Examination of the immunohistochemical localization and gene expression by RT-PCR of the oxytocin receptor in diabetic and non-diabetic mouse testis

Document Type: Original Article

Authors

1 Veterinary Faculty, Histology and Embryology Department, Kafkas University, 36100, Kars, Turkey

2 Medicine Faculty, Histology and Embryology Department, Kafkas University, 36100, Kars, Turkey

Abstract

Objective(s): The aim of this study was to determine Oxytocin receptor (OTR) gene expression and localization in diabetic and non-diabetic mouse testes by RT-PCR and immunohistochemistry, respectively.
Materials and Methods: In this study, 18 male BALB/c mice (8–12 weeks old) were used and divided into three groups: diabetic, sham, and control. Streptozotocin (STZ) was applied to the diabetic group and sodium citrate was administered to the sham group in the same way, however, the control group was left untouched. The testicular tissues were removed on the thirtieth day of testing; the right testis tissues were passed through a routine histologic process and sections were stained with H&E and PAS staining techniques. The avidin-biotin-peroxidase method was applied to determine OTR immunoreactivity, while the left testis tissues were used for RT-PCR.
Results: It was found that the body weight had decreased in the diabetic group and the diameter of the seminiferous tubules in the said group was shorter than those of the other groups. There were no obvious differences with regard to the histologic appearance between the groups. The immunohistochemical examination showed that the OTR immunoreactivity was strong in the control and sham groups but weak in the diabetic group, and the immunoreactivity was only seen in the Leydig cells. In addition, the OTR gene expression was lower in the diabetic group than in the other groups.
Conclusion: We concluded that diabetes reduces the OTR expression in the testis. It is suggested that OTR protection should be researched in diabetes for healthy reproduction and sexuality.

