Effect of Melatonin on the Expression of Apoptotic Genes in Vitrified-thawed Spermatogonia Stem Cells Type A of 6-Day-Old Mice

Authors

1 Department of Anatomy, Lorestan University of Medical Sciences, Khoramabad, Iran

2 Physiology Research Center, Ahvaz Jundishapur University of Medical Sciences (AJUMS), Ahvaz, Iran

3 Fertility, Infertility and Perinatology Research Center, Ahvaz Jundishapur University of Medical Sciences (AJUMS), Ahvaz, Iran

4 Department of Immunology, Ahvaz Jundishapur University of Medical Sciences (AJUMS), Ahvaz, Iran

5 Departmentof Medical Genetics, Ahvaz Jundishapur University of Medical Sciences(AJUMS), Ahvaz, Iran

Abstract

 Objective(s):  Being secreted by the pineal gland, melatonin induces cell proliferation in normal cells and induced apoptosis in cancer cells. The purpose of this study was to investigate effects of melatonin on main components and the expression of apoptotic genes in vitrified-thawed testicular germ cells of 6- day–old mice. 
Materials and Methods: Testes of neonate Balb/c mice were vitrified- thawed under standard condition with or without the addition of 100 μM melatonin to both vitrification and thawing solutions. Subsequently, Vitrified-thawed wholetestes were digested under standard condition and spermatogonial stem cells type A were separate in the suspension with CD90.1 (Thy1.1+) micro beads. Extraction of RNA and synthesis of cDNA was performed. Expression levels of apoptotic genes (Fas, P53, BCL-2 and BAX) were determined using Real-time PCR.
Results: With all genes being expressed, level of expression for Fas was higher and for that of P-53 was lower than the remaining genes.
Conclusion: Melatonin may cause apoptosis in cells being damaged under the influence of freezing thawing process. In order to examine the exact effects of melatonin on spermatogonia stem cells apoptosis, additional studies are required.

Keywords


1. Chiarini-Garcia H, Raymer AM, Russell LD. Non-random distribution of spermatogonia in rats: evidence of niches in the seminiferous tubules. Reproduction 2003; 126:669-680.

2. Brougham MF, Kelnar CJ, Sharpe RM, Wallace WH. Male fertility following childhood cancer: current concepts and future therapies. Asian J Androl 2003; 5:325-337.

3. Kamischke A, Jurgens H, Hertle L, Berdel WE, Nieschlag E. Cryopreservation of sperm from adolescents and adults with malignancies. J Androl  2004; 25:586-592.

4. Kelleher S, Wishart SM, Liu PY, Turner L, Di Pierro I, Conway AJ, et al. Long-term outcomes of elective human sperm cryostorage. Hum Reprod 2001; 16:2632-2639.

5. Honaramooz A, Behboodi E, Blash S, Megee SO, Dobrinski I. Germ cell transplantation in goats. Mol Reprod Dev   2003; 64:422-428.

6. Honaramooz A, Megee SO, Dobrinski I. Germ cell transplantation in pigs. Biol Reprod  2002; 66:21-28.

7. Schlatt S, Honaramooz A, Boiani M, Scholer HR, Dobrinski I. Progeny from sperm obtained after ectopic grafting of neonatal mouse testes. Biol Reprod  2003; 68:2331-2335.

8. Honaramooz A, Li MW, Penedo MC, Meyers S, Dobrinski I. Accelerated maturation of primate testis by xenografting into mice. Biol Reprod 2004; 70:1500-1503.

9. Dohle GR. Male infertility in cancer patients: Review of the literature. Int J Urol  2010; 17:327-331.

10. Wyns C, Curaba M, Petit S, Vanabelle B, Laurent P, Wese JF, et al. Management of fertility preservation in prepubertal patients: 5 years' experience at the Catholic University of Louvain. Hum Reprod  2011; 26:737-747.

