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24 February 2021 Carryover effect of atrazine and its metabolite—from treated bovine spermatozoa to the embryo's transcriptome
A. Komsky-Elbaz, D. Kalo, Z. Roth
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Abstract

Atrazine (ATZ) is an extensively used herbicide and ubiquitous environmental contaminant. ATZ and its metabolite, diaminochlorotriazine (DACT), cause several cellular and functional alterations in spermatozoa. We aimed to examine the effect of ATZ/DACT on spermatozoon DNA integrity, fertilization competence, embryonic development, and transcriptome profile of in vitro-produced embryos derived from fertilization with pre-exposed sperm. Bovine spermatozoa exposed to ATZ (0.1 or 1 µM) or DACT (1 or 10 µM) during in vitro capacitation were used for in vitro fertilization of untreated oocytes. Cleavage and blastocyst-formation rates were evaluated 42 h and 7 days postfertilization, respectively. The association between DNA fragmentation and apoptosis (annexin V kit) was determined. Fertilization competence of annexin-positive (AV+) and annexin-negative (AV–) spermatozoa was examined. Microarray analysis was performed for 7-day blastocysts. Intracytoplasmic sperm injection was performed with control (AV+, AV–) and DACT (AV+, AV–) spermatozoa. Cleavage rates did not differ between groups and blastocyst formation tended to be higher for AV– vs. AV+ in both control and DACT groups, suggesting that acrosome reaction, rather than DNA fragmentation, underlies the reduced cleavage. Transcriptomic analysis revealed 139 and 230 differentially expressed genes in blastocysts derived from ATZ- and DACT-exposed spermatozoa, respectively, relative to controls. Proteomic analysis shown differential expression of proteins in ATZ- or DACT-treated spermatozoa, in particular proteins related to cellular processes and biological pathways. Therefore, we assume that factors delivered by the spermatozoa, regardless of DNA fragmentation, are also involved. Overall, the current study reveals a deleterious carryover effect of ATZ/DACT from the spermatozoa to the developing embryo.

Summary sentence

Summary sentence: Atrazine and its metabolite, diaminochlorotriazine, induce multifactorial damage in sperm, which is carried over to the embryo's transcriptome.

© The Author(s) 2021. Published by Oxford University Press on behalf of Society for the Study of Reproduction. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com
A. Komsky-Elbaz, D. Kalo, and Z. Roth "Carryover effect of atrazine and its metabolite—from treated bovine spermatozoa to the embryo's transcriptome," Biology of Reproduction 104(5), 1162-1180, (24 February 2021). https://doi.org/10.1093/biolre/ioab027
Received: 22 September 2020; Accepted: 15 February 2021; Published: 24 February 2021
KEYWORDS
atrazine
diaminochlorotriazine
embryo development
microarray
proteomics
spermatozoa
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