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Vertebrate reproductive science and technology
RESEARCH ARTICLE

Seasonal variation in sperm freezability associated with changes in testicular germinal epithelium in domestic (Ovis aries) and wild (Ovis musimon) sheep

Lucía Martínez-Fresneda A B C , Emma O’Brien A , Rosario Velázquez A , Adolfo Toledano-Díaz A , Carlos M. Martínez-Cáceres D , Dawit Tesfaye B , Karl Schellander B , Francisco A. García-Vázquez https://orcid.org/0000-0001-7665-3858 C E and Julian Santiago-Moreno https://orcid.org/0000-0001-5551-8120 A F
+ Author Affiliations
- Author Affiliations

A Department of Animal Reproduction, Spanish National Institute for Agricultural and Food Research and Technology (INIA), Avda. Puerta de Hierro km 5.9, 28040 Madrid, Spain.

B Department of Animal Breeding and Husbandry, Institute of Animal Science, University of Bonn, Endenicher Allee 15, 53115 Bonn, Germany.

C Department of Physiology, Faculty of Veterinary Science, International Excellence Campus for Higher Education and Research ‘Campus Mare Nostrum’, University of Murcia, Campus de Espinardo, 30100 Murcia, Spain.

D Pathology Unit, Biomedical Research Institute of Murcia (IMIB-Arrixaca), Crta. Buenavista s/n, El Palmar 30120 Murcia, Spain.

E Reproductive Physiology and Assisted Reproduction Unit, Biomedical Research Institute of Murcia (IMIB-Arrixaca), Crta. Buenavista s/n, El Palmar 30120 Murcia, Spain.

F Corresponding author. Email: moreno@inia.es

Reproduction, Fertility and Development 31(10) 1545-1557 https://doi.org/10.1071/RD18511
Submitted: 12 December 2018  Accepted: 4 May 2019   Published: 11 June 2019

Abstract

The aim of this study was to examine ovine sperm cryoresistance during the rutting season (RS) and its association with sperm head area and seminiferous epithelium proliferation. Small ruminants show fluctuating testosterone levels throughout the year, which could interfere with spermatogenesis and sperm cryopreservation. Ejaculates, testicular biopsies and blood were collected during the middle and at the end of the RS (Middle-RS vs End-RS) during periods of high and low testosterone levels in Merino and Mouflon rams. Fresh and frozen–thawed sperm quality, sperm morphometry, seminiferous tubule morphometry and testicular proliferation markers (proliferating cell nuclear antigen, proliferation marker protein Ki-67 and transcription factor GATA-4) were evaluated. Post-thaw sperm viability was higher in the End-RS group in both Merino (69.9 ± 8.2 vs 41.6 ± 7.3%; P = 0.020) and Mouflon rams (40.9 ± 3.3 vs 24.2 ± 5.0%; P = 0.008). Mouflons had larger sperm head area at the End-RS (38.3 ± 0.2 vs 34.3 ± 0.1 µm2; P = 0.029), whereas there was no difference between Merino groups (35.7 ± 0.5 vs 34.8 ± 1.0 µm2). Seminiferous tubule morphometry and proliferation markers showed higher levels of germinal epithelium proliferation in the Middle-RS of both species. In conclusion, sperm freezability is affected during the RS in domestic and wild rams, which could be correlated with changes that occur during spermatogenesis, since there is an effect of season on cell proliferation in the testis.

Additional keywords: cryopreservation, Sertoli cell, spermatogenesis, spermatozoa, testis, testosterone.


References

Ambrosi, C. P., Rubio, N., Gimenez, G., Venturino, A., Aisen, E. G., and Lopez Armengol, M. F. (2018). Modeling of behavioral responses for successful selection of easy-to-train rams for semen collection with an artificial vagina. Anim. Reprod. Sci. 193, 90–97.
Modeling of behavioral responses for successful selection of easy-to-train rams for semen collection with an artificial vagina.Crossref | GoogleScholarGoogle Scholar | 29678566PubMed |

