Molecular cloning and tissue distribution of microsomal-1 and cytosolic prostaglandin E synthases in macaque

Prostaglandins & other Lipid Mediators 78 (2005) 27–37

Molecular cloning and tissue distribution of microsomal-1 and cytosolic prostaglandin E synthases in macaque Julie Parent a , Pierre Chapdelaine a , Michel A. Fortier a,b,∗ a Unit´ e de Recherche en Ontog´enie et Reproduction, Centre Hospitalier Universitaire de Qu´ebec (CHUL), Centre de Recherche en Biologie de la Reproduction (CRBR), Universit´e Laval, Ste-Foy, Que., Canada G1V 4G2 b D´ epartement d’Obst´etrique et Gyn´ecologie, Universit´e Laval, Ste-Foy, Que., Canada G1V 4G2

Received 6 October 2004; received in revised form 11 January 2005; accepted 14 February 2005 Available online 9 June 2005

Abstract Prostaglandins derived from arachidonic acid are involved in a wide variety of physiological and pathological processes. The primary enzymes involved in the production of PGE2 from arachidonic acid are cyclooxygenases and prostaglandin E synthases. These enzymes have been identified in human, but only partially in the monkey where microsomal PGES-1 and cytosolic PGES have not been characterized. The present study was undertaken to clone these enzymes and to study their tissue distribution, along with mPGES-2. The coding sequence of Macaque mPGES-1 is 98% homologous to human mPGES-1 at the nucleic acid level and the deduced amino acid sequence has 98% homology with the human protein. The Macaque cPGES cDNA is more than 99% homologous to the human and the deduced amino acids sequence is identical to that of the human cPGES. By Northern blot analysis, we found that mPGES-2 and cPGES mRNA were expressed in the endometrium, myometrium, ovary and oviduct, albeit at different levels, while mPGES-1 mRNA was detected at a weak level, mainly in the oviduct. Western Blot analysis revealed that mPGES-2, mPGES-1 and cPGES proteins were present in all tissues tested. These results suggest that production of PGE2 in Macaque may involve more than one PGES and that further studies will be needed to fully understand the conditions under which each PGES contributes to PGE2 production. © 2005 Elsevier Inc. All rights reserved. Keywords: Macaque; Tissue distribution; Cyclooxygenases; Prostaglandin E synthases ∗

Corresponding author. Tel.: +1 418 656 4141x46141; fax: + 1 418 654 2765. E-mail address: [email protected] (M.A. Fortier).

1098-8823/$ – see front matter © 2005 Elsevier Inc. All rights reserved. doi:10.1016/j.prostaglandins.2005.02.005

