Immunolocalisation and expression of oxytocin receptors and sex hormone-binding globulin in the testis and epididymis of dogs: correlation with sperm function
Andressa Dalmazzo A , João D. A. Losano A , Daniel S. R. Angrimani A , Isabel V. A. Pereira B , Marcelo D. Goissis A , Maria C. P. Francischini A , Everton Lopes A , Claudia K. Minazaki C , Marcel H. Blank A , Bruno Cogliati B , Ricardo J. G. Pereira A , Valquiria H. Barnabe A and Marcilio Nichi A DA Department of Animal Reproduction, Faculty of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, SP 05508-270, Brazil.
B Department of Pathology, Faculty of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, SP 05508-270, Brazil.
C Paulista University, Campinas, SP 18087-101, Brazil.
D Corresponding author. Email: mnichi@usp.br
Reproduction, Fertility and Development 31(9) 1434-1443 https://doi.org/10.1071/RD18452
Submitted: 11 May 2018 Accepted: 25 February 2019 Published: 3 May 2019
Abstract
The aim of this study was to confirm gene and protein expression of oxytocin receptor (OTR) and sex hormone-binding globulin (SHBG) in the testis and epididymis of dogs, correlating these data with sperm quality and production and testosterone concentrations. Positive correlations were found between OTR and SHBG expression in both the testis and epididymis. Testicular OTR expression was positively associated with plasma membrane and acrosome integrity in canine spermatozoa, whereas SHBG expression in the testis was positively correlated with various sperm characteristics, such as sperm concentration, total and progressive motility, plasma membrane integrity and acrosome integrity. Testicular expression of both OTR and SHBG was negatively correlated with low sperm mitochondrial activity. In the epididymis, SHBG expression was only positively correlated with plasma membrane integrity. Analysis of protein expression revealed that testicular OTR was positively correlated with testosterone concentrations and negatively correlated with the absence of sperm mitochondrial activity. In addition, SHBG expression in the testes was associated with epididymis SHBG expression and morphologically normal cells. Immunohistochemical (IHC) analysis revealed the presence of both OTR and SHBG in testicular smooth muscles and Leydig cells. However, in the epididymis, OTR was only located in smooth muscle cells, whereas neither IHC nor western blotting detected SHBG. Together, the results of this study suggest that OTR and SHBG play key roles in spermatogenesis and sperm maturation, being essential for male reproductive success.
Additional keywords: hormone receptor, spermatozoa, testosterone.
References
Angrimani, D. S. R., Nichi, M., Losano, J. D. A., Lucio, C. F., Veiga, G. A. L., Franco, M. V. J., and Vannucchi, C. I. (2017). Fatty acid content in epididymal fluid and spermatozoa during sperm maturation in dogs. J. Anim. Sci. Biotechnol. 8, 18.| Fatty acid content in epididymal fluid and spermatozoa during sperm maturation in dogs.Crossref | GoogleScholarGoogle Scholar |
Anthony, C. T., Danzo, B. J., and Orgebin-Crist, M. (1984). Investigations on the relationship between sperm fertilizing ability and androgen-binding protein in the restricted rat. Endocrinology 114, 1413–1418.
| Investigations on the relationship between sperm fertilizing ability and androgen-binding protein in the restricted rat.Crossref | GoogleScholarGoogle Scholar | 6538477PubMed |
Assinder, S. J., Carey, M., Parkinson, T., and Nicholson, H. D. (2000). Oxytocin and vasopressin expression in the ovine testis and epididymis: changes with the onset of spermatogenesis. Biol. Reprod. 63, 448–456.
| Oxytocin and vasopressin expression in the ovine testis and epididymis: changes with the onset of spermatogenesis.Crossref | GoogleScholarGoogle Scholar | 10906049PubMed |
Brito, M. M., Angrimani, D. D. S. R., Rui, B. R., Kawai, G. K. V., Losano, J. D. A., and Vannucchi, C. I. (2018). Effect of senescence on morphological, functional and oxidative features of fresh and cryopreserved canine sperm. Aging Male , .
| Effect of senescence on morphological, functional and oxidative features of fresh and cryopreserved canine sperm.Crossref | GoogleScholarGoogle Scholar | 29993302PubMed |
Caldwell, J. D., Song, Y., Englöf, I., Höfle, S., Key, M., and Morris, M. (2003). 5α-Reduced androgens block estradiol–BSA-stimulated release of oxytocin. Brain Res. 976, 259–261.
