Reproduction, Fertility and Development Reproduction, Fertility and Development Society
Vertebrate reproductive science and technology
RESEARCH ARTICLE

Caspase-3-mediated apoptosis and cell proliferation in the equine endometrium during the oestrous cycle

R. P. Roberto da Costa A , P. M. Serrão B , S. Monteiro B , P. Pessa C , J. Robalo Silva B and G. Ferreira-Dias B D

A Escola Superior Agrária, Instituto Politécnico de Coimbra, Bencanta, 3040-316 Coimbra, Portugal.

B CIISA, Faculdade de Medicina Veterinária, Universidade Técnica de Lisboa (TULisbon), 1300-477 Lisbon, Portugal.

C Hospitais da Universidade de Coimbra, 3000-075 Coimbra, Portugal.

D Corresponding author. Email: gmlfdias@fmv.utl.pt

Reproduction, Fertility and Development 19(8) 925-932 http://dx.doi.org/10.1071/RD06159
Submitted: 5 December 2006  Accepted: 22 July 2007   Published: 13 September 2007

Abstract

Cell proliferation and apoptosis are hormone-dependent physiological processes involved in endometrial growth and regression. The aims of the present study were: (1) to evaluate endometrial cell proliferation using proliferating cell nuclear antigen (PCNA) expression; (2) to evaluate the induction of endometrial cell death by the expression of active caspase-3 and the apoptotic phenotype visualised by DNA fragmentation; and (3) to relate these observations to endometrial tissue dynamics in the equine endometrium throughout the oestrous cycle. Endometria were assigned to follicular and luteal phases based on ovarian structures and plasma progesterone. Cell proliferation and active caspase-3-mediated apoptosis were expressed in both phases of the oestrous cycle. In the luteal phase, PCNA expression was higher than in the follicular phase. Highest PCNA activity was noted in the luminal and glandular structures. Active caspase-3 staining was increased in luminal epithelium and deep glandular cells during the luteal phase. However, in the follicular phase, stromal cells showed greater active caspase-3 expression. Only a few apoptotic endometrial cells were detected by terminal deoxyribonucleotidyl transferase-mediated dUTP–digoxigenin nick end-labelling (TUNEL) and these cells were mostly present in luminal and glandular structures. A simultaneous increase in DNA, cell proliferation and protein synthesis was observed in the endometrium during the mid-luteal phase. This suggests that cell hyperplasia occurs at the time the histotroph is needed for eventual embryo nourishment.

Additional keywords: apoptosis, mare, proliferating cell nuclear antigen, terminal deoxyribonucleotidyl transferase-mediated dUTP–digoxigenin nick end-labelling (TUNEL).


References

Aupperle H.Ozgen S. H. A.Schoon D.Hoppen H. O.Sieme H.Tannapfel A.2000Cyclical endometrial steroid hormone receptor expression and proliferation intensity in the mare.Equine Vet. J.32228232doi:10.2746/042516400776563554Pubmed Abstract

Ausubel F. M., Brent R., Kingston R. E., Moore D. D., Seidman J. G., Smith J. A., and Struhl K. (1992). ‘Current Protocols in Molecular Biology.’ (Greene Publishing Association, Wiley-Interscience: New York.)

Baserga R. (1985). Tissue growth. In ‘The Biology of Cell Reproduction’. (Ed. R. Baserga.) pp. 46–58. (Harvard University Press: Cambridge.)

Berkkanoglu M.Guzeloglu-Kayisli O.Kayisli U. A.Selam B. F.Arici A.2004Regulation of Fas ligand expression by vascular endothelial growth factor in endometrial stromal cells in vitro.Mol. Hum. Reprod.10393398doi:10.1093/MOLEHR/GAH060Pubmed Abstract

Blankenship T. N.King B. F.1994Developmental expression of Ki-67 antigen and proliferating cell nuclear antigen in macaque placentas.Dev. Dyn.201324333Pubmed Abstract

Brenner R. M., and Slayden O. D. (1994). Cyclic changes in the primate oviduct and endometrium. In ‘The Physiology of Reproduction’, vol. 1. (Eds E. Knobil and J. Neill.) pp. 541–569. (Raven Press: New York.)

Bursch W.Paffe S.Putz B.Barthel G.Schulte-Hermann R.1990Determination of the length of the histological stages of apoptosis in normal liver and in altered hepatic foci of rats.Carcinogenesis11847853doi:10.1093/CARCIN/11.5.847Pubmed Abstract

Celis J. E.Celis A.1985Cell cycle-dependent variations in the distribution of the nuclear protein cyclin proliferating cell nuclear antigen in cultured cells: subdivision of S phase.Proc. Natl Acad. Sci. USA8232623266doi:10.1073/PNAS.82.10.3262Pubmed Abstract

Chu P. Y.Wright P. J.Lee C. S.2002Apoptosis of endometrial cells in the bitch.Reprod. Fertil. Dev.14297305doi:10.1071/RD01108Pubmed Abstract

Chu P. Y., Lee C. S., and Wright P. J. (2006). Degeneration and apoptosis of endometrial cells in the bitch. Theriogenology 66, 1545–1549.

