![]() ![]() ![]() In response to osmotic stress, a distinct suite of modifications in gill epithelia is activated for functional adaptation, which involves cell proliferation and differentiation, changes in the activities, expressions and trafficking of different ion transporters/channels. Fish gills therefore possess great structural and functional plasticity in response to variations of water osmolality, to support the process of fluid and electrolyte homeostasis. The fish is euryhaline and actively engage physiological responses to oppose osmotic perturbations to stabilize body osmolality. The catadromous fish Japanese eels have a complex life cycle in freshwater and seawater environments. MicroRNA-messenger RNA interactome analysis of miR-29b-3p and miR-200b-3p revealed the gene targets are essential for osmotic stress responses. In this study, we have characterized the hypo-osmoregulatory responses and unraveled the modulation of miR-biogenesis factors/the dysregulation of miRs, using ex-vivo gill filament culture. The target-genes are known to regulate differentiation of gill ionocytes and cellular osmolality. Integrated miR-mRNA-omics analysis revealed the specific binding of miR-29b-3p on Klf4 and miR-200b-3p on slc17a5. Commonly upregulated gene transcripts from the hyperosmotic experiments and miR-inhibition studies, were overlaid, in which two miR-29b-3p target-genes and one miR-200b-3p target-gene (slc17a5) were identified. An inhibition of miR-29b-3p and miR-200b-3p in primary gill cell culture led to an upregulation of 100 and 93 gene transcripts, respectively. Transcriptome and miR-sequencing of gill filament samples at 4 and 8 h were conducted and two downregulated miRs, miR-29b-3p and miR-200b-3p were identified. drosha RNase III endonuclease, exportin-5, dicer ribonuclease III and argonaute-2) involved in miR biogenesis were dysregulated ( P < 0.05). In the hyperosmotic treatment, four key factors (i.e. The data illustrated that the ex-vivo gill filament culture exhibited distinctive responses to hyperosmotic challenge. Hypertonic responsive genes, including osmotic stress transcriptional factor, Na +/Cl −-taurine transporter, Na +/H + exchange regulatory cofactor, cystic fibrosis transmembrane regulator, inward rectifying K + channel, Na +/K +-ATPase, and calcium-transporting ATPase were significantly upregulated, while the hypo-osmotic gene, V-type proton ATPase was downregulated. ResultsĮx-vivo gill filament culture was exposed to Leibovitz’s L-15 medium (300 mOsmol l − 1) or the medium with an adjusted osmolality of 600 mOsmol l − 1 for 4, 8 and 24 h. The present study is to characterize an ex-vivo gill filament culture and using omics approach, to decipher the interaction between tonicity-responsive miRs and gene targets, in orchestrating the osmotic stress-induced responses. The osmoregulatory plasticity of gills provides an excellent model to study the role of microRNAs (miRs) in adaptive osmotic responses. Gills of euryhaline fishes possess great physiological and structural plasticity to adapt to large changes in external osmolality and to participate in ion uptake/excretion, which is essential for the re-establishment of fluid and electrolyte homeostasis. ![]()
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |