Prepubertal male rats with high rates of germ-cell apoptosis present exacerbated rates of germ-cell apoptosis after serotonin depletion
Néstor Méndez Palacios A , María Elena Ayala Escobar B , Maximino Méndez Mendoza A , Rubén Huerta Crispín A , Octavio Guerrero Andrade C , Javier Hernández Meléndez D and Andrés Aragón Martínez D E
+ Author Affiliations
- Author Affiliations
A Facultad de Medicina Veterinaria y Zootecnia, Benemérita Universidad Autónoma de Puebla, CP 75470, Tecamachalco, Puebla, México.
B Laboratorio de Pubertad, Unidad Multidisciplinaria de Investigación, Facultad de Estudios Superiores Zaragoza, Universidad Nacional Autónoma de México, AP 9-020, CP 15000, Distrito Federal, México.
C Universidad Autónoma Metropolitana Xochimilco, CP 04960, Distrito Federal, México.
D Laboratorio de Biología de la Reproducción, Facultad de Ingeniería y Ciencias, Universidad Autónoma de Tamaulipas, AP 337, CP 87149, Ciudad Victoria, Tamaulipas, México.
E Corresponding author. Email: armandres@gmail.com
Reproduction, Fertility and Development 28(6) 806-814 https://doi.org/10.1071/RD13382
Submitted: 11 November 2013 Accepted: 7 October 2014 Published: 20 November 2014
Abstract
Male germ-cell apoptosis occurs naturally and can be increased by exposure to drugs and toxic chemicals. Individuals may have different rates of apoptosis and are likely to also exhibit differential sensitivity to outside influences. Previously, we reported that p-chloroamphetamine (pCA), a substance that inhibits serotonin synthesis, induced germ-cell apoptosis in prepubertal male rats. Here, we identified prepubertal rats with naturally high or low rates of germ-cell apoptosis and evaluated gene expression in both groups. Bax and Shbg mRNA levels were higher in rats with high rates of germ-cell apoptosis. Rats were then treated with pCA and the neuro-hormonal response and gene expression were evaluated. Treatment with pCA induced a reduction in serotonin concentrations but levels of sex hormones and gonadotrophins were not changed. Rats with initially high rates of germ-cell apoptosis had even higher rates of germ-cell apoptosis after treatment with pCA. In rats with high rates of germ-cell apoptosis Bax mRNA expression remained high after treatment with pCA. On the basis of category, an inverse relationship between mRNA expression of Bax and Bcl2, Bax and AR and Bax and Hsd3b2 was found. Here we provide evidence that innate levels of germ-cell apoptosis could be explained by the level of mRNA expression of genes involved with apoptosis and spermatogenesis.
Additional keywords: mRNA expression, p-chloroamphetamine, prepuberty, serotonin synthesis, testis.
References
Aragón, M. A., Ayala, M. E., Marín, M., Avilés, A., Damián-Matzumura, P., and Domínguez, R. (2005). Serotoninergic system blockage in the prepubertal rat inhibits spermatogenesis development. Reproduction 129, 717–727.| Serotoninergic system blockage in the prepubertal rat inhibits spermatogenesis development.Crossref | GoogleScholarGoogle Scholar | 15923387PubMed |
Arïdoğan, I. A., Bayazït, Y., Yaman, M., Ersöz, C., and Doran, S. (2003). Comparison of fine-needle aspiration and open biopsy of testis in sperm retrieval and histopathologic diagnosis. Andrologia 35, 121–125.
| Comparison of fine-needle aspiration and open biopsy of testis in sperm retrieval and histopathologic diagnosis.Crossref | GoogleScholarGoogle Scholar | 12653787PubMed |
Ayala, M. E. (2009). Brain serotonin, psychoactive drugs and effects on reproduction. Cent. Nerv. Syst. Agents Med. Chem. 9, 258–276.
| Brain serotonin, psychoactive drugs and effects on reproduction.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXhsFWisLbJ&md5=0929a8cf36764d4575b2a52f70c1f3d0CAS | 20021359PubMed |
Ayala, M. E., Monroy, J., Morales, L., Castro, M. E., and Domínguez, R. (1998). Effects of a lesion in the dorsal raphe nuclei performed during the juvenile period of the female rat, on puberty. Brain Res. Bull. 47, 211–218.