Dallenbach-Hellweg G.1988The endometrium in natural and artificial luteal phases.Hum. Reprod.3165168Pubmed Abstract

Evans A. C. O.2003Characteristics of ovarian follicle development in domestic animals.Reprod. Domest. Anim.38240246doi:10.1046/J.1439-0531.2003.00439.XPubmed Abstract

Finn C. A.Martin L.1973Endocrine control of gland proliferation in the mouse uterus.Biol. Reprod.8585588Pubmed Abstract

Gargett C. E.Lederman F.Heryanto B.Gambino L. S.Rogers P. A.2001Focal vascular endothelial growth factor correlates with angiogenesis in human endometrium. Role of intravascular neutrophils.Hum. Reprod.1610651075doi:10.1093/HUMREP/16.6.1065Pubmed Abstract

Gerstenberg C.Allen W. R.Stewart F.1999Cell proliferation patterns in the equine endometrium throughout the non-pregnant reproductive cycle.J. Reprod. Fertil.116167175Pubmed Abstract

Ginther O. J. (Ed.) (1992a). Characteristics of the ovulatory season. In ‘Reproductive Biology of the Mare: Basic and Applied Aspects’, 2nd edn. pp. 173–232. (Equiservices: Cross Plains, WI.)

Ginther O. J. (Ed.) (1992b). Endocrinology of the ovulatory season. In ‘Reproductive Biology of the Mare: Basic and Applied Aspects’, 2nd edn. pp. 233–290. (Equiservices: Cross Plains, WI.)

Harada T.Kaponis A.Iwabe T.Taniguchi F.Makrydimas G.Sofikiti N.Paschopoulo M.Paraskevaidis E.Terakawa N.2004Apoptosis in human endometrium and endometriosis.Hum. Reprod. Update102938doi:10.1093/HUMUPD/DMH007Pubmed Abstract

Hayashi K.Carpenter K. D.Spencer T. E.2004Neonatal estrogen exposure disrupts uterine development in the postnatal sheep.Endocrinology14532473257doi:10.1210/EN.2004-0178Pubmed Abstract

Hengartner M. O.2000The biochemistry of apoptosis.Nature407770776doi:10.1038/35037710Pubmed Abstract

Heryanto B.Rogers P. A.2002Regulation of endometrial endothelial cell proliferation by oestrogen and progesterone in the ovariectomized mouse.Reproduction123107113doi:10.1530/REP.0.1230107Pubmed Abstract

Ito K.Sasano H.Watanabe K.Ozawa N.Sato S.Yajima A.1993Immunohistochemical study of PCNA (proliferating cell nuclear antigen) in normal and abnormal endometrium.Int. J. Gynecol. Cancer3122127doi:10.1046/J.1525-1438.1993.03020122.XPubmed Abstract

Johnson M. L.Redmer D. A.Reynolds L. P.1997Uterine growth, cell proliferation, and c-fos proto-oncogene expression throughout the estrous cycle in ewesBiol. Reprod.56393401doi:10.1095/BIOLREPROD56.2.393Pubmed Abstract

Jones R. K.Searle R. F.Stewart J. A.Turner S.Bulmer J. N.1998Apoptosis, bcl-2 expression, and proliferative activity in human endometrial stroma and endometrial granulated lymphocytes.Biol. Reprod.589951002doi:10.1095/BIOLREPROD58.4.995Pubmed Abstract

Kaeoket K.Persson E.Dalin A. M.2001The sow endometrium at different stages of the oestrous cycle: studies on morphological changes and infiltration by cells of the immune system.Anim. Reprod. Sci.6595114doi:10.1016/S0378-4320(00)00211-6Pubmed Abstract

Kayisli U. A.Mahutte N. G.Arici A.2002Uterine chemokines in reproductive physiology and pathology.Am. J. Reprod. Immunol.47213221doi:10.1034/J.1600-0897.2002.01075.XPubmed Abstract

Kokawa K.Shikone T.Nakano R.1996Apoptosis in the human uterine endometrium during the menstrual cycle.J. Clin. Endocrinol. Metab.8141444147doi:10.1210/JC.81.11.4144Pubmed Abstract

Lai M. D.Lee L. R.Cheng K. S.Wing L. Y.2000Expression of proliferating cell nuclear antigen in luminal epithelium during the growth and regression of rat uterus.J. Endocrinol.1668793doi:10.1677/JOE.0.1660087Pubmed Abstract

Lakhani S. A.Masud A.Kuida K.Porter G. A.JrBooth C. J.Mehal W. Z.Inayat I.Flavell R. A.2006Caspases 3 and 7: key mediators of mitochondrial events of apoptosis.Science311847851doi:10.1126/SCIENCE.1115035Pubmed Abstract

Lynch M. P.Nawaz S.Gerschenson L. E.1986Evidence for soluble factors regulating cell death and cell proliferation in primary cultures of rabbit endometrial cells grown on collagen.Proc. Natl Acad. Sci. USA8347844788doi:10.1073/PNAS.83.13.4784Pubmed Abstract

Martin L.Das R. M.Finn C. A.1973The inhibition by progesterone of uterine epithelial proliferation in the mouse.J. Endocrinol.57549554Pubmed Abstract

Masters R. A.Crean B. D.Yan W.Moss A. G.Ryan P. L.Wiley A. A.Bagnell C. A.Bartol F. F.2006Neonatal porcine endometrial development and epithelial proliferation affected by age and exposure to estrogen and relaxin.Domest. Anim. Endocrinol.Pubmed Abstract, In press.

