bluuebunny/crossref_metadata_2025_split huggingface dataset:
Phenotypic Plasticity in Morphological and Physiological Responses in Gills of Seawater‐ and Fresh water‐acclimated Asian Sea Bass, Lates calcarifer
Authors: Lee Tsung‐Han , Yang Wen‐Kai , Chao Tse‐Lih , Lee Sih‐Shien , Syu Jyun‐Jie , Chen Wei‐Zhu 
Published: June 2021

Osmoregulation is necessary for euryhaline teleosts in maintaining internal ion and water balance in response to environmental salinity changes. In the epithelia of teleostean gills, the ionocytes (i.e. mitochondrion‐rich cells) are thought to be the primary site of osmo/ionoregulation. In the ionocytes, the Na+, K+‐ATPase (NKA) plays a crucial role in triggering ion‐transporting systems. Asian sea bass (Lates calcarifer) is a marine euryhaline teleost and an important economic species in Taiwan, Southeast Asia, and Australia that can be cultured in environments from seawater (SW) to fresh water (FW). However, the modulatory mechanisms of osmoregulation in gills of euryhaline Asian sea bass were not clear to date. Hence the purposes of the present study were to investigate changes in physiological parameters, morphology of apical openings of gill ionoctyes, as well as expression and localization of branchial NKA in the SW‐ and FW‐acclimated Asian sea bass. The results showed that the sea bass was able to maintain their internal physiological parameters including plasma osmolality and muscle water content in a tolerated range. Observed by the scanning electron microscope, different types of apical surfaces of ionocytes with distinct features were found in the afferent epithelia of gills in FW and SW milkfish. On the other hand, the specific activity of branchial NKA was lower in the FW milkfish compared to the SW individuals. The immunohistological observation further revealed that most NKA‐immunoreactive cells in gills of the FW‐ and SW‐acclimated Asian sea bass were distributed in the afferent and interlamellar regions of the filaments. Taken together, these results illustrated that the Asian sea bass was an efficient osmoregulator with these salinity‐dependent alterations to maintain homeostasis of ions and water in environments of different salinities. These findings contributed to the knowledge of phenotypic plasticity of ionocytes and changes in activity of NKA, the crucial enzyme for ionoregulation, in gills of the Asian sea bass upon salinity challenge.Support or Funding InformationThis study was supported by a grant to THL from the Ministry of Science the Technology (MOST), Taiwan (105‐2313‐B‐005‐027‐MY3).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Walk the Walk: Parallels between Gait Parameters and Muscle Force Production in Turkeys
Authors: Vega Karina , Eng Carolyn , Roberts Thomas J. 
Published: June 2021

Running and walking involve regular patterns of limb kinematics, which can be characterized by simple parameters such as stride frequency and foot contact time. It is not clear whether these regular patterns of timing are mirrored at the level of muscle function, or whether variable sharing of force among different muscles might result in timing within individual muscles that does not reflect the timing of limb movements. We used direct measurements of force production in the lateral gastrocnemius (LG) muscle of wild turkeys during walking and running to test the hypothesis that stride parameters measured from limb movements would match those measured from muscle force production at all speeds of walking and running. Muscle force was measured via strain gauges attached to the bony tendon of the muscle, and stride parameters were measured with high‐speed video as animals ran on a treadmill at a range of walking and running speeds (1 to 3.5 m/s). Stride frequency and stride time values measured from video matched those determined from the onset of muscle force for all speeds. Time of foot contact measured from video decreased continuously with speed. Time of force development for the LG also decreased with speed, but the time spent in force development in the LG was significantly lower than the time the foot was on the ground for all speeds. The fraction of time the muscle was generating force was relatively constant across speed, with the muscle producing force for approximately 50% of the period of foot ground contact. This observation indicates that there is a temporal sharing of the task of force production between different muscles during the step, and that this sharing may remain relatively fixed across walking and running speeds. Such temporal sharing of force development should be considered in models of locomotor mechanics and energetics, and may be relevant to rehabilitative strategies for gait disorders.Support or Funding InformationThe Integrative Organismal Systems Physiology (IOSP) Undergraduate Summer Research Fellowship that made this research possible was supported by the American Physiological Society and a grant from the National Science Foundation (NSF) Integrative Organismal Systems (Award No. IOS‐1238831). Support from NIH grant AR‐055295 is also gratefully acknowledged.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Strategies to Assess Pharyngeal Pumping Variability in the Nematode C. elegans
Authors: Harris Michael B. , Berlemont Renaud , Ortiz Brianna , Taylor Barbara E. 
Published: June 2021