| 5α-Reduced androgens block estradiol–BSA-stimulated release of oxytocin.Crossref | GoogleScholarGoogle Scholar | 12763261PubMed |
Carmichael, M. S., Humbert, R., Dixen, J., Palmisano, G., Greenleaf, W., and Davidson, J. M. (1987). Plasma oxytocin increases in the human sexual response. J. Clin. Endocrinol. Metab. 64, 27–31.
| Plasma oxytocin increases in the human sexual response.Crossref | GoogleScholarGoogle Scholar | 3782434PubMed |
Carmichael, M. S., Warburton, V. L., Dixen, J., and Davidson, J. M. (1994). Relationships among cardiovascular, muscular, and oxytocin responses during human sexual activity. Arch. Sex. Behav. 23, 59–79.
| Relationships among cardiovascular, muscular, and oxytocin responses during human sexual activity.Crossref | GoogleScholarGoogle Scholar | 8135652PubMed |
Einspanier, A., and Ivell, R. (1997). Oxytocin and oxytocin receptor expression in reproductive tissues of the male marmoset monkey. Biol. Reprod. 56, 416–422.
| Oxytocin and oxytocin receptor expression in reproductive tissues of the male marmoset monkey.Crossref | GoogleScholarGoogle Scholar | 9116141PubMed |
Frayne, J., and Nicholson, H. D. (1994). Regulation of oxytocin production by purified adult rat Leydig cells in vitro: effects of LH, testosterone and lipoproteins. J. Endocrinol. 143, 325–332.
| Regulation of oxytocin production by purified adult rat Leydig cells in vitro: effects of LH, testosterone and lipoproteins.Crossref | GoogleScholarGoogle Scholar | 7829995PubMed |
Frayne, J., and Nicholson, H. D. (1995). Effect of oxytocin on testosterone production by isolated rat leydig cells is mediated via a specific oxytocin receptor. Biol. Reprod. 52, 1268–1273.
| Effect of oxytocin on testosterone production by isolated rat leydig cells is mediated via a specific oxytocin receptor.Crossref | GoogleScholarGoogle Scholar | 7632835PubMed |
Frayne, J., and Nicholson, H. D. (1998). Localization of oxytocin receptors in the human and macaque monkey male reproductive tracts: evidence for a physiological role of oxytocin in the male. Mol. Hum. Reprod. 4, 527–532.
| Localization of oxytocin receptors in the human and macaque monkey male reproductive tracts: evidence for a physiological role of oxytocin in the male. Crossref | GoogleScholarGoogle Scholar | 9665335PubMed |
Gimpl, G., and Fahrenholz, F. (2001). The oxytocin receptor system: structure, function, and regulation. Physiol. Rev. 81, 629–683.
| The oxytocin receptor system: structure, function, and regulation.Crossref | GoogleScholarGoogle Scholar | 11274341PubMed |
Hammond, G. L. (2011). Diverse roles for sex hormone-binding globulin in reproduction. Biol. Reprod. 85, 431–441.
| Diverse roles for sex hormone-binding globulin in reproduction.Crossref | GoogleScholarGoogle Scholar | 21613632PubMed |
Hashimoto, H., Uezono, Y., and Ueta, Y. (2012). Pathophysiological function of oxytocin secreted by neuropeptides: a mini review. Pathophysiology 19, 283–298.
| Pathophysiological function of oxytocin secreted by neuropeptides: a mini review.Crossref | GoogleScholarGoogle Scholar | 22902166PubMed |
Hau, M. (2007). Regulation of male traits by testosterone: implications for the evolution of vertebrate life histories. BioEssays 29, 133–144.
| Regulation of male traits by testosterone: implications for the evolution of vertebrate life histories.Crossref | GoogleScholarGoogle Scholar | 17226801PubMed |
Herbert, Z., Jirikowski, G. F., Petrusz, P., Englöf, I., and Caldwell, J. D. (2003). Distribution of androgen-binding protein in the rat hypothalamo-neurohypophyseal system, co-localization with oxytocin. Brain Res. 992, 151–158.
