The effects of long-term leptin administration on morphometrical changes of mice testicular tissue

Document Type: Original Article


1 Graduated Student of Veterinary Medicine, Shahid Bahonar University of Kerman, Kerman, Iran

2 Department of Clinical Sciences, Faculty of Veterinary Medicine, Shahid Bahonar University of Kerman, Kerman, Iran

3 Department of Pathobiology, Faculty of Veterinary Medicine, Shahid Bahonar University of Kerman, Kerman, Iran


Objective(s):Leptin is a novel and interesting hormone for anyone trying to lose weight, but its effects on male gonad structure in longitudinal study is unknown. The present study was designed to explore morphometrical changes of mouse testicular tissue after long-term administration of leptin.
Materials and Methods:Thirty healthy mature male mice were randomly assigned to either control (n=15) or treatment (n=15) groups. Leptin was intraperitoneally injected to the treatment group             (0.1 µg/100 µl of physiological saline) once a day for 30 consecutive days, and control animals received normal saline with the same volume and route. Five mice from each experimental group were sacrificed at 15, 30 and 60 days after the beginning of treatments. Left testes were removed, weighted and then fixed in 10% buffered formalin, and stained with hematoxylin and eosine for morphometrical assays.
Results:Except for sertoli cell nucleus diameter, which was affected from 30th day, evaluation of other morphometrical parameters such as Johnsen’s score, meiotic index, spermatogenesis, epithelial height, seminiferous tubules diameter and spermatogonial nucleus diameter revealed significant decrease from 15th day after leptin administration compare to those of the control group (P<0.05). Thus, meiotic index and spermatogonial cell nucleus diameter were two parameters that were further disturbed on 30th day compare to the day 15 (3.09±0.03 vs. 3.23±0.03, P=0.006 and 5.50±0.09 vs. 6.08±0.14, P=0.007, respectively).
Conclusion:Our results showed that long-term administration of leptin could disturb testicular tissue structure and delay spermatogenesis process.


1. Pelleymounter M, Cullen M, Baker M, Hecht R, Winters D, Boone T. Effects of the obese gene product on body weight regulation ob/ob mice. Science 1995; 269:540-543.

2. Zhang Y, Proenca R, Maffei M, Barone M, Leopold L, Friedman JM. Positional cloning of the mouse obese gene and its human homologue. Nature 1994; 372:425-432.

3. Campfield LA, Smith FJ, Guisez Y, Devos R, Burn P. Recombinant mouse OB protein: evidence for a peripheral signal linking adiposity and central neural networks. Science 1995; 269:546-549.

4. Halaas JL, Gajiwala KS, Maffei M, Cohen SL, Chait BT, Rabinowitz D, et al. Weight-reducing effects of

the plasma protein encoded by the obese gene. Science 1995; 269:543-546.

5. Pelleymounter MA, Cullen MJ, Baker MB, Hecht R, Winters D, Boone T, et al. Effects of the obese gene product on body weight regulation in ob/ob mice. Science 1995; 269:540-543.

6. Montague CT, Farooqi IS, Whitehead JP, Soos MA, Rau H, Wareham NJ, et al. Congenital leptin deficiency is associated with severe early-onset obesity in humans. Nature 1997; 387:903-908.

7. Strobel A, Issad T, Camoin L, Ozata M, Strosberg AD. A leptin missense mutation associated with hypogonadism and morbid obesity. Nat Genet 1998; 18:213-215.

8. Shimomura I, Hammer RE, Ikemoto S, Brown MS, Goldstein JL. Leptin reverses insulin resistance and diabetes mellitus in mice with congenital lipodystrophy. Nature 1999; 401:73-76.

9. Shimomura I, Hammer RE, Richardson JA, Ikemoto S, Bashmakov Y, Goldstein JL, et al. Insulin resistance and diabetes mellitus in transgenic mice expressing nuclear SREBP-1c in adipose tissue: model for congenital generalized lipodystrophy. Genes Dev 1998; 12:3182-3194.

10. Ramos CF, Zamoner A. Thyroid hormone and leptin in the testis. Front Endocrinol (Lausanne) 2014; 5:198.

11. Cunningham MJ, Clifton DK, Steiner RA. Leptin's actions on the reproductive axis: perspectives and mechanisms. Biol Reprod 1999; 60:216-222.

12. Smith JT, Mark PJ, Waddell BJ. Developmental increases in plasma leptin binding activity and tissue Ob-Re mRNA expression in the rat. J Endocrinol 2005; 184:535-541.

13. Lee GH, Proenca R, Montez JM, Carroll KM, Darvishzadeh JG, Lee JI, et al. Abnormal splicing of the leptin receptor in diabetic mice. Nature 1996; 379:632-635.