Argov-Argaman, N., Mahgrefthe, K., Zeron, Y., and Roth, Z. (2013). Season-induced variation in lipid composition is associated with semen quality in Holstein bulls. Reproduction 145, 479–489.
Season-induced variation in lipid composition is associated with semen quality in Holstein bulls.Crossref | GoogleScholarGoogle Scholar | 23630332PubMed |

Arruda, R. P., Ball, B. A., Gravance, C. G., Garcia, A. R., and Liu, I. K. M. (2002). Effects of extenders and cryoprotectants on stallion sperm head morphometry. Theriogenology 58, 253–256.
Effects of extenders and cryoprotectants on stallion sperm head morphometry.Crossref | GoogleScholarGoogle Scholar |

Bansode, F. W., Chowdhury, S. R., and Dhar, J. D. (2003). Seasonal changes in the seminiferous epithelium of rhesus and bonnet monkeys. J. Med. Primatol. 32, 170–177.
Seasonal changes in the seminiferous epithelium of rhesus and bonnet monkeys.Crossref | GoogleScholarGoogle Scholar | 12823627PubMed |

Barenton, B., and Pelletier, J. (1983). Seasonal changes in testicular gonadotropin receptors and steroid content in the ram. Endocrinology 112, 1441–1446.
Seasonal changes in testicular gonadotropin receptors and steroid content in the ram.Crossref | GoogleScholarGoogle Scholar | 6299695PubMed |

Barranco, I., Ortega, M. D., Martinez-Alborcia, M. J., Vazquez, J. M., Martinez, E. A., and Roca, J. (2013). Season of ejaculate collection influences the freezability of boar spermatozoa. Cryobiology 67, 299–304.
Season of ejaculate collection influences the freezability of boar spermatozoa.Crossref | GoogleScholarGoogle Scholar | 24045067PubMed |

Battle, M. A., Bondow, B. J., Iverson, M. A., Adams, S. J., Jandacek, R. J., Tso, P., and Duncan, S. A. (2008). GATA4 is essential for jejunal function in mice. Gastroenterology 135, 1676–1686.e1.
GATA4 is essential for jejunal function in mice.Crossref | GoogleScholarGoogle Scholar | 18812176PubMed |

Bench, C. J., Price, E. O., Dally, M. R., and Borgwardt, R. E. (2001). Artificial selection of rams for sexual performance and its effect on the sexual behavior and fecundity of male and female progeny. Appl. Anim. Behav. Sci. 72, 41–50.
Artificial selection of rams for sexual performance and its effect on the sexual behavior and fecundity of male and female progeny.Crossref | GoogleScholarGoogle Scholar | 11259825PubMed |

Blanco-Rodríguez, J., and Martínez-García, C. (1999). Apoptosis is physiologically restricted to a specialized cytoplasmic compartment in rat spermatids. Biol. Reprod. 61, 1541–1547.
Apoptosis is physiologically restricted to a specialized cytoplasmic compartment in rat spermatids.Crossref | GoogleScholarGoogle Scholar | 10570001PubMed |

Bravo, J. A., Montanero, J., Calero, R., and Roy, T. J. (2014). Influence of season and reproductive management on the morphometry of ram sperm head. Small Rumin. Res. 119, 114–119.
Influence of season and reproductive management on the morphometry of ram sperm head.Crossref | GoogleScholarGoogle Scholar |

Coloma, M. A., Toledano-Diaz, A., Castano, C., Velazquez, R., Gomez-Brunet, A., Lopez-Sebastian, A., and Santiago-Moreno, J. (2011). Seasonal variation in reproductive physiological status in the Iberian ibex (Capra pyrenaica) and its relationship with sperm freezability. Theriogenology 76, 1695–1705.
Seasonal variation in reproductive physiological status in the Iberian ibex (Capra pyrenaica) and its relationship with sperm freezability.Crossref | GoogleScholarGoogle Scholar | 21855981PubMed |

Curry, M. R. (2000). Cryopreservation of semen from domestic livestock. Rev. Reprod. 5, 46–52.
Cryopreservation of semen from domestic livestock.Crossref | GoogleScholarGoogle Scholar | 10711735PubMed |