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1. Introduction Prostaglandins (PG) are local mediators acting primarily through paracrine or autocrine mechanisms. Variations in the production of PG are involved in pathologies such as cancer, hypertension and arthritis [1]. Prostaglandin E2 (PGE2 ) is widely distributed and exhibits various biological activities such as smooth muscle dilatation or contraction, generation of fever, inhibition of immune response and promotion of angiogenesis [2–5]. PGE2 is an important regulator of reproductive processes such as ovulation, menstruation and dilation of the cervix in preparation for labor [1,6,7]. It also promotes uterine contractility at the time of labor in the myometrium [6]. Therefore, a tight regulation of PGE2 production is necessary for regulation of many physiological processes. Prostaglandins are derived from arachidonic acid (AA), converted into prostaglandin endoperoxide H2 (PGH2 ) by prostaglandin synthases (PGHS), also known as cyclooxygenases (COX). Two isoforms of the COX enzyme, type 1 and 2, are coded by different genes and catalyze the double oxygenation and reduction of AA. The next step for the production of PGE2 involves conversion into PGE2 via PGE synthases (PGES). Three different PGE synthases (PGES) have been characterized. Microsomal PGES-1 (mPGES-1) was the first to be identified and reported as an enzyme inducible by agents such as cytokines and LPS [7]. It is often coupled to COX-2 for delayed and sustained production of PGE2 [8]. A cytosolic PGES (cPGES), identical to p23, a ubiquitous chaperone protein weakly bound to the steroid hormone receptor/hsp90 complex [9,10], was characterized and found coupled to COX-1 for immediate production of PGE2 [11]. Enzymatic activity from a third PGES, microsomal PGES-2 (mPGES-2), was recently purified from the bovine heart and the corresponding human and Macaque enzymes were cloned [12]. This PGES was found associated with both isoforms of COX with a slight preference for COX-2 [13], but very little is known about its physiological functions. Morita et al. [8] have demonstrated in murine 3T3 cells that COX-1 is expressed equally in the endoplasmic reticulum and in the nuclear envelope, whereas COX-2 immunoreactivity is twice as high in the nuclear envelope compared to the endoplasmic reticulum. It was then postulated that different compartmentalization of COX-1 and COX-2, combined with their different sensitivity to arachidonic acid [9], may lead to the production of different PG. However, we now have to consider differential coupling between COX and PGES isoforms to explain PGE2 production. In the Macaque, sequences of mPGES-1 and cPGES are currently unknown, and no study has reported the simultaneous presence of the three PGE synthases in any species. Therefore, the objectives of the present study were (1) to clone and characterize Macaque mPGES-1 and cPGES and (2) to report tissue distribution mainly in female reproductive organs of the three Macaque PGE synthases. 2. Materials and methods 2.1. Materials Topo pEF6/V5 6xHis TA Cloning Kit, TRIzol and reagents used for the reversetranscription (RT) (dNTPs, Superscript II, buffer and random hexamers) were obtained

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from Invitrogen Corporation (Carlsbad, CA). Taq polymerase and buffer used for the polymerase chain-reaction (PCR), DNA labelling kit (-dCTP), Ficoll-Paque Plus and Hyperfilm ECL were purchased from Amersham Pharmacia Biotech (Baie d’Urf´e, Que., Canada). All restriction enzymes were purchased from New England Biolabs (Pickering, Ont., Canada). QIAquick gel extraction kit was purchased from Qiagen (Mississauga, Ont., Canada). BrightStar Plus membrane and ULTRAhyb solution were purchased from Ambion Inc. (Austin, TX). [␣-32 P] dCTP radioactivity and Western Lightning Chemiluminescence Reagent Plus were purchased from Perkin-Elmer Life Sciences (Boston, MA). Lipopolysaccharides (LPS) were purchased from Sigma (St. Louis, MO). Prestained protein markers were purchased from Mandel Scientific, New England Nuclear Life Science Products (Mississauga, Ont., Canada). Trans-Blot Transfer Medium (nitrocellulose membranes) was purchased from Bio-Rad Laboratories (Hercules, CA). Rabbit antibodies against sheep COX-1 and COX-2 were kindly provided by Dr. Stacia Kargman (Merck Frosst, Canada). Rabbit antibodies against human mPGES-1 and mPGES-2 were purchased from Cayman Chemical (Ann Arbor, MI). Mouse antibody against human cPGES was purchased from Affinity BioReagents Inc. (Golden, CO). The goat anti-rabbit and the goat anti-mouse antibodies conjugated to horseradish peroxidase were obtained from Jackson Immunoresearch Laboratories (West Grove, PA). BioMax films are from Eastman Kodak Corporation (New York, NY). 2.2. Isolation of RNA Different tissues were collected from one female Macaque (Macaca fascicularis) in the late proliferative phase of the cycle, frozen in liquid nitrogen and kept at −80 ◦ C until further processing. For RNA isolation, frozen samples were placed in liquid nitrogen and further pulverized in a mortar. The resulting powder was weighed and TRIzol was added at a 1 ml per 100 mg ratio. The suspension was homogenized three times on ice and RNA was extracted and processed according to the manufacturer’s instructions. RNA samples were resuspended in water containing DEPC (0.05%, v/v) and stored at −80 ◦ C. Before use, RNA was quantified by measurement of absorbance at 260 nm. 2.3. Platelet and neutrophilpurification Peripheral blood from 11 female Macaque at unknown stages of the cycle was collected on EDTA anticoagulant, pooled and platelets and neutrophils purified [14,15] to generate positive controls of cross reactivity of Macaque proteins with anti human COX-1 and COX2 antibodies. Neutrophils were incubated 1 h with LPS (100 ng/ml) at 37 ◦ C with gentle shaking to stimulate COX-2 expression. Cells were resuspended in PBS and then in SDSPAGE loading buffer (0.06 M Tris–HCl pH 6.8, 1% SDS, 1% 2-mercaptoethanol, 10% glycerol and 0.025% bromophenol blue). 2.4. Cloning and sequencing of Macaque mPGES-1 and cPGES RNA from Macaque endometrium was used as a template and was reverse transcribed using random primers and Superscript II reverse transcriptase. Full Macaque mPGES-

