11 research outputs found
Lack of <i>Sdccag8</i> expression in <i>Sdccag8</i><sup><i>SBT</i></sup> gene-trap mice.
<p><b>A)</b> Representative PCR genotyping results for <i>Sdccag8</i><sup><i>+/+</i></sup>, <i>Sdccag8</i><sup><i>+/SBT</i></sup>, <i>and Sdccag8</i><sup><i>SBT/SBT</i></sup> mice. Primer pairs Fw+Rw and Fm+Rm detect the presence of wild-type (WT) and mutant (Mut; SBT) alleles, respectively. <b>B)</b> qPCR results show the presence of <i>Sdccag8</i> mRNAs 5’ of the insertion site but their absence 3’ of the insertion. cDNAs from the brain, kidney, and lung were used for qPCR. Error bars represent standard errors. <b>C-E)</b> Immunoblot for SDCCAG8 shows loss of SDCCAG8 in the brain <b>(C)</b>, kidney <b>(D)</b>, and lung <b>(E)</b>. Arrowheads indicate full length SDCCAG8.</p
A region between rs3714172 and rs3141832 shows a significant association with survival.
<p>Comparison of allele distribution in mice that survive to P21 or later, analyzed using a Chi Square test with expected distribution.</p
Developmental defects in the <i>Sdccag8</i><sup><i>SBT/SBT</i></sup> mutant lung.
<p><b>A</b>) <i>Sdccag8</i><sup><i>SBT/SBT</i></sup> mice (right) are cyanotic at P0 (before death). A wild-type (WT) littermate is shown on the left. <b>B</b>) H&E staining of lung sections from a WT (left) and a mutant littermate (right). Scale bar = 20 μm.</p
Neonatal <i>Sdccag8</i><sup><i>SBT/SBT</i></sup> mice have secondary palate anomalies, pre-axial polydactyly, and brain abnormalities but not cystic kidney.
<p><b>A)</b> Alcian blue and Alizarin red staining of the secondary palate. Arrowheads show alterations to the basisphenoid (BS), presphenoid (PS), and premaxilla (PM) regions of the palate. <b>B)</b> Alcian blue and Alizarin red staining of the forelimb. <b>C)</b> Alcian blue and Alizarin red staining of the hind limb. <b>D)</b> H&E staining of kidney sections. Scale bar = 200 μm <b>E)</b> H&E-stained brain sections. Anterior commissures are circled and the dashed line highlights the white matter and corpus callosum. Scale bar = 500 μm. In all panels, wild-type (at P0) is shown on the left and <i>Sdccag8</i><sup><i>SBT/SBT</i></sup> mutant littermates are on the right.</p
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Prohibitin-1 Is a Dynamically Regulated Blood Protein With Cardioprotective Effects in Sepsis.
Background In sepsis, circulating cytokines and lipopolysaccharide elicit mitochondrial dysfunction and cardiomyopathy, a major cause of morbidity and mortality with this condition. Emerging research places the PHB1 (lipid raft protein prohibitin-1) at the nexus of inflammation, metabolism, and oxidative stress. PHB1 has also been reported in circulation, though its function in this compartment is completely unknown. Methods and Results Using a wide-ranging approach across multiple in vitro and in vivo models, we interrogated the functional role of intracellular and circulating PHB1 in the heart during sepsis, and elucidated some of the mechanisms involved. Upon endotoxin challenge or sepsis induction in rodent models, PHB1 translocates from mitochondria to nucleus in cardiomyocytes and is secreted into the circulation from the liver in a manner dependent on nuclear factor (erythroid-derived 2)-like 2, a key transcriptional regulator of the antioxidant response. Overexpression or treatment with recombinant human PHB1 enhances the antioxidant/anti-inflammatory response and protects HL-1 cardiomyocytes from mitochondrial dysfunction and toxicity from cytokine stress. Importantly, administration of recombinant human PHB1 blunted inflammation and restored cardiac contractility and ATP production in mice following lipopolysaccharide challenge. This cardioprotective, anti-inflammatory effect of recombinant human PHB1 was determined to be independent of nuclear factor (erythroid-derived 2)-like 2, but partially dependent on PI3K/AKTÂ signaling in the heart. Conclusions These findings reveal a previously unknown cardioprotective effect of PHB1 during sepsis, and illustrate a pro-survival, protective role for PHB1 in the circulation. Exploitation of circulating PHB1 as a biomarker and/or therapeutic could have widespread benefit in the clinical management of sepsis and other severe inflammatory disorders
Genotypic and phenotypic characterization of the <i>Sdccag8<sup>Tn(sb-Tyr)2161B.CA1C2Ove</sup></i> mouse model
<div><p>Nephronophthisis-related ciliopathies (NPHP-RC) are a group of disorders that present with end-stage renal failure in childhood/adolescence, kidney cysts, retinal degeneration, and cerebellar hypoplasia. One disorder that shares clinical features with NPHP-RC is Bardet-Biedl Syndrome (BBS). Serologically defined colon cancer antigen 8 (<i>SDCCAG8</i>; also known as NPHP10 and BBS16) is an NPHP gene that is also associated with BBS. To better understand the patho-mechanisms of NPHP and BBS caused by loss of SDCCAG8 function, we characterized an SDCCAG8 mouse model (<i>Sdccag8</i><sup><i>Tn(sb-Tyr)2161B</i>.<i>CA1C2Ove</i></sup>) generated by Sleeping Beauty Transposon (SBT)-mediated insertion mutagenesis. Consistent with the previously reported, independent SDCCAG8 mouse models, our mutant mice display pre-axial polydactyly in their hind limbs. In addition, we report patterning defects in the secondary palate, brain abnormalities, as well as neonatal lethality associated with developmental defects in the lung in our mouse model. The neonatal lethality phenotype is genetic background dependent and rescued by introducing 129S6/SvEvTac background. Genetic modifier(s) responsible for this effect were mapped to a region between SNPs rs3714172 and rs3141832 on chromosome 11. While determining the precise genetic lesion in our mouse model, we found that SBT insertion resulted in a deletion of multiple exons from both <i>Sdccag8</i> and its neighboring gene <i>Akt3</i>. We ascribe the patterning defects in the limb and the secondary palate as well as lung abnormalities to loss of SDCCAG8, while the developmental defects in the brain are likely due to the loss of AKT3. This mouse model may be useful to study features not observed in other SDCCAG8 models but cautions are needed in interpreting data.</p></div