Animal Reproduction (AR)
https://animal-reproduction.org/article/doi/10.1590/1984-3143-AR2023-0099
Animal Reproduction (AR)
ORIGINAL ARTICLE

Letrozole administration effects on the P450aromatase expression and the reproductive parameters in male sheep (Ovis aries)

Antônio Francisco da Silva Lisboa Neto; Túlio Teruo Yoshinaga; Antonio de Sousa Júnior; Marcílio Nichi; Alejandro Esteller-Vico; Antônio Chaves de Assis Neto

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Abstract

Letrozole comprises a non-steroid aromatase inhibitor that has been applied as a preventive for many uses, such as breast cancer prevention, treatment of hormonal dysfunction, and male infertility. Precisely in Northeast Brazil, ovine consist of the leading livestock produced, and their reproduction in captivity has been demonstrated difficult. Thus, we hypothesized whether the application of letrozole will improve male sheep reproduction. One group of 6 animals received a daily dosage of 0.5mg/Kg of letrozole for 60 days, while the other six animals were used as control. Samples were collected from control and treated animals after 30 and 60 days of the experiment. Blood samples were collected to measure the steroid hormone levels. Semen was collected from control and treated groups using an artificial vagina and cryopreserved for spermatozoa morphology and CASA analysis. The testicles were collected for histological analysis, gene expression, and immunohistochemistry of P450aromatase protein. Hormone levels demonstrate no differences in the estradiol/testosterone levels among the control and both treated groups. Immunohistochemistry analysis revealed the presence of P450aromatase protein in spermatogonia cells and Leydig cells in the control and treated groups in both periods analyzed. Moreover, there were no differences in the P450aromase gene expression in the control and treated group. Morphological analysis of the spermatozoa revealed that letrozole treatment did not affect mitochondrial activity or cause any deformities. In addition, motility parameters in the sperm from the treated group were not affected by letrozole treatment compared to the control group. Morphological analysis of the testis demonstrated that letrozole treatment increase the seminiferous tubule area but no signs of germ cell damage. Our results show that letrozole has a morphological effect on the testicles in the ovine model but no pathological or severe effect caused by hormone level imbalance. Overall, letrozole comprises a non-steroid aromatase inhibitor, which can be applied to ovine reproduction.

Keywords

steroidogenesis, estradiol, testosterone, aromatase inhibitors, male reproduction

References

Akça KT, Demirel MA, Süntar I. The role of aromatase enzyme in hormone related diseases and plant- based aromatase inhibitors as therapeutic regimens. Curr Top Med Chem. 2022;22(3):229-46. http://dx.doi.org/10.2174/1568026621666211129141631. PMid:34844542.

Arroyo MAM, Santos PRDS, De Oliveira MF, Neto ACDA. Prolonged use of letrozole causes morphological changes on gonads in Galea Spixii. Anim Reprod. 2021;18(2):e20200029. http://dx.doi.org/10.1590/1984-3143-ar2020-0029. PMid:34306212.

Arroyo MAM, Santos PRS, Oliveira MF, Conley AJ, Neto ACA. Immunolocalization of aromatase and estrogen receptor β in the testis and epididymis of Agoutis (Dasyprocta Leporina) during postnatal development. Anim Reprod Sci. 2016;169:130. http://dx.doi.org/10.1016/j.anireprosci.2016.03.081.

Barth AD, Oko RJ. Abnormal morphology of bovine spermatozoa [Internet]. Ames, Iowa USA: Iowa State Univesity Press; 1989 [cited 2023 June 25]. Available from: https://www.worldcat.org/title/abnormal-morphology-of-bovine-spermatozoa/oclc/19265200

Baufeld A, Vanselow J. A tissue culture model of estrogen-producing primary bovine granulosa cells. J Vis Exp. 2018;(139):1-7. PMid:30247464.

Bhatnagar AS. The discovery and mechanism of action of letrozole. Breast Cancer Res Treat. 2007;105(Suppl. 1):7-17. http://dx.doi.org/10.1007/s10549-007-9696-3. PMid:17912633.

Bourguiba S, Genissel C, Lambard S, Bouraïma H, Carreau S. Regulation of aromatase gene expression in leydig cells and germ cells. J Steroid Biochem Mol Biol. 2003;86(3–5):335-43. http://dx.doi.org/10.1016/S0960-0760(03)00343-1. PMid:14623530.

Carreau S, Bouraima-Lelong H, Delalande C. Estrogen, a female hormone involved in spermatogenesis. Adv Med Sci. 2012;57(1):31-6. http://dx.doi.org/10.2478/v10039-012-0005-y. PMid:22440937.

Carreau S, Bouraima-Lelong H, Delalande C. Estrogens in male germ cells. Spermatogenesis. 2011;1(2):90-4. http://dx.doi.org/10.4161/spmg.1.2.16766. PMid:22319655.

