Yonsei Med J.  2008 Oct;49(5):843-850. 10.3349/ymj.2008.49.5.843.

In Utero Exposure to 2,3,7,8-Tetrachlorodibenzo-p-Dioxin Affects the Development of Reproductive System in Mouse

• Jin MH
• Ko HK
• Hong CH
• Han SW

• KMID: 2158204
• DOI: http://doi.org/10.3349/ymj.2008.49.5.843

Abstract

PURPOSE
Exposure of male reproductive organs to 2,3,7,8-Tetrachlorodibenzo-p-Dioxin (TCDD) has been reported to cause developmental changes. In this study, we evaluated the effects of in utero TCDD exposure on male reproductive development. MATERIALS AND METHODS: Pregnant C57BL/6 mice were administered a single intraperitoneal injection of TCDD (1microgram/kg) on gestation day (GD) 15. The offspring were examined in the immature stage on postnatal day (PND) 30 and in the mature stage on PND 60. The testes were examined for histological changes, androgen receptor (AR), proliferating cell nuclear antigen (PCNA) and apoptosis following the measurement of morphological changes. RESULTS: Anogenital distance (AGD) and testis weights were reduced by TCDD exposure both on PND 30 and PND 60 while body weights and length of male offspring were not affected by TCDD. The regular sperm developmental stage was impaired with TCDD treatment on PND 30. However, no difference was found between the control group and TCDD groups on PND 60. Simultaneously, the expression of AR was also reduced on PND 30, while it was increased on PND 60 compared with the control group. The expression of PCNA was decreased whereas apoptosis was not affected by TCDD both on PND 30 and PND 60. CONCLUSION: These results suggest that in utero exposure to TCDD influences the development of testes by inhibiting the expression of AR and PCNA. Moreover, the adverse effects of TCDD on male offspring reduced over time.

Keyword

2,3,7,8-Tetrachlorodibenzo-p-Dioxin; androgen receptor; testis; apoptosis; proliferation

MeSH Terms

Animals
Apoptosis/drug effects
Cell Proliferation
Embryonic Development/*drug effects
Environmental Pollutants/*toxicity
Female
Male
*Maternal Exposure
Mice
Mice, Inbred C57BL
Organ Size/drug effects
Pregnancy
Receptors, Androgen/metabolism
Testis/*drug effects/embryology/pathology
Tetrachlorodibenzodioxin/*toxicity

Figure

• 

  Fig. 1 Histological assessment of testis from male offspring In utero exposure to TCDD. (A and C) Testes from control group on PND 30 and PND 60, respectively. (B and D) Testes from TCDD exposed group on PND 30 and PND 60. n = 9 - 10 for each group (Magnification: × 100). PND, postnatal day; TCDD, 2,3,7,8-Tetrachlorodibenzo-p-Dioxin.

• 

  Fig. 2 Effects of in utero TCDD exposure on expression of androgen receptor. Pregnant mice were injected with 1 µg/kg TCDD concentration on GD 15. Testes on PND 30 and PND 60 were used to examine the expression of androgen receptor (AR) by immunohistochemistry (n = 5, for each group). AR was present in Leydig cells (white arrow), Sertoli cells (black arrow) and peritubular myoid cells (arrow head) in testis. (A and C) Testes from control group on PND 30 and PND 60 respectively. (B and D) Testes from TCDD exposed group on PND 30 and PND 60 (Magnification: × 100). PND, postnatal day; TCDD, 2,3,7,8-Tetrachlorodibenzo-p-Dioxin.

• 

  Fig. 3 Effects of in utero TCDD exposure on cell proliferation. Pregnant mice were injected with 1 µg/kg TCDD concentration on GD 15. Testes on PND 30 and PND 60 were used to examine cell proliferation using specific anti-PCNA by immunohistochemistry. Three sections from different parts of each testis were chosen, and a total of 10 seminiferous tubules in each section were randomly chosen for counting PCNA-positive cells. (A and C) Testes from control group on PND 30 and PND 60, respectively. (B and D) Testes from TCDD exposed group on PND 30 and PND 60. (E) The quantification of PCNA positive germ cells. Data indicate means ± SEM. n = 5 for each group. PND, postnatal day; TCDD, 2,3,7,8-Tetrachlorodibenzo-p-Dioxin. *p < 0.01, **p < 0.01 (Magnification: × 100).

• 

  Fig. 4 Effects of in utero TCDD exposure on cell apoptosis. Pregnant mice were injected with 1 µg/kg TCDD concentration on GD 15. Testes from PND 30 and PND 60 were used to examine cell apoptosis TUNEL staining (arrow head) (n = 5 for each group). (A and C) Testis from control group on PND 30 and PND 60, respectively. (B and D) Testis from TCDD exposed group on PND 30 and PND 60 (Magnification × 200). PND, postnatal day; TCDD, 2,3,7,8-Tetrachlorodibenzo-p-Dioxin.

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