13 research outputs found
The first mitochondrial 5-methylcytosine map in a non-model teleost (Oreochromis niloticus) reveals extensive strand-specific and non-CpG methylation
DNA methylation is one of the main epigenetic mechanisms that regulate gene expression in a manner that depends on the genomic context and varies considerably across taxa. This DNA modification was first found in nuclear genomes of eukaryote several decades ago and it has also been described in mitochondrial DNA. It has recently been shown that mitochondrial DNA is extensively methylated in mammals and other vertebrates. Our current knowledge of mitochondrial DNA methylation in fish is very limited, especially in non-model teleosts. In this study, using whole-genome bisulfite sequencing, we determined methylation patterns within non-CpG (CH) and CpG (CG) contexts in the mitochondrial genome of Nile tilapia, a non-model teleost of high economic importance. Our results demonstrate the presence of mitochondrial DNA methylation in this species predominantly within a non-CpG context, similarly to mammals. We found a strand-specific distribution of methylation, in which highly methylated cytosines were located on the minus strand. The D-loop region had the highest mean methylation level among all mitochondrial loci. Our data provide new insights into the potential role of epigenetic mechanisms in regulating metabolic flexibility of mitochondria in fish, with implications in various biological processes, such as growth and development.publishedVersio
Micronutrient supplementation affects transcriptional and epigenetic regulation of lipid metabolism in a dose-dependent manner
Micronutrients (vitamins and minerals) have been less well studied compared to macronutrients (fats, proteins, and carbohydrates) although they play important roles in growth, metabolism, and maintenance of tissues. Hence, there is growing interest to understand the influence of micronutrients across various aspects in nutritional research. In the last two decades, aquaculture feeds have been shifted to containing more plant-based materials to meet the increasing demand and maintain the sustainability in the industry. In cultured fish, changing raw materials in feeds alters the requirement levels of micronutrients. A recent whole life cycle feeding trial of Atlantic salmon (Salmo salar) with graded levels of micronutrient packages has concluded that the levels of several B-vitamins and microminerals need to be increased from the current recommendation levels for optimal growth and fish welfare when plant-based diets are used. Here, we show the effect of micronutrient supplementation on transcriptional and epigenetic regulation by using liver samples from the same feeding trial. Specifically, our aim is to reveal the mechanisms of altered cell metabolism, which results in improved growth performance by micronutrient surpluses, at gene expression and DNA methylation levels. Our results strongly indicate that micronutrient supplementation suppresses gene expression in lipid metabolism in a dose-dependent manner and broadly affects DNA methylation in cell adhesion and cell signalling. In particular, it increases DNA methylation levels on the acetyl-CoA carboxylase alpha promoter in a concentration-dependent manner, which further suggests that acetyl-CoA carboxylase alpha is an upstream epigenetic regulator controlling its downstream lipid biosynthesis activities. This study demonstrates a comprehensive analysis to reveal an important role of micronutrients in lipid metabolism through epigenetic control of gene expression
The apoptosis-inducing activity towards leukemia and lymphoma cells in a cyanobacterial culture collection is not associated with mouse bioassay toxicity
Cyanobacteria (83 strains and seven natural populations) were screened for content of apoptosis (cell death)-inducing activity towards neoplastic cells of the immune (jurkat acute T-cell lymphoma) and hematopoetic (acute myelogenic leukemia) lineage. Apoptogenic activity was frequent, even in strains cultured for decades, and was unrelated to whether the cyanobacteria had been collected from polar, temperate, or tropic environments. The activity was more abundant in the genera Anabaena and Microcystis compared to Nostoc, Phormidium, Planktothrix, and Pseudanabaena. Whereas the T-cell lymphoma apoptogens were frequent in organic extracts, the cell death-inducing activity towards leukemia cells resided mainly in aqueous extracts. The cyanobacteria were from a culture collection established for public health purposes to detect toxic cyanobacterial blooms, and 54 of them were tested for toxicity by the mouse bioassay. We found no correlation between the apoptogenic activity in the cyanobacterial isolates with their content of microcystin, nor with their ability to elicit a positive standard mouse bioassay. Several strains produced more than one apoptogen, differing in biophysical or biological activity. In fact, two strains contained microcystin in addition to one apoptogen specific for the AML cells, and one apoptogen specific for the T-cell lymphoma. This study shows the potential of cyanobacterial culture collections as libraries for bioactive compounds, since strains kept in cultures for decades produced apoptogens unrelated to the mouse bioassay detectable bloom-associated toxins
Mitochondrial DNA methylation profiling in Nile tilapia (Oreochromis niloticus) through nanopore sequencing
6th International Symposium on Genomics in Aquaculture (GIA 2022), 4-6 May 2022, GranadaThe mitochondrion is a cellular organelle that plays a central role in the regulation of respiration and metabolic processes in the cell. Thus, it is implicated in growth, metabolism and aging of an organism. The first study on mitochondrial DNA (mtDNA) epigenetics have been published at the beginning of the 1970s (Nass, 1973). The identification of DNA methyltransferase 1 in mammalian mitochondria attracted researchers towards the study of the mitoepigenome (Shock et al., 2011). However, data about the methylation patterns of mtDNA in adult fish tissues is still absent, apart from a single report (Nedoluzhko et al., 2021). Mitochondria play a critical role in the production of reactive oxygen species, which in turn is essential for muscle function and growth. Growth is one of the most important traits in the aquaculture sector (Gjedrem, 2005). In the present study, mtDNA methylation profiles have been analysed in Nile tilapia (Oreochromis niloticus), fast muscle using Nanopore long-read sequencing. O. niloticus fast tissue was sampled, and used for genomic DNA isolation and construction of PCR-free libraries. The Nanopore MinION Mk1C sequencing workflow generated 673 Fast5 files, which were used for basecalling and methylation analysis with Megalodon v2.4.2 and Remora v0.1.2. The mean sequencing read length was 3275 bp and the coverage obtained for Nile tilapia mtDNA was 1082X. This methylation data analysis based on Nanopore long-read sequencing constitutes a valuable resource for future research on the mitoepigenome of farmed fishes.