Carreau S, Hess RA. Oestrogens and spermatogenesis. Philos Trans R Soc Lond B Biol Sci. 2010;365(1546):1517-35. http://dx.doi.org/10.1098/rstb.2009.0235. PMid:20403867.

Carreau S, Silandre D, Bourguiba S, Hamden K, Said L, Lambard S, Galeraud-Denis I, Delalande C. Estrogens and male reproduction: a new concept. Braz J Med Biol Res. 2007;40(6):761-8. http://dx.doi.org/10.1590/S0100-879X2007000600003. PMid:17581673.

Conley A, Hinshelwood M. Mammalian aromatases. Reproduction. 2001;121(5):685-95. http://dx.doi.org/10.1530/rep.0.1210685. PMid:11427156.

Cornil CA. On the role of brain aromatase in females: why are estrogens produced locally when they are available systemically? J Comp Physiol A Neuroethol Sens Neural Behav Physiol. 2018;204(1):31-49. http://dx.doi.org/10.1007/s00359-017-1224-2. PMid:29086012.

Crespilho AM, Chiaradia L, Cortez A, Dinelli M, Papa FO, Gomes GM, Peixoto KC. Sensitivity evaluation of the computer-assisted sperm analysis (CASA) in the determination of frozen-thawed bull semen concentration. Braz J Vet Res Anim Sci. 2017;54(3):247-52. http://dx.doi.org/10.11606/issn.1678-4456.bjvras.2017.127773.

Czajka-Oraniec I, Simpson ER. Aromatase research and its clinical significance. Endokrynol Pol. 2010;61(1):126-34. PMid:20205115.

Dent SF, Gaspo R, Kissner M, Pritchard KI. Aromatase inhibitor therapy: toxicities and management strategies in the treatment of postmenopausal women with hormone-sensitive early breast cancer. Breast Cancer Res Treat. 2011;126(2):295-310. http://dx.doi.org/10.1007/s10549-011-1351-3. PMid:21249443.

Habeeb HMH, Hazzard TM, Stormshak F, Kutzler MA. Effect of different dosages of PG-600 on ovulation and pregnancy rates in ewes during the breeding season. Transl Anim Sci. 2018;3(1):429-32. http://dx.doi.org/10.1093/tas/txy112. PMid:32704813.

Haynes BP, Dowsett M, Miller WR, Dixon JM, Bhatnagar AS. The pharmacology of letrozole. J Steroid Biochem Mol Biol. 2003;87(1):35-45. http://dx.doi.org/10.1016/S0960-0760(03)00384-4. PMid:14630089.

Hrudka F. Cytochemical and ultracytochemical demonstration of cytochrome c oxidase in spermatozoa and dynamics of its changes accompanying ageing or induced by stress. Int J Androl. 1987;10(6):809-28. http://dx.doi.org/10.1111/j.1365-2605.1987.tb00385.x. PMid:2828243.

Katiele T, De Jesus S, Rizzo H, Franco F. Caracterização do sistema de produção de caprinos e ovinos no estado de Sergipe, nordeste do Brasil. Acta Vet Brasilica. 2020;14(2):121-31. http://dx.doi.org/10.21708/avb.2020.14.2.9247.

Kondarewicz A, Kolasa A, Zawiślak B, Baranowska-Bosiacka I, Marchlewicz M, Wenda-Różewicka L, Wiszniewska B. Testis morphology in rats chronically treated with letrozole, an aromatase inhibitor. Folia Histochem Cytobiol. 2011;49(4):677-84. http://dx.doi.org/10.5603/FHC.2011.0091. PMid:22252764.

Larose H, Shami AN, Abbott H, Manske G, Lei L, Hammoud SS. Gametogenesis: a journey from inception to conception. Curr Top Dev Biol. 2019;132:257-310. http://dx.doi.org/10.1016/bs.ctdb.2018.12.006. PMid:30797511.

Lees JL. Functional Infertility in sheep. Vet Rec. 1978;102(11):232-6. http://dx.doi.org/10.1136/vr.102.11.232. PMid:644812.

Li X, Strauss L, Kaatrasalo A, Mayerhofer A, Huhtaniemi I, Santti R, Mäkelä S, Poutanen M. Transgenic mice expressing P450 aromatase as a model for male infertility associated with chronic inflammation in the testis. Endocrinology. 2006;147(3):1271-7. http://dx.doi.org/10.1210/en.2005-0654. PMid:16306085.

McManus C, Paiva SR, De Araújo RO. Genética e melhoramento de ovinos no Brasil. Rev Bras Zootec. 2010;39(Suppl. 1):236-46. http://dx.doi.org/10.1590/S1516-35982010001300026.