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1 (459 bp) and cPGES (483 bp) coding sequences were generated by RT-PCR using the following primers: mPGES-1: sense 5 -GAGATGCCTGCCC-3 and antisense 5 -AAACATATGTCACAGGTGGCGGGCCGCTTC-3 ; cPGES: sense 5 -ATGCAGCCTGCTTCTGCA-3 and antisense 5 -TTACTCCAGATCTGGCAT-3 . The primers were identical to the known mPGES-1 and cPGES sequences of human. The PCR conditions were 94 ◦ C for 30 s, 55 ◦ C for 30 s and 72 ◦ C for 30 s for 35 cycles. The RT-PCR products were cloned into TOPO cloning pEF6/V5 plasmid and sequenced. 2.5. DNA sequencing and sequence analyses The plasmid DNA was isolated using the QIAGEN plasmid purification system. After using the appropriate restrictions enzymes, the clones were sequenced by the sequencing service (CHUL, Quebec, Canada). Multiple sequence alignments were obtained using the BioEdit Sequence Alignment Editor program (North Carolina State University, Raleigh, NC). 2.6. Northern blot analysis Equal amounts (15 ␮g) of total RNA were loaded on a 1.2% (w/v) formaldehyde agarose gel, electrophoresed, and transferred onto a nylon membrane. cDNA probes for Macaque COX-1, COX-2, mPGES-1, mPGES-2 and cPGES were generated by PCR amplification with specific primers (COX-1: sense 5 -TCCAACCTTATCCCCAGTC-3 and antisense 5 -CATGGCAATGCGGTTGC-3 ; COX-2: sense 5 -TCCAGATCACATTTGATTGACA-3 and antisense 5 -TCTTTGACTGTGGGAGGATACA-3 ; mPGES-1: sense 5 -GAGATGCCTGCCC-3 and antisense 5 -AAACATATGTCACAGGTGGCGGGCGGCTTC-3 ; mPGES-2: sense 5 -GCAGGGCTGAGATCAAGTTC-3 and antisense 5 -GCCTTCATGGCTGGGTAGTA-3 ; cPGES: sense 5 -ATGCAGCCTGCTTCTGCA-3 and antisense 5 -TTACTCCAGATCTGGCAT-3 ). COX-1 and COX-2 primers were designed according to Kim et al. [16]. cDNA fragments of Macaque COX-1, COX-2, mPGES-1, mPGES-2 and cPGES were obtained by BamH1/EcoRV digestion of recombinant clones, thus liberating fragments ready to be labeled of 777, 449, 459, 175, and 483 bp, respectively. These recombinant clones were obtained by cloning using the pEF6/V5 plasmid of Topo Cloning kit (Invitrogen). Probes were labeled with DNA labelling kit (-dCTP) and [a32 P]-dCTP and purified by precipitation [17]. Prehybridization was performed at 45 ◦ C in UltraHyb solution for 4–5 h, then the labeled probe was added and hybridization was performed overnight at 45 ◦ C. Washings were done at room temperature 2× 10 min, and at 60 ◦ C 2× 15 min in SSC 0.2× supplemented with 0.1% SDS. Signals were detected by autoradiography on Kodak X-omat at −80 ◦ C, after exposure for 1–7 days before development. 2.7. Protein extraction Protein extraction and measurement were performed as described by Chapdelaine et al. [18]. Protein samples were suspended in 30 ␮l SDS-PAGE loading buffer and boiled for 5 min. Protein content was estimated using 1 ␮l of the sample.

