Skip to main content

Histomorphometric evaluation of cannabinoid receptor and anandamide modulating enzyme expression in the human endometrium through the menstrual cycle

Abstract

Plasma anandamide (AEA) levels fluctuate throughout the menstrual cycle and in early pregnancy in a pattern suggesting its involvement in implantation and early pregnancy maintenance through mechanisms that might involve its binding to cannabinoid receptors CB1 and CB2. Plasma AEA levels are maintained by the actions of the enzymes fatty acid amide hydrolase (FAAH) and N-acylphosphatidylethanolamine-phospholipase D (NAPE-PLD). All of these component parts of the ‘endocannabinoid system’ have been demonstrated in rodent but not in human uteri. This study aimed to demonstrate the presence of the endocannabinoid system in the human uterus and catalogue its modulation. Immunohistochemical techniques were employed to localise and determine the distribution of immunoreactive CB1, CB2, FAAH, and NAPE-PLD in well-characterised menstrual cycle biopsy samples. Immunoreactive CB1 and CB2 were widely distributed throughout the uterine tissue. In the myometrium and endometrium, smooth muscle cells were immunoreactive, although the vascular smooth muscle cells in both tissues were more so. In the endometrium, CB1 and CB2 immunoreactivity was primarily restricted to the glandular epithelium and expression was unrelated to the phase of the cycle. FAAH immunoreactivity in the endometrium was highest in the mid-proliferative gland and mid-secretory stroma, whilst NAPE-PLD immunoreactivity was down-regulated in the secretory epithelial gland compared to the proliferative epithelial gland and unaffected in the stroma. These data indicate that elements of the ‘endocannabinoid system’ coexist in many cell types within the uterus and may provide insight into the sites of action of endogenous and exogenous cannabinoids during endometrial transformation.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3

References

  • Acone G, Trabucco E, Colacurci N, Cobellis L, Mackie K, Meccariello R, Cacciola G, Chioccarelli T, Fasano S, Pierantoni R, Cobellis G (2009) Low type I cannabinoid receptor levels characterize placental villous in labouring delivery. Placenta 30:203–205

    Article  CAS  PubMed  Google Scholar 

  • Battista N, Pasquariello N, Di Tommaso M, Maccarrone M (2008) Interplay between endocannabinoids, steroids and cytokines in the control of human reproduction. J Neuroendocrinol 20:82–89

    Article  CAS  PubMed  Google Scholar 

  • Deutsch DG, Ueda N, Yamamoto S (2002) The fatty acid amid hydrolase (FAAH). Prostaglandins Leukot Essent Fatty Acids 66:201–210

    Article  CAS  PubMed  Google Scholar 

  • Di Marzo V (2008) Targeting the endocannabinoid system: to enhance or reduce? Nat Rev Drug Discov 7:438–455

    Article  CAS  PubMed  Google Scholar 

  • El-Talatini MR, Taylor AH, Spurling SM, Konje JC (2008) Variation of plasma anandamide levels and fatty acid amide hydrolase expression throughout the menstrual cycle. Reprod Sci 15:167A

    Google Scholar 

  • El-Talatini MR, Taylor AH, Konje JC (2009a) Fluctuation in anandamide levels from ovulation to early pregnancy in in vitro fertilization-embryo transfer women, and its hormonal regulation. Hum Reprod 24:1989–1998

    Article  CAS  PubMed  Google Scholar 

  • El-Talatini MR, Taylor AH, Elson JC, Brown L, Davidson AC, Konje JC (2009b) Localisation and function of the endocannabinoid system in the human ovary. PLoS ONE 4(2):e4579. doi:10.1371/journal.pone.0004579

    Article  PubMed  CAS  Google Scholar 

  • El-Talatini MR, Taylor AH, Konje JC (2009c) The relationship between plasma levels of the endocannabinoid, anandamide, sex steroids, and gonadotrophins during the menstrual cycle. Fertil Steril. doi:10.1016/j.fertnstert.2008.12.033