| Distribution of androgen-binding protein in the rat hypothalamo-neurohypophyseal system, co-localization with oxytocin.Crossref | GoogleScholarGoogle Scholar | 14625054PubMed |
Herbert, Z., Weigel, S., Sendemir, E., Marshall, A., Caldwell, J., Petrusz, P., Peuckert, C., and Jirikowski, G. (2005). Androgen-binding protein is co-expressed with oxytocin in the male reproductive tract. Anat. Histol. Embryol. 34, 286–293.
| Androgen-binding protein is co-expressed with oxytocin in the male reproductive tract.Crossref | GoogleScholarGoogle Scholar | 16159369PubMed |
Hughes, A. M., Everitt, B. J., Lightman, S. L., and Todd, K. (1987). Oxytocin in the central nervous system and sexual behaviour in male rats. Brain Res. 414, 133–137.
| Oxytocin in the central nervous system and sexual behaviour in male rats.Crossref | GoogleScholarGoogle Scholar | 3620914PubMed |
Kirchhoff, C. (2002). The dog as a model to study human epididymal function at a molecular level. Mol. Hum. Reprod. 8, 695–701.
| The dog as a model to study human epididymal function at a molecular level.Crossref | GoogleScholarGoogle Scholar | 12149399PubMed |
Koenig, E. M., Fisher, C., Bernard, H., Wolenski, F. S., Gerrein, J., Carsillo, M., and Meehan, A. (2016). The beagle dog MicroRNA tissue atlas: identifying translatable biomarkers of organ toxicity. BMC Genomics 17, 649.
| The beagle dog MicroRNA tissue atlas: identifying translatable biomarkers of organ toxicity.Crossref | GoogleScholarGoogle Scholar | 27535741PubMed |
Lazaros, L., Xita, N., Kaponis, A., Zikopoulos, K., Sofikitis, N., and Georgiou, I. (2008). Evidence for association of sex hormone-binding globulin and androgen receptor genes with semen quality. Andrologia 40, 186–191.
| Evidence for association of sex hormone-binding globulin and androgen receptor genes with semen quality.Crossref | GoogleScholarGoogle Scholar | 18477206PubMed |
Lucio, C. F., Brito, M. M., Angrimani, D. S. R., Belaz, K. R. A., Morais, D., Zampieri, D., Losano, J. D. A,, Assumpção, M. E. O. A., Nichi, M., Eberlin, M. N., and Vannucchi, C. I. (2017). Lipid composition of the canine sperm plasma membrane as markers of sperm motility. Reprod. Domest. Anim. 52, 208–213.
| Lipid composition of the canine sperm plasma membrane as markers of sperm motility.Crossref | GoogleScholarGoogle Scholar | 27807900PubMed |
Lui, C., Cui, X.-g., Wang, Y.-x., You, Z.-d., and Xu, D.-f. (2010). Association between neuropeptide oxytocin and male infertility. J. Assist. Reprod. Genet. 27, 525–531.
| Association between neuropeptide oxytocin and male infertility.Crossref | GoogleScholarGoogle Scholar | 20711752PubMed |
Murphy, M. R., Seckl, J. R., Burton, S., Checkley, S. A., and Lightman, S. L. (1987). Changes in oxytocin and vasopressin secretion during sexual activity in men. J. Clin. Endocrinol. Metab. 65, 738–741.
| Changes in oxytocin and vasopressin secretion during sexual activity in men.Crossref | GoogleScholarGoogle Scholar | 3654918PubMed |
Ogawa, S., Kudo, S., Kitsunai, Y., and Fukuchi, S. (1980). Increase in oxytocin secretion at ejaculation in male. Clin. Endocrinol. (Oxf) 13, 95–97.
| Increase in oxytocin secretion at ejaculation in male.Crossref | GoogleScholarGoogle Scholar | 7438466PubMed |
Ostrander, E. A., Galibert, F., and Patterson, D. F. (2000). Canine genetics comes of age. Trends Genet. 16, 117–124.
| Canine genetics comes of age.Crossref | GoogleScholarGoogle Scholar | 10689352PubMed |
Pfaffl, M. W. (2001). A new mathematical model for relative quantification in real-time RT–PCR Nucleic Acids Res. , .