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2.8. Western blot analysis Aliquots (10 ␮g) of total protein were loaded in each lane and electrophoresed through 13.5% SDS-polyacrylamide gels followed by electro transfer onto a nitrocellulose membrane. Prestained protein standards were used as molecular weight standard for each analysis. After staining with Ponceau Red to ensure that the same amount of protein was transferred onto the membrane, blocking was done in 5% fat free dry milk powder in PBS and 0.05% Tween-20 (PBS-T) overnight at 4 ◦ C. The membrane was then incubated with the antibody raised against COX-2 or COX-1 (dilution 1/3000), or mPGES-1 (dilution 1/500), or mPGES-2 (dilution 1/200), or cPGES (dilution 1/1500 in 2% fat free dry milk powder in PBS-T) for 1 h at room temperature. Washings were done for 30 min in PBS-T. The second antibodies, goat anti-rabbit (COX-1, COX-2, mPGES-1 and mPGES-2 analyses) or goat anti-mouse (cPGES analysis) conjugated to horse radish peroxidase (dilution 1/10,000 in 2% fat free dry milk powder in PBS-T) were then incubated for 45 min at room temperature. The membrane was washed for another 30 min in PBS-T. Bands were revealed by addition of a chemiluminescent substrate applied according to the manufacturer’s instructions (Western Lightning, Perkin-Elmer). The blots were exposed to Hyperfilm ECL with intensifying screen.

3. Results 3.1. Molecular cloning of Macaque microsomal prostaglandin E synthase-1 and cytosolic prostaglandin E synthase The complete coding sequence of Macaque mPGES-1 (GenBank/EBI accession no. AY573809) is shown in Fig. 1. The amplified sequence of Macaque mPGES-1, consists

Fig. 1. PCR cloning and sequencing of Macaque mPGES-1 cDNA. Human nucleic acids sequence is shown at the top and differences with Macaque sequence are noted. Differences in amino acids sequence for Macaque are noted with asterisk (*). Numbers on the left refer to the first nucleic acid on that line. The Macaque mPGES-1 sequence was cloned as described in Section 2.

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Fig. 2. PCR cloning and sequencing of Macaque cPGES cDNA. Human nucleic acids sequence is shown at the top and differences with Macaque sequence are noted. Deduced amino acids sequence for human and Macaque cPGES is shown at the bottom. Numbers on the left refer to the first nucleic acid on that line. The Macaque cPGES sequence was cloned as described in Section 2.

of 459 bp, with an homology of 98% with human mPGES-1 at the nucleic acids level. The deduced amino acids sequence of 153 amino acids is 98% homologous to the human mPGES-1. In comparison with the human sequence, only three amino acids are different in Macaque: alanine7 , lysine112 and valine114 . The coding sequence and deduced amino acids for Macaque mPGES-1 share common features to mPGES-1 in other species: conserved arginine110 is involved in catalytic function of mPGES-1, and the consensus sequence ERXXXAXXNXXE, necessary for arachidonic acid liaison and/or oxygenation products, is also present in Macaque at positions 66–77. The complete coding sequence of Macaque cPGES (GenBank/EBI accession no. AY573810) is presented in Fig. 2. For this sequence, consisting of 483 bp, the homology is more than 99% with human sequence in nucleic acids, and deduced amino acids sequence is 100% identical to the human cPGES protein. It is worth noting that tyrosine9 involved in enzymatic activity and conserved among the species, is also present in the Macaque cPGES sequence. 3.2. Tissue distribution of cyclooxygenases and prostaglandin E synthases As shown in Fig. 3, all three PGE synthase mRNA were detected by Northern blot analysis at various levels in female reproductive tissues. mPGES-1 mRNA is present in oviduct and faint signals are present in myometrium and ovary; mPGES-2 mRNA is expressed in all tissues (liver > oviduct > endometrium = myometrium = ovary = lung); cPGES mRNA is expressed everywhere without any variation. COX-2 mRNA is expressed in liver, lung and oviduct: the expression was undetectable in other tissues. COX-1 mRNA is present in oviduct > lung > ovary = endometrium. As shown in Fig. 4A, protein expression assayed by Western blot analysis was also different from one tissue to another for each PGE synthase. mPGES-1 protein was most abundant in myometrium, ovary and lung, but was also detected at lower levels in endometrium, oviduct and

