scholarly journals Endurance exercise ameliorates phenotypes in Drosophila models of Spinocerebellar Ataxias

2021 ◽  
Author(s):  
Alyson Sujkowski ◽  
Kristin Richardson ◽  
Matthew V. Prifti ◽  
R. J. Wessells ◽  
Sokol V. Todi
Keyword(s):  
Endurance Exercise ◽  
Neural Plasticity ◽  
Disease Incidence ◽  
Early Death ◽  
Autophagic Flux ◽  
Chronic Exercise ◽  
Spinocerebellar Ataxias ◽  
Protein Levels ◽  
Disease Protein ◽  
Inducible Protein

AbstractEndurance exercise is a potent intervention with widespread benefits proven to reduce disease incidence and impact across species. While endurance exercise supports neural plasticity, enhanced memory, and reduced neurodegeneration, less is known about the effect of chronic exercise on the progression of movement disorders such as ataxias. Here, we focused on three different types of ataxias, Spinocerebellar Ataxias Type (SCAs) 2, 3, and 6, belonging to the polyglutamine (polyQ) family of neurodegenerative disorders. In Drosophila models of these SCAs, flies progressively lose motor function. Here, we observe marked protection of speed and endurance in exercised SCA2 flies and modest protection in exercised SCA6 models, while no benefit is observed in SCA3 flies. Causative protein levels are reduced in SCA2 flies after chronic exercise, but not in SCA3 models, linking protein levels to exercise-based benefits. Additional investigations indicate that the exercise-inducible protein, Sestrin (Sesn) suppresses mobility decline and improves early death in SCA2 flies, even without exercise, coincident with disease protein level reduction and increased autophagic flux. These improvements depend on previously established functions of Sesn that reduce oxidative damage and modulate mTOR activity. Our study suggests differential responses of polyQ SCAs to exercise, highlighting the potential for more extensive application of exercise-based therapies in the prevention of polyQ neurodegeneration. Defining the mechanisms by which endurance exercise suppresses polyQ SCAs will open the door for more effective treatment for these diseases.

scholarly journals Identification of Keratin 23 as a Hepatitis C Virus-Induced Host Factor in the Human Liver

Cells ◽  
2019 ◽  
Vol 8 (6) ◽  
pp. 610 ◽  
Author(s):  
Volker Kinast ◽  
Stefan L. Leber ◽  
Richard J. P. Brown ◽  
Gabrielle Vieyres ◽  
Patrick Behrendt ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Liver Disease ◽  
Hepatitis C ◽  
Ectopic Expression ◽  
Host Factor ◽  
Hcv Infection ◽  
Mrna Levels ◽  
Structural Support ◽  
Protein Levels ◽  
Inducible Protein

Keratin proteins form intermediate filaments, which provide structural support for many tissues. Multiple keratin family members are reported to be associated with the progression of liver disease of multiple etiologies. For example, keratin 23 (KRT23) was reported as a stress-inducible protein, whose expression levels correlate with the severity of liver disease. Hepatitis C virus (HCV) is a human pathogen that causes chronic liver diseases including fibrosis, cirrhosis, and hepatocellular carcinoma. However, a link between KRT23 and hepatitis C virus (HCV) infection has not been reported previously. In this study, we investigated KRT23 mRNA levels in datasets from liver biopsies of chronic hepatitis C (CHC) patients and in primary human hepatocytes experimentally infected with HCV, in addition to hepatoma cells. Interestingly, in each of these specimens, we observed an HCV-dependent increase of mRNA levels. Importantly, the KRT23 protein levels in patient plasma decreased upon viral clearance. Ectopic expression of KRT23 enhanced HCV infection; however, CRIPSPR/Cas9-mediated knockout did not show altered replication efficiency. Taken together, our study identifies KRT23 as a novel, virus-induced host-factor for hepatitis C virus.

