bluuebunny/crossref_metadata_2025_split huggingface dataset:
Characterization of TSSK6 Activating Co‐chaperone (TSACC) in Skeletal Muscle
Authors: Tello Tala Shourbagi , Deluna Jonnaly , Waddell David 
Published: June 2021

Skeletal muscle atrophy is a physiological condition that is caused by a variety of conditions, including immobilization, denervation, spinal cord injury and aging and results in decreased muscle size and strength. To better characterize the molecular genetic events of neurogenic atrophy, a previous study isolated gastrocnemius muscle from mice following 3 days and 14 days of sciatic nerve denervation. The gene expression profile in the denervated muscle tissue was then analyzed by microarray and compared to control muscle tissue to identify novel neurogenic atrophy‐induced genes. The microarray data revealed for the first time that TSSK6 Activating Co‐chaperone (TSACC) is expressed in skeletal muscle and is significantly induced in response to denervation. To confirm that TSACC is expressed in muscle, the predicted TSACC cDNA was cloned from cultured muscle cells, as was a novel TSACC splice variant that contains an additional exon. Quantitative PCR (qPCR) was then used to assess TSACC expression levels of the full‐length and novel splice variant in proliferating and differentiated muscle cells. The results demonstrate that full‐length TSACC expression levels are relatively low in proliferating myoblasts but show significantly elevated expression in differentiated myotubes, whereas the novel splice variant is expressed relatively evenly in proliferating and differentiated muscle cells. In addition, characterization of the transcriptional regulation of TSACC was assessed by fusing fragments of the proximal promoter located immediately upstream of the start of transcription with a reporter gene. The reporter plasmids were then transfected into C2C12 mouse muscle cells in combination with myogenic regulatory factor (MRF) expression plasmids, which resulted in significant activation of reporter gene activity. Interestingly, there are several predicted E‐box elements in the proximal promoter region of the TSACC gene further suggesting that TSACC may be regulated by muscle‐specific transcription factors. Finally, to determine a possible role for TSACC in skeletal muscle, sub‐cellular localization was determined by fusing the cDNA with GFP and expressing the fusion protein in cultured muscle cells. The discovery that TSACC is induced in response to neurogenic atrophy helps further our understanding of the molecular genetic events of muscle atrophy and may eventually lead to the identification of new therapeutic targets for the treatment of muscle atrophy.Support or Funding InformationThe work was support by University of North Florida Transformational Learning Opportunity grants and a University of North Florida Foundation Board Grant to D.W.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Characterization of the Regulation and Function of Family with Sequence Similarity 83D (Fam83d) in Skeletal Muscle
Authors: Kavanaugh Jack , Sanchez Victoria , Cooper Lisa , Waddell David 
Published: June 2021

Skeletal muscle atrophy is a serious condition that can arise due to aging, cancer, corticosteroid exposure, and denervation. To better characterize the molecular genetic events of neurogenic atrophy, a previous study isolated mouse gastrocnemius muscle from mice at 3 and 14 days following sciatic nerve denervation. The gene expression profile in the denervated muscle tissue was then analyzed by microarray and compared to control muscle tissue to identify novel neurogenic atrophy‐induced genes. The microarray data revealed for the first time that Fam83d is expressed in skeletal muscle and is significantly induced in response to denervation. Quantitative PCR (qPCR) analysis revealed that Fam83d is more highly expressed in proliferating myoblasts compared to differentiated myotubes. Characterization of the transcriptional regulation of Fam83d was analyzed by cloning fragments of the proximal promoter region located immediately upstream from the start of transcription and fusing them with a reporter gene. The reporter plasmids were then transfected into C2C12 mouse muscle cells alone or in combination with myogenic regulatory factor expression plasmids, resulting in a differential effect on reporter activity. Analysis of the promoter of Fam83d revealed conserved E‐box elements in the proximal regulatory region, which are known myogenic regulatory factor (MRF) binding sites. To assess where Fam83d is localized in the cell, we fused Fam83d with green fluorescent protein (GFP), transfected the Fam83d‐GFP expression plasmid into C2C12 mouse myoblasts cells, visualized by confocal microscopy, and observed that Fam83d is localized exclusively to the cytoplasm. It has recently been determined that Fam83d contains a putative phospholipase domain, is mis‐regulated in several different cancer types, and may modulate MAP Kinase signaling. Therefore, Fam83d was ectopically expressed in cultured muscle cells and markers of muscle cell differentiation and MAP Kinase signaling were analyzed. Fam83d overexpression resulted in significantly repressed levels of myosin heavy chain and myogenin expression, while levels of phosphorylated ERK were dramatically repressed. The discovery that Fam83d is expressed in skeletal muscle combined with the observation that Fam83d, a potential modulator of MAP Kinase signaling, is induced in response to neurogenic atrophy helps further our understanding of the molecular and cellular events of skeletal muscle wasting.Support or Funding InformationThe work was support by University of North Florida Transformational Learning Opportunity grants and a University of North Florida Foundation Board Grant to D.W.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Identification and Characterization of Ovarian Tumor Protease Domain Containing 1 (OTUD1) in Skeletal Muscle
Authors: Wortherly Keri , Waddell David 
Published: June 2021

