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International Journal of Molecular... Aug 2022Acute kidney injury (AKI) is commonly associated with severe human diseases, and often worsens the outcome in hospitalized patients. The mammalian kidney has the ability...
Acute kidney injury (AKI) is commonly associated with severe human diseases, and often worsens the outcome in hospitalized patients. The mammalian kidney has the ability to recover spontaneously from AKI; however, little progress has been made in the development of supportive treatments. Increasing evidence suggest that histone deacetylases (HDAC) and NF-κB promote the pathogenesis of AKI, and inhibition of Hdac activity has a protective effect in murine models of AKI. However, the role of HDAC at the early stages of recovery is unknown. We used the zebrafish pronephros model to study the role of epigenetic modifiers in the immediate repair response after injury to the tubular epithelium. Using specific inhibitors, we found that the histone deacetylase Hdac2, Hdac6, and Hdac8 activities are required for the repair via collective cell migration. We found that , , and expression levels were upregulated in the repairing epithelial cells shortly after injury. Depletion of , , or with morpholino oligonucleotides impaired the repair process, whereas the combined depletion of all three genes synergistically suppressed the recovery process. Furthermore, time-lapse video microscopy revealed that the lamellipodia and filopodia formation in the flanking cells was strongly reduced in -depleted embryos. Our findings suggest that Hdac activity and NF-κB are synergistically required for the immediate repair response in the zebrafish pronephros model of AKI, and the timing of HDAC inhibition might be important in developing supportive protocols in the human disease.
Topics: Acute Kidney Injury; Animals; Histone Deacetylase 6; Histone Deacetylase Inhibitors; Histone Deacetylases; Humans; Mice; NF-kappa B; Pronephros; Repressor Proteins; Zebrafish; Zebrafish Proteins
PubMed: 36076983
DOI: 10.3390/ijms23179582 -
Journal of the American Society of... Nov 2011Unbiased transcriptome profiling and functional genomics approaches identified glucocorticoid-induced transcript 1 (GLCCI1) as being a transcript highly specific for the...
Unbiased transcriptome profiling and functional genomics approaches identified glucocorticoid-induced transcript 1 (GLCCI1) as being a transcript highly specific for the glomerulus, but its role in glomerular development and disease is unknown. Here, we report that mouse glomeruli express far greater amounts of Glcci1 protein compared with the rest of the kidney. RT-PCR and Western blotting demonstrated that mouse glomerular Glcci1 is approximately 60 kD and localizes to the cytoplasm of podocytes in mature glomeruli. In the fetal kidney, intense Glcci1 expression occurs at the capillary-loop stage of glomerular development. Using gene knockdown in zebrafish with morpholinos, morphants lacking Glcci1 function had collapsed glomeruli with foot-process effacement. Permeability studies of the glomerular filtration barrier in these zebrafish morphants demonstrated a disruption of the selective glomerular permeability filter. Taken together, these data suggest that Glcci1 promotes the normal development and maintenance of podocyte structure and function.
Topics: Animals; Cytoplasm; Dexamethasone; Disease Models, Animal; Gene Expression Regulation, Developmental; Glucocorticoids; Kidney Glomerulus; Male; Mice; Mice, Inbred ICR; Oligonucleotides, Antisense; Podocytes; Pronephros; Proteinuria; Rabbits; Receptors, Glucocorticoid; Zebrafish; Zebrafish Proteins
PubMed: 21949092
DOI: 10.1681/ASN.2010111147 -
Journal of Anatomy Aug 2009Cystathionine gamma-lyase (CSE) is a key enzyme in the trans-sulphuration pathway for the biosynthesis of cysteine from methionine and catalyses the hydrolysis of...
Cystathionine gamma-lyase (CSE) is a key enzyme in the trans-sulphuration pathway for the biosynthesis of cysteine from methionine and catalyses the hydrolysis of cystathionine into cysteine. It has been reported to be expressed in mammalian liver and kidney but so far no comprehensive developmental expression analysis of CSE has been available. We cloned a 600 bp fragment of chick CSE cDNA and analysed its expression pattern during avian embryonic development until embryonic day 13. We found CSE expression in various developing organs including the notochord, eye, neural tube, limb bud mesenchyme and sclerotomal compartment of the somites. Notably, prominent expression was found in renal epithelia throughout kidney development, i.e. in the tubular structures of pronephros, mesonephros and metanephros. Our data introduce CSE as a novel marker gene to study avian kidney development.