| Effects of a lesion in the dorsal raphe nuclei performed during the juvenile period of the female rat, on puberty.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaK1M%2FotFCmsg%3D%3D&md5=ab94ce424ed61f2fe4c1bad452b0c4d3CAS | 9865852PubMed |
Barone, F., Aguanno, S., and D’Agostino, A. (2005). Modulation of MAA-induced apoptosis in male germ cells: role of Sertoli cell P/Q-type calcium channels. Reprod. Biol. Endocrinol. 3, 13.
| Modulation of MAA-induced apoptosis in male germ cells: role of Sertoli cell P/Q-type calcium channels.Crossref | GoogleScholarGoogle Scholar | 15840169PubMed |
Billig, H., Furuta, I., Rivier, C., Tapanainen, J., Parvinen, M., and Hsueh, A. J. (1995). Apoptosis in testis germ cells: developmental changes in gonadotrophin dependence and localisation to selective tubule stages. Endocrinology 136, 5–12.
| Apoptosis in testis germ cells: developmental changes in gonadotrophin dependence and localisation to selective tubule stages.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK2MXjt1Knsbw%3D&md5=1f716b369845ebeb34e11573a090b9b9CAS | 7828558PubMed |
Brinkworth, M. H., Weinbauer, G. F., Schlatt, S., and Nieschlag, E. (1995). Identification of male germ cells undergoing apoptosis in adult rats. J. Reprod. Fertil. 105, 25–33.
| Identification of male germ cells undergoing apoptosis in adult rats.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK2MXptVKmtrY%3D&md5=478e512270c86552f5e19583cd9fa990CAS | 7490711PubMed |
Chen, L.-Y., Huang, Y.-L., Liu, M.-Y., Leu, S.-F., and Huang, B.-M. (2003). Effects of amphetamine on steroidogenesis in MA-10 mouse Leydig tumour cells. Life Sci. 72, 1983–1995.
| Effects of amphetamine on steroidogenesis in MA-10 mouse Leydig tumour cells.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3sXht1Glsrk%3D&md5=f8e225d5130f802ae609d398d0ef6073CAS | 12597997PubMed |
Collin, O., Damber, J. E., and Bergh, A. (1996). 5-Hydroxytryptamine – a local regulator of testicular blood flow and vasomotion in rats. J. Reprod. Fertil. 106, 17–22.
| 5-Hydroxytryptamine – a local regulator of testicular blood flow and vasomotion in rats.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK28XmtVCnsA%3D%3D&md5=7356be44bf651b3fbca0df22d3d7b6adCAS | 8667341PubMed |
Committee for the Update of the Guide for the Care and Use of Laboratory Animals (2011). Guide for the Care and Use of Laboratory Animals [Online]. Eighth edition. The National Academic Press. Washington, DC. Available at: http://grants.nih.gov/grants/olaw/Guide-for-the-care-and-use-of-laboratory-animals.pdf [Verified 6 November 2014]
Foulkes, N. S., Mellström, B., Benusiglio, E., and Sassone-Corsi, P. (1992). Developmental switch of CREM function during spermatogenesis: from antagonist to activator. Nature 355, 80–84.
| Developmental switch of CREM function during spermatogenesis: from antagonist to activator.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK3sXhvF2isA%3D%3D&md5=d3f9e280de3eb4d47e44e1ca8ba2d291CAS | 1370576PubMed |
Furu, K., and Klungland, A. (2013). Tzfp represses the androgen receptor in mouse testis. PLoS ONE 8, e62314.
| Tzfp represses the androgen receptor in mouse testis.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3sXntlClsLk%3D&md5=45633b2a729b1e24a0339a0925fcaf6dCAS | 23634227PubMed |
Henriksén, K., Hakovirta, H., and Parvinen, M. (1995). Testosterone inhibits and induces apoptosis in rat seminiferous tubules in a stage-specific manner: in situ quantification in squash preparations after administration of ethane dimethane sulfonate. Endocrinology 136, 3285–3291.