Morohashi K, Baba T, Tanaka M. Steroid hormones and the development of reproductive organs. Sex Dev. 2013;7(1–3):61-79. http://dx.doi.org/10.1159/000342272. PMid:22986257.

Ni FD, Shuang LH, Wan XY. Multiple signaling pathways in sertoli cells: recent findings in spermatogenesis. Cell Death Dis. 2019;10(8):541. http://dx.doi.org/10.1038/s41419-019-1782-z. PMid:31316051.

Nichi M, Bols PE, Züge RM, Barnabe VH, Goovaerts IG, Barnabe RC, Cortada CN. Seasonal variation in semen quality in Bos Indicus and Bos Taurus bulls raised under tropical conditions. Theriogenology. 2006;66(4):822-8. http://dx.doi.org/10.1016/j.theriogenology.2006.01.056. PMid:16529802.

Nunes FAP, Farias MLFD, Oliveira FP, Vieira L No, Lima LFC, Paranhos FDP No, et al. Use of aromatase inhibitors in patients with breast cancer is associated with deterioration of bone microarchitecture and density. Arch Endocrinol Metab. 2021;65(4):505-11. http://dx.doi.org/10.20945/2359-3997000000385. PMid:34283901.

Ohkawa H, Ohishi N, Yagi K. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem. 1979;95(2):351-8. http://dx.doi.org/10.1016/0003-2697(79)90738-3. PMid:36810.

Pilutin A, Misiakiewicz-Has K, Kolasa A, Baranowska-Bosiacka I, Marchlewicz M, Wiszniewska B. The immunoexpression of androgen receptor, estrogen receptors α and β, Vanilloid Type 1 receptor and cytochrome P450 aromatase in rats testis chronically treated with letrozole, an aromatase inhibitor. Folia Histochem Cytobiol. 2014;52(3):206-17. http://dx.doi.org/10.5603/FHC.2014.0024. PMid:25308736.

Pistelli M, Della Mora A, Ballatore Z, Berardi R. Aromatase inhibitors in premenopausal women with breast cancer: the state of the art and future prospects. Curr Oncol. 2018;25(2):168-75. http://dx.doi.org/10.3747/co.25.3735. PMid:29719441.

Pope CE, Zhang YZ, Dresser BL. A simple staining method for evaluating acrosomal status of cat spermatozoa. J Zoo Wildl Med. 1991 [cited 2023 June 25];22(1):87-95. Available from: http://www.jstor.org/stable/20095123

Ribeiro FA, Pereira CSB, Chi RJ, Yokomizo PL, Fregnani JHTG, Rocha RM. Comparative assessment between objective and subjective methods in slides stained by immunohistochemistry. Rev Bras Otorrinolaringol (Engl Ed). 2013;79(6):704-8. http://dx.doi.org/10.5935/1808-8694.20130129. PMid:24474481.

Ronde W, Jong FH. Aromatase inhibitors in men: effects and therapeutic options. Reprod Biol Endocrinol. 2011;9(1):93. http://dx.doi.org/10.1186/1477-7827-9-93. PMid:21693046.

Rosati L, Agnese M, Di Fiore MM, Andreuccetti P, Prisco M. P450 aromatase: a key enzyme in the spermatogenesis of the Italian Wall Lizard, Podarcis Sicula. J Exp Biol. 2016;219(Pt 15):2402-8. http://dx.doi.org/10.1242/jeb.135996. PMid:27489219.

Rosati L, Di Fiore MM, Prisco M, Di Giacomo Russo F, Venditti M, Andreuccetti P, Chieffi Baccari G, Santillo A. Seasonal expression and cellular distribution of star and steroidogenic enzymes in quail testis. J Exp Zoolog B Mol Dev Evol. 2019;332(6):198-209. v http://dx.doi.org/10.1002/jez.b.22896. PMid:31433565.

Rosati L, Prisco M, Di Lorenzo M, De Falco M, Andreuccetti P. Immunolocalization of aromatase P450 in the epididymis of podarcis sicula and Rattus Rattus. Eur J Histochem. 2020;64(1). http://dx.doi.org/10.4081/ejh.2020.3080. PMid:31988532.

Rosenfeld CS, Shay DA, Vieira-Potter VJ. Cognitive effects of aromatase and possible role in memory disorders. Front Endocrinol. 2018;9:610. http://dx.doi.org/10.3389/fendo.2018.00610. PMid:30386297.

Santolaria P, Palacin I, Yaniz J. Management factors affecting fertility in sheep. In Manafi M, editor. Artificial insemination in farm animals. London, UK: InTech; 2011. http://dx.doi.org/10.5772/18013.

Santos A, Oliveira M, Viana D, Miglino M, Neto AA. Immunolocalization of the cytochrome P450 enzymes: C17, NADPH reductase and aromatase in the vaginal mucosa of the spix’s yellow-toothed cavy (Galea Spixii) during the estrous cycle. Placenta. 2014;35(9):A22. http://dx.doi.org/10.1016/j.placenta.2014.06.074.