Darin-Bennett, A., and White, I. G. (1977). Influence of the cholesterol content of mammalian spermatozoa on susceptibility to cold-shock. Cryobiology 14, 466–470.
Influence of the cholesterol content of mammalian spermatozoa on susceptibility to cold-shock.Crossref | GoogleScholarGoogle Scholar | 560945PubMed |

Domínguez, M. P., Falcinelli, A., Hozbor, F., Sánchez, E., Cesari, A., and Alberio, R. H. (2008). Seasonal variations in the composition of ram seminal plasma and its effect on frozen–thawed ram sperm. Theriogenology 69, 564–573.
Seasonal variations in the composition of ram seminal plasma and its effect on frozen–thawed ram sperm.Crossref | GoogleScholarGoogle Scholar | 18242682PubMed |

Esteso, M. C., Fernandez-Santos, M. R., Soler, A. J., and Garde, J. J. (2003). Head dimensions of cryopreserved red deer spermatozoa are affected by thawing procedure. Cryo Lett. 24, 261–268.

Esteso, M. C., Soler, A. J., Fernandez-Santos, M. R., Quintero-Moreno, A. A., and Garde, J. J. (2006). Functional significance of the sperm head morphometric size and shape for determining freezability in Iberian red deer (Cervus elaphus hispanicus) epididymal sperm samples. J. Androl. 27, 662–670.
Functional significance of the sperm head morphometric size and shape for determining freezability in Iberian red deer (Cervus elaphus hispanicus) epididymal sperm samples.Crossref | GoogleScholarGoogle Scholar | 16728722PubMed |

Esteso, M. C., Rodriguez, E., Toledano-Diaz, A., Castano, C., Pradiee, J., Lopez-Sebastian, A., and Santiago-Moreno, J. (2015). Descriptive analysis of sperm head morphometry in Iberian ibex (Capra pyrenaica): optimum sampling procedure and staining methods using Sperm-Class Analyzer®. Anim. Reprod. Sci. 155, 42–49.
Descriptive analysis of sperm head morphometry in Iberian ibex (Capra pyrenaica): optimum sampling procedure and staining methods using Sperm-Class Analyzer®.Crossref | GoogleScholarGoogle Scholar | 25721563PubMed |

Gravance, C. G., Vishwanath, R., Pitt, C., Garner, D. L., and Casey, P. J. (1998). Effects of cryopreservation on bull sperm head morphometry. J. Androl. 19, 704–709.
| 9876021PubMed |

Hales, D. B. (2001). Editorial: gonadal-specific transcription factors – gata (go) 4 it! Endocrinology 142, 974–976.
Editorial: gonadal-specific transcription factors – gata (go) 4 it!Crossref | GoogleScholarGoogle Scholar | 11181508PubMed |

He, Y., Wang, K., Zhao, X., Zhang, Y., Ma, Y., and Hu, J. (2016). Differential proteome association study of freeze–thaw damage in ram sperm. Cryobiology 72, 60–68.
Differential proteome association study of freeze–thaw damage in ram sperm.Crossref | GoogleScholarGoogle Scholar | 26617253PubMed |

Hochereau-de Reviers, M. T., Monet-Kuntz, C., and Courot, M. (1987). Spermatogenesis and Sertoli cell numbers and function in rams and bulls. J. Reprod. Fertil. Suppl. 34, 101–114.
| 3305912PubMed |

Hodgson, Y. M., Irby, D. C., Kerr, J. B., and de Kretser, D. M. (1979). Studies of the structure and function of the Sertoli cell in a seasonally breeding rodent. Biol. Reprod. 21, 1091–1098.
Studies of the structure and function of the Sertoli cell in a seasonally breeding rodent.Crossref | GoogleScholarGoogle Scholar | 574778PubMed |

Holt, W. V. (2001). Germplasm cryopreservation in elephants and wild ungulates. In ‘Cryobanking the Genetic Resource. Wildlife Conservation for the Future?’. (Eds P. F. Watson and W. V. Holt.) pp. 319–348. (Taylor and Francis: London.)

