TY - JOUR
T1 - Methamidophos alters sperm function and DNA at different stages of spermatogenesis in mice
AU - Urióstegui-Acosta, Mayrut
AU - Hernández-Ochoa, Isabel
AU - Sánchez-Gutiérrez, Manuel
AU - Piña-Guzmán, Belem
AU - Rafael-Vázquez, Leticia
AU - Solís-Heredia, M. J.
AU - Martínez-Aguilar, Gerardo
AU - Quintanilla-Vega, Betzabet
N1 - Funding Information:
The authors want to thank Ricardo Gaxiola Centeno and Rafael Leyva Muñoz for their technical assistance with the animals, Víctor H. Rosales García for his assistance with flow cytometry, Alberto Hernández Alcántara for his help with the comet assay, and Dr. Leticia Hernández-Cadena for her critical discussion of statistical analyses. Also we want to acknowledge the generosity of Dr. Manuel Hernández-Hernández from the Department of Cell Biology (CINVESTAV-IPN) for the gift of the anti-actin primary antibody. This study was supported by CONACYT -Mexico (Grant # 58213 given to BQV). MUA was a recipient of a scholarship from CONACYT-Mexico.
PY - 2014/9/15
Y1 - 2014/9/15
N2 - Methamidophos (MET) is a highly toxic organophosphate (OP) pesticide that is widely used in developing countries. MET has male reproductive effects, including decreased fertility. We evaluated MET effects on sperm quality, fertilization and DNA integrity, exploring the sensitivity of different stages of spermatogenesis. Adult male mice received MET (3.75 or 5. mg/kg-bw/ip/day/4. days) and were euthanized 1, 28 or 45. days post-treatment (dpt) to evaluate MET's effects on epididymal maturation, meiosis or mitosis, respectively. Spermatozoa were obtained from the cauda epididymis-vas deferens and were evaluated for sperm quality, acrosome reaction (AR; Coomassie staining), mitochondrial membrane potential (by JC-1), DNA damage (comet assay), oxidative damage (malondialdehyde (MDA) production), in vitro fertilization and protein phosphorylation (immunodetection), and erythrocyte acetylcholinesterase (AChE) activity. At 1-dpt, MET inhibited AChE (43-57%) and increased abnormal cells (6%). While at 28- and 45-dpt, sperm motility and viability were significantly reduced with an increasing MET dose, and abnormal morphology increased at 5. mg/kg/day/4. days. MDA and mitochondrial activity were not affected at any dose or time. DNA damage (OTM and %DNA) was observed at 5. mg/kg/day/4. days in a time-dependent manner, whereas both parameters were altered in cells from mice exposed to 3.75. mg/kg/day/4. days only at 28-dpt. Depending on the time of collection, initial-, spontaneous- and induced-AR were altered at 5. mg/kg/day/4. days, and the fertilization capacity also decreased. Sperm phosphorylation (at serine and tyrosine residues) was observed at all time points. Data suggest that meiosis and mitosis are the more sensitive stages of spermatogenesis for MET reproductive toxicity compared to epididymal maturation.
AB - Methamidophos (MET) is a highly toxic organophosphate (OP) pesticide that is widely used in developing countries. MET has male reproductive effects, including decreased fertility. We evaluated MET effects on sperm quality, fertilization and DNA integrity, exploring the sensitivity of different stages of spermatogenesis. Adult male mice received MET (3.75 or 5. mg/kg-bw/ip/day/4. days) and were euthanized 1, 28 or 45. days post-treatment (dpt) to evaluate MET's effects on epididymal maturation, meiosis or mitosis, respectively. Spermatozoa were obtained from the cauda epididymis-vas deferens and were evaluated for sperm quality, acrosome reaction (AR; Coomassie staining), mitochondrial membrane potential (by JC-1), DNA damage (comet assay), oxidative damage (malondialdehyde (MDA) production), in vitro fertilization and protein phosphorylation (immunodetection), and erythrocyte acetylcholinesterase (AChE) activity. At 1-dpt, MET inhibited AChE (43-57%) and increased abnormal cells (6%). While at 28- and 45-dpt, sperm motility and viability were significantly reduced with an increasing MET dose, and abnormal morphology increased at 5. mg/kg/day/4. days. MDA and mitochondrial activity were not affected at any dose or time. DNA damage (OTM and %DNA) was observed at 5. mg/kg/day/4. days in a time-dependent manner, whereas both parameters were altered in cells from mice exposed to 3.75. mg/kg/day/4. days only at 28-dpt. Depending on the time of collection, initial-, spontaneous- and induced-AR were altered at 5. mg/kg/day/4. days, and the fertilization capacity also decreased. Sperm phosphorylation (at serine and tyrosine residues) was observed at all time points. Data suggest that meiosis and mitosis are the more sensitive stages of spermatogenesis for MET reproductive toxicity compared to epididymal maturation.
KW - DNA damage
KW - Male fertility
KW - Methamidophos
KW - Organophosphate pesticides
KW - Protein phosphorylation
KW - Sperm function
UR - http://www.scopus.com/inward/record.url?scp=84907169808&partnerID=8YFLogxK
U2 - 10.1016/j.taap.2014.06.017
DO - 10.1016/j.taap.2014.06.017
M3 - Artículo
SN - 0041-008X
VL - 279
SP - 391
EP - 400
JO - Toxicology and Applied Pharmacology
JF - Toxicology and Applied Pharmacology
IS - 3
ER -