ACRYLAMIDE INFLUENCE ON THE ACTIVITY OF CHOLINESTERASES, OXIDATIVE STRESS MARKERS, CASPASE-3 EXPRESSION AND MICROSTRUCTURE OF THE FOREBRAIN OF CHICKEN EMBRYOS

Marta Batoryna,Marta Kopańska,Grzegorz Formicki,Agnieszka Leśniak

doi:10.15414/jmbfs.2019/20.9.3.643-647

Abstract

The aim of this study was to examine the influence of acrylamide (ACR), a common neurotoxin, on the neuroepithelial cells utilizing in ovo model for embryonic brain development. On day five (E5), 75 eggs with living embryos were randomly divided into three groups: one control and two experimental (n=25) and injected with 0.7% NaCl and/or ACR saline solution at the dose of 30 μg or 300 μg per egg , respectively. The injections were performed directly into the amniotic fluid, and after 7 h of incubation, tissues the brain was collected Additionally, for the brain microstructure evaluation brain tissues were obtained from 10th day embryos. Immunohistochemical localization of caspase-3 and histochemical staining demonstrated deterioration and significant damage to neuroepithelial cells and affected caspase-3 distribution/expression in brain. Results of biochemical assays showed increased activity of enzymes (SOD, AChE, BuChE) whereas ACR had no impact on the MDA concentration. This suggests that ACR has no direct impact on the caspases, and its action is an effect of SOD response to free radicals probably. ACR has an impact on the activity of cholinergic nerves and brain cytoarchitecture of the developing embryos. All of the observed effects suggests serious influence of ACR on the developing brain by several different mechanisms.

Highlights

Acrylamide (ACR) and its polymer- polyacrylamide, are broadly applied in laboratories and industrial processes
In control embryos (Fig. 5A), caspase-3 was evenly distributed throughout all the layers of neuroepithelial cells, whereas in embryos exposed to ACR (Fig. 5B, 5C) the enzyme positive cells concentrated in the mantle layer of the forebrain vesicle
In our studies inhibition of caspases at E5 resulted in increased number of neural cells in brain hemispheres at E10, which points to the disturbances in the selection of the neuron cells in embryos exposed to ACR (Fig. 6A, 6B)

Summary

Introduction

Acrylamide (ACR) and its polymer- polyacrylamide, are broadly applied in laboratories and industrial processes. Previous studies indicate that ACR influences human astrocytoma cells and nerve terminal degeneration of the central nervous system (Yu et al, 2005; Chen et al, 2013). Another study indicated declined activity of AChE in the mice brain after ACR administration (Kopańska et al, 2015). Taking the above into consideration, our study has been focusing on the ACR impact on the neuroepithelial (NE) cells, which undergo intensive proliferation in order to produce more progenitor, neuronal and macroglial cells within the central nervous system. To recognize the possible mechanism of ACR toxicity in the brain of animal embryos, we have performed measurements of antioxidative enzymes activity, cholinesterases activity, and analysis of caspase-3 expression. In ovo model has been chosen as a wellrecognized and suitable model for animal embryo development studies (Stern, 2005)

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