| Testosterone inhibits and induces apoptosis in rat seminiferous tubules in a stage-specific manner: in situ quantification in squash preparations after administration of ethane dimethane sulfonate.Crossref | GoogleScholarGoogle Scholar | 7628362PubMed |
Kalia, S., and Bansal, M. P. (2008). Diethyl maleate-induced oxidative stress leads to testicular germ-cell apoptosis involving Bax and Bcl-2. J. Biochem. Mol. Toxicol. 22, 371–381.
| Diethyl maleate-induced oxidative stress leads to testicular germ-cell apoptosis involving Bax and Bcl-2.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXlsFahtw%3D%3D&md5=454046b9cc6b421ab4a33ff6c8c0b257CAS | 19110998PubMed |
Kosir, R., Juvan, P., Perse, M., Budefeld, T., Majdic, G., Fink, M., Sassone-Corsi, P., and Rozman, D. (2012). Novel insights into the downstream pathways and targets controlled by transcription factors CREM in the testis. PLoS ONE 7, e31798.
| Novel insights into the downstream pathways and targets controlled by transcription factors CREM in the testis.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC38XjsVejurg%3D&md5=78247a97195c9fe8319cb93e65780bfcCAS | 22384077PubMed |
Lysiak, J. J., Turner, S. D., and Turner, T. T. (2000). Molecular pathway of germ-cell apoptosis following ischemia/reperfusion of the rat testis. Biol. Reprod. 63, 1465–1472.
| Molecular pathway of germ-cell apoptosis following ischemia/reperfusion of the rat testis.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3cXnslCktb4%3D&md5=627de838b4627edae4f882f104064d34CAS | 11058553PubMed |
Marshall, O. J. (2004). PerlPrimer: cross-platform, graphical primer design for standard, bisulphite and real-time PCR. Bioinformatics 20, 2471–2472.
| PerlPrimer: cross-platform, graphical primer design for standard, bisulphite and real-time PCR.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2cXos1Gisrc%3D&md5=28ce05652449a988e95d5335e89a6e26CAS | 15073005PubMed |
Meena, R., Misro, M. M., Ghosh, D., and Nandan, D. (2012). Complete sperm suppression induced by dienogest plus testosterone undecanoate is associated with down-regulation in the expression of upstream steroidogenic enzyme genes in rat testis. Contraception 86, 163–171.
| Complete sperm suppression induced by dienogest plus testosterone undecanoate is associated with down-regulation in the expression of upstream steroidogenic enzyme genes in rat testis.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC38XpvFOltrY%3D&md5=c4d7b64696f0af96c9adec5d97491c45CAS | 22264663PubMed |
Pareek, T. K., Joshi, A. R., Sanyal, A., and Dighe, R. R. (2007). Insights into male germ-cell apoptosis due to depletion of gonadotrophins caused by GnRH antagonists. Apoptosis 12, 1085–1100.
| Insights into male germ-cell apoptosis due to depletion of gonadotrophins caused by GnRH antagonists.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXkvFKit7g%3D&md5=d7507eaf0ef8c7c09da53e56f436a4faCAS | 17268770PubMed |
Paxinos, G., and Watson, C. (2007). ‘The Rat Brain in Stereotaxic Coordinates’. (Academic Press: New York.)
Piner, J., Sutherland, M., Millar, M., Turner, K., Newall, D., and Sharpe, R. M. (2002). Changes in vascular dynamics of the adult rat testis leading to transient accumulation of seminiferous tubule fluid after administration of a novel 5-hydroxytryptamine (5-HT) agonist. Reprod. Toxicol. 16, 141–150.
| Changes in vascular dynamics of the adult rat testis leading to transient accumulation of seminiferous tubule fluid after administration of a novel 5-hydroxytryptamine (5-HT) agonist.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD38XivVSms7Y%3D&md5=0648ad8c8e7a6f88b0c3fa0d0014916eCAS | 11955945PubMed |
Rasband, W. S. (2005). ‘ImageJ (computer program)’. (US National Institutes of Health: Bethesda, MD, USA.)