Topics: Amino Acid Sequence; Animals; Chick Embryo; Cystathionine gamma-Lyase; Embryonic Development; Gene Expression Regulation, Developmental; Gene Expression Regulation, Enzymologic; Kidney; Molecular Sequence Data; Notochord; Sequence Alignment; Species Specificity
PubMed: 19486201
DOI: 10.1111/j.1469-7580.2009.01092.x -
BMC Genomics Nov 2022Infectious Salmon Anaemia virus (ISAV) is an orthomyxovirus responsible for large losses in Atlantic salmon (Salmo salar) aquaculture. Current available treatments and...
BACKGROUND
Infectious Salmon Anaemia virus (ISAV) is an orthomyxovirus responsible for large losses in Atlantic salmon (Salmo salar) aquaculture. Current available treatments and vaccines are not fully effective, and therefore selective breeding to produce ISAV-resistant strains of Atlantic salmon is a high priority for the industry. Genomic selection and potentially genome editing can be applied to enhance the disease resistance of aquaculture stocks, and both approaches can benefit from increased knowledge on the genomic mechanisms of resistance to ISAV. To improve our understanding of the mechanisms underlying resistance to ISAV in Atlantic salmon we performed a transcriptomic study in ISAV-infected salmon with contrasting levels of resistance to this virus.
RESULTS
Three different tissues (gills, head kidney and spleen) were collected on 12 resistant and 12 susceptible fish at three timepoints (pre-challenge, 7 and 14 days post challenge) and RNA sequenced. The transcriptomes of infected and non-infected fish and of resistant and susceptible fish were compared at each timepoint. The results show that the responses to ISAV are organ-specific; an important response to the infection was observed in the head kidney, with up-regulation of immune processes such as interferon and NLR pathways, while in gills and spleen the response was more moderate. In addition to immune related genes, our results suggest that other processes such as ubiquitination and ribosomal processing are important during early infection with ISAV. Moreover, the comparison between resistant and susceptible fish has also highlighted some interesting genes related to ubiquitination, intracellular transport and the inflammasome.
CONCLUSIONS
Atlantic salmon infection by ISAV revealed an organ-specific response, implying differential function during the infection. An immune response was observed in the head kidney in these early timepoints, while gills and spleen showed modest responses in comparison. Comparison between resistance and susceptible samples have highlighted genes of interest for further studies, for instance those related to ubiquitination or the inflammasome.
Topics: Animals; Isavirus; Head Kidney; Salmo salar; Spleen; Gills; Transcriptome; Inflammasomes
PubMed: 36443659
DOI: 10.1186/s12864-022-09007-4 -
Frontiers in Cell and Developmental... 2017Polycystic kidney disease (PKD) proteins are trans-membrane proteins that have crucial roles in many aspects of vertebrate development and physiology, including the...
Polycystic kidney disease (PKD) proteins are trans-membrane proteins that have crucial roles in many aspects of vertebrate development and physiology, including the development of many organs as well as left-right patterning and taste. They can be divided into structurally-distinct PKD1-like and PKD2-like proteins and usually one PKD1-like protein forms a heteromeric polycystin complex with a PKD2-like protein. For example, PKD1 forms a complex with PKD2 and mutations in either of these proteins cause Autosomal Dominant Polycystic Kidney Disease (ADPKD), which is the most frequent potentially-lethal single-gene disorder in humans. Here, we identify the complete family of genes in zebrafish and other teleosts. We describe the genomic locations and sequences of all seven genes: , and . are likely to be ohnologs of , preserved from the whole genome duplication that occurred at the base of the teleosts. However, in contrast to mammals and cartilaginous and holostei fish, teleosts lack , and genes, suggesting that these have been lost in the teleost lineage. In addition, teleost, and holostei fish have only a partial sequence, suggesting that this gene may be in the process of being lost in the ray-finned fish lineage. We also provide the first comprehensive description of the expression of zebrafish genes during development. In most structures we detect expression of one -like gene and one -like gene, consistent with these genes encoding a heteromeric protein complex. For example, we found that and are expressed in Kupffer's vesicle and and are expressed in the developing pronephros. In the spinal cord, we show that and are co-expressed in KA cells. We also identify potential co-expression of and in the floor-plate. Interestingly, and in contrast to mouse, we observe expression of all seven genes in regions that may correspond to taste receptors. Taken together, these results provide a crucial catalog of genes in an important model system for elucidating cell and developmental processes and modeling human diseases and the most comprehensive analysis of embryonic gene expression in any vertebrate.
PubMed: 28271061
DOI: 10.3389/fcell.2017.00005 -
Scientific Reports Feb 2021A wide variety of environmental contaminants has been shown to disrupt immune functions of fish and may compromise their defense capability against pathogens....