References:
Gjedrem, T. (2005). Selection and Breeding Programs in Aquaculture: Springer Netherlands.
Nass, M.M. (1973). Differential methylation of mitochondrial and nuclear DNA in cultured mouse, hamster and virus-transformed hamster cells in vivo and in vitro methylation. Journal of Molecular Biology, 80(1), 155-175.
Nedoluzhko, A., Mjelle, R., Renström, M., Skjærven, K.H., Piferrer, F., and Fernandes, J.M.O. (2021). The first mitochondrial 5-methylcytosine map in a non-model teleost (Oreochromis niloticus) reveals extensive strand-specific and non-CpG methylation. Genomics, 113, 3050-3057.
Shock, L.S., Thakkar, P.V., Peterson, E.J., Moran, R.G., and Taylor, S. M. (2011). DNA methyltransferase 1, cytosine methylation, and cytosine hydroxymethylation in mammalian mitochondria. Proceedings of the National Academy of Sciences, 108(9), 3630-3635This work was supported by the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation programme (grant agreement no 683210) and by the Research Council of Norway under the Toppforsk programme (grant agreement no 250548/F20), with additional support from Notur/NorStore (project NN9337K)Peer reviewe
Nanopore sequencing reveals growth rates-dependent differential methylation of the mitogenome in Nile tilapia (Oreochromis niloticus) muscle
Aquaculture Europe 22, Innovative solutions in a changing world, 27-30 September 2022, Rimini, ItalyThe mitochondrion is a cellular organelle that plays a central role in the regulation of respiration and metabolic processes in the cell, which are essential for muscle function and growth. Mitochondrial efficiency is directly linked with growth rate and varies among individuals, but the molecular basis of these differences is currently unknown. The aim of this study was to investigate a potential link between mtDNA methylation and growth rates in Nile tilapia (Oreochromis niloticus) muscle, since we have reported that the liver mitogenome is 20 % methylated (25 % CpGs) in this species. (Nedoluzhko et al., 2021). Fast muscle was sampled from slow and fast-growing Nile tilapia males and females, and used for genomic DNA isolation and construction of PCR-free libraries. In a representative library, the Nanopore MinION Mk1C sequencing workflow generated 673 Fast5 files, which were used for basecalling and methylation analysis with Megalodon v2.4.2 and Remora v0.1.2. The mean sequencing read length was 3275 bp and the coverage obtained for Nile tilapia mtDNA was 1082X. Methylation data generated from Nanopore long-read sequencing constitute a valuable resource for future research on the mitoepigenome related to muscle growth, metabolism and oxidation status of farmed fishesReferences: Nedoluzhko, A., Mjelle, R., Renström, M., Skjærven, K.H., Piferrer, F., and Fernandes, J.M.O. (2021). The first mitochondrial 5-methylcytosine map in a non-model teleost (Oreochromis niloticus) reveals extensive strand-specific and non-CpG methylation. Genomics, 113, 3050-3057This work was supported by the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation programme (grant agreement no 683210) and by the Research Council of Norway under the Toppforsk programme (grant agreement no 250548/F20), with additional support from Notur/NorStore (project NN9337K)Peer reviewe
The mitochondrial 5-methylcytosine profile in Nile tilapia (Oreochromis niloticus)
6th International Symposium on Genomics in Aquaculture (GIA), 5-7 May 2021Growth and metabolism are closely linked to mitochondrion, since this organelle¿s primary function is generation of cellular energy by production of ATP (adenosine triphosphate). Mitochondrion has a compact circular genome (16,5 Kb in average), which is related to its bacterial origin. The mitogenome in Nile tilapia (Oreochromis niloticus) is 16,625 bp long and contains 13 protein-coding genes (involved in oxidative phosphorylation), two rRNA genes, 22 tRNA genes and the D-loop (putative control region). The number and the order of mitochondrial genes in Nile tilapia matches those in other cichlids. DNA methylation is one of several epigenetic regulatory mechanisms that control gene expression. In fish, epigenetic changes associated with mtDNA and their potential relation to functional effects on growth is still a question that remains unanswered. In this study, we isolated mtDNA from liver of 5 full-sib Nile tilapia females. A whole genome bisulfite sequencing (Illumina NextSeq) approach was used for identifying methylation patterns within CHH, CHG and CpG context in the mitochondrial genome. We described and compared methylation rate among mitochondrial genes to create the first methylome map of mtDNA in fish. Our results demonstrated the presence of mtDNA methylation in fish, predominantly within non-CpG context. We found a strand-specific distribution of mtDNA methylation, where most of methylated cytosines were located on the light (minus) strand. The D-loop region had the highest mean methylation value among all mitochondrial genes. These data provide a new insight into mitochondrial epigenetics in fish and can serve as a background for further functional investigation of potential epigenetic markers involved in shaping metabolic flexibility and growth in Nile tilapi