Medh R. D.Thompson E. B.2000Hormonal regulation of physiological cell turnover and apoptosis.Cell Tissue Res.301101124doi:10.1007/S004419900159Pubmed Abstract

Morris G. F.Mathews M. B.1989Regulation of proliferating cell nuclear antigen during the cell cycle.J. Biol. Chem.26413 85613 864Pubmed Abstract

Narkar M.Kholkute S.Chitlange S.Nandedkar T.2006Expression of steroid hormone receptors, proliferation and apoptotic markers in primate endometrium.Mol. Cell. Endocrinol.246107113doi:10.1016/J.MCE.2005.11.029Pubmed Abstract

Nawaz S.Lynch M. P.Galand P.Gerschenson L. E.1987Hormonal regulation of cell death in rabbit uterine epithelium.Am. J. Pathol.1275159Pubmed Abstract

Otsuki Y.2001Apoptosis in human endometrium: apoptotic detection methods and signaling.Med. Electron Microsc.34166173doi:10.1007/S007950100011Pubmed Abstract

Reynolds L. P.Killilea S. D.Redmer D. A.1992Angiogenesis in the female reproductive system.FASEB J.6886892Pubmed Abstract

Roberto da Costa R. P.Branco V.Pessa P.Robalo Silva J.Ferreira-Dias G.2005Progesterone receptors and proliferating cell nuclear antigen expression in the equine luteal tissue.Reprod. Fertil. Dev.17659666doi:10.1071/RD05024Pubmed Abstract

Roberto da Costa R. P.Ferreira-Dias G.Mateus L.Korzekwa A.Andronowska A.Platek R.Skarzynski D. J.2007Endometrial nitric oxide production and nitric oxide synthases in the equine endometrium: relationship with microvascular density during the estrous cycle.Domest. Anim. Endocrinol.34287302

Selam B.Kayisli U. A.Mulayim N.Arici A.2001Regulation of Fas ligand expression by estradiol and progesterone in human endometrium.Biol. Reprod.65979985
doi:10.1095/BIOLREPROD65.4.979Pubmed Abstract

Shikone T.Kokawa K.Yamoto M.Nakano R.1997Apoptosis of human ovary and uterine endometrium during the menstrual cycle.Horm. Res.482734Pubmed Abstract

Sukjumlong S.Persson E.Kaeoket K.Dalin A. M.2004Immunohistochemical studies on oestrogen receptor alpha (ERalpha) and the proliferative marker Ki-67 in the sow uterus at oestrus and early pregnancy.Reprod. Domest. Anim.39361369doi:10.1111/J.1439-0531.2004.00529.XPubmed Abstract

Sukjumlong S.Dalin A. M.Sahlin L.Persson E.2005Immunohistochemical studies on the progesterone receptor (PR) in the sow uterus during the oestrous cycle and in inseminated sows at oestrus and early pregnancy.Reproduction129349359doi:10.1530/REP.1.00514Pubmed Abstract

Taylor K. M.Gray C. A.Joyce M. M.Stewart M. D.Bazer F. W.Spencer T. E.2000Neonatal ovine uterine development involves alterations in expression of receptors for estrogen, progesterone, and prolactin.Biol. Reprod.6311921204doi:10.1095/BIOLREPROD63.4.1192Pubmed Abstract

Van Cruchten S.Van den Broeck W.Duchateau L.Simoens P.2003Apoptosis in the canine endometrium during the estrous cycle.Theriogenology6015951608doi:10.1016/S0093-691X(03)00178-XPubmed Abstract

Vaskivuo T. E.Stenback F.Karhumaa P.Risteli J.Dunkel L.Tapanainen J. S.2000Apoptosis and apoptosis-related proteins in human endometrium.Mol. Cell. Endocrinol.1657583doi:10.1016/S0303-7207(00)00261-6Pubmed Abstract

Vaux D. L.1999Caspases and apoptosis: biology and terminology.Cell Death Differ.6493494doi:10.1038/SJ.CDD.4400523Pubmed Abstract

Vaux D. L.Korsmeyer S. J.1999Cell death in development.Cell96245254doi:10.1016/S0092-8674(00)80564-4Pubmed Abstract

Waga S.Hannon G. J.Beach D.Stillman B.1994The p21 inhibitor of cyclin-dependent kinases controls DNA replication by interaction with PCNA.Nature369574578doi:10.1038/369574A0Pubmed Abstract

Wasowska B.Ludkiewicz B.Stefanczyk-Krzymowska S.Grzegorzewski W.Skipor J.2001Apoptotic cell death in the porcine endometrium during the oestrous cycle.Acta Vet. Hung.497179doi:10.1556/AVET.49.2001.1.9Pubmed Abstract



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