14. Murakami T, Yamashita T, Iida M, Kuwajima M, Shima K. A short form of leptin receptor performs signal transduction. Biochem Biophys Res Commun 1997; 231:26-29.

15. Dieterich KD, Lehnert H. Expression of leptin receptor mRNA and the long form splice variant in human anterior pituitary and pituitary adenoma. Exp Clin Endocrinol Diabetes 1998; 106:522-525.

16. Tartaglia LA. The leptin receptor. J Biol Chem 1997; 272:6093-6096.

17. Glasow A, Haidan A, Hilbers U, Breidert M, Gillespie J, Scherbaum WA, et al. Expression of Ob receptor in normal human adrenals: differential regulation of adrenocortical and adrenomedullary function by leptin. J Clin Endocrinol Metab 1998; 83:4459-4466.

18. Breidert M, Miehlke S, Glasow A, Orban Z, Stolte M, Ehninger G, et al. Leptin and its receptor in normal human gastric mucosa and in Helicobacter pylori-associated gastritis. Scand J Gastroenterol 1999; 34:954-961.

19. Margetic S, Gazzola C, Pegg GG, Hill RA. Leptin: a review of its peripheral actions and interactions. Int J Obes Relat Metab Disord 2002; 26:1407-14033.

20. El-Hefnawy T, Ioffe S, Dym M. Expression of the leptin receptor during germ cell development in the mouse testis. Endocrinology 2000; 141:2624-2630.

21. Caprio M, Fabbrini E, Ricci G, Basciani S, Gnessi L, Arizzi M, et al. Ontogenesis of leptin receptor in rat Leydig cells. Biol Reprod 2003; 68:1199-1207.

22. Hatami-Baroogh L, Razavi S, Zarkesh-Esfahani H, Tavalaee M, Tanhaei S, Ghaedi K, et al. Evaluation of the leptin receptor in human spermatozoa. Reprod Biol Endocrinol 2010; 8:17.

23. Soyupek S, Armagan A, Serel TA, Hoscan MB, Perk H, Karaoz E, et al. Leptin expression in the testicular tissue of fertile and infertile men. Arch Androl 2005; 51:239-246.

24. Jope T, Lammert A, Kratzsch J, Paasch U, Glander HJ. Leptin and leptin receptor in human seminal plasma and in human spermatozoa. Int J Androl 2003; 26:335-341.

25. Aquila S, Gentile M, Middea E, Catalano S, Morelli C, Pezzi V, et al. Leptin secretion by human ejaculated spermatozoa. J Clin Endocrinol Metab 2005; 90:4753-61.

26. De Ambrogi M, Spinaci M, Galeati G, Tamanini C. Leptin receptor in boar spermatozoa. Int J Androl 2007; 30:458-461.

27. Aquila S, Rago V, Guido C, Casaburi I, Zupo S, Carpino A. Leptin and leptin receptor in pig spermatozoa: evidence of their involvement in sperm capacitation and survival. Reproduction 2008; 136:23-32.

28. Haron MN, D'Souza UJ, Jaafar H, Zakaria R, Singh HJ. Exogenous leptin administration decreases sperm count and increases the fraction of abnormal sperm in adult rats. Fertil Steril 2010; 93:322-324.

29. Babaei H, Sivandi S. Caprine cauda epididymal sperm motility pattern in the capacitation medium supplemented with leptin. Comp Clin Pathol 2013; 22:655-660.

30. Joo JK, Joo BS, Kim SC, Choi JR, Park SH, Lee KS. Role of leptin in improvement of oocyte quality by regulation of ovarian angiogenesis. Anim Reprod Sci 2010; 119:329-334.

31. Johnsen SG. Testicular biopsy score count--a method for registration of spermatogenesis in human testes: normal values and results in 335 hypogonadal males. Hormones 1970; 1:2-25.

32. Azizollahi S, Babaei H, Derakhshanfar A, Oloumi MM. Effects of co-administration of dopamine and vitamin C on ischaemia-reperfusion injury after experimental testicular torsion-detorsion in rats. Andrologia 2011; 43:100-105.

33. Kheirandish R, Askari N, Babaei H. Zinc therapy improves deleterious effects of chronic copper administration on mice testes: histopathological evaluation. Andrologia 2014; 46:80-85.

34. Kamohara S, Burcelin R, Halaas JL, Friedman JM, Charron MJ. Acute stimulation of glucose metabolism in mice by leptin treatment. Nature 1997; 389(6649):374-377.

35. Rossetti L, Massillon D, Barzilai N, Vuguin P, Chen W, Hawkins M, et al. Short term effects of leptin on hepatic gluconeogenesis and in vivo insulin action. J Biol Chem 1997; 272:27758-27763.

