bluuebunny/crossref_metadata_2025_split huggingface dataset:
Diverse nephron cell type‐specific adaptation to furosemide by morphometry and single cell RNA sequencing
Authors: Schaffert Steven , Krauson Aram , Walczak Elisabeth , Nizar Jonathan , Holdgate Gwendolyn , Iyer Sonali , Elsayed Ragwa , Qiao Yanli , Khatri Purvesh , Bhalla Vivek 
Published: June 2021

Furosemide, a commonly prescribed diuretic to treat hypertension and edema‐forming states, inhibits sodium reabsorption in the thick ascending limb (TAL) of the loop of Henle. Tubular adaptation to diuretics has been documented but not fully explored. Using morphometry and single cell RNA sequencing in furosemide vs. vehicle‐treated mice, we found that furosemide induces renal tubular epithelial hyperplasia selectively in distal, but not proximal, nephron segments after 1 week that subsides by 3 weeks. We measured an increase in kidney size in furosemide‐treated mice, but in contrast to hyperplasia, we measured cellular hypertrophy in both proximal and distal nephron segments, after 1 week treatment that progresses at 3 weeks. We detected an increase in receptor tyrosine kinase pathways and IGF‐1 protein in whole kidney with furosemide, and inducible deletion of renal tubular Igf1r, diminished overall kidney growth and proximal but not distal tubular hypertrophy. In TAL, furosemide enhanced TUNEL‐positive nuclei with selective depletion of Cldn10(+) / Cldn16(−) cells that were enriched for pro‐apoptotic genes Hist1h1c and Fxyd2. Each of the 15 identified nephron segments / cell‐types had diverse changes in relative gene expression with a small cluster of precursor Wnt4(+) cells demonstrating the largest reactivity and a transition toward a sodium reabsorptive phenotype. We demonstrate that furosemide‐treated mice undergo cell type‐specific forms of tubular remodeling including selective changes in hyperplasia, distinct IGF‐1 receptor‐dependent and independent hypertrophy, Cldn10(+) cell death, and significant perturbations of the transcriptional programme of Wnt4(+) cells. These findings highlight mechanisms of diuretic resistance and greatly expand the implications of long‐term diuretic use for human disease.Support or Funding InformationNIH / NIDDKThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Polycystin 1 is an atypical adhesion GPCR that responds to non‐canonical WNT signals and inhibits GSK3β
Authors: Gresko Nikolay , Merrick David M , Mistry Kavita M , Caplan Michael J 
Published: June 2021

Autosomal dominant polycystic kidney disease (ADPKD) is caused by mutations in the PKD1 and PKD2 genes, which encode PC1 and PC2, respectively. PC1 is a 460kD multi‐spanning membrane protein. PC1 undergoes multiple proteolytic cleavages, one of which is an autocatalytic cleavage that produces a large N terminal fragment (NTF) that remains non‐covalently attached to the membrane‐embedded C terminal fragment (CTF). A recent study demonstrates that PC1 serves as a receptor for Wnt ligands through the direct binding of these ligands to the PC1 NTF. We find that PC1 responds to Wnt9b by shedding its 350kD NTF, which bears a striking resemblance to the activation mechanism of an adhesion GPCR. For aGPCRs, NTF removal liberates a short peptide sequence near the GPS cleavage site called stachel, which then interacts with an internal binding site and engages a G protein signalling cascade. We find that Wnt9b induces a PC1‐dependent phosphorylation and inhibition of the GSK3β kinase through the RhoA GTPase‐ROCK kinase pathway. Expression of a “constitutively active” aGPCR form of PC1 that lacks its NTF leads to profound suppression of GSK3β activity that is dependent upon RhoA and ROCK. Interestingly, GSK3β is an important negative regulator of the HIPPO/non‐canonical Wnt signaling target TAZ(WWTR1), lack of which leads to the development severe renal cysts. Furthermore, recent data show that pharmacological inhibition of GSK3β is beneficial in mouse models of ADPKD. We find that exogenous expression of an active form of TAZ in PC1 null cells or in PKD1a/b morphant zebrafish suppresses the development of relevant phenotypes. We also demonstrate that pharmacological inhibition of GSK3β leads to accumulation of the TAZ protein and to reduced cystogenesis in a 3D matrigel assay employing PC1 null cells. TAZ abundance and activity have been shown to be upregulated through the non‐canonical Wnt signalling pathway. We find that HEK293 cells that express PC1 and PC2 respond to Wnt9b treatment by significantly increasing TAZ abundance as compared to the wild type HEK293 cells treated in the same manner. Taken together, our data suggest that PC1 is an aGPCR‐like receptor for non‐canonical Wnt ligands that participates in a novel signalling pathway by linking non‐canonical Wnt ligands to the GSK3β‐dependent regulation of TAZ, a multifaceted signalling molecule whose absence is sufficient to induce renal cystic disease.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Ibuprofen Treatment Results in Gender‐Specific Proteasome Dysfunction in Murine Kidneys
Authors: Georgieva Anna Y , Tiwari Shuchita , Gomes Aldrin V 
Published: June 2021