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Fig. 3. Expression of cyclooxygenases and prostaglandin E synthases mRNA in. Macaque tissues. Tissues were collected from one cyclic female Macaque in the proliferative or secretory phase and were frozen in liquid nitrogen. Total RNA was extracted with Trizol reagent, submitted to electrophoresis and transferred onto a nylon membrane. Northern blot hybridizations were done with specific Macaque COX-1, COX-2, mPGES-1, mPGES-2 and cPGES probes. Blots were exposed for 3–14 days. 18S is shown as a control of RNA quantity as seen on the gel. E: endometrium; M: myometrium; Ovi: oviduct; Ova: ovary; Li: liver; Lu: lung.

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Fig. 4. Expression of cyclooxygenases and prostaglandin E synthases protein in Macaque tissues. (A) Tissues were collected from one Macaque and were frozen in liquid nitrogen. (B) Platelets and neutrophils were isolated as described in Section 2. Proteins were then extracted, submitted to electrophoresis and transferred onto a nitrocellulose membrane. Western blot hybridizations were done with antibodies raised against COX-1, COX-2, mPGES-1, mPGES-2 and cPGES. Blots were exposed for 30–120 s. E: endometrium; M: myometrium; Ovi: oviduct; Ova: ovary; Li: liver; Lu: lung; P: platelets; N: neutrophils; C+: cultured human endometrial cells.

the liver. mPGES-2 protein is mainly expressed in liver, followed by the endometrium, myometrium, ovary, lung and oviduct. cPGES protein was expressed at higher level in the lung > endometrium = ovary > myometrium = oviduct > liver. Both mPGES-1 and mPGES2 were expressed at higher level in the human cultured cells, and cPGES protein was also detected. COX-1 and COX-2 were not detected at the basal level in any tissues tested, but were detected in the positive control (cultured human endometrial cells). To eliminate the possibility that our COX-1 and COX-2 antibodies did not cross-react with Macaque proteins, we performed controls in tissues known to possess a good amount of COX-1 or COX-2. As shown in Fig. 4B, signals are detected with the COX-1 antibody in Macaque platelets and with the COX-2 antibody in Macaque neutrophils stimulated with LPS.

4. Discussion In the present report, we cloned Macaque microsomal prostaglandin E synthase-1 and Macaque cytosolic prostaglandin E synthase and we studied tissue distribution of PGES and cyclooxygenases. To the best of our knowledge, the present study is the first to report simultaneous expression of the three known PGE synthases. Since the mPGES-2 sequence was already known in monkey (GenBank/EBI accession no. AB046026) [12], we cloned the other two PGES, for which homologous sequences were