Keywords

Main Subjects


1. Kimura T, Makino Y, Saji F, Takemura M, Inoue T, Kikuchi T, et al. Molecular characterization of a cloned human oxytocin receptor. Eur J Endocrinol 1994; 131:385-390.
2. Anjum S, Anuradha A, Krishna A. A possible direct action of oxytocin on spermatogenesis and steroidogenesis in pre-pubertal mouse. Andrologia 2018; e12958: 1-11.
3. Lockard MA, Ebert MS, Bargmann CI. Oxytocin mediated behavior in invertebrates: an evolutionary perspective. Dev Neurobiol 2017; 77: 128-142.
4. Nicholson HD and Jenkın L. Oxytocin and prostatic function. Adv Exp Med Biol 1995; 395:529-538.
5. Roth KI. Diabetes treatment-bridging the divide. N Engl J Med 2007; 356:1499-1501.
6. Gurka MJ, Golden SH, Musani SK, Sims M, Vishnu A, Guo Y, et al. Independent associations between a metabolic syndrome severity score and future diabetes by sex and race: the atherosclerosis risk in communities study and Jackson heart study. Diabetologia 2017; 60:1261-1270.
7. Agbaje IM, Rogers DA, Mcvicar CM, Mcclure N, Atkinson AB, Mallidis C, et al. Insulin dependent diabetes mellitus: implications for male reproductive function. Hum Reprod 2007; 22:1871-1877.
8. Kort HI, Massey JB, Elsner CW, Mitchell-Leef D, Shapiro DB, Witt MA, et al. Impact of body mass index values on sperm quantity and quality. J Androl 2006; 27:450-452.
9. Bingol SA ve Kocamıs H. The gene expression profile by RT-PCR and Immunohistochemical expression pattern of catalase in the kidney tissue of both health and diabetic mice. Kafkas Univ Vet Fak Derg  2010; 16:825-834.
10.    Kanitkar M, Bhonde R. Existence of islet regenerating factors within the pancreas. The Rev Diabet Stud 2004; 1:185-192.
11.    Shu S, Ju G, Fan L. The glucose oxidase-DAB–nickel in peroxidase histochemistry of the   nervous system.  Neurosci Lett 1988; 85:169-171.
12.    Kasahara Y, Sato K, Takayanagi Y, Mizukami H, Ozawa K, Hidema S,  et al. Oxytocin receptor in the hypothalamus is sufficient to rescue normal thermoregulatory function in male oxytocin receptor knockout mice. Endocrinology 2013; 154:4305-4315.
13.    Göçmen C, Seçilmiş A, Kumcu EK, Ertuğ PU, Önder S, Dikmen A, et al. Effects of  vitamin E and sodium selenate on neurogenic and endothelial relaxation of corpus cavernosum in the diabetic mouse. Eur J Pharmacol 2000; 398:93-98.
14.    Wada J, Zhang H, Tsuchiyama Y, Hiragushi K, Hıda K, Shikata K, et al. Gene exspression profile in streptozotosin-induced diabetic mice kidneys undergoing glomerulosclerosis. Kidney Int 2001; 59:1363-1373.
15.    Elis Yildiz S, Deprem T, Karadag Sari E, Bingol SA, Koral Tasci S, Aslan S, et al. Immunohistochemical distribution of leptin in kidney tissues of melatonin treated diabetic rats. Biotech  Histochem 2015; 90:270-277.
16.    Kim NK, Stankovic M, Cushman TT, Goldstein I, Munarrız R, Traish AM.  Streptozotosin- induced diabetes in the rat is associated with changes in vaginal hemodynamics, morphology and biochemikal markers. BMC Physiol 2006; 6:1-9.
17.    Orman D, Vardi N, Ates B, Taslidere E, Elbe H. Aminoguanidine mitigates apoptosis, testicular seminiferous tubules damage, and oxidative stress in streptozotocin-induced diabetic rats. Tissue Cell 2015; 47:284-290.
18.    Haan JB, Stefanovic  N, Paterson DN, Scurr  LL, Croft KD, Mori TA, et al. Kidney expression of glutathione peroxidase-1 is not protective against streptozotocin-induced diabetic nephropathy. Am J Physiol Renal Physiol 2005; 289:F544-F551.
19.    Jelodar G, Khaksar Z, Pourahmadi M. Endocrine profile and testicular histomorphometry in  adult rat offspring of diabetic mothers. J Physiol Sci 2009; 59:377-382.
20.    Cai L, Chen S, Evans T, Deng DX, Mukherjee K, Chakrabarti S. Apoptotic germ-cell death and testicular damage in experimental diabetes: prevention by endothelin antagonism. Urol res 2000; 28:342-347.  
21.    Guneli E, Tugyan K, Ozturk H, Gumustekin M, Cilaker S, Uysal N. Effect of melatonin on testicular damage in   streptozotocin-induced diabetes rats. Eur Surg Res 2008; 40:354-360.
22.    Akkoc H, Kelle I, Tunik S, Erdinc M, Erdinc L, Nergiz Y. Effects of Ethyl Pyruvate on Testicular Damage in Rats with Stretozotocin-Induced Diabetes. Acta Endocrinol 2012; 8:35-45.
23.    Kianifard D, Sadrkhanlou RA, Hasanzadeh S. The ultrastructural changes of the sertoli and leydig cells following streptozotocin induced diabetes. Iran J Basic Med Sci 2012; 15:623-635.
24.    Koroglu P, Erkanli Senturk G, Yucel D, Bingol Ozakpinar O, Uras F, Arbak S. The effect of exogenous oxytocin on streptozotocin (STZ)-induced diabetic adult rat testes. Peptides 2015; 63:47-54.
25.    Erbas O, Korkmaz AH, Oltulu F, Aktug H, Yavasoglu A, Akman L, et al. Oxytocin alleviates cisplatin-induced renal damage in rats. Iran J Basic Med Sci 2014; 17:747-752.
26.    Veening JG, de Jong TR, Waldinger MD, Korte SM, Oliver B. The role of oxytocin in male and female reproductive behavior. Eur J Pharmacol 2015; 753.209-228.
27.    Gossen A, Hahn A, Westphal S, Prinz S, Schultz RT, Gründer G, et al. Oxytocin plasma concentrations after single intranasal oxytocin administration - a study in healthy men. Neuropeptides 2012; 46:211-215.
28.    Frayne J and Nıcholson HD. 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 1998; 4:527-532.
29.    Einspanier A, Ivell R. Oxytocin and receptor expression in reproductive tissues of the male marmoset monkey. Biol Reprod  1997; 56:416-422.
30.    Sanna F, Bratzu J, Argiolas A, Melis MR. Oxytocin induces penile erection and yawning when injected into the bed nucleus of the stria terminalis: Involment of glutamic acid, dopamine, and nitric oxide. Horm Behav 2017; 96:52-61.
31.    Li M, Wang T, Guo S, Rao K, Liu J, Ye Z. Expression of oxcytocin receptor in diabetic rat penis. Andrologia 2012; 44:716-720.
32.    Whittington K, Assinder SJ, Parkinson T, Lapwood KR, Nicholson HD. Function and localization of oxytocin receptors in the reproductive tissue of rams. Reproduction 2001; 122:317-325.
33.    Zhang XH, Filippi S, Vignozzi L, Morelli A, Mancina R, Luconi M, et al. Identification, localization and functional in vitro and in vivo activity of oxytocin receptor in the rat penis. J Endocrinol 2005; 184:567-576.
34.    Prapaiwan N, Manee-in S, Moonarmart W, Srisuwatanasagul S. The expressions in oxytocin and sex steroid receptors in the reproductive tissues of normal and unilateral cryptorchid dogs. Theriogenology 2017; 100:59-65.