Impact of increased expression of testicular caspase-3 in early ages of rats exposed to Ivermectin
DOI:
https://doi.org/10.5380/avs.v30i4.97634Keywords:
Avermectins; Morphometry; Testis histological analysis; Seminiferous epithelium; ApoptosisAbstract
Studies from our group have revealed that treatment with a therapeutic dose of ivermectin in adult rats increases the expression of caspase-3 and the number of tubules with labeled cells in the tubular lumen, thereby affecting the testicular natural homeostasis process. These effects resulted in impairments of spermatogenesis and spermiogenesis. The present study investigated whether exposure to a therapeutic dose of ivermectin at young and peripubertal ages affected spermatogenesis, spermiogenesis, and the expression of caspase-3 in the testes of rats. Methods: Wistar rats received two doses of 1.0 mg/kg IVM or 0.9% saline solution at 29 and 44 days of age. At 45 days of age, these groups were euthanized, and the rats and testicles were weighed. We conducted a macroscopic evaluation of the animals and a microscopic morphometry evaluation of the seminiferous epithelium. The testis tissue apoptosis was made using Caspase's methodology. The rat's body weight, testicular volume, relative weight, the major and minor axes, and the frequency of Leydig cells did not differ between groups. The epithelium tubular height increased in ivermectin-treated rats compared to the control group. In the IVM group, histopathological analysis revealed disorganization of the germinal epithelium, vacuolation, desquamation of cells into the tubular lumen, and cell nuclei with pyknotic aspects. Ivermectin increased the expression of caspase-3 in median and strongly labeled tubules, suggesting possible testicular damage at young and peripubertal ages in male rats.
References
Bernardi MM, Kirsten TB, Spinosa HS, et al. Ivermectin impairs sexual behavior in sexually naïve, but not sexually experienced male rats. Res Vet Sci 91:77–81,2011. https://doi.org/10.1016/j.rvsc.2010.07.026
Cordeiro F, Ceglio FL, Galvão NA, et al.Does a therapeutical dose of ivermectin impairs testicular homeostasis of rats via excessive apoptosis? Vet Ital 59:145–152,2023. https://doi.org/10.12834/VetIt.2692.19722.2
Cordeiro F, Gonçalves V, Moreira N, et al. Ivermectin acute administration impaired the spermatogenesis and spermiogenesis of adult rats. Res Vet Sci 117:178–186. 2018. https://doi.org/10.1016/j.rvsc.2017.12.013
Gajić Bojić M, Aranđelović J, Škrbić R,et al.Peripheral GABAA receptors - Physiological relevance and therapeutic implications. Pharmacol Ther 266:108759,2025. https://doi.org/10.1016/J.PHARMTHERA.2024.108759
Geigerseder C, Doepner R, Thalhammer A, et al Evidence for a GABAergic System in Rodent and Human Testis: Local GABA Production and GABA Receptors. Neuroendocrinology 77:314–323,2003. https://doi.org/10.1159/000070897
Geigerseder C, Doepner RF, Thalhammer A, et al. Stimulation of TM3 Leydig cell proliferation via GABA(A) receptors: a new role for testicular GABA. Reprod Biol Endocrinol. 24;2:13,2004. https://doi: 10.1186/1477-7827-2-13.
Jurjen, G.; Oosterhuis, E.; Mulder, A. B.; et al. Measuring apoptosis in human spermatozoa: a biological assay for semen quality? 74(2) 2000.10.1016/S0015-0282(00)00623-3
Gürgen, D. G.; Çilingir, Ö. T. Sirvanci, S. Gamma-amino butyric acid immunoreactivity in the testis tissue of Wistar and genetic absence epilepsy rats. Int. J. Morphol. 39(6):1709-1718, 2021.https://intjmorphol.com/abstract/?art_id=8354
Hauet T, Yao Z-X, Bose HS, et al. Peripheral-type benzodiazepine receptor-mediated action of steroidogenic acute regulatory protein on cholesterol entry into Leydig cell mitochondria. Mol Endocrinol 19:540–554,2005. https://doi.org/10.1210/me.2004-0307.
He X, Zhang Y, Yan Y et al. Identification of GABABR2 in rat testis and sperm. J Reprod Dev. Oct;49(5):397-402,2003. doi: 10.1262/jrd.49.397. PMID: 14967916.
Holstein AF, Schulze W, Davidoff M. Understanding spermatogenesis is a prerequisite for treatment. Reprod Biol Endocrinol. 14;1:107,2003. doi: 10.1186/1477-7827-1-107.
Hu Y-T, Tan Z-L, Hirjak D, et al. Brain-wide changes in excitation-inhibition balance of major depressive disorder: a systematic review of topographic patterns of GABA- and glutamatergic alterations. Mol Psychiatry 28:3257–3266,2023. https://doi.org/10.1038/s41380-023-02193-x
Jittamala P, Monteiro W, Smit MR, et al .A systematic review and an individual patient data meta-analysis of ivermectin use in children weighing less than fifteen kilograms: Is it time to reconsider the current contraindication? PLoS Negl Trop Dis 15:2021. https://doi.org/10.1371/journal.pntd.0009144
Johnston GAR, Chebib M, Hanrahan JR, et al. Neurochemicals for the investigation of GABA(C) receptors. Neurochem Res 35:1970–7,2010. https://doi.org/10.1007/s11064-010-0271-7
Oosterhuis GJ, Mulder AB, Kalsbeek-Batenburg E,et al. Measuring apoptosis in human spermatozoa: a biological assay for semen quality? Fertil Steril. 74(2):245-50,2000. doi: 10.1016/s0015-0282(00)00623-3.