BRCA1 modulates sensitivity to 5F-203 by regulating xenobiotic stress-inducible protein levels and EROD activity

2007 ◽  
Vol 62 (4) ◽  
pp. 689-697 ◽  
Author(s):  
Hyo Jin Kang ◽  
Hee Jeong Kim ◽  
Sang Hoon Kwon ◽  
Brian DongHoon Kang ◽  
Thomas E. Eling ◽  
...  
Keyword(s):  
Erod Activity ◽  
Xenobiotic Stress ◽  
Protein Levels ◽  
Inducible Protein

scholarly journals (Pro)renin receptor regulates autophagy and apoptosis in podocytes exposed to high glucose

2015 ◽  
Vol 309 (3) ◽  
pp. E302-E310 ◽  
Author(s):  
Caixia Li ◽  
Helmy M. Siragy
Keyword(s):  
Signaling Pathway ◽  
High Glucose ◽  
Caspase 3 ◽  
Mtor Signaling ◽  
Autophagic Flux ◽  
Bafilomycin A1 ◽  
Mtor Signaling Pathway ◽  
Autophagosome Formation ◽  
Podocyte Injury ◽  
Protein Levels

High glucose reduces autophagy and enhances apoptosis of podocytes. Previously, we reported that high glucose induced podocyte injury through upregulation of the (pro)renin receptor (PRR). We hypothesized that increasing PRR reduces autophagy and increases apoptosis of mouse podocytes exposed to high glucose via activation of the PI3K/Akt/mTOR signaling pathway. Mouse podocytes were cultured in normal (5 mmol/l) or high (25 mmol/l) d-glucose for 48 h. High glucose significantly increased mRNA and protein levels of PRR, phosphorylation of PI3K/Akt/mTOR, and p62. In contrast, high glucose decreased activation of UNC-51-like kinase-1 (ULK1) by phosphorylating Ser757 and protein levels of microtubule-associated protein-1 light chain 3B (LC3B)-II and Lamp-2. Bafilomycin A1 increased LC3BII and p62 accumulation in high-glucose-treated cells. High glucose reduced the autophagic flux. Confocal microscopy studies showed significant reduction in the protein level of LC3B in response to high glucose. Cyto-ID autophagy staining showed a significant decrease in autophagosome formation with high glucose. In the absence of PRR, activation of Akt with sc-79 or mTOR with MHY-1485 increased p62 accumulation. Caspase-3/7 activity and apoptosis monitored by TUNEL assay were significantly increased in podocytes treated with high glucose. PRR siRNA significantly reversed the effects of high glucose. Based on these data, we conclude that high glucose decreases autophagy and increases apoptosis in mouse podocytes through the PRR/PI3K/Akt/mTOR signaling pathway.

scholarly journals Loss of Profilin3 Impairs Spermiogenesis by Affecting Acrosome Biogenesis, Autophagy, Manchette Development and Mitochondrial Organization

2021 ◽  
Vol 9 ◽  
Author(s):  
Naila Umer ◽  
Lena Arévalo ◽  
Sharang Phadke ◽  
Keerthika Lohanadan ◽  
Gregor Kirfel ◽  
...  
Keyword(s):  
Actin Polymerization ◽  
Regulatory Proteins ◽  
Autophagic Flux ◽  
Nuclear Fraction ◽  
Rna Seq ◽  
Head Shape ◽  
Male Factor ◽  
Protein Levels ◽  
Acrosome Formation ◽  
Actin Regulatory Proteins

Profilins (PFNs) are key regulatory proteins for the actin polymerization in cells and are encoded in mouse and humans by four Pfn genes. PFNs are involved in cell mobility, cell growth, neurogenesis, and metastasis of tumor cells. The testes-specific PFN3 is localized in the acroplaxome–manchette complex of developing spermatozoa. We demonstrate that PFN3 further localizes in the Golgi complex and proacrosomal vesicles during spermiogenesis, suggesting a role in vesicle transport for acrosome formation. Using CRISPR/Cas9 genome editing, we generated mice deficient for Pfn3. Pfn3–/– males are subfertile, displaying a type II globozoospermia. We revealed that Pfn3–/– sperm display abnormal manchette development leading to an amorphous sperm head shape. Additionally, Pfn3–/– sperm showed reduced sperm motility resulting from flagellum deformities. We show that acrosome biogenesis is impaired starting from the Golgi phase, and mature sperm seems to suffer from a cytoplasm removal defect. An RNA-seq analysis revealed an upregulation of Trim27 and downregulation of Atg2a. As a consequence, mTOR was activated and AMPK was suppressed, resulting in the inhibition of autophagy. This dysregulation of AMPK/mTOR affected the autophagic flux, which is hallmarked by LC3B accumulation and increased SQSTM1 protein levels. Autophagy is involved in proacrosomal vesicle fusion and transport to form the acrosome. We conclude that this disruption leads to the observed malformation of the acrosome. TRIM27 is associated with PFN3 as determined by co-immunoprecipitation from testis extracts. Further, actin-related protein ARPM1 was absent in the nuclear fraction of Pfn3–/– testes and sperm. This suggests that lack of PFN3 leads to destabilization of the PFN3–ARPM1 complex, resulting in the degradation of ARPM1. Interestingly, in the Pfn3–/– testes, we detected increased protein levels of essential actin regulatory proteins, cofilin-1 (CFL1), cofilin-2 (CFL2), and actin depolymerizing factor (ADF). Taken together, our results reveal the importance for PFN3 in male fertility and implicate this protein as a candidate for male factor infertility in humans.