  • Gasperi V, Fezza F, Spagnuolo P, Pasquariello N, Maccarrone M (2005) Further insights into the regulation of human FAAH by progesterone and leptin implications for endogenous levels of anandamide and apoptosis of immune and neuronal cells. Neurotoxicology 26:811–817

    Article  CAS  PubMed  Google Scholar 

  • Guney M, Oral B, Karahan N, Mungan T (2007) Expression of leukaemia inhibitory factor (LIF) during the window of implantation in copper T380A intrauterine device users. Eur J Contracept Reprod Health Care 12:212–219

    Article  CAS  PubMed  Google Scholar 

  • Guo Y, Wang H, Okamoto Y, Ueda N, Kingsley PJ, Marnett LJ, Schmid HHO, Das SK, Dey SK (2005) N-Acylphosphatidylethanolamine-hydrolyzing phospholipase D is an important determinant of uterine anandamide levels during implantation. J Biol Chem 280:23429–23432

    Article  CAS  PubMed  Google Scholar 

  • Habayeb OM, Taylor AH, Evans MD, Cooke MS, Taylor DJ, Bell SC, Konje JC (2004) Plasma levels of the endocannabinoid anandamide in women—a potential role in pregnancy maintenance and labor? J Clin Endocrinol Metab 89:5482–5487

    Article  CAS  PubMed  Google Scholar 

  • Habayeb OM, Taylor AH, Bell SC, Taylor DJ, Konje JC (2008a) Expression of the endocannabinoid system in human first trimester placenta and its role in trophoblast proliferation. Endocrinology 149:5052–5060

    Article  CAS  PubMed  Google Scholar 

  • Habayeb OM, Taylor AH, Finney M, Evans MD, Konje JC (2008b) Plasma anandamide concentration and pregnancy outcome in women with threatened miscarriage. JAMA 299:1135–1136

    Article  CAS  PubMed  Google Scholar 

  • Hamilton PW (1995) Designing a morphometric study. In: Allen DC (ed) Quantitative clinical pathology. Blackwell Science, Oxford, pp 311–315

    Google Scholar 

  • Horne AW, Phillips JA 3rd, Kane N, Lourenco PC, McDonald SE, Williams AR, Simon C, Dey SK, Critchley HO (2008) CB1 expression is attenuated in Fallopian tube and decidua of women with ectopic pregnancy. PLoS ONE 3:e3969. doi:10.1371/journal.pone.0003969

    Article  PubMed  CAS  Google Scholar 

  • Jones RL, Findlay JK, Salamonsen LA (2006) The role of activins during decidualisation of human endometrium. Aust N Z J Obstet Gynaecol 46:245–249

    Article  PubMed  Google Scholar 

  • Lam PMW, Marczylo TH, El-Talatini M, Finney M, Nallendran V, Taylor AH, Konje JC (2008) Ultra performance liquid chromatography tandem mass spectrometry method for the measurement of anandamide in human plasma. Anal Biochem 380:195–201

    Article  CAS  PubMed  Google Scholar 

  • Lazzarin N, Valensise H, Bari M, Ubaldi F, Battista N, Finazzi-Agro A, Maccarrone M (2004) Fluctuations of fatty acid amide hydrolase and anandamide levels during the human ovulatory cycle. Gynecol Endocrinol 18:212–218

    Article  CAS  PubMed  Google Scholar 

  • Liu WM, Duan EK, Cao YJ (2002) Effects of anandamide on embryo implantation in the mouse. Life Sci 71:1623–1632

    Article  CAS  PubMed  Google Scholar 

  • Ludwig M, Klein HH, Diedrich K, Ortmann O (2000) Serum leptin concentrations throughout the menstrual cycle. Arch Gynecol Obstet 263:99–101

    Article  CAS  PubMed  Google Scholar 

  • Maccarrone M (2008) CB2 receptors in reproduction. Br J Pharmacol 153:189–198

    Article  CAS  PubMed  Google Scholar 

  • Maccarrone M (2009) Endocannabinoids: friends and foes of reproduction. Prog Lipid Res 48:344–354

