Oestrous cycle-dependent expression of Fas and Bcl2 family gene products in normal canine endometrium
S. Henriques A , E. Silva A , S. Cruz A , M. F. Silva A , G. Ferreira-Dias A , L. Lopes-da-Costa A and L. Mateus A B
+ Author Affiliations
- Author Affiliations
A CIISA, Faculty of Veterinary Medicine, University of Lisbon, Avenida da Universidade Técnica, 1300-477 Lisboa, Portugal.
B Corresponding author. Email: lmateus@fmv.ulisboa.pt
Reproduction, Fertility and Development 28(9) 1307-1317 https://doi.org/10.1071/RD14245
Submitted: 10 July 2014 Accepted: 10 January 2015 Published: 24 February 2015
Abstract
During the oestrous cycle canine endometrium undergoes cyclical cellular proliferation, apoptosis and differentiation. To study the regulation of endometrial apoptosis and proliferation events the expression of apoptosis-related genes was analysed by real-time polymerase chain reaction and cellular expression of their proteins was identified through immunohistochemistry. Cellular apoptosis and proliferation events were detected by TdT-mediated dUTP-biotin nick end labeling (TUNEL) and proliferation marker Ki67 immunostaining, respectively. The highest proliferative index was observed in the follicular phase (all endometrial cellular components) and at early dioestrus (basal glands). This was associated with a low apoptotic index and a strong expression of anti- (Bcl2) and pro-apoptotic proteins (Fas, FasL, Bax). Subsequently (Days 11–45 of dioestrus), basal glandular epithelium experienced the highest apoptotic index, coincidental with a decrease of Bcl2 expression and a low ratio of Bcl2/Bax transcription. An increase in the apoptotic index of crypts, stromal and endothelial cells was observed at late dioestrus and the beginning of anoestrus. These results indicate that pro- and anti-apoptotic proteins regulate the balance between cell proliferation and death in the canine endometrium during the oestrous cycle. High Bcl2 expression in both the follicular and early dioestrous phases stimulate glandular proliferation and prevent apoptosis but, in the non-pregnant uterus, a decrease in Bcl2 expression together with an increase in pro-apoptotic proteins induces apoptosis of basal glandular epithelium cells.
Additional keywords: apoptosis, bitch, proliferation.
References
Applied Biosystems (2003). Creating standard curves with genomic DNA or plasmid DNA templates for use in quantitative PCR. Available from: http://www6.appliedbiosystems.com/support/tutorials/pdf/quant_pcr.pdf [Verified 6 February 2015].Chu, P.-Y., Lee, C. S., and Wright, P. J. (2006). Degeneration and apoptosis of endometrial cells in the bitch. Theriogenology 66, 1545–1549.
| Degeneration and apoptosis of endometrial cells in the bitch.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD28Xps1Oit7s%3D&md5=60e3e15dd45a3af0de4f45a2aaf965e4CAS | 16473400PubMed |
Fernandes, P. A., Bowen, R. A., Sawyer, H. R., Nett, T. M., and Gorell, T. A. (1989). Concentration of receptors for oestradiol and progesterone in canine endometrium during oestrus and dioestrus. Am. J. Vet. Res. 50, 64–67.
| 1:CAS:528:DyaL1MXmvFKjsw%3D%3D&md5=25e666b4186e9c6140a6fdca729458b7CAS | 2919829PubMed |
Galabova, G., Egerbacher, M., Aurich, J. E., Leitner, M., and Walter, I. (2003). Morphological changes of endometrial epithelium in the bitch during metoestrus and anoestrus. Reprod. Domest. Anim. 38, 415–420.
| Morphological changes of endometrial epithelium in the bitch during metoestrus and anoestrus.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BD3svosVSluw%3D%3D&md5=7f75d45fdd3e43d673758f401b52e4e0CAS | 12950696PubMed |
Gerstenberg, C., Allen, W. R., and Stewart, F. (1999). Cell proliferation patterns in the equine endometrium throughout the non-pregnant reproductive cycle. J. Reprod. Fertil. 116, 167–175.
| Cell proliferation patterns in the equine endometrium throughout the non-pregnant reproductive cycle.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1MXjvVemsbc%3D&md5=dd189b527d26726d729a3dbffafabba9CAS | 10505067PubMed |
Giulietti, A., Overbergh, L., Valckx, D., Decallonne, B., Bouillon, R., and Mathieu, C. (2001). An overview of real-time quantitative PCR: applications to quantify cytokine gene expression. Methods 25, 386–401.
| An overview of real-time quantitative PCR: applications to quantify cytokine gene expression.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD38XhtFelt7o%3D&md5=292b5c172944f6fe58818afaa4282bbaCAS | 11846608PubMed |
Groppetti, D., Pecile, A., Arrighi, S., Di Giancamillo, A., and Cremonesi, F. (2010). Endometrial cytology and computerised morphometric analysis of epithelial nuclei: a useful tool for reproductive diagnosis in the bitch. Theriogenology 73, 927–941.