A wide variety of environmental contaminants has been shown to disrupt immune functions of fish and may compromise their defense capability against pathogens. Immunotoxic effects, however, are rarely considered in ecotoxicological testing strategies. The aim of this study was to systematically evaluate the suitability of an in vitro immuno-assay using selected fish immune parameters to screen for chemicals with known immunotoxic potential and to differentiate them from non-immunotoxicants. Non-stimulated and lipopolysaccharide-stimulated head kidney leukocytes of rainbow trout (Oncorhynchus mykiss) were exposed for 3 h or 19 h to chemicals with different modes of action. As immune parameters, phagocytosis activity, oxidative burst activity and cytokine transcription (IL-1β, TNFα, IL-10) were examined, accompanied by in silico modelling. The immunotoxicants dexamethasone, benzo(a)pyrene, ethinylestradiol and bisphenol A significantly altered the immune parameters at non-cytotoxic concentrations whereas diclofenac had only weak effects. However, the two baseline chemicals with no known immunotoxic potential, butanol and ethylene glycol, caused significant effects, too. From our results it appears that the in vitro fish leukocyte assay as performed in the present study has only a limited capacity for discriminating between immunotoxicants and non-immunotoxicants.
Topics: Animals; Benzhydryl Compounds; Benzo(a)pyrene; Butanols; Dexamethasone; Diclofenac; Ethinyl Estradiol; Ethylene Glycol; Female; Fish Proteins; Gene Expression Regulation; Head Kidney; Immunotoxins; Interleukin-10; Interleukin-1beta; Leukocytes; Oncorhynchus mykiss; Phagocytosis; Phenols; Primary Cell Culture; Respiratory Burst; Transcription, Genetic; Tumor Necrosis Factor-alpha; Water Pollutants, Chemical
PubMed: 33542403
DOI: 10.1038/s41598-021-82711-5 -
Uchu Seibutsu Kagaku Mar 2000The Amphibia bridge the phyletic gap between the aquatic fishes and the terrestrial vertebrates. This transition has involved many interesting changes of metabolisms. In... (Review)
Review
The Amphibia bridge the phyletic gap between the aquatic fishes and the terrestrial vertebrates. This transition has involved many interesting changes of metabolisms. In this short review, we have attempted to summarize the kidney structure and functions on the osmoregulations in the Amphibia. Amphibians excrete the water absorbed through their skin as a dilute urine. Pronephros of tadpoles may start to work in the hatching stages and metanephros is well developed and functions. Glomerular filtration rate is relatively large and glomerular intermittency is important to regulate urine production. The proximal tubule reabsorbs approximately 20-45% of filtered water and sodium. Absorption is driven by the basolateral Na+, K(+)-ATPase common to all tubular cells. The diluting segment, early parts of distal nephron, highly develops basolateral interdigitation and reabsorbs approximately 40% of filtered Na+, K+, and Cl-, but is impermeable to water, thus this part results in the formation of hypo-osmotic tubular fluid. In the late distal tubule, the primary mechanism of reabsorption may be via a luminal NaCl synporter, driven by the ubiquitous Na+, K(+)-ATPase on basolateral membrane. In collecting tubule, there are two types of cells, the principal cells and the intercalated cells. Many hormonal and nervous regulations are involved in the glomerular filtration rate and reabsorptions in the amphibian nephrons.
Topics: Adaptation, Biological; Amphibians; Animals; Anura; Glomerular Filtration Rate; Kidney; Kidney Tubules; Microscopy, Electron; Nephrons; Water-Electrolyte Balance
PubMed: 11543150
DOI: 10.2187/bss.14.22 -
The International Journal of... 2012The translation initiation factor Eif6 has been implicated as a regulator of ribosome assembly, selective mRNA translation and apoptosis. Many of these activities depend...
The translation initiation factor Eif6 has been implicated as a regulator of ribosome assembly, selective mRNA translation and apoptosis. Many of these activities depend upon the phosphorylation of eif6 Serine 235 by protein kinase C (PKC). Eif6-60S is probably part of the RNA-induced silencing complex (RISC). eif6 over-expression in Xenopus embryos causes aberrant eye development. kermit2/gipc2 morphants have an eye phenotype similar to that of the eif6 overexpressors. Eye formation is regulated by insulin growth factor (IGF) signalling. eif6 interacts with the IGF receptor (IGFR) and kermit2/gipc2, which also binds to igfr. eif6 over-expression in Xenopus causes also the formation of antero-ventral oedema, suggesting a malfunction of the excretory system. Here we evaluated the pronephros phenotype. The oedema grows into the nephrocoel, expanding its boundary and is accompanied by a strong reduction of the pronephros. The three main components of the pronephros are severely impaired in eif6 over-expressors, while are not affected in eif6 morphants. Conversely, gipc2 depletion induces the oedema phenotype and reduction of the pronephros, while gipc2 overexpression does not. p110*, a constitutively active p110 subunit of the PI3 kinase partially recovers the oedema phenotype. We also determined that PKC-dependent phosphorylation of Ser235 in eif6 is not required to produce defective pronephroi. These results indicate that the levels of eif6 are highly regulated during development and instrumental for proper morphogenesis of the pronephros. Moreover, it appears that for proper pronephros development the gipc2 level should be kept within or over the physiological range and that the oedema phenotype is partly due to the inhibition of IGF signalling.