Holt, W. V., and Pickard, A. R. (1999). Role of reproductive technologies and genetic resource banks in animal conservation. Rev. Reprod. 4, 143–150.
Role of reproductive technologies and genetic resource banks in animal conservation.Crossref | GoogleScholarGoogle Scholar | 10521151PubMed |

Hötzel, M. J., Markey, C. M., Walkden-Brown, S. W., Blackberry, M. A., and Martin, G. B. (1998). Morphometric and endocrine analyses of the effects of nutrition on the testis of mature Merino rams. J. Reprod. Fertil. 113, 217–230.
Morphometric and endocrine analyses of the effects of nutrition on the testis of mature Merino rams.Crossref | GoogleScholarGoogle Scholar | 9861162PubMed |

Janett, F., Thun, R., Bettschen, S., Burger, D., and Hassig, M. (2003). Seasonal changes of semen quality and freezability in Franches–Montagnes stallions. Anim. Reprod. Sci. 77, 213–221.
Seasonal changes of semen quality and freezability in Franches–Montagnes stallions.Crossref | GoogleScholarGoogle Scholar | 12695055PubMed |

Jiménez, R., Burgos, M., and Barrionuevo, F. J. (2015). Circannual testis changes in seasonally breeding mammals. Sex Dev. 9, 205–215.
Circannual testis changes in seasonally breeding mammals.Crossref | GoogleScholarGoogle Scholar | 26375035PubMed |

Jones, R. E. (1997). Synthesis of ether lipids and phosphatidylethanolamine by ejaculated human spermatozoa. Arch. Androl. 38, 181–189.
Synthesis of ether lipids and phosphatidylethanolamine by ejaculated human spermatozoa.Crossref | GoogleScholarGoogle Scholar | 9140614PubMed |

Jones, R. (1998). Plasma membrane structure and remodelling during sperm maturation in the epididymis. J. Reprod. Fertil. Suppl. 53, 73–84.
| 10645268PubMed |

Koivisto, M. B., Costa, M. T. A., Perri, S. H. V., and Vicente, W. R. R. (2009). The effect of season on semen characteristics and freezability in Bos indicus and Bos taurus bulls in the southeastern region of Brazil. Reprod. Domest. Anim. 44, 587–592.
The effect of season on semen characteristics and freezability in Bos indicus and Bos taurus bulls in the southeastern region of Brazil.Crossref | GoogleScholarGoogle Scholar | 19090817PubMed |

Kus, I., Akpolat, N., Oner, H., Ayar, A., Pekmez, H., Ozen, O. A., and Sarsilmaz, M. (2003). The effects of photoperiod on testes in rat: a morphometric and immunohistochemical study. Neuroendocrinol. Lett. 24, 209–214.
| 14523359PubMed |

Ladha, S. (1998). Lipid heterogeneity and membrane fluidity in a highly polarized cell, the mammalian spermatozoon. J. Membr. Biol. 165, 1–10.
Lipid heterogeneity and membrane fluidity in a highly polarized cell, the mammalian spermatozoon.Crossref | GoogleScholarGoogle Scholar | 9705977PubMed |

Leahy, T., and Gadella, B. M. (2011). Sperm surface changes and physiological consequences induced by sperm handling and storage. Reproduction 142, 759–778.
Sperm surface changes and physiological consequences induced by sperm handling and storage.Crossref | GoogleScholarGoogle Scholar | 21964828PubMed |

Ledesma, A., Fernandez-Alegre, E., Cano, A., Hozbor, F., Martinez-Pastor, F., and Cesari, A. (2016). Seminal plasma proteins interacting with sperm surface revert capacitation indicators in frozen–thawed ram sperm. Anim. Reprod. Sci. 173, 35–41.
Seminal plasma proteins interacting with sperm surface revert capacitation indicators in frozen–thawed ram sperm.Crossref | GoogleScholarGoogle Scholar | 27570190PubMed |

Lincoln, G. A. (1998). Reproductive seasonality and maturation throughout the complete life-cycle in the mouflon ram (Ovis musimon). Anim. Reprod. Sci. 53, 87–105.
Reproductive seasonality and maturation throughout the complete life-cycle in the mouflon ram (Ovis musimon).Crossref | GoogleScholarGoogle Scholar | 9835369PubMed |