scholarly journals Effect of Deferent Type of Training on C-reactive protein Level in Healthy Middle Age Men

2016 ◽  
Vol 12 (20) ◽  
pp. 136
Author(s):  
Mohammad Al Hindawi ◽  
Majed Mjallie
Keyword(s):  
Exercise Training ◽  
Endurance Exercise ◽  
Protein Level ◽  
Middle Age ◽  
Exercise Program ◽  
C Reactive Protein ◽  
Protein Levels ◽  
Reactive Protein ◽  
Type Of Exercise ◽  
The University

Objective: The goal of our study was to investigate the effect of different type of exercise training on C-reactive protein level in middle age men. Methods: A total of 20 male subjects’ mean age 45±5 years were investigated at the University of Jordan Hospital to evaluate CRP level after training. Subjects were divided randomly into two groups they participated in supervise exercise training resistance and endurance exercise for 11 weeks. CRP level was measured pre-post training. Results: Mean C-reactive protein level measured before training program was 1.82 mg/L. And 1.81 mg/L for endurance and resistance groups respectively. Measurement of the CRP after the completion of the exercise program was 1.654 mg/L and 1.764 mg/L for both groups the endurance and the resistance respectively. TC was significantly reduced from 199.1 ml/dl to 176.1 ml/dl, for the endurance group and from 193.50 ml/dl to 181.60 ml/dl, for the resistance group. TG reduced significantly from 172.2 ml/dl to 161.50 ml/dl for the endurance group, and from 163.50 ml/dl to 159.20 ml/dl for the resistance group. HR at rest reduced from 79.63 b.pm to 74.47 b.pm for the endurance group, and from 77.50 b.pm to 74.6 b.pm for the resistance group. Fat % reduced significantly from 25.25 to 23.22 for the endurance group and from 23.03 to 21.60 for the resistance group. BMI reduced from 28.82 to 27.41 for the endurance group and from 27.63 to 27.40 for the resistance group.VO2max increased significantly from 37.13 to 43.30 ml.kg.min for the endurance group, and from 37.67 to 39.3 for the resistance group. HDL also increased from 46.12 mi/dl to 48.25 mi/dl for the endurance group, and increased from 42.70 mi/dl to 44.30 mi/dl for the resistance group Training gropes had lower CRP, weight, BMI, and body fat after the completion of the exercise program. Conclusions: Plasma C-reactive protein levels are reduced in response to both strength and endurance exercise training in sedentary healthy men (there was no different in the type of exercise in reducing CRP level.

Abstract 30: Endurance Exercise Induces Cardiac Hypertrophy in Aged Nrf2-/- Mice

2012 ◽  
Vol 111 (suppl_1) ◽  
Author(s):  
Rajasekaran Namakkal Soorappan ◽  
Dinesh Devdoss ◽  
Snkaranarayanan Kannan ◽  
Curtis Olsen ◽  
Sellamuthu Subbanna Gounder ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Cardiac Hypertrophy ◽  
Endurance Exercise ◽  
Weight Ratio ◽  
The Elderly ◽  
Exercise Stress ◽  
Aging Heart ◽  
Protein Levels ◽  
Nuclear Levels ◽  
Endurance Stress