Non‐steroidal anti‐inflammatory drugs (NSAIDs) are some of the most commonly consumed drugs currently available. In the United States alone, more than 30 billion over the counter NSAIDs are consumed annually, and more than 600 different products contain NSAIDs. These drugs are popular among all generations due to their ability to relieve pain, inflammation, and fevers. The enzymes cyclooxygenase 1 and 2 (COX‐1 and COX‐2) synthesize prostaglandins, which are associated with the pain, swelling and fevers accompanying tissue damage or infection. NSAIDs lessen these effects by inhibiting COX‐1 and/or COX‐2 and reducing the synthesis of prostaglandins. However, previous studies show that there is a significant correlation between chronic NSAID use and cardiovascular disease, with a 1.3–3.36‐fold increase for risk of stroke. Diclofenac, another common NSAID, has been linked to a >95% population decline of the wild Oriental white‐backed vulture due to renal failure. Previously published data from our lab shows that mice treated with Diclofenac have proteasome dysfunction in heart, liver, and kidney tissues. The 26S proteasome is an ATP‐dependent multi‐catalytic protease, which is responsible for degrading 60–80% of intracellular proteins in all tissues. The proteasome degrades poly‐ubiquitinated proteins through the ubiquitin‐proteasome system (UPS). We hypothesize that proteasome dysfunction occurs in murine kidneys when the commonly used NSAID, ibuprofen, is administered to mice. Male and female eight‐week old mice were treated with ibuprofen (100mg/kg/day) for seven days, and then their kidneys were harvested. The activity of the three catalytic sites on the 26S proteasome, β1, β2, and β5, were measured in control vs ibuprofen kidney samples using biochemical activity assays. To determine protein expression levels of proteasome subunits and poly‐ubiquitinated proteins, western blots were carried out using the respective antibodies. Both male and female samples showed a significant decrease in β1 (caspase‐like) activity. Interestingly, female samples had a significant decrease in β2 (trypsin‐like) activity, while males did not, but males show a significant decrease in β5 (chymotrypsin‐like) activity, while females show no significant difference. We found no difference in free ubiquitin or poly‐ubiquitinated proteins in both males or females, as well as no changes in proteasome subunit expression in both males or females. Our results suggest that ibuprofen treatment causes proteasome dysfunction in murine kidneys in a gender dependent manner.Support or Funding InformationResearch was funded by 5P42ES004966 NIH grant and AHA16GRNT31350040 grant to Aldrin Gomes. A.Y.G. was supported by PREP@UCD, NIH 5R25GM116690‐03 to UC Davis.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Activation of G Protein‐coupled Estrogen Receptor Prevents High Salt‐induced Hypertension
Authors: Aceves Jeffrey O. , Gohar Eman Y. , Daugherty Elizabeth , Pollock David M. 
Published: June 2021