Lincoln, G. A., Lincoln, C. E., and McNeilly, A. S. (1990). Seasonal cycles in the blood plasma concentration of FSH, inhibin and testosterone, and testicular size in rams of wild, feral and domesticated breeds of sheep. J. Reprod. Fertil. 88, 623–633.
Seasonal cycles in the blood plasma concentration of FSH, inhibin and testosterone, and testicular size in rams of wild, feral and domesticated breeds of sheep.Crossref | GoogleScholarGoogle Scholar | 2109070PubMed |

Liu, M., Cao, G., Zhang, Y., Qu, J., Li, W., Wan, X., Li, Y.-X., Zhang, Z., Wang, Y.-L., and Gao, F. (2016). Changes in the morphology and protein expression of germ cells and Sertoli cells in plateau pikas testes during non-breeding season. Sci. Rep. 6, 22697.
Changes in the morphology and protein expression of germ cells and Sertoli cells in plateau pikas testes during non-breeding season.Crossref | GoogleScholarGoogle Scholar | 26939551PubMed |

Manjunath, P., Bergeron, A., Lefebvre, J., and Fan, J. (2007). Seminal plasma proteins: functions and interaction with protective agents during semen preservation. Soc. Reprod. Fertil. Suppl. 65, 217–228.
| 17644964PubMed |

McClusky, L. M. (2005). Stage and season effects on cell cycle and apoptotic activities of germ cells and Sertoli cells during spermatogenesis in the spiny dogfish (Squalus acanthias). Reproduction 129, 89–102.
Stage and season effects on cell cycle and apoptotic activities of germ cells and Sertoli cells during spermatogenesis in the spiny dogfish (Squalus acanthias).Crossref | GoogleScholarGoogle Scholar | 15615901PubMed |

Mendis-Handagama, S. M., Zirkin, B. R., and Ewing, L. L. (1988). Comparison of components of the testis interstitium with testosterone secretion in hamster, rat, and guinea pig testes perfused in vitro. Am. J. Anat. 181, 12–22.
Comparison of components of the testis interstitium with testosterone secretion in hamster, rat, and guinea pig testes perfused in vitro.Crossref | GoogleScholarGoogle Scholar | 3348144PubMed |

Mocé, E., Blanch, E., Tomás, C., and Graham, J. K. (2010). Use of cholesterol in sperm cryopreservation: present moment and perspectives to future. Reprod. Domest. Anim. 45, 57–66.
Use of cholesterol in sperm cryopreservation: present moment and perspectives to future.Crossref | GoogleScholarGoogle Scholar | 20591066PubMed |

Molkentin, J. D., Lin, Q., Duncan, S. A., and Olson, E. N. (1997). Requirement of the transcription factor GATA4 for heart tube formation and ventral morphogenesis. Genes Dev. 11, 1061–1072.
Requirement of the transcription factor GATA4 for heart tube formation and ventral morphogenesis.Crossref | GoogleScholarGoogle Scholar | 9136933PubMed |

Mortimer, S. T. (2000). CASA – practical aspects. J. Androl. 21, 515–524.
| 10901437PubMed |

Mortimer, S. T., and Maxwell, W. M. (1999). Kinematic definition of ram sperm hyperactivation. Reprod. Fertil. Dev. 11, 25–30.
Kinematic definition of ram sperm hyperactivation.Crossref | GoogleScholarGoogle Scholar | 10680999PubMed |

Nakanishi, Y., and Shiratsuchi, A. (2004). Phagocytic removal of apoptotic spermatogenic cells by Sertoli cells: mechanisms and consequences. Biol. Pharm. Bull. 27, 13–16.
Phagocytic removal of apoptotic spermatogenic cells by Sertoli cells: mechanisms and consequences.Crossref | GoogleScholarGoogle Scholar | 14709891PubMed |

Pelletier, J., Garnier, D. H., de Reviers, M. M., Terqui, M., and Ortavant, R. (1982). Seasonal variation in LH and testosterone release in rams of two breeds. J. Reprod. Fertil. 64, 341–346.
Seasonal variation in LH and testosterone release in rams of two breeds.Crossref | GoogleScholarGoogle Scholar | 7069655PubMed |