| Endometrial cytology and computerised morphometric analysis of epithelial nuclei: a useful tool for reproductive diagnosis in the bitch.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BC3c3gsVWgtA%3D%3D&md5=7b0e3ddbf60cdc3e3feff0f252f083c7CAS | 20116837PubMed |
Hanada, M., Aimé-Sempe, C., Sato, T., and Reed, J. C. (1995). Structure–function analysis of Bcl2 protein. Identification of conserved domains important for homodimerisation with Bcl2 and heterodimerisation with Bax. J. Biol. Chem. 270, 11 962–11 969.
| Structure–function analysis of Bcl2 protein. Identification of conserved domains important for homodimerisation with Bcl2 and heterodimerisation with Bax.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK2MXlvVWktr8%3D&md5=af121d4c70c24f9d3f38d4080b420c2fCAS |
Harada, T., Kaponis, A., Iwabe, T., Taniguchi, F., Makrydimas, G., Sofikitis, N., Paschopoulos, M., Paraskevaidis, E., and Terakawa, N. (2004). Apoptosis in human endometrium and endometriosis. Hum. Reprod. Update 10, 29–38.
| Apoptosis in human endometrium and endometriosis.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2cXislygsbs%3D&md5=3a51b56dcfdd0a6c0d12980b7655ee45CAS | 15005462PubMed |
Kida, K., Maezono, Y., Kawate, N., Inaba, T., Hatoya, S., and Tamada, H. (2010). Epidermal growth factor, transforming growth factor-α and epidermal growth factor receptor expression and localisation in the canine endometrium during the oestrous cycle and in bitches with pyometra. Theriogenology 73, 36–47.
| Epidermal growth factor, transforming growth factor-α and epidermal growth factor receptor expression and localisation in the canine endometrium during the oestrous cycle and in bitches with pyometra.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXhsVyntb%2FI&md5=b1a9644f40d394d0f0868023d4e43693CAS | 19853901PubMed |
Liman, N., Alan, E., Bayram, G. K., and Gürbulak, K. (2013). Expression of survinin, Bcl2 and Bax proteins in the domestic cat (Felis catus) endometrium during the oestrus cycle. Reprod. Domest. Anim. 48, 33–45.
| Expression of survinin, Bcl2 and Bax proteins in the domestic cat (Felis catus) endometrium during the oestrus cycle.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3sXktVOhtrk%3D&md5=e339811adb7e575eb89c5598b0dabde7CAS | 22471444PubMed |
Okano, A., Ogawa, H., Takahashi, H., and Geshi, M. (2007). Apoptosis in the porcine uterine endometrium during the oestrous cycle, early pregnancy and post partum. J. Reprod. Dev. 53, 923–930.
| Apoptosis in the porcine uterine endometrium during the oestrous cycle, early pregnancy and post partum.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXhtFGgsL3O&md5=b665312db44cc8a217b21223c12862e1CAS | 17380038PubMed |
Oltvai, Z. N., Milliman, C. L., and Korsmeyer, S. J. (1993). Bcl-2 heterodimerises in vivo with a conserved homolog BAX, that accelerates programmed cell death. Cell 74, 609–619.
| 1:CAS:528:DyaK3sXms1eqtbs%3D&md5=bf76e131b27b46659cc82d09eb95686aCAS | 8358790PubMed |
Onclin, K., Murphy, B., and Verstegen, J. P. (2002). Comparisons of oestradiol, LH and FSH patterns in pregnant and non-pregnant beagle bitches. Theriogenology 57, 1957–1972.
| Comparisons of oestradiol, LH and FSH patterns in pregnant and non-pregnant beagle bitches.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD38XltFans7c%3D&md5=307bf6eff6f879c8dbd1f27cf5e531beCAS | 12066857PubMed |
Otsuki, Y. (2001). Apoptosis in human endometrium: apoptotic detection methods and signalling. Med. Electron Microsc. 34, 166–173.
| Apoptosis in human endometrium: apoptotic detection methods and signalling.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD38XhtFShtb0%3D&md5=3c0d8b7edb42819a604ed8b455a453f4CAS | 11793192PubMed |
Otsuki, Y., Misaki, O., Sugimoto, O., Ito, Y., Tsujimoto, Y., and Akao, Y. (1994). Cyclic Bcl2 gene expression in human uterine endometrium during menstrual cycle. Lancet 344, 27–29.
