Research Progress in Area 2 方向 ( 二 ) 課題進展 123 Abstract Benzo[a]pyrene (BaP) is a typical polycyclic aromatic hydrocarbon (PAH) pollutant in the environment. Direct exposure to BaP/PAHs causes skeletal deformity in fish and impair bone healing in mammals. A deformed spine will affect the fish’s swimming ability, compromising the individual’s survival fitness on feeding and mating. BaP-mediated transgenerational inheritance of skeletal deformity indeed poses a significant threat to the sustainability of fish populations. In this project, Prof. Kong proposes to identify and validate deregulated genes in fish (F1-F3) descended from F0 ancestors exposed to BaP using transcriptome, bioinformatics and real time PCR; to investigate altered DNA methylation pattern in the deregulated genes using bifulfite sequencing and whole-genome sequencing; to examine gene-specific histone modifications in the differentially expressed genes (DEGs); and to unravel the molecular-gene pathways and epigenetic marks underlying BaP induced transgenerational bone impairment in fish. Research Activities and Progress • Conducted whole-genome bisulfite sequencing on F3 medaka vertebrae and sperms; • Completed integrated analysis on the mRNA transcriptome and small RNA transcriptome; • Quantified osteoblast abundance in F3 medaka vertebrae using bone histomorphometry; • Validated expression of genes and miRNA in F3 medaka vertebrae using real time PCR. Key Findings • Ancestral BaP exposure reduced osteoblast abundance in F3 male fish vertebrae; • Functional analysis of differentially-methylated genes (DMGs) in F3 vertebrae revealed they serve roles in osteoblastogenesis, chondrogenesis, and osteoclastogenesis; • frzb, c-fos, gstr and ola-miR-1-3p were identified as biomarker candidates of transgenerational BaP osteotoxicity; • DMGs ( cxcr3, sparc, col12a1) in F3 vertebrae may have escaped reprogramming and likely inherited from F1 sperm; • Methylation marks in sperm also suggested a neurotoxic effect in offspring as a result of parental/ancestral BaP exposure. Research Output Publication 1 Trained personnel 3 Ancestral Exposure to Environmental Benzo[a]pyrene Causes Transgenerational Bone Impairment in Fish: Underlying Mechanisms at the Genetic and Epigenetic Level Prof. Richard Yuen Chong Kong City University of Hong Kong Fig 1. Benzo[a]pyrene osteotoxicity and the regulatory roles of genetic and epigenetic factors. (Mo et al., 2021 CREST).
RkJQdWJsaXNoZXIy NDk5Njg=