Pintus, E., Ros-Santaella, J. L., and Garde, J. J. (2015). Variation of spermatogenic and Sertoli cell number detected by fine needle aspiration cytology (FNAC) in Iberian red deer during and out of the breeding season. Reprod. Fertil. Dev. 27, 812–822.
Variation of spermatogenic and Sertoli cell number detected by fine needle aspiration cytology (FNAC) in Iberian red deer during and out of the breeding season.Crossref | GoogleScholarGoogle Scholar | 25483787PubMed |

Pradiee, J., O’Brien, E., Esteso, M. C., Castano, C., Toledano-Diaz, A., Lopez-Sebastian, A., Marcos-Beltran, J. L., Vega, R. S., Guillamon, F. G., Martinez-Nevado, E., Guerra, R., and Santiago-Moreno, J. (2016). Effect of shortening the prefreezing equilibration time with glycerol on the quality of chamois (Rupicapra pyrenaica), ibex (Capra pyrenaica), mouflon (Ovis musimon) and aoudad (Ammotragus lervia) ejaculates. Anim. Reprod. Sci. 171, 121–128.
Effect of shortening the prefreezing equilibration time with glycerol on the quality of chamois (Rupicapra pyrenaica), ibex (Capra pyrenaica), mouflon (Ovis musimon) and aoudad (Ammotragus lervia) ejaculates.Crossref | GoogleScholarGoogle Scholar | 27346588PubMed |

Ramaswamy, S., and Weinbauer, G. F. (2014). Endocrine control of spermatogenesis: role of FSH and LH/ testosterone. Spermatogenesis 4, e996025.
Endocrine control of spermatogenesis: role of FSH and LH/ testosterone.Crossref | GoogleScholarGoogle Scholar | 26413400PubMed |

Ramos-Vara, J. A., and Miller, M. A. (2009). Immunohistochemical evaluation of GATA-4 in canine testicular tumors. Vet. Pathol. 46, 893–896.
Immunohistochemical evaluation of GATA-4 in canine testicular tumors.Crossref | GoogleScholarGoogle Scholar | 19429994PubMed |

Salmon, V. M., Leclerc, P., and Bailey, J. L. (2016). Cholesterol-loaded cyclodextrin increases the cholesterol content of goat sperm to improve cold and osmotic resistance and maintain sperm function after cryopreservation. Biol. Reprod. 94, 85.
Cholesterol-loaded cyclodextrin increases the cholesterol content of goat sperm to improve cold and osmotic resistance and maintain sperm function after cryopreservation.Crossref | GoogleScholarGoogle Scholar | 26888968PubMed |

Sancho, M., Perez-Sanchez, F., Tablado, L., de Monserrat, J. J., and Soler, C. (1998). Computer assisted morphometric analysis of ram sperm heads: evaluation of different fixative techniques. Theriogenology 50, 27–37.
Computer assisted morphometric analysis of ram sperm heads: evaluation of different fixative techniques.Crossref | GoogleScholarGoogle Scholar | 10734471PubMed |

Sanford, L. M., Price, C. A., Leggee, D. G., Baker, S. J., and Yarney, T. A. (2002). Role of FSH, numbers of FSH receptors and testosterone in the regulation of inhibin secretion during the seasonal testicular cycle of adult rams. Reproduction 123, 269–280.
Role of FSH, numbers of FSH receptors and testosterone in the regulation of inhibin secretion during the seasonal testicular cycle of adult rams.Crossref | GoogleScholarGoogle Scholar | 11866694PubMed |

Santiago-Moreno, J., Lopez-Sebastian, A., Gonzalez-Bulnes, A., Gomez-Brunet, A., and Chemineau, P. (2000). Seasonal changes in ovulatory activity, plasma prolactin, and melatonin concentrations, in mouflon (Ovis gmelini musimon) and Manchega (Ovis aries) ewes. Reprod. Nutr. Dev. 40, 421–430.
Seasonal changes in ovulatory activity, plasma prolactin, and melatonin concentrations, in mouflon (Ovis gmelini musimon) and Manchega (Ovis aries) ewes.Crossref | GoogleScholarGoogle Scholar | 11140814PubMed |

