Dataset Information

Response of the hepatic transcriptome to aflatoxin B1 in domestic turkey (Meleagris gallopavo).

ABSTRACT: Dietary exposure to aflatoxin B1 (AFB1) is detrimental to avian health and leads to major economic losses for the poultry industry. AFB1 is especially hepatotoxic in domestic turkeys (Meleagris gallopavo), since these birds are unable to detoxify AFB1 by glutathione-conjugation. The impacts of AFB1 on the turkey hepatic transcriptome and the potential protection from pretreatment with a Lactobacillus-based probiotic mixture were investigated through RNA-sequencing. Animals were divided into four treatment groups and RNA was subsequently recovered from liver samples. Four pooled RNA-seq libraries were sequenced to produce over 322 M reads totaling 13.8 Gb of sequence. Approximately 170,000 predicted transcripts were de novo assembled, of which 803 had significant differential expression in at least one pair-wise comparison between treatment groups. Functional analysis linked many of the transcripts significantly affected by AFB1 exposure to cancer, apoptosis, the cell cycle or lipid regulation. Most notable were transcripts from the genes encoding E3 ubiquitin-protein ligase Mdm2, osteopontin, S-adenosylmethionine synthase isoform type-2, and lipoprotein lipase. Expression was modulated by the probiotics, but treatment did not completely mitigate the effects of AFB1. Genes identified through transcriptome analysis provide candidates for further study of AFB1 toxicity and targets for efforts to improve the health of domestic turkeys exposed to AFB1.

SUBMITTER: Monson MS

PROVIDER: S-EPMC4076218 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature

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Response of the hepatic transcriptome to aflatoxin B1 in domestic turkey (Meleagris gallopavo).

Monson Melissa S MS Settlage Robert E RE McMahon Kevin W KW Mendoza Kristelle M KM Rawal Sumit S El-Nezami Hani S HS Coulombe Roger A RA Reed Kent M KM

PloS one 20140630 6

Dietary exposure to aflatoxin B1 (AFB1) is detrimental to avian health and leads to major economic losses for the poultry industry. AFB1 is especially hepatotoxic in domestic turkeys (Meleagris gallopavo), since these birds are unable to detoxify AFB1 by glutathione-conjugation. The impacts of AFB1 on the turkey hepatic transcriptome and the potential protection from pretreatment with a Lactobacillus-based probiotic mixture were investigated through RNA-sequencing. Animals were divided into four ...[more]

PMID: 24979717

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Project description:Domestic turkeys (Meleagris gallopavo) are one of the most susceptible animals known to the toxic effects of the mycotoxin aflatoxin B1 (AFB1), a potent human hepatocarcinogen, and universal maize contaminant. We have demonstrated that such susceptibility is associated with the inability of hepatic glutathione S-transferases (GSTs) to detoxify the reactive electrophilic metabolite exo-AFB1-8,9-epoxide (AFBO). Unlike their domestic counterparts, wild turkeys, which are relatively AFB1-resistant, possess hepatic GST-mediated AFBO conjugating activity. Here, we characterized the molecular and functional properties of hepatic alpha-class GSTs (GSTAs) from wild and domestic turkeys to shed light on the differences in resistance between these closely related strains. Six alpha-class GST genes (GSTA) amplified from wild turkeys (Eastern and Rio Grande subspecies), heritage breed turkeys (Royal Palm) and modern domestic (Nicholas strain) turkeys were sequenced, and catalytic activities of heterologously-expressed recombinant enzymes determined. Alpha-class identity was affirmed by conserved GST domains and four signature motifs. All GSTAs contained single nucleotide polymorphisms (SNPs) in their coding regions: GSTA1.1 (5 SNPs), GSTA1.2 (7), GSTA1.3 (3), GSTA2 (3), GSTA3 (1) and GSTA4 (2). E. coli-expressed GSTAs possessed varying activities toward GST substrates 1-chloro-2,4-dinitrobenzene (CDNB), 1,2-dichloro-4-nitrobenzene (DCNB), ethacrynic acid (ECA), cumene hydroperoxide (CHP). As predicted by their relative resistance, livers from domestic turkeys lacked detectable GST-mediated AFBO detoxification activity, whereas those from wild and heritage birds possessed this critical activity, suggesting that intensive breeding and selection resulted in loss of AFB1-protective alleles during domestication. Our observation that recombinant tGSTAs detoxify AFBO, whereas their hepatic forms do not, implies that the hepatic forms of these enzymes are down-regulated, silenced, or otherwise modified by one or more mechanisms. These data may inform of possible molecular mechanisms of resistance to AFB1, and may also have the benefit of identifying genetic markers which could be used to enhance AFB1 resistance in modern domestic strains.