| Cyclic Bcl2 gene expression in human uterine endometrium during menstrual cycle.Crossref | GoogleScholarGoogle Scholar |
Reed, J. C. (1996). A day in the life of the Bcl-2 protein: does the turnover rate of Bcl-2 serve as a biological clock for cellular lifespan regulation? Leuk. Res. 20, 109–111.
| A day in the life of the Bcl-2 protein: does the turnover rate of Bcl-2 serve as a biological clock for cellular lifespan regulation?Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK28Xis1OltLk%3D&md5=0ab1b9a5d9c21b573a8f3a2bef3e4eb6CAS | 8628008PubMed |
Rehm, S., Stanislaus, D. J., and Williams, A. M. (2007). Oestrous cycle-dependent histology and review of sex steroid receptor expression in dog reproductive tissues and mammary gland and associated hormone levels. Birth Defects Res. B Dev. Reprod. Toxicol. 80, 233–245.
| Oestrous cycle-dependent histology and review of sex steroid receptor expression in dog reproductive tissues and mammary gland and associated hormone levels.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXnslyiurk%3D&md5=55d4079448fe1bec76c2246a9be7a51dCAS | 17570128PubMed |
Schäfer-Somi, S., Sabitzer, S., Klein, D., Tomaszewski, C., Kanca, H., Beceriklisoy, H. B., Kucukaslan, I., Agaoglu, R., Kaya, D., Ay, S., and Aslan, S. (2012). Is apoptosis a regulatory mechanism during early canine pregnancy? Reprod. Domest. Anim. 47, 169–172.
| Is apoptosis a regulatory mechanism during early canine pregnancy?Crossref | GoogleScholarGoogle Scholar | 23279491PubMed |
Sedlak, T. W., Oltvai, Z. N., Yang, E., Wang, K., Boise, L. H., Thompson, C. B., and Korsmeyer, S. J. (1995). Multiple Bcl2 family members demonstrate selective dimerisations with BAX. Proc. Natl. Acad. Sci. USA 92, 7834–7838.
| Multiple Bcl2 family members demonstrate selective dimerisations with BAX.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK2MXnsFOnt74%3D&md5=3f596061558589ac95f3bed572b2bdb1CAS | 7644501PubMed |
Selam, B., Kayisli, U. A., Mulayim, N., and Arici, A. (2001). Regulation of Fas ligand expression by oestradiol and progesterone in human endometrium. Biol. Reprod. 65, 979–985.
| Regulation of Fas ligand expression by oestradiol and progesterone in human endometrium.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3MXnt1Cnu7s%3D&md5=255ad2b37a33e97d9a1cd8a611f6d858CAS | 11566716PubMed |
Shia, W.-Y., Chu, P.-C., Chou, C.-C., and Lee, W.-M. (2011). Investigation of the relationship between endometrial MMP activities and sex hormones in mature bitches. Thai. J. Vet. Med. 41, 289–297.
Shimizu, S., Eguchi, Y., Kamiike, W., Matsuda, H., and Tsujimoto, Y. (1996). Bcl2 expression prevents activation of the ICE protease cascade. Oncogene 12, 2251–2257.
| 1:CAS:528:DyaK28XjvFCmt70%3D&md5=f583e6a60e7013f0a799fd476974ea71CAS | 8649764PubMed |
Silva, E., Henriques, S., Brito, S., Ferreira-Dias, G., Lopes-da-Costa, L., and Mateus, L. (2012). Canine endometrial changes in toll-like receptors and prostaglandin production throughout the oestrous cycle. J. Reprod. Immunol. 96, 45–57.
| Canine endometrial changes in toll-like receptors and prostaglandin production throughout the oestrous cycle.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC38XhvVSmsLrJ&md5=b15ab39bb940b700ba0303ef662ed6e9CAS | 22959486PubMed |
Song, J., Rutherford, T., Naftolin, F., Brown, S., and Mor, G. (2002). Hormonal regulation of apoptosis and Fas and Fasl system in human endometrial cells. Mol. Hum. Reprod. 8, 447–455.
| Hormonal regulation of apoptosis and Fas and Fasl system in human endometrial cells.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD38XkvVGqur0%3D&md5=3a87945ce654c785688194e35fbd67e7CAS | 11994542PubMed |
Tan, K. H., and Hunziker, W. (2003). Compartmentalisation of Fas and Fas ligand may prevent auto- or paracrine apoptosis in epithelial cells. Exp. Cell Res. 284, 283–290.
| Compartmentalisation of Fas and Fas ligand may prevent auto- or paracrine apoptosis in epithelial cells.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3sXitVylu7s%3D&md5=cdab9163be0c61b55c1460d2829605aeCAS | 12651160PubMed |
Tanaka, M., Suda, T., Haze, K., Nakamura, N., Sato, K., Kimura, F., Motoyoshi, K., Tagawa, S., Ohga, S., Hatake, K., Drummond, A. H., and Nagata, S. (1996). Fas ligand in human serum. Nat. Med. 2, 317–322.