Santiago-Moreno, J., Gomez-Brunet, A., Gonzalez-Bulnes, A., Toledano-Diaz, A., Malpaux, B., and Lopez-Sebastian, A. (2005). Differences in reproductive pattern between wild and domestic rams are not associated with inter-specific annual variations in plasma prolactin and melatonin concentrations. Domest. Anim. Endocrinol. 28, 416–429.
Differences in reproductive pattern between wild and domestic rams are not associated with inter-specific annual variations in plasma prolactin and melatonin concentrations.Crossref | GoogleScholarGoogle Scholar | 15826776PubMed |

Santiago-Moreno, J., Castano, C., Toledano-Diaz, A., Esteso, M. C., Lopez-Sebastian, A., Guerra, R., Ruiz, M. J., Mendoza, N., Luna, C., Cebrian-Perez, J. A., and Hildebrandt, T. B. (2013). Cryopreservation of aoudad (Ammotragus lervia sahariensis) sperm obtained by transrectal ultrasound-guided massage of the accessory sex glands and electroejaculation. Theriogenology 79, 383–391.
Cryopreservation of aoudad (Ammotragus lervia sahariensis) sperm obtained by transrectal ultrasound-guided massage of the accessory sex glands and electroejaculation.Crossref | GoogleScholarGoogle Scholar | 23158213PubMed |

Sarli, G., Benazzi, C., Preziosi, R., and Marcato, P. S. (1994). Proliferative activity assessed by anti-PCNA and Ki67 monoclonal antibodies in canine testicular tumours. J. Comp. Pathol. 110, 357–368.
Proliferative activity assessed by anti-PCNA and Ki67 monoclonal antibodies in canine testicular tumours.Crossref | GoogleScholarGoogle Scholar | 7914524PubMed |

Schanbacher, B. D., and Ford, J. J. (1979). Photoperiodic regulation of ovine spermatogenesis: relationship to serum hormones. Biol. Reprod. 20, 719–726.
Photoperiodic regulation of ovine spermatogenesis: relationship to serum hormones.Crossref | GoogleScholarGoogle Scholar | 454762PubMed |

Smith, J. F., Parr, J., Murray, G. R., McDonald, R. M., and Lee, R.-F. (1999). Seasonal changes in the protein content and composition of ram seminal plasma. Proc. N.Z. Soc. Anim. Prod. 59, 223–225.

Steger, K., Aleithe, I., Behre, H., and Bergmann, M. (1998). The proliferation of spermatogonia in normal and pathological human seminiferous epithelium: an immunohistochemical study using monoclonal antibodies against Ki-67 protein and proliferating cell nuclear antigen. Mol. Hum. Reprod. 4, 227–233.
The proliferation of spermatogonia in normal and pathological human seminiferous epithelium: an immunohistochemical study using monoclonal antibodies against Ki-67 protein and proliferating cell nuclear antigen.Crossref | GoogleScholarGoogle Scholar | 9570268PubMed |

Tarulli, G. A., Stanton, P. G., Lerchl, A., and Meachem, S. J. (2006). Adult Sertoli cells are not terminally differentiated in the Djungarian hamster: effect of FSH on proliferation and junction protein organization. Biol. Reprod. 74, 798–806.
Adult Sertoli cells are not terminally differentiated in the Djungarian hamster: effect of FSH on proliferation and junction protein organization.Crossref | GoogleScholarGoogle Scholar | 16407497PubMed |

Thurston, L. M., Watson, P. F., Mileham, A. J., and Holt, W. V. (2001). Morphologically distinct sperm subpopulations defined by Fourier shape descriptors in fresh ejaculates correlate with variation in boar semen quality following cryopreservation. J. Androl. 22, 382–394.
| 11330638PubMed |

Toledano-Díaz, A., Santiago-Moreno, J., Gómez-Brunet, A., Pulido-Pastor, A., and López-Sebastián, A. (2007). Horn growth related to testosterone secretion in two wild Mediterranean ruminant species: the Spanish ibex (Capra pyrenaica hispanica) and European mouflon (Ovis orientalis musimon). Anim. Reprod. Sci. 102, 300–307.
Horn growth related to testosterone secretion in two wild Mediterranean ruminant species: the Spanish ibex (Capra pyrenaica hispanica) and European mouflon (Ovis orientalis musimon).Crossref | GoogleScholarGoogle Scholar | 17123755PubMed |