Project description:The application of transcriptomics to the study of the reproductive tract in male turkeys can significantly increase our current knowledge regarding the specifics of bird reproduction. To characterize the complex transcriptomic changes that occur in the testis, epididymis, and ductus deferens, deep sequencing of male turkey RNA samples (n = 6) was performed, using Illumina RNA-Seq. The obtained sequence reads were mapped to the turkey genome, and relative expression values were calculated to analyze differentially expressed genes (DEGs). Statistical analysis revealed 1,682; 2,150; and 340 DEGs in testis/epididymis, testis/ductus deferens, and epididymis/ductus deferens comparisons, respectively. The expression of selected genes was validated using quantitative real-time reverse transcriptase-polymerase chain reaction. Bioinformatics analysis revealed several potential candidate genes involved in spermatogenesis, spermiogenesis and flagellum formation in the testis, and in post-testicular sperm maturation in the epididymis and ductus deferens. In the testis, genes were linked with the mitotic proliferation of spermatogonia and the meiotic division of spermatocytes. Histone ubiquitination and protamine phosphorylation were shown to be regulatory mechanisms for nuclear condensation during spermiogenesis. The characterization of testicular transcripts allowed a better understanding of acrosome formation and development and flagellum formation, including axoneme structures and functions. Spermatozoa motility during post-testicular maturation was linked to the development of flagellar actin filaments and biochemical processes, including Ca2+ influx and protein phosphorylation/dephosphorylation. Spermatozoa quality appeared to be controlled by apoptosis and antioxidant systems in the epididymis and ductus deferens. Finally, genes associated with reproductive system development and morphogenesis were identified. To the best of our knowledge, this is the first genome-wide functional investigation of genes associated with tissue-specific processes in turkey reproductive tract. A catalog of genes worthy of further studies to understand the avian reproductive physiology and regulation was provided.

Project description:BackgroundThe turkey (Meleagris gallopavo) is an important agricultural species and the second largest contributor to the world's poultry meat production. Genetic improvement is attributed largely to selective breeding programs that rely on highly heritable phenotypic traits, such as body size and breast muscle development. Commercial breeding with small effective population sizes and epistasis can result in loss of genetic diversity, which in turn can lead to reduced individual fitness and reduced response to selection. The presence of genomic diversity in domestic livestock species therefore, is of great importance and a prerequisite for rapid and accurate genetic improvement of selected breeds in various environments, as well as to facilitate rapid adaptation to potential changes in breeding goals. Genomic selection requires a large number of genetic markers such as e.g. single nucleotide polymorphisms (SNPs) the most abundant source of genetic variation within the genome.ResultsAlignment of next generation sequencing data of 32 individual turkeys from different populations was used for the discovery of 5.49 million SNPs, which subsequently were used for the analysis of genetic diversity among the different populations. All of the commercial lines branched from a single node relative to the heritage varieties and the South Mexican turkey population. Heterozygosity of all individuals from the different turkey populations ranged from 0.17-2.73 SNPs/Kb, while heterozygosity of populations ranged from 0.73-1.64 SNPs/Kb. The average frequency of heterozygous SNPs in individual turkeys was 1.07 SNPs/Kb. Five genomic regions with very low nucleotide variation were identified in domestic turkeys that showed state of fixation towards alleles different than wild alleles.ConclusionThe turkey genome is much less diverse with a relatively low frequency of heterozygous SNPs as compared to other livestock species like chicken and pig. The whole genome SNP discovery study in turkey resulted in the detection of 5.49 million putative SNPs compared to the reference genome. All commercial lines appear to share a common origin. Presence of different alleles/haplotypes in the SM population highlights that specific haplotypes have been selected in the modern domesticated turkey.

Dataset Information

Response of the hepatic transcriptome to aflatoxin B1 in domestic turkey (Meleagris gallopavo).

Publications

Response of the hepatic transcriptome to aflatoxin B1 in domestic turkey (Meleagris gallopavo).

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