Santos AC, Conley AJ, Oliveira MF, Oliveira GB, Viana DC, Assis AC No. Immunolocalization of steroidogenic enzymes in the vaginal mucous of Galea Spixii during the estrous cycle. Reprod Biol Endocrinol. 2017;15(1):30. http://dx.doi.org/10.1186/s12958-017-0248-3. PMid:28438170.

Schlegel PN. Aromatase inhibitors for male infertility. Fertil Steril. 2012;98(6):1359-62. http://dx.doi.org/10.1016/j.fertnstert.2012.10.023. PMid:23103016.

Shelton M, Figueiredo EAP. Genetic aspects of adaptation of animals to tropical environments with special reference to sheep and goats in northeast Brazil. In: Simpósio Internacional De Bioclimatologia Animal nos Trópicos: Pequenos e Grandes Ruminantes. Brasília, DF: EMBRAPA; 1990 [cited 2023 June 25]. p. 46-61. Available from: https://www.embrapa.br/busca-de-publicacoes/-/publicacao/515345/genetic-aspects-of-adaptation-of-animals-to-tropical-environments-with-special-reference-to-sheep-and-goats-in-northeast-brazil.

Shen Z-G, Fan QX, Yang W, Zhang YL, Wang HP. Effects of 17α-methyltestosterone and aromatase inhibitor letrozole on sex reversal, gonadal structure, and growth in yellow catfish Pelteobagrus Fulvidraco. Biol Bull. 2015;228(2):108-17. http://dx.doi.org/10.1086/BBLv228n2p108. PMid:25920714.

Shuling L, Sie Kuei ML, Saffari SE, Jiayun Z, Yeun TT, Leng JPW, Viardot-Foucault V, Nadarajah S, Chan JKY, Hao TH. Do men with normal testosterone–oestradiol ratios benefit from letrozole for the treatment of male infertility? Reprod Biomed Online. 2019;38(1):39-45. http://dx.doi.org/10.1016/j.rbmo.2018.09.016. PMid:30449700.

Solar II, Donaldson EM. Studies on genetic and hormonal sex control in a domesticated rainbow trout. 2. Use of methyltestosterone for masculinization and sterilization in cultured rainbow trout (Salmo Gairdneri Richardson). Can Tech Rep Fish Aquat Sci. 1985 [cited 2023 June 25];1380. Available from: https://www.researchgate.net/publication/308519254_Studies_on_genetic_and_hormonal_sex_control_in_domesticated_rainbow_trout_II_Use_of_methyltestosterone_for_masculinization_and_sterilization_in_cultured_rainbow_trout_Salmo_gairdneri_Richardson

Ulhaq ZS. Brain aromatase modulates cardiac functions in embryonic zebrafish. Int J Vet Sci Med. 2019;7(1):31-4. http://dx.doi.org/10.1080/23144599.2019.1675287. PMid:31692872.

Van Emon ML, Vonnahme KA, Berg PT, Redden RR, Thompson MM, Kirsch JD, Schauer CS. Influence of level of dried distillers grains with solubles on feedlot performance, carcass characteristics, serum testosterone concentrations, and spermatozoa motility and concentration of growing rams1. J Anim Sci. 2013;91(12):5821-8. http://dx.doi.org/10.2527/jas.2013-6369. PMid:24146156.

Vanderstichel R, Forzán MJ, Pérez GE, Serpell JA, Garde E. Changes in blood testosterone concentrations after surgical and chemical sterilization of male free-roaming dogs in southern Chile. Theriogenology. 2015;83(6):1021-7. http://dx.doi.org/10.1016/j.theriogenology.2014.12.001. PMid:25557187.

Yang C, Li P, Li Z. Clinical application of aromatase inhibitors to treat male infertility. Hum Reprod Update. 2021;28(1):30-50. http://dx.doi.org/10.1093/humupd/dmab036. PMid:34871401.

Yaron Z, Levavi-Sivan B. 2011. Endocrine regulation of fish reproduction. In Farrell AP, editor. Encyclopedia of fish physiology. London: Elsevier, p. 1500–1508. http://dx.doi.org/10.1016/B978-0-12-374553-8.00058-7.

Zhao H, Zhou L, Shangguan AJ, Bulun SE. Aromatase expression and regulation in breast and endometrial cancer. J Mol Endocrinol. 2016;57(1):R19-33. http://dx.doi.org/10.1530/JME-15-0310. PMid:27067638.

Zirkin BR, Papadopoulos V. Leydig cells: formation, function, and regulation. Biol Reprod. 2018;99(1):101-11. http://dx.doi.org/10.1093/biolre/ioy059. PMid:29566165.
 


Submitted date:
06/25/2023

Accepted date:
10/24/2023

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