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cloned in human, namely mPGES-1 and cPGES (Figs. 1 and 2). The sequences of those three PGES are highly homologous to their human counterparts (98%, 97% and 99% for mPGES-1, mPGES-2 and cPGES, respectively). One may expect that cyclooxygenases have the same degree of homology between the Macaque and human. It is therefore surprising that rabbit antibodies raised against sheep COX-1 and COX-2 do not react with Macaque tissues but do with human endometrial cells in culture (Fig. 4A). One explanation could be that the basal proteins levels in the tissues tested are too low to be detected, even if we detected mRNA expression (Fig. 3). In support of that hypothesis, results presented in Fig. 4B show that the two antibodies do cross-react with Macaque in tissues known to express significant levels of COX-1 protein (platelets) or COX-2 protein (neutrophils stimulated with LPS). Another explanation may result from the nature of cyclooxygenases known to be suicide enzymes that disappear as they catalyze the formation of PGH2 . Thus, the amount of COX1 and COX-2 proteins may have decreased rapidly in absence of sufficient messenger or translation activity to replenish the pool. We have already demonstrated that in bovine endometrium, COX-2 protein and mRNA but not COX-1 protein and mRNA are present during the estrous cycle [19]. The primary focus of our laboratory being to elucidate the role of PG in recognition and maintenance of pregnancy, mainly female reproductive tissues, such as the endometrium, myometrium, oviduct and ovary were used in the present study. Liver and lung were used as controls because those tissues can have abundant expression of PGs biosynthetic enzymes as we have previously shown in bovine [20,21]. RNase protection assay already demonstrated that cPGES mRNA is expressed at a very high level in the mouse testis, and at lower levels in the ovary and uterus [22]. Our results also show the presence of cPGES in the ovary and uterus (myometrium and endometrium) but do not show any variation in expression levels. In support of the present results for mPGES-1 mRNA expression, a study in the rat also found no expression in the lung and liver while there was expression in kidney and testis [23]. Female reproductive tissues such as those reported here were not tested in that study. In the human, among the tissues tested for mPGES-2 expression by Tanikawa et al. [12], only the placenta was related to female reproduction and they showed a faint expression; by contrast, in some tissues such as skeletal muscle, brain, heart and kidney, mPGES-1 was expressed at high levels. Very few studies were published on PG biosynthesis in the primate, especially in reproduction. Lipocalin-type PGD synthase (PGDS) was reported in the epididymidis [24]. Kim et al. [16] demonstrated COX-1 mRNA and COX-2 mRNA and protein expression in the baboon endometrium during the menstrual cycle and pregnancy. Duffy and Stouffer [25] showed that COX-2 mRNA but not COX-1 mRNA, is stimulated by human chorionic gonadotropin (hCG) in ovarian follicles of rhesus monkey. However, in the monkey, the majority of studies are restricted to evaluation of COX and PG receptor expression, and are focused on late pregnancy and parturition (see [26] for a review). PGs catabolism by 15-hydroxy PG dehydrogenase (PGDH) has also been studied in late gestation and parturition in the baboon: its presence was reported in the chorion, decidua, lower uterine segment, fundal myometrium and cervix [27]. While the present manuscript was in preparation, another study reported the presence and the regulation of mPGES-1 and cPGES, along with COX-1 and COX-2, in the endometrium of the rhesus monkey [28]. In that study, immunohistochemical analysis suggested that all four