Skeletal muscle atrophy is a physiological condition that is caused by an array of conditions, including immobilization, denervation, spinal cord injury and corticosteroid use and results in decreased muscle size and strength. The molecular genetic events of neurogenic atrophy were analyzed in a previous study using gastrocnemius muscle isolated from mice following 3 days and 14 days of sciatic nerve denervation. The gene expression profile in the denervated muscle tissue was analyzed by microarray and compared to control muscle tissue to identify novel neurogenic atrophy‐induced genes. The microarray data revealed for the first time that Ovarian Tumor Protease Domain OTUD1 is expressed in skeletal muscle and is differentially regulated in response to denervation. OTUD1 is a deubiquitinase that has been linked to TGF‐β signaling in cancer via deubiquitination of Smad7. To confirm that OTUD1 is expressed in muscle, quantitative PCR (qPCR) was used to assess OTUD1 expression levels in both proliferating and differentiated muscle cells and the results demonstrate that expression levels appear to remain relatively constant in proliferating myoblasts and differentiated myotubes. Furthermore, Western blot analysis supported the qPCR data and showed that OTUD1 protein levels remain relatively constant as cultured myoblasts differentiate to myotubes. The discovery that OTUD1 is differentially regulated in response to neurogenic atrophy helps further our understanding of the molecular genetic events of muscle wasting and may eventually lead to the identification of new therapeutic targets for the treatment and prevention of atrophy.Support or Funding InformationThe work was support by University of North Florida Transformational Learning Opportunity grants and a University of North Florida Foundation Board Grant to D.W.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Identification and Characterization of 1700029J07RIK, a Novel Gene Expressed in Skeletal Muscle
Authors: Lynch Sarah , Waddell David 
Published: June 2021

Skeletal muscle atrophy is the loss of muscle mass that results from a wide range of conditions including denervation, corticosteroid exposure, and aging. 1700029J07RIK is a putative novel gene suggested to be involved in skeletal muscle atrophy. The 1700029J07RIK gene was found to be induced following denervation in mice and is differentially expressed in Muscle RING Finger 1 (MuRF1) knockout mice compared to wild‐type mice. A previous microarray analysis of muscle tissue isolated from wild‐type control mice and MuRF1 knockout mice revealed that the expression of the 1700029J07RIK gene is unchanged at 3 days post‐denervation, while expression is significantly induced in wild‐type mice at 14 days post denervation but remains unchanged in the MuRF1‐null animals. It has been well established that muscle cells go through a distinct developmental timeline in which they proliferate as single cell myoblasts and then differentiate and fuse into multinucleated myotubes. Quantitative PCR was used to confirm that the 1700029J07RIK gene is expressed in proliferating cells and increases in expression as muscle cells differentiate. To characterize the transcriptional regulation of 1700029J07RIK, promoter fragments were cloned into a reporter plasmid, transfected into muscle cells and found to have significant transcriptional activity, while ectopic expression of myogenic regulatory factors appear to modulate the activity of the 1700029J07RIK reporter genes. In addition, the 1700029J07RIK cDNA was cloned from muscle cells and fused with green fluorescent protein and visualized by confocal fluorescent microscopy revealing an interesting perinuclear and cytoplasmic localization of the 1700029J07RIK protein in muscle cells. Finally, it was observed that overexpression of 1700029J07RIK resulted in inhibition of muscle cell differentiation and attenuation of the ERK branch of the MAP Kinase signaling pathway. While there is virtually nothing known about this novel gene in the context of skeletal muscle, the results presented here suggest 1700029J07RIK may play a role in muscle cell differentiation and in the skeletal muscle atrophy cascade.Support or Funding InformationThe work was support by University of North Florida Transformational Learning Opportunity grants and a University of North Florida Foundation Board Grant to D.W.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Skeletal Muscle Mitochondrial Function in Goto‐Kakizaki Rat Model of Type 2 Diabetes
Authors: Lewis Matthew T , Kasper Jonathan D , Bazil Jason N , Frisbee Jefferson C , Meyer Ronald A , Wiseman Robert W 
Published: June 2021

