Evaluation of Effects of Methamphetamine Repeated Dosing on Proliferation and Apoptosis of Rat Germ Cells

Abstract

Methamphetamine (MAMP) is a central nervous system stimulant that is increasingly abused especially by teenagers and young adults, a group in its reproductive age. MAMP effects on the male reproductive system are not clear. In this experimental study, we evaluated the effects of MAMP administration on proliferation and apoptosis in seminiferous tubules of rat testis. Methamphetamine hydrochloride was synthesized by iodination of norephedrine hydrochloride and reduction to methamphetamine. Mature male rats were randomly divided into four groups (n=7) and were injected intraperitoneally with MAMP (1, 5 or 10 mg/kg) or saline at the same time, once daily for 14 consecutive days. Twenty four hs after the last injection, perfused testis were fixed, sectioned and stained by TUNEL labeling or proliferating cell nuclear antigen (PCNA) immunostaining. Apoptotis and proliferation indices were calculated and ratios of proliferation/apoptosis in the seminiferous tubules were obtained. Cell proliferation and the ratio of proliferation to apoptosis decreased significantly in all experimental groups compared to the control group. Conversely, apoptosis was increased in these groups. Such differences were observed in both spermatogonia and primary spermatocytes. In the control group, more than 95% of spermatogonia were PCNA-positive. However, 10 mg/kg of MAMP caused a reduction to approximately 75% PCNA staining in spermatogonia. In some tubules of the experimental groups, more than 10 TUNEL-positive germ cells were seen, although in the control group, the tubules with 3 TUNEL-positive germ cells were rarely observed and the majority of tubules were without such cells. There were significant differences in the indices between the 1 mg/kg group and the higher dose groups, but there was no such difference between the 5 mg/kg and 10 mg/kg groups. In some tubules of the experimental groups, significant gaps in the epithelium between the spermatogonia layer and other cell layers were observed. These results show that repeated administration of MAMP, especially at higher doses, may cause a decrease in cellular proliferation, induce apoptosis and change the proliferation/apoptosis ratio in testis. This might explain the MAMP effect on the spermatogenesis process. It is suggested that studies on the consequence of MAMP consumption on male fertility is warranted.

Administration of high dose of methamphetamine has detrimental effects on sperm parameters and DNA integrity in mice

“…The effects of MA on sperm chromatin also appear to be dose-depended as similar drugs which had more negative effects in high doses. Our findings were similar to the study by Alavi and colleagues which have demonstrated that frequent consumption of high dosage of MA may reduce the number of sperm in the epididymis tail and has negative effects on reproductive system in male rats (34). Additionally, several experimental studies showed that administration of different dose of MA caused decreasing in the sperm count and normal morphology (35, 36).…”

Methamphetamine induces abnormal sperm morphology, low sperm concentration and apoptosis in the testis of male rats

“…Recently, repeated methamphetamine administration caused a decrease in cell proliferation, induce apoptosis and reduce the proliferation/apoptosis ratio in male rat germ cells. Apoptotic cells were increased in both spermatogonia and primary spermatocytes after repeated administration of methamphetamine (Alavi et al, 2008). These findings may reflect that the genotoxic effect of methamphetamine seems to be involved especially in spermatogenesis (Yamamoto et al, 2002).…”

The Effect of Alcoholic Extract of Ginger on Apoptosis and Inflammation of Germ Cells in Streptozotocin-Induced Diabetic Rats

“…In this context, the effect of antioxidants in preventing damages resulting from free radicals has been studies in diabetic diseases [3]. Since the oxidative stress results from an imbalance between the production of free radicals and reactive oxygen species, antioxidant defence mechanism must be able to reduce the harmful effects of these aggressive agents [15].…”

“…Positive control sections were incubated with micrococcal nuclease or recombinant DNase1 to induce DNA strand breaks prior to labelling. Negative control sections were processed with label solution (without terminal transferase) [14,15].…”

“…The quantity of cytokines mRNA in the testis was expressed as units relative to the amount of ß-actin mRNA. The dissociation stages, melting curves and quantitative analyses of the data were performed using CFX manager software version 1.1.308.111 (Bio-Rad, USA) [15][16][17].…”