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Dibutyl phthalate disrupts glycogen synthase kinase 3α essential for sperm motility.
Park, Seung Hyun; Gye, Myung Chan.
Afiliación
  • Park SH; Department of Life Science and Institute for Natural Sciences, Hanyang University, Seoul 04763, Republic of Korea.
  • Gye MC; Department of Life Science and Institute for Natural Sciences, Hanyang University, Seoul 04763, Republic of Korea. Electronic address: mcgye@hanyang.ac.kr.
Ecotoxicol Environ Saf ; 271: 115977, 2024 Feb.
Article en En | MEDLINE | ID: mdl-38242044
ABSTRACT
To unravel the toxic mechanism of phthalate ester plasticizer endocrine disruptor in spermatozoa, we examined the effect of dibutyl phthalate (DBP) on the stability and inhibitory phosphorylation of glycogen synthase kinase 3α (GSK3α), a protein kinase crucial for sperm motility in mice. In DBP-treated spermatozoa, reactive oxygen species (ROS) and lipid peroxide were significantly increased. In computer-assisted sperm analysis, DBP at concentrations of 10 - 100 µg/mL significantly decreased total motility and progressive motility of spermatozoa. On western blots, DBP decreased p-GSK3α(Ser21) and increased p-GSK3α(Tyr279) in spermatozoa. Similarly, hydrogen peroxide decreased p-GSK3α(Ser21) but not p-GSK3α(Tyr279) in spermatozoa. Immunofluorescent labeling demonstrated that DBP markedly decreased immunoreactivities of GSK3α and p-GSK3α(Ser21) but increased immunoreactivity of p-GSK3α(Tyr279) in spermatozoa. DBP at a concentration of 100 µg/mL significantly increased phosphatase activity in spermatozoa. Calyculin A, a protein phosphatase 1 and 2 A inhibitor, markedly increased p-GSK3α(Ser21) and sperm motility and attenuated a DBP-induced decrease of p-GSK3α(Ser21) and sperm motility. On western blot, 1-100 µg/mL DBP decreased GSK3α in spermatozoa. On immunoprecipitation western blot, DBP at 10 - 100 µg/mL increased polyubiquitinated sperm proteins including GSK3α. The MG115, proteasome inhibitor attenuated degradation of GSK3α in DBP-treated spermatozoa. Hydrogen peroxide at 10 µM increased polyubiquitinated sperm proteins, suggesting that DBP may increase ubiquitination of GSK3α via ROS induction. Together, DBP may decrease the cellular amount of GSK3α through the ubiquitin-proteasome pathway and p-GSK3α(Ser21) through ROS generation and activation of protein phosphatases, impairing sperm motility.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Motilidad Espermática / Dibutil Ftalato Límite: Animals Idioma: En Revista: Ecotoxicol Environ Saf Año: 2024 Tipo del documento: Article Pais de publicación: Países Bajos

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Motilidad Espermática / Dibutil Ftalato Límite: Animals Idioma: En Revista: Ecotoxicol Environ Saf Año: 2024 Tipo del documento: Article Pais de publicación: Países Bajos