Estrogen protects premenopausal women against cardiovascular and renal diseases. G protein‐coupled estrogen receptor (GPER) is a membrane bound protein that prompts protective effects throughout the cardiovascular and renal systems. Accumulating evidence suggests that GPER activation contributes to the maintenance of cardiovascular and renal health in women and may even be a potential therapeutic approach for post‐menopausal hypertension. The goal of the current study was to test the hypothesis that GPER activation reduces the elevation in blood pressure in ovariectomized (OVX) rats in response to high salt (HS) diet. In our experiment, female Sprague Dawley rats were implanted with telemetry transmitters, subjected to bilateral ovariectomy, and treated with vehicle or G1, a GPER agonist (400 μg/kg/day, osmotic minipump). Two weeks later, rats were placed into metabolic cages to obtain 24‐hour urine samples under normal salt (NS = 0.49% NaCl), and HS (4% NaCl) diets. Vehicle treated rats had significantly higher mean arterial blood pressures after OVX and HS challenge compared to their baseline (113±1 vs. 105±1 mmHg, respectively, n=6, p<0.05). This hypertensive response was prevented by G1 (106±3 vs. 104±2 mmHg, respectively, n=6). Similar effects were observed in both diastolic and systolic blood pressures. Given that the renal endothelin‐1 (ET‐1) plays an important role the regulation of blood pressure and Na+ homeostasis, we tested whether GPER activation regulate the expression of ET‐1 receptors in the kidney. We found that renal inner medulla obtained from HS‐fed OVX rats treated with G1 for 3 weeks elicited a significant reduction in the mRNA expression of ET‐1 receptor subtype A (ETAreceptor) relative to vehicletreated rats (0.62±0.05 vs 1.00±0.13 fold change, respectively, n=5, p<0.05). A separate group of intact female rats were treated with G15, a GPER antagonist (400 μg/kg/day, osmotic minipump). After being challenged with a HS diet, a significant increase in mean arterial pressure was observed compared to baseline (113±1 vs. 109±1 mmHg, respectively, n=6, p<0.05) in rats given G15. These data supports the hypothesis that GPER plays an important role in the maintenance of blood pressure in females fed HS diet.Support or Funding InformationThis project was supported by a STRIDE fellowship from the APS to JOA, AHA 18CDA34110010 and 15POST25090329 to EYG, and P01 HL 069999 and P01 HL136267 to DMP.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Assessing Myo‐mechanicals Characteristic of Rectus Abdominis Muscle after Stem Cell Therapy
Authors: Quiroz Sofia , Barbosa Angélica , Floriano Juliano , Graeff Carlos , Marcondes João , Rudge Marilza 
Published: June 2021

Hyperglycemia during gestation have the deleterious effect in the skeletal muscle leading to occurrence of urinary incontinence and pelvic floor muscle dysfunctions in women. Induces functional, metabolic and structural changes in skeletal muscle; decreasing the functional capacity of the muscle, currently there is not therapies in diabetic gestational myopathy and the new proposal of muscle regeneration is based in stem cell therapy in biodevice use. To evaluate the functionality of the muscle in the new therapies to investigate the effectiveness of the muscle regeneration by assessing myo‐mechanicals characteristic of rectus abdominis muscle after therapeutic application of biodevice with mesenchymal stem cells in experimental model in diabetic and non‐diabetic pregnant rats. 20 postpartum rats of the Sprague Dawley® specific pathogen free strain, composing IV groups as follows: I. Diabetic group without biodevice (GD), II. Diabetic group with biodevice (GD ‐Bio), III. Non‐diabetic group without biodevice (GND) and IV. Non‐diabetic group with biodevice (GND‐Bio). The study was approved by the Botucatu Medical School Ethics Committee on the Use of Animals (CEUA) No. 1234/2017. After the euthanasia of the animals, the rectus abdominis samples will be removed and immediately mounted on the supports inside the isolated organ chambers of the Myograph equipment (Model 820MS; Danish Myo Technology®), the segments will be attached and connected to isometric transducers, submerged in solution physiological composition of Krebs [(mM): NaCl, 130; NaHCO3, 14.9; dextrose, 5.5; KCl, 4.7; KH2PO4, 1.18; MgSO47H2O, 1.17 e CaCl22H2O, 1.6]. Maintained at 37 ° C and constantly bubbled with 95% O2 / 5% CO2 mixture being fixed between a force transducer and a fixed unit. Application of stabilization protocol and then electrical stimulation. The parcial results show that the treated group has an increase of 3mN more than the untreated group; this indicates a better myo‐mechanicals condition in non‐diabetic rats. The Biodevice treatment is effective for the development of muscle regeneration therapy, being able to increase the contractile capacity of the rectus abdominis muscle in the model of diabetic and non‐diabetic pregnant rats.Support or Funding InformationThis project is funded for Sao Paulo Research Foundation, FAPESP (number grant 2016/01743‐5)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Macrophage BACH1, a heme regulated transcriptional repressor, controls HMOX1 and skeletal muscle regeneration
Authors: Patsalos Andreas , Tzerpos Petros , Halasz Laszlo , Nagy Gergely , Giannakis Nikolas , Pap Attila , Lyroni Konstantina , Koliaraki Vasiliki , Kollias George , Spilianakis Charalambos G. , Nagy Laszlo 
Published: June 2021

