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Mono-(2-ethylhexyl) phthalate (MEHP) induces spermatogenic cell apoptosis in guinea pig testes at prepubertal stage
M A Awal1, M Kurohmaru, M Ishii
1Department of Anatomy and Histology, Bangladesh Agricultural University, Mymensingh, Bangladesh. awal1963@yahoo.com
Abstract:
The effects of mono-(2-ethylhexyl) phthalate (MEHP), an active metabolite of di-(2-ethylhexyl) phthalate (DEHP), on prepubertal guinea pig testes in vitro were investigated. The testes of 35-day-old guinea pigs were surgically excised. They were seeded in a defined medium containing antibiotics and administered MEHP at concentrations of 1, 10, and 100 nmol/ml, respectively. The control groups were administered a similar volume of corn oil vehicle. The tissues were incubated for 3, 6, and 9 h. The specimens were collected at 3, 6, and 9 h after treatment. They were fixed in 4% paraformaldehyde or 5% glutaraldehyde. For quantitation of the apoptotic spermatogenic cells, the terminal dUTP nick end-labeling (TUNEL) staining was performed by light microscopy. Detachment and displacement of spermatogenic cells, thin seminiferous epithelia, and Sertoli cell vacuolization were observed. Maximal testicular damage was recognized at 100 nmol/ml and 9 h after MEHP treatment. The percentage (%) of apoptotic spermatogenic cells significantly increased at 3, 6, and 9 h after treatment, compared to the control groups. Because the loss of spermatogenic cells by MEHP treatment varies among species, the present study, using guinea pigs, was designed and conducted to obtain further information.
Insights
Mono-(2-ethylhexyl) phthalate (MEHP) exposure in vitro significantly increased apoptotic spermatogenic cells in prepubertal guinea pig testes. This phthalate metabolite causes testicular damage, impacting reproductive health.
Area of Science:
- Reproductive Toxicology
- Developmental Biology
- Environmental Health
Background:
- Di-(2-ethylhexyl) phthalate (DEHP) is a common plasticizer.
- Mono-(2-ethylhexyl) phthalate (MEHP) is its active metabolite.
- Phthalates are endocrine disruptors with potential reproductive toxicity.
Purpose of the Study:
- To investigate the in vitro effects of MEHP on prepubertal guinea pig testes.
- To quantify MEHP-induced apoptosis in testicular cells.
- To assess the dose- and time-dependent testicular damage caused by MEHP.
Main Methods:
- Surgical excision and in vitro incubation of 35-day-old guinea pig testes.
- Administration of MEHP at 1, 10, and 100 nmol/ml concentrations.
- TUNEL staining for apoptotic cell quantitation and light microscopy for tissue damage assessment.
Main Results:
- MEHP exposure led to detachment and displacement of spermatogenic cells.
- Thinning of seminiferous epithelia and Sertoli cell vacuolization were observed.
- A significant increase in apoptotic spermatogenic cells was noted at 3, 6, and 9 hours post-treatment, with maximal damage at 100 nmol/ml and 9 hours.
Conclusions:
- MEHP induces significant testicular damage and apoptosis in prepubertal guinea pigs in vitro.
- The findings highlight MEHP's potential as a reproductive toxicant.
- Guinea pig models provide valuable insights into species-specific phthalate toxicity.
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