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NF-B Is Activated in the Rat Testis Following Exposure to Mono-(2-Ethylhexyl) Phthalate

Department of Pathology and Laboratory Medicine, Brown University, Providence, Rhode Island 02912

The process of spermatogenesis requires a delicate balance of proapoptotic and antiapoptotic signaling to maintain optimal sperm output. A major transcription factor known to regulate numerous apoptosis-related genes is NF-B. Here we show that mono-(2-ethylhexyl) phthalate (MEHP, 1 g/kg) induces translocation of NF-B subunits (p65, p50, and c-Rel) to germ cell nuclei in young rats (Postnatal Day 28) as early as 1 h after exposure. Immunohistochemistry of rat testes exposed to MEHP showed increased p50 and c-Rel presence in spermatocytes and spermatogonia. In addition, there was increased p65 nuclear positivity in Sertoli cells and germ cells after MEHP, while Rel-B localization was unchanged. These alterations correlated with increased nuclear NF-B-binding activity after MEHP exposure, as shown by electrophoretic mobility shift assays of whole-testis nuclear protein extracts. The increased activity of this transcription factor was associated with a transient protection of the seminiferous epithelium manifested as a decreased number of germ cell apoptotic nuclei measured by TUNEL assay 6 h after MEHP exposure. These results suggest that NF-B is involved in the testicular response to MEHP-induced injury.

¹ Correspondence: Kim Boekelheide, Department of Pathology and Laboratory Medicine, 175 Meeting Street, Brown University, Providence, RI 02912. FAX: 401 863 9008; Kim_Boekelheide{at}brown.edu

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