Type 2 diabetes (T2D) presents with significant comorbidities and reported exercise intolerance affecting over 30 million people in the US alone. Aerobic metabolism is the predominant pathway for fueling exercise and utilizing glucose, therefore mitochondrial oxidative phosphorylation (MOP) is important in diabetes and the future of diabetic interventions. However, studies in T2D have conflicting reports regarding the functional status of the mitochondrial pool suggesting deficits are an inherent cause of the disease but a comprehensive study has yet to be done. The Goto‐Kakizaki (GK) rat model of T2D was used to measure both mitochondrial function and density in vitro and in vivo over a broad range of metabolic challenges. Mitochondrial densities were similar in red (RG) and white gastrocnemius (WG) muscles of the GK rat (Cyt c oxidase, RG: 22.2 ± 1.6 versus 23.3 ± 1.7 μmol/min/g, WG: 10.8 ± 1.1 versus 12.1 ± 0.9 μmol/min/g; n=7–8). Mitochondria isolated from the muscles of GK rats and their Wistar controls showed no difference in mitochondrial respiration in vitro as measured by high resolution respirometry. Mitochondrial function in vivo, measured by 31Phosphorus magnetic resonance spectroscopy, was similar between GK and Wistar controls determined from phosphocreatine dynamics reaching similar exercise steady states at submaximal workloads with no difference in recovery time constant (72 ± 6 s GK vs. 71 ± 2 s Wistar). However, at exercise intensities beyond what can be sustained aerobically, the GK rat exhibited bioenergetic deficits suggesting something other than mitochondrial function is responsible at these workloads. The current work demonstrates normal mitochondrial function in a non‐obese non‐sedentary rat model of T2D in vitro and in vivo and no change in mitochondrial content. Taken together these results show when considering both density and function of mitochondria no deficit in MOP is present in the GK T2D rat model and may not be in T2D patients but these studies remain to be done.Support or Funding InformationNIH DK 095210, NIH R00HL121160, CIHR R5521A02This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Chronic Dexamethasone Exposure Induced Mucosal Injuries by Activating Epithelial Apoptosis and Inflammatory Response in The Colon of Goats
Authors: Cai Liuping 
Published: June 2021