    Article  CAS  PubMed  Google Scholar 

  • Maccarrone M, Finazzi-Agro A (2004) Anandamide hydrolase: a guardian of human reproduction? Trends Pharmacol Sci 25:353–357

    Article  CAS  PubMed  Google Scholar 

  • Maccarrone M, De Felici M, Bari M, Klinger F, Siracusa G, Finazzi-Agro A (2000a) Down-regulation of anandamide hydrolase in mouse uterus by sex hormones. Eur J Biochem 267:2991–2997

    Article  CAS  PubMed  Google Scholar 

  • Maccarrone M, Valensise H, Bari M, Lazzarin N, Romanini C, Finazzi-Agro A (2000b) Relation between decreased anandamide hydrolase concentrations in human lymphocytes and miscarriage. Lancet 355:1326–1329

    Article  CAS  PubMed  Google Scholar 

  • Maccarrone M, Bari M, Di Rienzo M, Finazzi-Agro A, Rossi A (2003) Progesterone activates fatty acid amide hydrolase (FAAH) promoter in human T lymphocytes through the transcription factor Ikaros. Evidence for a synergistic effect of leptin. J Biol Chem 278:32726–32732

    Article  CAS  PubMed  Google Scholar 

  • Maccarrone M, DeFelici M, Klinger FG, Battista N, Fezza F, Dainese E, Siracusa G, Finazzi-Agro A (2004) Mouse blastocytes release a lipid which activates anandamide hydrolase in intact uterus. Mol Hum Reprod 10:215–221

    Article  CAS  PubMed  Google Scholar 

  • Noyes RW, Hertig AT, Rock J (1950) Dating the endometrial biopsy. Fertil Steril 1:3–25

    Google Scholar 

  • Pacher P, Batkai S, Kunos G (2006) The endocannabinoid system as an emerging target of pharmacology. Pharmacol Rev 58:389–462

    Article  CAS  PubMed  Google Scholar 

  • Paria BC, Deutsch DD, Dey SK (1996) The uterus is a potential site for anandamide synthesis and hydrolysis: differential profiles of anandamide synthase and hydrolase activities in the mouse uterus during the periimplantation period. Mol Reprod Develop 45:183–192

    Article  CAS  Google Scholar 

  • Paria BC, Zhao X, Wang J, Das SK, Dey SK (1999) Fatty-acid amide hydrolase is expressed in the mouse uterus and embryo during the peri-implantation period. Biol Reprod 60:1151–1157

    Article  CAS  PubMed  Google Scholar 

  • Paria BC, Song H, Wang X, Schmid PC, Krebsbach RJ, Schmid HHO, Bonner TI, Zimmer A, Dey SK (2001) Dysregulated cannabinoid signaling disrupts uterine receptivity for embryo implantation. J Biol Chem 276:20523–20528

    Article  CAS  PubMed  Google Scholar 

  • Schmid PC, Paria BC, Krebsbach RJ, Schmid HHO, Dey SK (1997) Changes in anandamide levels in mouse uterus are associated with uterine receptivity for embryo implantation. Proc Natl Acad Sci USA 94:4188–4192

    Article  CAS  PubMed  Google Scholar 

  • Sharkey AM, Smith SK (2003) The endometrium as a cause of implantation failure. Best Pract Res Clin Obstet Gynaecol 17:289–307

    Article  PubMed  Google Scholar 

  • Sun X, Dey SK (2008) Aspects of endocannabinoid signaling in periimplantation biology. Mol Cell Endocrinol 286S:S2–S11

    Google Scholar 

  • Taylor CR, Shi SR, Chaiwun B, Young L, Imam SA, Cote RJ (1994) Strategies for improving the immunohistochemical staining of various intranuclear prognostic markers in formalin-paraffin sections: androgen receptor, estrogen receptor, progesterone receptor, p53 protein, proliferating cell nuclear antigen, and Ki-67 antigen revealed by antigen retrieval techniques. Hum Pathol 25:263–270