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enzymes studied varied during the menstrual cycle, and that most of the time COX-1 and cPGES were co-located to allow basal PGE2 production. mPGES-1 and COX-2 were not always co-located, by contrast with what was found in the majority of studies in other systems [8]; however, consistent with the results presented in Figs. 3 and 4A. Interestingly, it has been reported recently that knock-out mice for mPGES-1 were fertile [29]. The results shown in the present study raise the hypothesis that lack of PGE2 production by mPGES-1 in those mice can be compensated by production through mPGES-2 and cPGES. Because both enzymes are expressed in the endometrium of the Macaque at the mRNA level and also at the protein level, we suppose that this also may be the case in the mouse. In conclusion, we report here the cloning of two Macaque PGE synthases, mPGES-1 and cPGES. Their tissue distribution was studied, along with mPGES-2, COX-1 and COX-2. The high homology of the Macaque PGES enzymes with the human strongly supports the use of this species for functional studies relevant to human physiology. However, more studies will be needed to fully understand the specific role and implication of each PGE synthase in PG biosynthesis in various physiological processes and pathologies. Acknowledgments The authors would like to thank Dr. Stacia Kargman from Merck Frosst (Kirkland, Que., Canada) for kindly providing antibodies raised against COX-1 and COX-2, Eric Madore for his help with sequence analyses, and Dr. Sylvain Bourgoin and his team for their help with platelet and neutrophil isolation. References [1] Dubois RN, Abramson SB, Crofford L, et al. Cyclooxygenase in biology and disease. FASEB J 1998;12:1063–73. [2] Chang SH, Liu CH, Conway R, et al. Role of prostaglandin E2-dependent angiogenic switch in cyclooxygenase 2-induced breast cancer progression. Proc Natl Acad Sci USA 2004;101:591–6. [3] Narumiya S, Sugimoto Y, Ushikubi F. Prostanoid receptors: structures, properties, and functions. Physiol Rev 1999;79:1193–226. [4] Serhan CN, Levy B. Success of prostaglandin E2 in structure–function is a challenge for structure-based therapeutics. Proc Natl Acad Sci USA 2003;100:8609–11. [5] Coceani F, Akarsu ES. Prostaglandin E2 in the pathogenesis of fever. An update. Ann NY Acad Sci 1998;856:76–82. [6] Poyser NL. The control of prostaglandin production by the endometrium in relation to luteolysis and menstruation. Prostaglandins Leukot Essent Fatty Acids 1995;53:147–95; Challis JRG, Matthews SG, Gibb W, Lye SJ. Endocrine and paracrine regulation of birth at term and preterm. Endocrinol Rev 2000;21:514–50. [7] Sales KJ, Jabbour HN. Cyclooxygenase enzymes and prostaglandins in reproductive tract physiology and pathology. Prostaglandins Other Lipid Mediat 2003;71:97–117; Jakobsson PJ, Thor´en S, Morgenstern R, Samuelsson B. Identification of human prostaglandin E synthase: a microsomal, glutathione-dependent, inducible enzyme, constituting a potential novel drug target. Proc Natl Acad Sci USA 1999;96:7220–5. [8] Morita I, Schindler M, Regier MK, et al. Different intracellular locations for prostaglandin endoperoxide H synthase-1 and -2. J Biol Chem 1995;270:10902–8;

J. Parent et al. / Prostaglandins & other Lipid Mediators 78 (2005) 27–37

[9]

[10]

[11]

[12] [13] [14] [15] [16] [17] [18]

[19] [20]

[21]

[22] [23] [24]

[25] [26] [27]

[28] [29]

