Relative gene expression, micronuclei formation, and ultrastructure alterations induced by heavy metal contamination in Pimelia latreillei (Coleoptera: Tenebrionidae) in an urban-industrial area of Alexandria, Egypt

埃及亚历山大市城市工业区重金属污染对拟步甲(鞘翅目:拟步甲科)相对基因表达、微核形成和超微结构改变的影响

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作者:Lamia M El-Samad,Saeed El-Ashram,Dalia A Kheirallah,Karolin K Abdul-Aziz,Noura A Toto,El Hassan M Mokhamer

Abstract

The present research aims to evaluate the impact of industrial processes and anthropogenic activities on the beetle Pimelia latreillei inhabiting the polluted site at Zawya Abd El- Qader, Alexandria, Egypt. Beetles were collected from the vicinity of five factories. The genotoxic effects of environmental exposures to industrial heavy metals were monitored using a broad range of assays, including energy-dispersive X ray microanalysis and X-ray diffraction (SEM and EDX)), qRT-PCR gene expression assay, micronuclei formation, and transmission electron microscope (TEM). Energy dispersive X-ray microanalysis for the soil and testicular tissues of beetles collected from the polluted site revealed a higher percentage of heavy metals than the beetles collected from the reference site (Sidi Kirier, Alexandria, Egypt). To analyze/monitor genotoxicity in P. latreillei sampled from the polluted site, the transcription levels of levels of heat shock proteins (Hsps) and accessory gland seminal fluid protein (AcPC01) in testicular tissues were recorded. The incidence of micronuclei (MN) formation in the testicular cells was also observed. Quantitative RT-PCR (RT-qPCR) analysis was carried out to detect the changes in the gene expression of the aforementioned proteins. Genes encoding heat shock proteins (Hsp60, Hsp70, and Hsp90) were significantly overexpressed (> 2-fold) in specimens sampled from the polluted site; however, AcPC01 gene expression was under-expressed (<1.5-folds). The incidence of MN was significantly increased in specimens sampled from the polluted site. Ultrastructure anomalies (nuclear and cytoplasmic disruption) were also observed in the testicular cells of the beetles sampled from the polluted site compared to those sampled from the unpolluted site. Our results, therefore, advocate a need for adequate measures to reduce increasing environmental pollution in the urban-industrial areas.

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