BackgroundChronic stress has a profound effect on health in both animals and humans. Chronic stress can influence the level of glucocorticoid. Dexamethasone (Dex), a synthetic glucocorticoid, is used to induce chronic stress. However, the aim of the present study was to investigate the effect of chronic stress caused by long term injection of Dex on changes on colonic epithelium in goats.ResultsThe results show that chronic stress increased the number of TUNEL‐apoptosis‐positive cells, upregulated caspase‐3 enzyme activity, and decreased the proteins expression of cell proliferation markers proliferating cell nuclear antigen (PCNA) and Cyclin D2 (CCND2) in the colonic epithelium. The gene expression of claudin‐1, claudin‐4 and ZO‐1 were upregulated by Dex (P < 0.05). But claudin‐4 protein expression was increased by Dex (P < 0.05). The gene expression of cell cycle such as CDKs (cyclin‐dependent kinases), CCND1, and CCND2 were all increased by Dex (P < 0.05). The gene expression of inflammtory cytokines interlukin‐10 (IL‐10), interlukin‐1β (IL‐1β), inducible nitric oxide synthase 2 (iNOS2), and inflammatory‐related transcriptional factors mitogen‐activated protein kinase (MAPK), signal transducer and activator of transcription 3 (STAT3), and nuclear factor kappa B (NFκB) were all upregulated by Dex (P < 0.05). The protein expression of NFκB and Toll‐like receptors (TLR4) were increased by Dex (P < 0.05). Moreover, the results showed that the methylation levels of total DNA and TLR4, caspase‐3 promoter were significantly decreased in the colonic epithelium compared to control group.ConclusionsThese results indicated that long‐term injection of a low dosage of Dex caused damage to the colon epithelium and inhibition of cell proliferation, and result in the activation of the inflammatory response and decreased methylation of the total DNA.Support or Funding Information1. National Nature Science Foundation of China (project no.31572433)2. The Program for New Century Excellent Talents in University (NCET‐13‐0862) and the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Surface Functionalization of Nanocrystalline and Nanostructured Diamond to Form Interface with Newborn and Adult Neurons
Authors: Krusek Jan , Dittert Ivan , Smejkalova Tereza , Korinek Miloslav , Gottftriedova Kristyna , Freislebenova Hana , Neuhoferova Eva , Taylor Andrew , Mortet Vincent , Petrak Vaclav , Benson Veronika , Petrakova Vladimira 
Published: June 2021

Boron doped diamonds (BDD) are a promising material for fabrication of neuroelectrodes and formation of neurointerfaces. Surface properties of BDD thin films can be adjusted by variation of grain sizes, patterning, lithography techniques and coating by different biopolymers to further promote performance of neuroelectrode and enhance the neuron adhesion.We used 3T3 fibroblasts and rat hippocampal neurons (newborn and adult) to estimate planar and nanostructured BDD surface biocompatibility. Optical microscopy, lactate dehydrogenase assay and calcium imaging were used to check cell adhesion and longterm survival.3T3 fibroblasts are able to grow on BDD without any coating. No signs of cytotoxicity and changes in cell cycle were found. Adhesion and longterm survival of newborn rat hippocampal neurons were observed only on poly‐L‐lysine coated planar and nanostructured BDD surface. Calcium imaging of longterm cultures of newborn neurons grown on poly‐L‐lysine coated planar and nanostructured BDD showed functionality and synaptic activity of hippocampal neurons. Nanodiamond surface without poly‐L‐lysine coating do not support adhesion of newborn neurons.Adult rat hippocampal neurons preferred poly‐D‐lysine coated planar nanodiamonds but were able to grow on nanostructured diamonds without any coating.Support or Funding InformationThis work was supported by Czech Science Foundation grant 17–15319S.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 Onset of Exercise‐induced Microvascular Hyperpermeability is Delayed in Diabetic Skeletal Muscle: In Vivo Imaging Using Two‐Photon Laser Scanning Microscopy
Authors: Hotta Kazuki , Shimotsu Rie , Masamoto Kazuto , Yagishita Kazuyoshi , Behnke Brad J , Poole David C , Kano Yutaka 
Published: June 2021

