Silencing of PPARαBb mRNA in brown trout primary hepatocytes: effects on molecular and morphological targets under the influence of an estrogen and a PPARα agonist
Autor: | Ivone Pinheiro, Ralph Urbatzka, Fernanda Malhão, Tânia Vieira Madureira, L. Filipe C. Castro, Eduardo Rocha |
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Rok vydání: | 2019 |
Předmět: |
Fish Proteins
0301 basic medicine Gene isoform Trout Physiology medicine.drug_class Primary Cell Culture Biology Biochemistry Mediator Complex Subunit 1 03 medical and health sciences 0302 clinical medicine medicine Animals Humans Gene silencing PPAR alpha Gene Silencing Receptor Molecular Biology Gene Oxidase test Messenger RNA Estrogens Peroxisome Cell biology Pyrimidines 030104 developmental biology Estrogen 030220 oncology & carcinogenesis Hepatocytes |
Zdroj: | Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology. 229:1-9 |
ISSN: | 1096-4959 |
Popis: | The crosstalk between peroxisome proliferator-activated receptor α (PPARα) and estrogenic pathways are shared from fish to humans. Salmonid fish had an additional genome duplication, and two PPARα isoforms (PPARαBa and PPARαBb) were previously identified. Since a negative regulation between estrogen signaling and PPARα was described, a post-transcriptional gene silencing for PPARαBb was designed in primary brown trout hepatocytes. The aims of the study were to: (i) decipher the effects of PPARαBb knock-down on peroxisome morphology and on mRNA expression of potential target genes, and (ii) to assess the cross-interferences caused by an estrogenic compound (17α-ethinylestradiol - EE2) and a PPARα agonist (Wy-14,643 - Wy) using the established knock-down model. A knock-down efficiency of 70% was achieved for PPARαBb and its silencing significantly reduced the volume density of peroxisomes, but did not alter mRNA levels of the studied genes. Exposure to Wy did not change peroxisome morphology or mRNA expression, but under silencing conditions Wy rescued the volume density of peroxisomes to control levels, and increased acyl-coenzyme A oxidase 1-3l (Acox1-3l) mRNA. Exposure to EE2 caused a reduction of peroxisome volume density, but under silencing conditions this effect was abolished and ApoA1 mRNA level was diminished. The morphological alterations of peroxisomes by WY and EE2 demonstrated that obtained results are PPARαBb dependent, and suggest the regulation of unknown downstream targets of PPARαBb. In summary, PPARαBb is involved in the control of peroxisome size and/or number, which opens future opportunities to explore its regulation and molecular targets. |
Databáze: | OpenAIRE |
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