11. Wyns C, Curaba M, Vanabelle B, Van Langendonckt A, Donnez J. Options for fertility preservation in prepubertal boys. Hum Reprod Update 2010; 16:312-328.

12. Abrishami M, Anzar M, Yang Y, Honaramooz A. Cryopreservation of immature porcine testis tissue to maintain its developmental potential after xenografting into recipient mice. Theriogenology  2009; 73:86-96.

13. Amorim CA, Curaba M, Van Langendonckt A, Dolmans MM, Donnez J. Vitrification as an alternative means of cryopreserving ovarian tissue. Reprod Biomed Online 2011; 23:160-186.

14. Curaba M, Poels J, van Langendonckt A, Donnez J, Wyns C. Can prepubertal human testicular tissue be cryopreserved by vitrification? Fertil Steril  2011; 95:2123 e9-12.

15. Abrishami M. Crypreservation and xenografting of testis tissue Saskatchewan: University of Saskatchewan, Saskatoon; 2009.

16. Bagchi A, Woods EJ, Critser JK. Cryopreservation and vitrification: recent advances in fertility preservation technologies. Expert Rev Med Devices 2008; 5:359-370.

17. Bank HL, Brockbank KG. Basic principles of cryobiology. J Card Surg 1987; 2:137-143.

18. Fuller BJ. Cryoprotectants: the essential antifreezes to protect life in the frozen state. Cryo Letters 2004; 25:375-388.

19. Pegg DE. The history and principles of cryopreservation. Semin Reprod Med  2002; 20:5-13.

20. Pegg DE. Principles of cryopreservation. Methods Mol Biol 2007; 368:39-57.

21. Vaux DI, Korsmyer SJ. Cell death in development. Cell  1999; 96:245-254.

22. Gholami MR, Saki G, Hemadi M,  Khodadadi A, Mohammadi-Asl J. Supplementation vitrified-thawed media with melatonin do not protecting immature mouse testicular tissue from vitrified-thawed induced injury. Asian J Anim Vet Adv 2012; 7:940-949.

23. Hardeland R, Cardinali DP, Srinivasan V, Spence DW, Brown GM, Pandi-Perumal SR. Melatonin--a pleiotropic, orchestrating regulator molecule. Prog Neurobiol  2011; 93:350-384.

24. Joo SS, Yoo YM. Melatonin induces apoptotic death in LNCaP cells via p38 and JNK pathways: therapeutic implications for prostate cancer. J Pineal Res 2009; 47:8-14.

25. Millan-Plano S, Piedrafita E, Miana-Mena FJ, Fuentes-Broto L, Martinez-Ballarin E, Lopez-Pingarron L, et al. Melatonin and structurally-related compounds protect synaptosomal membranes from free radical damage. Int J Mol Sci  2010; 11:312-328.

26. Reiter RJ, Tan DX, Manchester LC, Paredes SD, Mayo JC, Sainz RM. Melatonin and reproduction revisited. Biol Reprod  2009; 81:445-456.

27. Reiter RJ, Tan DX, Terron MP, Flores LJ, Czarnocki Z. Melatonin and its metabolites: new findings regarding their production and their radical scavenging actions. Acta Biochim Pol  2007; 54:1-9.

28. Paternoster L, Radogna F, Accorsi A, Cristina Albertini M, Gualandi G, Ghibelli L. Melatonin as a modulator of apoptosis in B-lymphoma cells. Ann N Y Acad Sci  2009; 1171:345-349.

29. Radogna F, Paternoster L, Albertini MC, Accorsi A, Cerella C, D'Alessio M, et al. Melatonin as an apoptosis antagonist. Ann N Y Acad Sci  2006; 1090:226-233.

30. Reiter RJ, Korkmaz A, Paredes SD, Manchester LC, Tan DX. Melatonin reduces oxidative/nitrosative stress due to drugs, toxins, metals, and herbicides. Neuro Endocrinol Lett  2008; 29:609-613.