| A new mathematical model for relative quantification in real-time RT–PCRCrossref | GoogleScholarGoogle Scholar | 11328886PubMed |
Prapaiwan, N., Manee-in, S., Moonarmart, W., and Srisuwatanasagul, S. (2017). The expressions in oxytocin and sex steroid receptors in the reproductive tissues of normal and unilateral cryptorchid dogs. Theriogenology 100, 59–65.
| The expressions in oxytocin and sex steroid receptors in the reproductive tissues of normal and unilateral cryptorchid dogs.Crossref | GoogleScholarGoogle Scholar | 28708534PubMed |
Selva, D. M., Hogeveen, K. N., Seguchi, K., Tekpetey, F., and Hammond, G. L. (2002). A human sex hormone-binding globulin isoform accumulates in the acrosome during spermatogenesis. J. Biol. Chem. 277, 45291–45298.
| A human sex hormone-binding globulin isoform accumulates in the acrosome during spermatogenesis.Crossref | GoogleScholarGoogle Scholar | 12235141PubMed |
Sendemir, E., Herbert, Z., Sivukhina, E., Zermann, D. H., Arnold, R., and Jirikowski, G. (2008). Colocalization of androgen binding protein, oxytocin receptor, caveolin 1 and proliferation marker p21 in benign prostate hyperplasia. Anat. Histol. Embryol. 37, 325–331.
| Colocalization of androgen binding protein, oxytocin receptor, caveolin 1 and proliferation marker p21 in benign prostate hyperplasia.Crossref | GoogleScholarGoogle Scholar | 18312604PubMed |
Sharma, O. P., and Hays, R. L. (1973). Release of an oxytocic substance following genital stimulation in bulls. J. Reprod. Fertil. 35, 359–362.
| Release of an oxytocic substance following genital stimulation in bulls.Crossref | GoogleScholarGoogle Scholar | 4752149PubMed |
Sinervo, B., Miles, D. B., Frankino, W. A., Klukowski, M., and DeNardo, D. F. (2000). Testosterone, endurance, and Darwinian fitness: natural and sexual selection on the physiological bases of alternative male behaviors in side-blotched lizards. Horm. Behav. 38, 222–233.
| Testosterone, endurance, and Darwinian fitness: natural and sexual selection on the physiological bases of alternative male behaviors in side-blotched lizards.Crossref | GoogleScholarGoogle Scholar | 11104640PubMed |
Soler, C., Yeung, C. H., and Cooper, T. G. (1994). Development of sperm motility patterns in the murine epididymis. Int. J. Androl. 17, 271–278.
| Development of sperm motility patterns in the murine epididymis.Crossref | GoogleScholarGoogle Scholar | 7698853PubMed |
Tablado, L., Pérez-Sánchez, F., and Soler, C. (1996). Is sperm motility maturation affected by static magnetic fields? Environ. Health Perspect. 104, 1212–1216.
| Is sperm motility maturation affected by static magnetic fields?Crossref | GoogleScholarGoogle Scholar | 8959411PubMed |
Thackare, H., Nicholson, H. D., and Whittington, K. (2006). Oxytocin – its role in male reproduction and new potential therapeutic uses. Hum. Reprod. Update 12, 437–448.
| Oxytocin – its role in male reproduction and new potential therapeutic uses.Crossref | GoogleScholarGoogle Scholar | 16436468PubMed |
Walsh, F. (2009). Human–animal bonds I: the relational significance of companion animals. Fam. Process 48, 462–480.
| Human–animal bonds I: the relational significance of companion animals.Crossref | GoogleScholarGoogle Scholar | 19930433PubMed |
Whittington, K., Assinder, S., Parkinson, T., Lapwood, K., and Nicholson, H. (2001). Function and localization of oxytocin receptors in the reproductive tissue of rams. Reproduction 122, 317–325.
| Function and localization of oxytocin receptors in the reproductive tissue of rams.Crossref | GoogleScholarGoogle Scholar | 11467983PubMed |
Whittington, K., Assinder, S., Gould, M., and Nicholson, H. (2004). Oxytocin, oxytocin-associated neurophysin and the oxytocin receptor in the human prostate. Cell Tissue Res. 318, 375–382.
| Oxytocin, oxytocin-associated neurophysin and the oxytocin receptor in the human prostate.Crossref | GoogleScholarGoogle Scholar | 15459766PubMed |