37

Murakami M, Naraba H, Tanioka T, et al. Regulation of prostaglandin E2 biosynthesis by inducible membrane-associated prostaglandin E2 synthase that acts in concert with cyclooxygenase-2. J Biol Chem 2000;275:32783–92. Morita I. Distinct functions of COX-1 and COX-2. Prostaglandins Other Lipid Mediat 2002;68/69:165–75; Johnson JL, Beito TG, Krco CJ, Toft DO. Characterization of a novel 23-kilodalton protein of unactive progesterone receptor complexes. Mol Cell Biol 1994;14:1956–63. Hutchison KA, Stancato LF, Owens-Grillo JK, et al. The 23 kDa acidic protein in reticulocyte lysate is the weakly bound component of the hsp foldosome that is required for assembly of the glucocorticoid receptor into a functional heterocomplex with hsp90. J Biol Chem 1995;270:18841–7. Tanioka T, Nakatani Y, Semmyo N, Murakami M, Kudo I. Molecular identification of cytosolic prostaglandin E2 synthase that is functionally coupled with cyclooxygenase-1 in immediate prostaglandin E2 biosynthesis. J Biol Chem 2000;275:32775–82. Tanikawa N, Ohmiya Y, Ohkubo H, et al. Identification and characterization of a novel type of membraneassociated prostaglandin E synthase. Biochem Biophys Res Commun 2002;291:884–9. Murakami M, Nakashima K, Kamei D, et al. Cellular prostaglandin E2 production by membrane-bound prostaglandin E synthase-2 via both cyclooxygenases-1 and -2. J Biol Chem 2003;278:37937–47. Lagarde M, Bryon PA, Guichardant M, Dechavanne M. A simple and efficient method for platelet isolation from their plasma. Thromb Res 1980;17:581–8. Pouliot M, Gilbert C, Borgeat P, et al. Expression and activity of prostaglandin endoperoxide synthase-2 in agonist-activated human neutrophils. FASEB J 1998;12:1109–23. Kim JJ, Wang J, Bambra C, Das SK, Dey SK, Fazleabas AT. Expression of cyclooxygenase-1 and -2 in the baboon endometrium during the menstrual cycle and pregnancy. Endocrinology 1999;140:2672–8. Chapdelaine P, Ho-Kim MA, Tremblay RR, Dub´e JY. Southern blot analysis with synthetic oligonucleotides Application to prostatic protein genes. Int J Biochem 1990;22:75–82. Chapdelaine P, Vignola K, Fortier MA. Protein estimation directly from SDS-PAGE loading buffer for standardization of samples from cell lysates or tissue homogenates before Western blot analysis. Biotechniques 2001;31:478, 480, 482. Arosh JA, Parent J, Chapdelaine P, Sirois J, Fortier MA. Expression of cyclooxygenases 1 and 2 and prostaglandin E synthase in bovine endometrial tissue during the estrous cycle. Biol Reprod 2002;67:161–9. Parent J, Chapdelaine P, Sirois J, Fortier MA. Expression of microsomal prostaglandin E synthase in bovine endometrium: coexpression with cyclooxygenase type 2 and regulation by interferon-tau. Endocrinology 2002;143:2936–43. Madore E, Harvey N, Parent J, Chapdelaine P, Arosh JA, Fortier MA. An aldose reductase with 20 alpha-hydroxysteroid dehydrogenase activity is most likely the enzyme responsible for the production of prostaglandin F2 alpha in the bovine endometrium. J Biol Chem 2003;278:11205–12. Guan Y, Zhang Y, Schneider A, et al. Urogenital distribution of a mouse membrane-associated prostaglandin E(2) synthase. Am J Physiol Renal Physiol 2001;281:F1173–7. Satoh K, Nagano Y, Shimomura C, Suzuki N, Saeki Y, Yokota H. Expression of prostaglandin E synthase mRNA is induced in beta-amyloid treated rat astrocytes. Neurosci Lett 2000;283:221–3. Fouchecourt S, Chaurand P, DaGue BB, et al. Epididymal lipocalin-type prostaglandin D2 synthase: identification using mass spectrometry, messenger RNA localization, and immunodetection in mouse, rat, hamster, and monkey. Biol Reprod 2002;66:524–33. Duffy DM, Stouffer RL. The ovulatory gonadotrophin surge stimulates cyclooxygenase expression and prostaglandin production by the monkey follicle. Mol Hum Reprod 2001;7:731–9. Nathanielsz PW, Smith G, Wu W. Topographical specialization of prostaglandin function in late pregnancy and at parturition in the baboon. Prostaglandins Leukot Essent Fatty Acids 2004;70:199–206. Wu WX, Ma XH, Smith GC, Mecenas CA, Koenen SV, Nathanielsz PW. Prostaglandin dehydrogenase mRNA in baboon intrauterine tissues in late gestation and spontaneous labor. Am J Physiol Regul Integr Comp Physiol 2000;279:R1082–90. Sun T, Li SJ, Diao HL, Teng CB, Wang HB, Yang ZM. Cyclooxygenases and prostaglandin E synthases in the endometrium of the rhesus monkey during the menstrual cycle. Reproduction 2004;127:465–73. Trebino CE, Stock JL, Gibbons CP, et al. Impaired inflammatory and pain responses in mice lacking an inducible prostaglandin E synthase. Proc Natl Acad Sci USA 2003;100:9044–9.