Kanbara K, Okamoto K, Nomura S, et al.Cellular localization of GABA and GABAB receptor subunit proteins during spermiogenesis in rat testis. J Androl 26:485–493,2005. https://doi.org/10.2164/jandrol.04185
Kanbara K, Okamoto K, Nomura S, et al.The cellular expression of GABA(A) receptor alpha1 subunit during spermatogenesis in the mouse testis. Histol Histopathol 25:1229–38,2010. https://doi: 10.14670/HH-25.1229.
Kim JJ, Hibbs RE. Direct Structural Insights into GABAA Receptor Pharmacology. Trends Biochem Sci 46:502–517,2021.https://doi: 10.1016/j.tibs.2021.01.011.
Li S, Zhang Y, Liu H, et al Identification and expression of GABAC receptor in rat testis and spermatozoa. Acta Biochim Biophys Sin (Shanghai) 40:761–767,2008. https://doi.org/10.1111/j.1745-7270.2008.00453.x
Lirdi LC, Stumpp T, Sasso-Cerri E, et.al.Amifostine protective effect on cisplatin-treated rat testis. Anatomical Record 291:797–808,2008. https://doi.org/10.1002/ar.20693
Liu J, Zhang K, Cheng L, et al.Progress in understanding the molecular mechanisms underlying the antitumour effects of ivermectin. Drug Des Devel Ther 14:285–296,2020.https://doi: 10.2147/DDDT.S237393.
Lobo Y, Wheller L. A narrative review of the roles of topical permethrin and oral ivermectin in the management of infantile scabies. Australas J Dermatol 62:267–277,2021. https://doi.org/10.1111/ajd.13654
Michałowski MA, Kłopotowski K, Wiera G, et al.Molecular mechanisms of the GABA type A receptor function. Q Rev Biophys 58:e3,2025. https://doi.org/10.1017/S0033583524000179
Miraglia SM, Hayashi H .Histomorphometry of immature rat testis after heating. J Morphol 217:65–74,1993.. https://doi.org/10.1002/jmor.1052170106
Moreira N, Bernardi MM, Spinosa HS. Ivermectin reduces sexual behavior in female rats. Neurotoxicol Teratol 43:33–38,2014. https://doi.org/10.1016/j.ntt.2014.03.003
Moreira N, Sandini TM, Reis-Silva TM, et al. Ivermectin reduces motor coordination, serum testosterone, and central neurotransmitter levels but does not affect sexual motivation in male rats. Reproductive Toxicology 74:195–203,2017. https://doi.org/10.1016/j.reprotox.2017.10.002
Moreira N, Vicente FL, Sandini TM, et al.Effects of ivermectin treatment during prepubertal and pubertal period on sexual parameters and sexual behavior in adulthood in rats. Res Vet Sci 129:21–27,2020. https://doi.org/10.1016/j.rvsc.2019.12.013
Santos R (1999) Morte celular no testículo. Revista Brasileira de Reprodução Animal 23:486–499
Sears SMS, Hewett SJ. Influence of glutamate and GABA transport on brain excitatory/inhibitory balance. Exp Biol Med 246:1069–1083,2021.https://doi: 10.1177/1535370221989263
Shaha C, Tripathi R, Mishra DP. Male germ cell apoptosis: regulation and biology. Philosophical Transactions of the Royal Society B: Biological Sciences 365:1501–1515,2010. https://doi.org/10.1098/rstb.2009.0124
Tawfeek SE, Domouky AM, Abdel-Kareem RH .Protective effect of vitamin C against ivermectin induced nephrotoxicity in different age groups of male Wistar rats: bio-histopathological study. Anat Cell Biol 54:501–517.2021. https://doi.org/10.5115/ACB.21.124
Wilkins AL, Steer AC, Cranswick N et al. Question 1: Is it safe to use ivermectin in children less than five years of age and weighing less than 15 kg? Arch Dis Child 103:514–519,2018. https://doi.org/10.1136/archdischild-2017-314505
Xie W, Li Y, Wang X, Blokhina E et al. GABAB Receptor: Structure, Biological Functions, and Therapy for Diseases. MedComm (2020). 2025 Apr 16;6(5):e70163. doi: 10.1002/mco2.70163
Yin Y, Stahl BC, DeWolf WC, Morgentaler A. p53-mediated germ cell quality control in spermatogenesis. Dev Biol. 1998 ,204(1):165-71. doi: 10.1006/dbio.1998.9074.
Zhang X, Yamamoto N, Soramoto S et al. (2001) Cisplatin-induced germ cell apoptosis in mouse testes. Arch Androl 46:43–9. https://doi.org/10.1080/01485010150211146
Zhang Y, Luo M, Xu W, et al.Avermectin Confers Its Cytotoxic Effects by Inducing DNA Damage and Mitochondria-Associated Apoptosis. J Agric Food Chem 64:6895–902,2016. https://doi.org/10.1021/acs.jafc.6b02812
Downloads
Published
How to Cite
Issue
Section
License

This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.
Authors that wish to publish in AVS agree with the following conditions:
- To keep copyright of the article and allow the AVS to publish the first time. The article will be licensed by Creative Commons - Atribuição 4.0 Internacional allowing the sharing of their work.
- Authors may distribute their work by other channel of distribution (ex.: local or public repository).
- Authors have the permission to publish their work online, using different channels (similar to above), even before the final editorial process.