Tuli, R. K., and Holtz, W. (1995). Effect of season on the freezability of Boer goat semen in the northern temperate zone. Theriogenology 43, 1359–1363.
Effect of season on the freezability of Boer goat semen in the northern temperate zone.Crossref | GoogleScholarGoogle Scholar |

van Tilburg, M. F., Salles, M. G. F., Silva, M. M., Moreira, R. A., Moreno, F. B., Monteiro-Moreira, A. C. O., Martins, J. A. M., Candido, M. J. D., Araujo, A. A., and Moura, A. A. A. (2015). Semen variables and sperm membrane protein profile of Saanen bucks (Capra hircus) in dry and rainy seasons of the northeastern Brazil (3 degrees S). Int. J. Biometeorol. 59, 561–573.
Semen variables and sperm membrane protein profile of Saanen bucks (Capra hircus) in dry and rainy seasons of the northeastern Brazil (3 degrees S).Crossref | GoogleScholarGoogle Scholar | 25086569PubMed |

Vincent, J. N., McQuown, E. C., and Notter, D. R. (2000). Duration of the seasonal anestrus in sheep selected for fertility in a fall-lambing system. J. Anim. Sci. 78, 1149–1154.
Duration of the seasonal anestrus in sheep selected for fertility in a fall-lambing system.Crossref | GoogleScholarGoogle Scholar | 10834566PubMed |

Wang, H., Wang, H., Xiong, W., Chen, Y., Ma, Q., Ma, J., Ge, Y., and Han, D. (2006). Evaluation on the phagocytosis of apoptotic spermatogenic cells by Sertoli cells in vitro through detecting lipid droplet formation by Oil Red O staining. Reproduction 132, 485–492.
Evaluation on the phagocytosis of apoptotic spermatogenic cells by Sertoli cells in vitro through detecting lipid droplet formation by Oil Red O staining.Crossref | GoogleScholarGoogle Scholar | 16940289PubMed |

Yanagimachi, R. (1970). The movement of golden hamster spermatozoa before and after capacitation. J. Reprod. Fertil. 23, 193–196.
The movement of golden hamster spermatozoa before and after capacitation.Crossref | GoogleScholarGoogle Scholar | 5472441PubMed |

Yang, C., Zhang, J., Ding, M., Xu, K., Li, L., Mao, L., and Zheng, J. (2018). Ki67 targeted strategies for cancer therapy. Clin. Transl. Oncol. 20, 570–575.
Ki67 targeted strategies for cancer therapy.Crossref | GoogleScholarGoogle Scholar | 29058263PubMed |

Yániz, J. L., Soler, C., and Santolaria, P. (2015). Computer assisted sperm morphometry in mammals: a review. Anim. Reprod. Sci. 156, 1–12.
Computer assisted sperm morphometry in mammals: a review.Crossref | GoogleScholarGoogle Scholar | 25802026PubMed |

Young, K. A., and Nelson, R. J. (2001). Mediation of seasonal testicular regression by apoptosis. Reproduction 122, 677–685.
Mediation of seasonal testicular regression by apoptosis.Crossref | GoogleScholarGoogle Scholar | 11690527PubMed |

Zamiri, M. J., Khalili, B., Jafaroghli, M., and Farshad, A. (2010). Seasonal variation in seminal parameters, testicular size, and plasma testosterone concentration in Iranian Moghani rams. Small Rumin. Res. 94, 132–136.
Seasonal variation in seminal parameters, testicular size, and plasma testosterone concentration in Iranian Moghani rams.Crossref | GoogleScholarGoogle Scholar |

Zhang, Z., Shao, S., and Meistrich, M. L. (2007). The radiation-induced block in spermatogonial differentiation is due to damage to the somatic environment, not the germ cells. J. Cell. Physiol. 211, 149–158.
The radiation-induced block in spermatogonial differentiation is due to damage to the somatic environment, not the germ cells.Crossref | GoogleScholarGoogle Scholar | 17167785PubMed |