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Malathion-Induced Granulosa Cell Apoptosis in Caprine Antral Follicles: An Ultrastructural and Flow Cytometric Analysis

Published online by Cambridge University Press:  20 November 2014

Jitender K. Bhardwaj  and
Priyanka Saraf
Jitender K. Bhardwaj*
Affiliation:
Reproductive Physiology Laboratory, Department of Zoology, Kurukshetra University, Kurukshetra-136119, Haryana, India
Priyanka Saraf
Affiliation:
Reproductive Physiology Laboratory, Department of Zoology, Kurukshetra University, Kurukshetra-136119, Haryana, India
*
*Corresponding author. [email protected]
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Abstract

Organophosphate pesticides (OPs) like malathion interfere with normal ovarian function resulting in an increased incidence of atresia and granulosa cell apoptosis that plays a consequential role in the loss of ovarian follicles or follicular atresia. The aim of present study was to assess malathion-induced (100 nM) reproductive stress, ultrastructural damage and changes in apoptosis frequency in ovarian granulosa cells of antral follicles. Transmission electron microscopy (TEM) was employed for ultrastructural characterization, oxidative stress was evaluated using thiobarbituric acid reactive substances (TBARS) assay to measure lipid peroxidation, and apoptosis was quantified via flow cytometry. By TEM, apoptosis was identified by the presence of an indented nuclear membrane with blebbing, pyknotic crescent-shaped fragmented nuclei, increased vacuolization, degenerating mitochondria, and lipid droplets. The results indicate a significant increase in malondialdehyde (MDA) level (nmols/g wet tissue) at a 100 nM dose of malathion i.e. 7.57±0.033*, 8.53±0.12*, and 12.87±0.78** at 4, 6, or 8 h, respectively, as compared with controls (6.07±0.033, p<0.01*, p<0.05**) showing a positive correlation between malathion-induced lipid peroxidation and percentage of granulosa cell apoptosis (r=1; p<0.01). The parallel use of these three methods enabled us to determine the role of malathion in inducing apoptosis as a consequence of cytogenetic damage and oxidative stress generated in granulosa cells of antral follicles.

Keywords

apoptosismalathionoxidative stressultrastructure
Type
Biological Applications
Information
Microscopy and Microanalysis , Volume 20 , Issue 6 , December 2014 , pp. 1861 - 1868
Copyright
© Microscopy Society of America 2014 

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