Basal Expression Profile of Seven Glutathione-S-Transferases (GSTs) Genes in Six Tissues of Tilapia (Oreochromis niloticus) for Polymerase Chain Reaction Array
DOI:
https://doi.org/10.56919/usci.2434.015Keywords:
PCR, Tilapia, thin films, Quantitative polymerase chain reaction, Glutathione-s-transferases, Basal expression, XenobioticAbstract
Study’s Excerpt
- The differential expression patterns of Glutathione-S-transferase (GST) isoforms in six tissues of Nile tilapia are investigated.
- Seven specific GST genes are examined across six tissues, and it is determined that the liver, spleen, and intestines are key sites of significant gene expression.
- GST isoforms could be potentially used as biomarkers for assessing xenobiotic stress in aquatic environments.
Full Abstract
Because aquatic habitats are being destroyed and biodiversity is declining, xenobiotic pollution of water is a critical environmental problem that has garnered a lot of attention in the past few decades. This study looked at the expression patterns of Glutathione-S-transferases isoforms (GSTMA, GSTO1LA, GSTA, GSTR1, GSTK, GSTT1, and MGST), which are genes involved in xenobiotic metabolism, in Nile tilapia. From adult Nile tilapia, six tissues—liver, spleen, intestines, gills, heart, and muscle—were chosen, and seven genes that were optimised via quantitative polymerase chain reaction (qPCR) were used for validation. Gene expressions were assessed using a PCR array that included duplicate tissues in 96-well qPCR plates, with each gene/plate being assayed six times. Water is used in the remaining wells as no template control (NTC). As reference genes, Pan Ribosomal Protein L3 (RPL3) and Pan 18S ribosomal RNA (18S RNA) were employed. The genes’ basal tissue mRNA expression was measured in each tissue. The muscle was utilised to renormalise the expression levels of all genes throughout the remaining tissues since, under assay’s circumstances the muscle showed very low signal for most of the genes. Using quantitative real-time PCR (qPCR), the result demonstrated widespread differential expression patterns of the seven GST mRNA in the liver, spleen, intestine, gills, heart, and muscle of an untreated fish. However, it was clear that the highest significant expression level of most genes (different GST’s isoforms) was in the liver, followed by the spleen and gut, then the heart and gills compared to muscle (P<0.05). The findings demonstrated widespread differential expression of the GST’s isoforms at basal level and highlighted their utility as biomarkers for xenobiotic stress in aquatic environments.
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