31. Sanchez-Hidalgo M, Lu Z, Tan DX, Maldonado MD, Reiter RJ, Gregerman RI. Melatonin inhibits fatty acid-induced triglyceride accumulation in ROS17/2.8 cells: implications for osteoblast differentiation and osteoporosis. Am J Physiol Regul Integr Comp Physiol 2007; 292:R2208-15.

32. Winczyk K, Lawnicka H, Pawlikowski M, Kunert-Radek J, Karasek M. Growth-inhibitory action of melatonin and thiazolidinedione derivative CGP 52608 on murine 16/C breast cancer cells. Neuro Endocrinol Lett  2006; 27:351-354.

33. Hemadi M, Zargar M, Sobhani A. Assessment of morphological and functional changes in neonate vitrified testis grafts after host treatment with melatonin. Folia Morphol (Warsz)  2011; 70:95-102.

 

34. Milazzo JP, Vaudreuil L, Cauliez B, Gruel E, Masse L, Mousset-Simeon N, et al. Comparison of conditions for cryopreservation of testicular tissue from immature mice. Hum Reprod  2008; 23:17-28.

35. Kerr JF, Wyllie AH, Currie AR. Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer  1972; 26:239-257.

36. Lanoix D, Lacasse AA, Reiter RJ, Vaillancourt C. Melatonin: the smart killer: the human trophoblast as a model. Mol Cell Endocrinol  2012; 348:1-11.

37. Deigner HP, Haberkorn U, Kinscherf R. Apoptosis modulators in the therapy of neurodegenerative diseases. Expert Opin Investig Drugs  2000; 9:747-764.

38. Oberholzer C, Oberholzer A, Clare-Salzler M, Moldawer LL. Apoptosis in sepsis: a new target for therapeutic exploration. FASEB J 2001; 15:879-892.

39. Asadi E, Jahanshahi M, Golalipour MJ. Effect of Vitamin E on Oocytes Apoptosis in Nicotine-Treated Mice. Iran J Basic Med Sci  2012; 15:880-884.

40. Babaei-Balderlou F, Zare S, Heidari R, Farrokhi F. Effects of Melatonin and Vitamin E on Peripheral Neuropathic Pain in Streptozotocin-Induced Diabetic Rats. Iran J Basic Med Sci 2010; 13:118-123.

41. Eck KM, Yuan L, Duffy L, Ram PT, Ayettey S, Chen I, et al. A sequential treatment regimen with melatonin and all-trans retinoic acid induces apoptosis in MCF-7 tumour cells. Br J Cancer  1998; 77:2129-2137.

42. Post A, Holsboer F, Behl C. Induction of NF-kappaB activity during haloperidol-induced oxidative toxicity in clonal hippocampal cells: suppression of NF-kappaB and neuroprotection by antioxidants.  J Neurosci 1998; 18:8236-8246.

43. Das A, Belagodu A, Reiter RJ, Ray SK, Banik NL. Cytoprotective effects of melatonin on C6 astroglial cells exposed to glutamate excitotoxicity and oxidative stress. J Pineal Res 2008; 45:117-124.

44. Das A, McDowell M, Pava MJ, Smith JA, Reiter RJ, Woodward JJ, et al. The inhibition of apoptosis by melatonin in VSC4.1 motoneurons exposed to oxidative stress, glutamate excitotoxicity, or TNF-alpha toxicity involves membrane melatonin receptors. J Pineal Res  2010; 48:157-169.

45. Mediavilla MD, Sanchez-Barcelo EJ, Tan DX, Manchester L, Reiter RJ. Basic mechanisms involved in the anti-cancer effects of melatonin. Curr Med Chem 2010; 17:4462-4481.

46. Mauduit C, Siah A, Foch M, Chapet O, Clippe S, Gerard JP, Benahmed M. Differential expression of growth factors in irradiated mouse testes. Int J Radiat Oncol Biol Phys 2001; 50: 203-212.