatsuguchi, K. Shinoki, M. Kusunoki, K. Suzuki, T. Hiratsuka, K. Wada, S. Futagami, K. Miyake, et al.
Induced microsomal PGE synthase-1 is involved in cyclooxygenase-2-dependent PGE2 production in gastric fibroblasts
Am J Physiol Gastrointest Liver Physiol, February 1, 2005; 288(2): G308 - G315.
[Abstract] [Full Text] [PDF]

Home page
 FASEB J.Home page
T. D. WARNER and J. A. MITCHELL
Cyclooxygenases: new forms, new inhibitors, and lessons from the clinic
FASEB J, May 1, 2004; 18(7): 790 - 804.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
D. Kamei, M. Murakami, Y. Nakatani, Y. Ishikawa, T. Ishii, and I. Kudo
Potential Role of Microsomal Prostaglandin E Synthase-1 in Tumorigenesis
J. Biol. Chem., May 23, 2003; 278(21): 19396 - 19405.
[Abstract] [Full Text] [PDF]

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Purchase Article
Right arrow View Shopping Cart
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 Request Copyright Permission
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Lazarus, M.
Right arrow Articles by Urade, Y.
Right arrow Search for Related Content