| Fas ligand in human serum.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK28XhsVGkt7o%3D&md5=ee67ca14cb054c871bc30eb396b63decCAS | 8612231PubMed |
Tao, X.-J., Tilly, K. I., Maravei, D. V., Shifren, J. L., Krajewski, S., Reed, J. C., Tilly, J. L., and Isaacson, K. B. (1997). Differential expression of members of the Bcl-2 gene family in proliferative and secretory human endometrium: glandular epithelial cell apoptosis is associated with increased expression of Bax. J. Clin. Endocrinol. Metab. 82, 2738–2746.
| 1:CAS:528:DyaK2sXltFGlsrg%3D&md5=2c89724e9b8bb0718d4d6e42fb897e21CAS | 9253363PubMed |
Van Cruchten, S., Van den Broeck, W., Duchateau, L., and Simoens, P. (2003). Apoptosis in the canine endometrium during the oestrous cycle. Theriogenology 60, 1595–1608.
| Apoptosis in the canine endometrium during the oestrous cycle.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3sXot12js7g%3D&md5=40b1432bfacad8c33f7007115ed13089CAS | 14580643PubMed |
Van Cruchten, S., Van den Broeck, W., D`haeseleer, M., and Simoens, P. (2004). Proliferation patterns in the canine endometrium during the oestrous cycle. Theriogenology 62, 631–641.
| Proliferation patterns in the canine endometrium during the oestrous cycle.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BD2czisF2nuw%3D%3D&md5=efed6e48c0a1d12412dd26fb2cad9eb3CAS | 15226018PubMed |
Vaskivuo, T. E., Stenbäck, F., Karhumaa, P., Risteli, J., Dunkel, L., and Tapanainen, J. S. (2000). Apoptosis and apoptosis-related proteins in human endometrium. Mol. Cell. Endocrinol. 165, 75–83.
| Apoptosis and apoptosis-related proteins in human endometrium.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3cXlsFCjs7Y%3D&md5=50de3b402a53e973e319c73764f78533CAS | 10940486PubMed |
Vermeirsch, H., Simoens, P., Lauwers, H., and Coryn, M. (1999). Immunohistochemical detection of oestrogen receptors in the canine uterus and their relation to sex-steroid hormone levels. Theriogenology 51, 729–743.
| Immunohistochemical detection of oestrogen receptors in the canine uterus and their relation to sex-steroid hormone levels.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1MXjsFegtbk%3D&md5=1a331e532220b83926ce4589e98bc77cCAS | 10728998PubMed |
Vermeirsch, H., Simoens, P., Hellemans, A., Coryn, M., and Lauwers, H. (2000). Immunohistochemical detection of progesterone receptors in the canine uterus and their relation to sex-steroid hormone levels. Theriogenology 53, 773–788.
| Immunohistochemical detection of progesterone receptors in the canine uterus and their relation to sex-steroid hormone levels.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3cXhvVyhs7k%3D&md5=a0e2a68bc96bc9a0bbf815ee915ce673CAS | 10735043PubMed |
Watanabe, H., Kanzaki, H., Narukawa, S., Inoue, T., Katsuragawa, H., Kaneko, Y., and Mori, T. (1997). Bcl2 and Fas expression in eutopic and ectopic human endometrium during the menstrual cycle in relation to endometrial cell apoptosis. Am. J. Obstet. Gynecol. 176, 360–368.
| Bcl2 and Fas expression in eutopic and ectopic human endometrium during the menstrual cycle in relation to endometrial cell apoptosis.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaK2s3gsFCmtg%3D%3D&md5=ec5e2173f4e495445d3a129a811931bcCAS | 9065182PubMed |
Wei, P., Jin, X., Tao, S.-T., Han, C.-S., and Liu, Y.-X. (2005). Fas, FasL, Bcl2 and Bax in the endometrium of Rhesus monkey during the menstrual cycle. Mol. Reprod. Dev. 70, 478–484.
| Fas, FasL, Bcl2 and Bax in the endometrium of Rhesus monkey during the menstrual cycle.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2MXit1Sls7g%3D&md5=f5d99d44768fba6b5bc9cecee51aa8ceCAS | 15685630PubMed |
Yamashita, H., Otsuki, Y., Matsumoto, K., Ueki, K., and Ueki, M. (1999). Fas ligand, Fas antigen and Bcl2 expression in human endometrium during the menstrual cycle. Mol. Hum. Reprod. 5, 358–364.
| Fas ligand, Fas antigen and Bcl2 expression in human endometrium during the menstrual cycle.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1MXisFakurk%3D&md5=d5a9a2d0cb9b5cefeff72e29fdc53f5fCAS | 10321808PubMed |