36. Harris RB, Mitchell TD, Hebert S. Leptin-induced changes in body composition in high fat-fed mice. Exp Biol Med (Maywood) 2003; 228:24-32.

37. Pelleymounter MA, Cullen MJ, Healy D, Hecht R, Winters D, McCaleb M. Efficacy of exogenous recombinant murine leptin in lean and obese 10- to 12-mo-old female CD-1 mice. Am J Physiol 1998; 275:R950-959.

38. Yuan M, Huang G, Li J, Zhang J, Li F, Li K, et al. Hyperleptinemia directly affects testicular matu-ration at different sexual stages in mice, and suppressor of cytokine signaling 3 is involved in this process. Reprod Biol Endocrinol 2014; 12:15.

39. Yamashita T, Murakami T, Otani S, Kuwajima M, Shima K. Leptin receptor signal transduction: OBRa and OBRb of fa type. Biochem Biophys Res Commun 1998; 246:752-759.

40. Caprio M, Isidori AM, Carta AR, Moretti C, Dufau ML, Fabbri A. Expression of functional leptin receptors in rodent Leydig cells. Endocrinology 1999; 140:4939-4947.

41. Zamorano PL, Mahesh VB, De Sevilla LM, Chorich LP, Bhat GK, Brann DW. Expression and localization of the leptin receptor in endocrine and neuroendo-crine tissues of the rat. Neuroendocrinology 1997; 65:223-228.

42. Tena-Sempere M, Pinilla L, Gonzalez LC, Dieguez C, Casanueva FF, Aguilar E. Leptin inhibits testosterone secretion from adult rat testis in vitro. J Endocrinol 1999; 161:211-218.

43. Tena-Sempere M, Pinilla L, Gonzalez LC, Navarro J, Dieguez C, Casanueva FF, et al. In vitro pituitary and testicular effects of the leptin-related synthetic peptide leptin(116-130) amide involve actions both similar to and distinct from those of the native leptin molecule in the adult rat. Eur J Endocrinol 2000; 142:406-410.

44. Kus I, Sarsilmaz M, Canpolat S, Yilmaz B, Kelestimur H, Akpolat N, et al. Immunohistochemical, histological and ultrastructural evaluation of the effects of leptin on testes in mice. Arch Androl 2005; 51:395-405.

45. Banks WA, McLay RN, Kastin AJ, Sarmiento U, Scully S. Passage of leptin across the blood-testis barrier. Am J Physiol 1999; 276(6 Pt 1):E1099-104.

46. Herrid M, O'Shea T, McFarlane JR. Ontogeny of leptin and its receptor expression in mouse testis during the postnatal period. Mol Reprod Dev 2008; 75:874-880.

47. Tena-Sempere M, Pinilla L, Zhang FP, Gonzalez LC, Huhtaniemi I, Casanueva FF, et al. Developmental and hormonal regulation of leptin receptor (Ob-R) messenger ribonucleic acid expression in rat testis. Biol Reprod 2001; 64:634-643.

48. Ishikawa T, Fujioka H, Ishimura T, Takenaka A, Fujisawa M. Expression of leptin and leptin receptor in the testis of fertile and infertile patients. Andrologia 2007; 39:22-27.

 49. Boggio V, Cutrera R, Carbone S, Scacchi P, Ponzo OJ. Leptin inhibits the reproductive axis in adult male Syrian hamsters exposed to long and short photoperiod. Reprod Biol 2013; 13:203-208.

50. O'Donnell L, Robertson KM, Jones ME, Simpson ER. Estrogen and spermatogenesis. Endocr Rev 2001; 22:289-318.

51. Amann RP, Schanbacher BD. Physiology of male reproduction. J Anim Sci 1983; 57 Suppl 2:380-403.

52. Fombonne J, Charrier C, Goddard I, Moyse E, Krantic S. Leptin-mediated decrease of cyclin A2 and increase of cyclin D1 expression: relevance for the control of prepubertal rat Leydig cell division and differentiation. Endocrinology 2007; 148:2126-2137.

53. Landry D, Cloutier F, Martin LJ. Implications of leptin in neuroendocrine regulation of male reproduction. Reprod Biol 2013; 13:1-14.

54. Gottsch ML, Cunningham MJ, Smith JT, Popa SM, Acohido BV, Crowley WF, et al. A role for kisspeptins in the regulation of gonadotropin secretion in the mouse. Endocrinology 2004; 145:4073-4077.

55. Tena-Sempere M. GPR54 and kisspeptin in reproduction. Hum Reprod Update 2006; 12:631-639.