To investigate whether Type I diabetes (DIA) alters the temporal characteristics of muscle microvascular function and fiber damage incurred by eccentric contraction (ECC)‐induced muscle injury, male Wistar rats (n=53, 10–14 wks old) were randomly assigned to control (CON) or Streptozocin (45 mg/kg, i.p. DIA) groups. Four weeks later (blood glucose, 354±46 mg/dL), the tibialis anterior (TA) muscle of 1 leg was injured by 200 times ECC. Microvascular permeability was assessed by in vivo imaging using two‐photon laser scanning microscopy on 1, 3, 7 and 14 days post‐ECC. The TA microcirculation was visualized after intravenous infusion of fluorescent dye; extravasation of which was quantified to assess microvascular permeability. CON and ECC TA muscles were dissected and frozen after in vivo imaging, and serial cryosections were co‐stained with vascular endothelial growth factor A (VEGFA) antibody and hematoxylin‐eosin. Compared with CON rats, non‐ECC TA microvascular permeability was slightly but significantly enhanced in DIA with higher expression of VEGFA in myocytes. In CON rats, compared with non‐ECC TA, injured myocytes and microvascular hyper‐permeability were observed simultaneously on 1 and 3 days (injured myocyte 48, 73%, extravasation 262, 266 μm3×106xmin, 1, 3 days post‐ECC, respectively) and were repaired by 7 days after ECC (no injured myocytes, regenerated myocyte 29%, extravasation 101 μm3×106xmin). DIA TA exhibited delayed muscle injury onset and microvascular hyper‐permeability from 1 to between 3 and 7 days (injured 8, 81, 35%, extravasation 33, 183, 180 μm3×106xmin, 1, 3, 7 days post‐ECC, respectively). Regenerated myocyte was observed between 7 and 14 days post‐ECC (regenerated 47, 23%, 7, 14 days, respectively, extravasation 105 μm3×106xmin, 14 days post‐ECC). Regenerated cell size was smaller in DIA compared to CON (3214 vs. 1377 μm2, CON vs. DIA 14 days post‐ECC, P<0.01). Percentage of VEGFA‐positive myocytes to injured myocytes was increased in both CON (28%, 43%, 1 and 3 days post‐ECC, respectively) and DIA (44%, 43%, 65%, 1, 3, 7 days post‐ECC) rats with a temporal profile corresponding to that of increased dye extravasation. These results suggest that DIA retards the onset of microvascular hyper‐permeability after ECC thereby delaying skeletal muscle injury and recovery. VEGF may regulate microvascular permeability in both CON and DIA skeletal muscle and therefore play a commanding role in post‐ECC damage and regeneration.Support or Funding InformationThis work was supported by Japan Society for the Promotion of Science KAKENHI Grants JP17K13185, 16700450, 18700526, and 23650403.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Exosomes derived from adipose tissue stem cells regenerate ischemic myocardium following intravenous administration
Authors: Wang Min , Xu Jie , Liu Min , Wang YiGang , Ashraf Muhammad , Xu Meifeng 
Published: June 2021

ObjectiveExosomes (EXO) are nanosized bi‐lipid‐layered vesicles that contain proteins, mRNAs, and miRs. Increased evidence indicates that EXO play an important role in intercellular communication and cardiac repair. They have long circulating half‐life and cellular effects. EXO can be administered intravenously (iv) without risk of microvasculature entrapment. Our previous studies indicated that mesenchymal stem cells (MSCs) isolated from adipose tissue (AD‐MSCs) proliferated faster and secreted more EXO compared to those derived from bone marrow (BM‐MSCs). In this study, we investigated if intravenous administration of AD‐EXO can improve cardiac function and regenerate ischemic myocardium via native cardiomyocytes (CMs) protection.Methods and ResultsMSCs were harvested from adipose tissue of 6–8 weeks old C57BL/6 mice. EXO were collected from MSC conditional medium by ultracentrifugation, and were characterized by morphology and surface markers. Mouse myocardial infarction (MI) was created by ligation of left anterior descending (LAD) coronary artery. EXO (50 μg) were injected through tail vein every two days for a total of 2 or 4 times, respectively. Primary CMs were obtained from ventricles of 2‐day‐old rats. Ischemic injury was mimicked by exposing CMs to hypoxic culture (1% O2, 5%CO2, and 94% N2). We found that 1) Left ventricular function following LAD ligation was significantly improved and infarction size was significantly reduced in mice injected with AD‐EXO (50 μg) for a total of 4 times. 2) Distribution of AD‐EXO in heart was clearly observed following injection of pre‐labeled EXO with PKH26. 3) AD‐EXO significantly protected CMs against hypoxia induced injury.ConclusionsEXO derived from AD‐MSCs can be successfully delivered to myocardium following intravenous administration, which results in improved cardiac function and reduced infarction size in mouse MI model.Support or Funding InformationThis work was supported by NIH Grants HL114654, and HL140962 (M. Xu).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.