Background: Radical forms of oxygen and nitrogen species (ROS/RNS) are highly reactive with nucleic acids, proteins and lipids and promote their oxidation. Normally, cellular ROS/RNS concentrations are tightly controlled by the inducible antioxidant system, which is predominantly regulated by the transcription factor Nrf2 (nuclear erythroid-2 like factor-2) and its cytosolic repressor protein, Keap1. We hypothesized that a decrease or an abrogation of Nrf2 impairs cardiac function and induce hypertrophy upon endurance stress in aging heart. Methods: Age-matched wild-type (WT) and Nrf2-/- (KO) mice (n=12/gp) at 2 and >20 months were subjected to endurance exercise stress (EES; 20 meter/min, 12% grade) and assessed the activation of Nrf2/ARE-dependent transcriptional mechanisms in the heart. Cardiac hypertrophy was determined by echocardiography, heart/body weight ratio and biochemical/molecular marker analyses. Results: Interestingly, both old-WT and Nrf2-/- mice exhibited oxidative stress on EES due to significant decrease or abrogation of Nrf2 nuclear levels, respectively, suggesting that aged-WT is equally susceptible to stress as Nrf2-/- mice. Age-dependent loss of Nrf2 decreased the transcription of Nrf2-dependent antioxidants and thereby elevated ROS levels to cause a more oxidized intracellular environment. Importantly, the loss of Nrf2 induced cardiac hypertrophy upon endurance stress in the aged (>20 mon) mice. At the end of 2-weeks of endurance stress, both the old-WT and Nrf2-/- mice had developed cardiac hypertrophy. Also, qPCR analysis showed significant (p<0.05) upregulation of hypertrophy markers (ANF and BNF) in the old-WT or Nrf2-/- when compared to sedentary WT mice confirmed cardiac hypertrophy due to loss of Nrf2. These results indicate that either a decrease or an abrogation of Nrf2 can increase susceptibility to stress induced hypertrophy in an aging heart. Conclusions: We conclude that enhancing protein levels and stability of nuclear Nrf2 could activate the transcription of major antioxidant enzymes and biosynthesis of key antioxidants. Enhancing protective mechanisms against oxidative stress in the elderly is expected to prevent or delay the onset of age-associated cardiac hypertrophy and cardiomyopathy.

Abstract WP265: Interferon-inducible Protein 10 Increases following Stroke

Stroke ◽  
2013 ◽  
Vol 44 (suppl_1) ◽  
Author(s):  
Hilary A Seifert ◽  
Lisa A Collier ◽  
Stanley A Benkovic ◽  
Alison E Willing ◽  
Keith R Pennypacker
Keyword(s):  
Time Course ◽  
Neutralizing Antibody ◽  
Artery Occlusion ◽  
T Helper ◽  
Protein Levels ◽  
Organ Systems ◽  
Inducible Protein ◽  
Cerebral Artery Occlusion ◽  
The Brain ◽  
Interferon Inducible Protein

Objective: The splenic response to injury furthers cellular degeneration as its removal is protective in ischemic injuries to several organ systems including the brain. Previously, we have shown that the proinflammatory cytokine interferon gamma (IFNg), which activates microglia/macrophages, is elevated in the spleen and the brain following permanent middle cerebral artery occlusion (MCAO). IFNg induces the production of interferon-inducible protein 10 (IP-10) which further propagates the inflammatory response. Therefore, we investigated the expression of IP-10 in the brain and spleen following ischemic stroke. Hypothesis: IFNg production in the brain and the spleen results in elevated levels of IP-10 in the same tissues post-MCAO. Methods: A time course was conducted to investigate splenic and brain protein levels of IP-10 in rats over time following MCAO and sham-MCAO (n≥3). In a second experiment, rats were administered an IFNg neutralizing antibody following MCAO with a survival time of 96 h: vehicle control (n=4), goat IgG 5μg (n=7), and IFNg antibody 5μg (n=7). Spleens and brains were collected for all groups. Results: IP-10 levels were significantly elevated in the brain at 72 and 96 h post-MCAO (p<0.01) compared to naïve brains. In the spleen IP-10 levels become significantly elevated at 24 h and remain elevated out to 96 h post-MCAO (p<0.0007) compared to naïve spleens. Administration of a neutralizing antibody directed against IFNg significantly decreased IP-10 levels in the brain (p<0.009) but did not affect IP-10 levels in the spleen. Conclusion: These results demonstrate that increased production of IFNg results in elevated levels of IP-10 in both the spleen and the brain following stroke. However, administration of a neutralizing antibody against IFNg decreased the amount of IP-10 in the brain. Levels of IFNg and IP-10 in the brain increase at the same time following stroke. Based on these data, IFNg propagates a proinflammatory T helper cell response to stroke through IP-10. Inhibition of this signaling could reduce neuroinflammation thereby improving stroke outcome.