    Article  CAS  PubMed  Google Scholar 

  • Taylor AH, Guzail M, Wahab M, Thompson JR, Al-Azzawi F (2005) Quantitative histomorphometric analysis of gonadal steroid receptor distribution in the normal human endometrium through the menstrual cycle. Histochem Cell Biol 123:463–474

    Article  CAS  PubMed  Google Scholar 

  • Taylor AH, Ang C, Bell SC, Konje JC (2007) The role of the endocannabinoid system in gametogenesis, implantation and early pregnancy. Hum Reprod Update 13:501–513

    Article  CAS  PubMed  Google Scholar 

  • Taylor AH, Guzail M, Al-Azzawi F (2008) Evidence of differential expression of oestrogen receptor isoforms in the fibromuscular layer of the vulva and reduction of androgen receptor levels in the fibrovascular layer of vulvae lichen sclerosus but not chronic vaginitis. Br J Dermatol 158:319–328

    CAS  PubMed  Article  Google Scholar 

  • Trabucco E, Acone G, Marenna A, Pierantoni R, Cacciola G, Chioccarelli T, Mackie K, Fasano S, Colacurci N, Meccariello R, Cobellis G, Cobellis L (2009) Endocannabinoid system in first trimester placenta: low FAAH and high CB1 expression characterize spontaneous miscarriage. Placenta 30:516–522

    Article  CAS  PubMed  Google Scholar 

  • Ueda N, Puffenbarger RA, Yamamoto S, Deutsch DG (2000) The fatty acid amide hydrolase (FAAH). Chem Phys Lipids 108:107–121

    Article  CAS  PubMed  Google Scholar 

  • Ueda N, Okamoto Y, Morishita J (2005) N-acylphosphatidylethanolamine-hydrolyzing phospholipase D: a novel enzyme of the β-lactamase fold family releasing anandamide and other N-acylethanolamines. Life Sci 77:1750–1758

    Article  CAS  PubMed  Google Scholar 

  • Wahab M, Thompson J, Hamid B, Deen S, Al-Azzawi F (1999) Endometrial histomorphometry of trimegestone-based sequential hormone replacement therapy: a weighted comparison with the endometrium of the natural cycle. Hum Reprod 14:2609–2618

    Article  CAS  PubMed  Google Scholar 

  • Wang H, Dey SK, Maccarrone M (2006a) Jekyll and hyde: two faces of cannabinoid signaling in male and female fertility. Endocr Rev 27:427–448

    Article  CAS  PubMed  Google Scholar 

  • Wang H, Xie H, Dey SK (2006b) Endocannabinoid signaling directs periimplantation events. AAPS J 8:E425–E432

    PubMed  Google Scholar 

  • Wang H, Xie H, Sun X, Kingsley PJ, Marnett LJ, Cravatt BF, Dey SK (2007) Differential regulation of endocannabinoid synthesis and degradation in the uterus during embryo implantation. Prostaglandins Other Lipid Mediat 83:62–74

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

The authors thank Mrs Susan Spurling for cutting the sections, Dr Lawrence Brown (Gynaecology Histopathologist, Pathology Department, Leicester Royal Infirmary, Leicester) and Dr Suha Deen (Consultant Histopathologist, Pathology Department, Queen’s Medical Centre, Nottingham) for accurate dating of the biopsy samples. This work was funded in part by the BUPA Foundation and by the Reproductive Sciences Section, University of Leicester.

Conflict of interest statement

The authors declare that they have no conflicts of interest.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Anthony H. Taylor.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Taylor, A.H., Abbas, M.S., Habiba, M.A. et al. Histomorphometric evaluation of cannabinoid receptor and anandamide modulating enzyme expression in the human endometrium through the menstrual cycle. Histochem Cell Biol 133, 557–565 (2010). https://doi.org/10.1007/s00418-010-0695-9

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00418-010-0695-9

Keywords

  • Cannabinoid receptors
  • Fatty acid amide hydrolase
  • Menstrual cycle
  • Human
  • Uterus