Topics: Adaptor Proteins, Signal Transducing; Animals; Carrier Proteins; Edema; Embryo, Nonmammalian; Female; Immunoenzyme Techniques; In Situ Hybridization; Morphogenesis; Nerve Tissue Proteins; Peptide Initiation Factors; Phosphatidylinositol 3-Kinases; Pronephros; Receptor, IGF Type 2; Xenopus Proteins; Xenopus laevis
PubMed: 22689378
DOI: 10.1387/ijdb.120009nd -
Scientific Reports Apr 2019The genetic regulation of nephron patterning during kidney organogenesis remains poorly understood. Nephron tubules in zebrafish are composed of segment populations that...
The genetic regulation of nephron patterning during kidney organogenesis remains poorly understood. Nephron tubules in zebrafish are composed of segment populations that have unique absorptive and secretory roles, as well as multiciliated cells (MCCs) that govern fluid flow. Here, we report that the transcription factor iroquois 2a (irx2a) is requisite for zebrafish nephrogenesis. irx2a transcripts localized to the developing pronephros and maturing MCCs, and loss of function altered formation of two segment populations and reduced MCC number. Interestingly, irx2a deficient embryos had reduced expression of an essential MCC gene ets variant 5a (etv5a), and were rescued by etv5a overexpression, supporting the conclusion that etv5a acts downstream of irx2a to control MCC ontogeny. Finally, we found that retinoic acid (RA) signaling affects the irx2a expression domain in renal progenitors, positioning irx2a downstream of RA. In sum, this work reveals new roles for irx2a during nephrogenesis, identifying irx2a as a crucial connection between RA signaling, segmentation, and the control of etv5a mediated MCC formation. Further investigation of the genetic players involved in these events will enhance our understanding of the molecular pathways that govern renal development, which can be used help create therapeutics to treat congenital and acquired kidney diseases.
Topics: Animals; Cell Differentiation; Organogenesis; Pronephros; Transcription Factors; Zebrafish; Zebrafish Proteins
PubMed: 31015532
DOI: 10.1038/s41598-019-42943-y -
Kidney International Oct 2022The zebrafish is an important animal system for modeling human diseases. This includes kidney dysfunction as the embryonic kidney (pronephros) shares considerable...
The zebrafish is an important animal system for modeling human diseases. This includes kidney dysfunction as the embryonic kidney (pronephros) shares considerable molecular and morphological homology with the human nephron. A key clinical indicator of kidney disease is proteinuria, but a high-throughput readout of proteinuria in the zebrafish is currently lacking. To remedy this, we used the Tol2 transposon system to generate a transgenic zebrafish line that uses the fabp10a liver-specific promoter to over-express a nanoluciferase molecule fused with the D3 domain of Receptor-Associated Protein (a type of molecular chaperone) which we term NL-D3. Using a luminometer, we quantified proteinuria in NL-D3 zebrafish larvae by measuring the intensity of luminescence in the embryo medium. In the healthy state, NL-D3 is not excreted, but when embryos were treated with chemicals that affected either proximal tubular reabsorption (cisplatin, gentamicin) or glomerular filtration (angiotensin II, Hanks Balanced Salt Solution, Bovine Serum Albumin), NL-D3 is detected in fish medium. Similarly, depletion of several gene products associated with kidney disease (nphs1, nphs2, lrp2a, ocrl, col4a3, and col4a4) also induced NL-D3 proteinuria. Treating col4a4 depleted zebrafish larvae (a model of Alport syndrome) with captopril reduced proteinuria in this system. Thus, our findings validate the use of the NL-D3 transgenic zebrafish as a robust and quantifiable proteinuria reporter. Hence, given the feasibility of high-throughput assays in zebrafish, this novel reporter will permit screening for drugs that ameliorate proteinuria, thereby prioritizing candidates for further translational studies.
Topics: Angiotensin II; Animals; Animals, Genetically Modified; Captopril; Cisplatin; Gentamicins; Humans; Kidney Glomerulus; Nephritis, Hereditary; Nephrotic Syndrome; Proteinuria; Serum Albumin, Bovine; Zebrafish
PubMed: 35716957
DOI: 10.1016/j.kint.2022.05.019