scholarly journals Assessing methods to quantitatively validate TGFβ-dependent autophagy

Biology Open ◽  
2020 ◽  
Vol 9 (11) ◽  
pp. bio055103
Author(s):  
Charles B. Trelford ◽  
Gianni M. Di Guglielmo
Keyword(s):  
Cell Lines ◽  
Transforming Growth Factor Beta ◽  
Epithelial To Mesenchymal Transition ◽  
Transforming Growth Factor ◽  
Nsclc Cell ◽  
Serum Starvation ◽  
Autophagic Flux ◽  
Mesenchymal Transition ◽  
Protein Levels ◽  
Lc3 Puncta

ABSTRACTTransforming growth factor beta (TGFβ) promotes tumorigenesis by suppressing immune surveillance and inducing epithelial to mesenchymal transition (EMT). TGFβ may augment tumorigenesis by activating autophagy, which protects cancer cells from chemotherapy and promotes invasive and anti-apoptotic properties. Here, we assess how TGFβ1 modulates autophagy related (ATG) gene expression and ATG protein levels. We also assessed microtubule-associated protein light chain 3 (LC3) lipidation, LC3 puncta formation and autophagosome-lysosome co-localization in non-small cell lung cancer (NSCLC) cell lines. These experimental approaches were validated using pharmacological autophagy inhibitors (chloroquine and spautin-1) and an autophagy activator (MG132). We found that TGFβ1, chloroquine and MG132 had little effect on ATG protein levels but increased LC3 lipidation, LC3 puncta formation and autophagosome-lysosome co-localization. Since similar outcomes were observed using chloroquine and MG132, we concluded that several techniques employed to assess TGFβ-dependent autophagy may not differentiate between the activation of autophagy versus lysosomal inhibition. Thus, NSCLC cell lines stably expressing a GFP-LC3-RFP-LC3ΔG autophagic flux probe were used to assess TGFβ-mediated autophagy. Using this approach, we observed that TGFβ, MG132 and serum starvation increased autophagic flux, whereas chloroquine and spautin-1 decreased autophagic flux. Finally, we demonstrated that ATG5 and ATG7 are critical for TGFβ-dependent autophagy in NSCLC cells. The application of this model will fuel future experiments to characterize TGFβ-dependent autophagy, which is necessary to understand the molecular processes that link, TGFβ, autophagy and tumorigenesis.

Effect of Endurance Exercise Training on Liver Gene Expression in Male and Female Mice

2020 ◽  
Author(s):  
Luma Melo ◽  
Karen Tilmant ◽  
Amit Hagar ◽  
JAMES E KLAUNIG
Keyword(s):  
Gene Expression ◽  
Metabolic Disease ◽  
Endurance Exercise ◽  
Metabolic Diseases ◽  
Molecular Pathways ◽  
Hepatic Gene Expression ◽  
Chronic Exercise ◽  
Male And Female ◽  
Female Mice ◽  
Liver Gene

Chronic endurance exercise is a therapeutic strategy in the treatment of many chronic diseases in humans, including the prevention and treatment of metabolic diseases such as diabetes mellitus. Metabolic, cardiorespiratory and endocrine pathways targeted by chronic endurance exercise have been identified. In the liver however, the cellular and molecular pathways that are modified by exercise and have preventive or therapeutic relevance to metabolic disease remain unresolved. The mouse model used in the current study allows for the quantification of a human-relevant exercise “dosage”. In this study we show hepatic gene expression differences between sedentary female and sedentary male mice, and that chronic exercise modifies the transcription of hepatic genes related to metabolic disease and steatosis in both male and female mice. Chronic exercise induces molecular pathways involved in glucose tolerance, glycolysis and gluconeogenesis while producing a decrease in pathways related to insulin resistance, steatosis, fibrosis, and inflammation. Given these findings, this mouse exercise model has potential to dissect the cellular and molecular hepatic changes following chronic exercise with application to understanding the role that chronic exercise plays in preventing human diseases. Novelty Bullets: • Exercise modifies the hepatic gene expression and hepatic pathways related to metabolic disease in male and female mice. • Gender differences were seen in hepatic gene expression between sedentary and exercised mice. • The mouse exercise model used in this study allows for application and evaluation of exercise effects in human disease

Sign in / Sign up

Export Citation Format

Share Document