Skeletal muscle regeneration following injury is a complex process that involves inflammatory components. Macrophage infiltration to damaged tissues upon injuries and their in‐situ differentiation into effector/inflammatory and repair macrophages is indispensable for necrotic debris clearance, and for coordination of tissue repair. Regulatory events controlling the highly dynamic macrophage phenotype switch are not well understood. Here, by using an acute sterile skeletal muscle injury model, and by studying macrophage chromatin accessibility (ATAC‐seq) and gene expression (RNA‐seq) dynamics during inflammation and tissue regeneration, we identified a heme‐ and MARE motif binding transcription factor, BTB Domain And CNC Homolog 1 (BACH1), as a novel regulatory molecule of the macrophage phenotype switch following tissue injury. We show that Bach1 deletion impairs muscle regeneration, alters the dynamics of the macrophage phenotype transition, impacts myoblast proliferation and leads to transcriptional deregulation of critical inflammatory and repair‐related genes. We also find that BACH1 directly binds and regulates distal regulatory elements of these genes, suggesting a role of BACH1 in controlling a spectrum of the inflammatory and tissue repair response genes in macrophages upon tissue injury. As proof of concept, inactivation of heme oxygenase 1 (Hmox1), a known downstream BACH1 target and the most deregulated gene in the Bach1 KO, in macrophages, impairs muscle regeneration by changing the dynamics of the macrophage phenotype switch. Collectively, our data suggest the existence of a regulatory axis, involving BACH1 and HMOX1 that control the phenotype and function of the infiltrating myeloid cells upon tissue damage, shaping the overall tissue repair kinetics.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Expression of Vitamin D Receptor and Local Vitamin D Metabolism at the Different Stages of Skeletal Muscle Growth
Authors: Srikuea Ratchakrit , Hirunsai Muthita , Charoenphandhu Narattaphol 
Published: June 2021

Vitamin D metabolism is an important regulatory process for vitamin D3 action on the target cells. Nevertheless, expression of vitamin D receptor (VDR) and its association with the local regulation of vitamin D metabolism in skeletal muscle at the different stages of growth are currently unknown. In this study, hindlimb muscle of male C57BL/6 mice at the different growth stages including development (1‐month‐old), maturation (6‐month‐old), and aging (18‐month‐old) were investigated. Histological features of skeletal muscle were examined to evaluate an alteration of skeletal muscle plasticity across ages. Protein expression levels of VDR and vitamin D metabolizing enzymes (CYP24A1 and CYP27B1) at all ages were analyzed. Serum 25(OH)D3 was quantified to determine its relationship with VDR, CYP24A1, and CYP27B1 protein expression in skeletal muscle. The results revealed a significant increase of muscle fiber cross‐sectional area but not fiber number during developmental to maturation stage of skeletal muscle growth. In contrast, aging muscle presented a significant increase of centronucleated muscle fibers. These changes of skeletal muscle plasticity in mature and aged muscles were accompanied with the increase of serum 25(OH)D3 and VDR protein expression, respectively. In contrast, CYP24A1 and CYP27B1 protein expression in skeletal muscle were not changed at any age investigated. Collectively, these findings demonstrated the associations of 25(OH)D3 and VDR protein expression levels with the stages of skeletal muscle growth. However, the alterations of circulating 25(OH)D3 and VDR protein expression did not affect local vitamin D metabolism in skeletal muscle.Support or Funding InformationThis work was supported by grants(to RS) from the Thailand Research Fund and Office of the Higher Education Commission (MRG6080090) and the Faculty of Science, Mahidol University.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Loss of Endogenous Neuregulin‐4 Increases Intestinal Epithelial Permeability and Apoptosis
Authors: Bernard Jessica Kathleen , Schumacher Michael A , Frey Mark R 
Published: June 2021