Ruwanpura, S. M., McLachlan, R. I., and Meachem, S. J. (2010). Hormonal regulation of male germ-cell development. J. Endocrinol. 205, 117–131.
| Hormonal regulation of male germ-cell development.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3cXmvVCgs7s%3D&md5=ca69640b3c479797f54f687c6096f837CAS | 20144980PubMed |
Sadate-Ngatchou, P. I., Pouchnik, D. J., and Griswold, M. D. (2004). Follicle-stimulating hormone induced changes in gene expression of murine testis. Mol. Endocrinol. 18, 2805–2816.
| Follicle-stimulating hormone induced changes in gene expression of murine testis.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2cXpslKksb8%3D&md5=b9a7819bbc2d974849230cc7f222ee2cCAS | 15297604PubMed |
Sanders-Bush, E., Bushing, J. A., and Sulser, F. (1972). p-Chloroamphetamine inhibition of cerebral tryptophan hydroxylase. Biochem. Pharmacol. 21, 1501–1510.
| p-Chloroamphetamine inhibition of cerebral tryptophan hydroxylase.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaE387pt1Citw%3D%3D&md5=c317cde7c4dad8eac97f464853464f68CAS | 4260485PubMed |
Sassone-Corsi, P. (1998). Regulating the balance between differentiation and apoptosis: role of CREM in the male germ cells. J. Mol. Med. 76, 811–817.
| Regulating the balance between differentiation and apoptosis: role of CREM in the male germ cells.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1MXitVGh&md5=a33e860afb0ebd431c9a76d843e4d4aeCAS | 9846951PubMed |
Shastry, B. S. (2006). Pharmacogenetics and the concept of individualised medicine. Pharmacogenomics J 6, 16–21.
| Pharmacogenetics and the concept of individualised medicine.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD28XjtVyltw%3D%3D&md5=7a8220045e27a621fb93f52247388c7cCAS | 16302022PubMed |
Sinha Hikim, A. P., and Swerdloff, R. S. (1999). Hormonal and genetic control of germ-cell apoptosis in the testis. Rev. Reprod. 4, 38–47.
| Hormonal and genetic control of germ-cell apoptosis in the testis.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaK1M7lvVerug%3D%3D&md5=983bf87b77f7c00ad32ad16063df6606CAS | 10051101PubMed |
Sinha Hikim, A. P., Lue, Y., Diaz-Romero, M., Yen, P. H., Wang, C., and Swerdloff, R. S. (2003). Deciphering the pathways of germ-cell apoptosis in the testis. J. Steroid Biochem. Mol. Biol. 85, 175–182.
| Deciphering the pathways of germ-cell apoptosis in the testis.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3sXms1Orurg%3D&md5=341f33bfc6d3876f9df27cfdeebb6c40CAS | 12943702PubMed |
Team, R. D. C. (2011). R: A Language and Environment for Statistical Computing. Vienna, Austria. Available at http://www.R-project.org/ [Verified 11 October 2014].
Tinajero, J. C., Fabbri, A., Ciocca, D. R., and Dufau, M. L. (1993). Serotonin secretion from rat Leydig cells. Endocrinology 133, 3026–3029.
| Serotonin secretion from rat Leydig cells.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK2cXks1CltA%3D%3D&md5=3fbf62bf904c3613c4389a366f67ed38CAS | 8243331PubMed |
Tsai, S. C., Chiao, Y. C., Lu, C. C., Doong, M. L., Chen, Y. H., Shih, H. C., Liaw, C., Wang, S. W., and Wang, P. S. (1996). Inhibition by amphetamine of testosterone secretion through a mechanism involving an increase of cyclic AMP production in rat testes. Br. J. Pharmacol. 118, 984–988.
| Inhibition by amphetamine of testosterone secretion through a mechanism involving an increase of cyclic AMP production in rat testes.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK28Xkt1Oqtbw%3D&md5=e28f0ede9c29921b5d1883cbe19869a6CAS | 8799572PubMed |
Tsai, S. C., Chen, J. J., Chiao, Y. C., Lu, C. C., Lin, H., Yeh, J. Y., Lo, M. J., Kau, M. M., Wang, S. W., and Wang, P. S. (1997). The role of cyclic AMP production, calcium channel activation and enzyme activities in the inhibition of testosterone secretion by amphetamine. Br. J. Pharmacol. 122, 949–955.
| The role of cyclic AMP production, calcium channel activation and enzyme activities in the inhibition of testosterone secretion by amphetamine.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK2sXnt1Wlsbw%3D&md5=cd7dd48592d795d8705712e76784ed1cCAS | 9384514PubMed |