Growth factor receptors such as the ErbB receptor tyrosine kinases can preserve intestinal barrier integrity in the face of inflammation by activating anti‐apoptotic pathways and stabilizing epithelial tight junction components. Activation of ErbB4 with pharmacological administration of its specific ligand neuregulin‐4 (NRG4) promotes epithelial cell survival by anti‐apoptotic pathways (Src and PI3K/Akt) and reduces intestinal inflammation. However, it is unclear whether ErbB4 is an essential regulator of intestinal permeability and if endogenous NRG4 is required for the response to intestinal inflammation. In this study, we investigated the role of endogenous NRG4 in intestinal barrier function in vivo.METHODSBaseline intestinal permeability in 11–17 week old wild type and NRG4 knockout mice was determined by oral gavage with FITC‐dextran (4kDa) followed by analysis of blood collected by cardiac puncture 4 h later. Barrier recovery from bacterial antigen challenge was determined 24 h after FITC‐dextran gavage and LPS‐induced injury. Ileal epithelial apoptosis was assessed by TUNEL and cleaved caspase‐3 staining of formalin‐fixed paraffin embedded tissue.RESULTSNRG4−/−mice had compromised barrier function compared to wild type littermates at baseline, as shown by an increase in FITC‐dextran leakage to the blood (1.4‐fold increase). Furthermore, NRG4 deletion reduced the ability to recover barrier 24 hours after LPS‐induced injury (2.4‐fold increase, p<0.05). NRG4−/− mice had exaggerated ileal epithelial cell apoptosis 24 h after LPS as well, measured by TUNEL (32 positive cells per 100 crypts (KO) vs 14.58 positive cells per 100 crypts (WT), p<0.05) and cleaved caspase‐3 staining.CONCLUSIONEndogenous NRG4 promotes integrity of the intestinal epithelial barrier. Further investigation of the role of NRG4 within the intestinal mucosa may lead to more effective strategies for targeting barrier dysfunction seen in diseases such as inflammatory bowel disease or necrotizing enterocolitis.Support or Funding InformationSupported by NIH award DK095004 and a Senior Research Award from The Crohn's and Colitis Foundation.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Cell Signaling by Vitamin E Nicotinate
Authors: Suzuki Yuichiro Justin , Marcocci Lucia , Shults Nataliia V. 
Published: June 2021

Vitamin E nicotinate is an ester of two vitamins, tocopherol (vitamin E) and niacin (vitamin B3), in which niacin is linked to the hydroxyl group of active vitamin E. This vitamin E ester can be chemically synthesized and has been used for supplementation. However, whether vitamin E nicotinate is formed in the biological system is unclear. In rats, vitamin E nicotinate was found in the heart and its level was determined to be 30‐times lower in heart failure. The diet used in these experiments contained vitamin E acetate and niacin separately, but not in the form of vitamin E nicotinate. Thus, vitamin E and niacin are esterified to form vitamin E nicotinate in the biological system. Since other forms of vitamin E were not found to be decreased, we propose that the vitamin E nicotinate structure elicits specific biologic functions, independently of serving as a source of active antioxidant. Consistent with this idea, we found that the treatment of cultured human cells with vitamin E nicotinate, but not vitamin E plus niacin, promoted the formation of anandamide within 10 min. Vitamin E nicotinate also activated MAP kinase signaling. These results suggest that vitamin E nicotinate can endogenously be formed in the biological system and the vitamin E nicotinate structure elicits cell signaling.Support or Funding InformationNIHThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.