In C. elegans, the feeding behavior of pharyngeal pumping occurs spontaneously in proportion to metabolism and in response to stimuli. Age‐ and health‐related changes in tissue morphology and function correlate with declines in the frequency of pumping. As such, pumping frequency is an established index of C. elegans “health”, and pumping changes illustrate and quantify functional decline. Timing of pharyngeal pumping is controlled by two pairs of pharyngeal motor neurons (MC and M3). Each pump cycle corresponds to the propagation of a single pharyngeal muscle action potential, initiated by MC and terminated by M3. We currently model this system with MC acting as an oscillator and synaptically linked to the pharyngeal muscle. Pumping can change by influences on the pace and regularity of the oscillator and/or of the fidelity of pharyngeal muscle response to neuronal inputs.Traditionally, this phenomenon has been reported as an occurrence frequency, describing the number of pumping events observed per unit time over a period of observation. We have found variation in pumping frequency between subjects in an experimental group, and the relative coarseness of measures of occurrence frequency may interfere with the resolution of subtle, yet significant, treatment effects. Recently developed technologies facilitate precise recording of the electrical signatures of pharyngeal pumping in the form of an electropharyngeogram (EPG), allowing specific identification of timing of individual pumping events. This precision allows not only characterization of the occurrence frequency, but also the variability of pumping. We are considering that changes in variability may be a more sensitive measure of treatment effects.We have developed a strategy to normalize pumping occurrence in EPG recordings of individual subjects, allowing relevant comparisons between subjects. We have applied a series of variability assessments to quantify pumping patterns and identify variation stemming from changes in fidelity of drive from MC to the pharyngeal muscle. Finally, we have applied assessments that we speculate can distinguish influences at or above the MC oscillator. This presentation describes the theory and practice of normalization and variability assessments. The use of this analysis strategy on an experimental dataset is described in a companion presentation.Support or Funding InformationWork reported in this publication was supported by the National Heart Lung and Blood Institute and National Institute of General Medical Sciences of the National Institutes of Health under awards 1R15HL126105 and 1SC2GM112570. The work is solely the responsibility of the authors and does not necessarily represent the official view of the National Institutes of Health or any other funding body.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Muscle DRP‐1 and FIS‐1 Regulation by IL‐6 Signaling
Authors: Fix Dennis , VanderVeen Brandon N. , Hardee Justin , Carson James A. 
Published: June 2021

IntroductionThe IL‐6 cytokine family signals through the ubiquitously expressed glycoprotein 130 (gp130) receptor to induce signal transducer and activator of transcription 3 (STAT3) and extracellular signal‐regulated kinase 1/2 (ERK1/2). Evidence suggests that IL‐6 can negatively regulate muscle mitochondrial quality control and disrupt muscle oxidative metabolism. Mitochondrial fission is a critical component of skeletal muscle mitochondria maintenance and oxidative metabolism, which is regulated impart by the expression of FIS‐1 and DRP‐1 proteins. The purpose of this study is to examine muscle DRP‐1 and FIS‐1 expression by IL‐6 signaling.MethodsFully differentiated C2C12 myotubes were treated with 100ng of IL‐6 for 24 hours in the presence of gp130siRNA, C188‐9 (STAT3 inhibitor), or PD98059 (ERK1/2 inhibitor). Additionally, male C57BL/6 (B6) and muscle specific gp130 knockout mice (KO) were electroporated in the quadriceps muscles with 50ng of IL‐6 and monitored for two weeks.ResultsIL‐6 induced DRP‐1 and FIS‐1 expression in myotubes and skeletal muscle. Knock down of gp130 in myotubes and KO in skeletal muscle inhibited the IL‐6 induction of DRP‐1 and FIS‐1. STAT3 inhibition in myotubes did not alter IL‐6 induced DRP‐1 and FIS‐1 expression, while ERK 1/2 inhibited the IL‐6 induction. Collectively, these results provide evidence for skeletal muscle mitochondrial fission regualtion through an IL‐6/gp130/ERK 1/2 signaling axis.Support or Funding InformationThis work was supported by National Institutes of Health grant # NCI‐R01CA121249 (JAC).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
ADGRG6/GPR126 as a Novel Therapeutic Target in Pulmonary Arterial Hypertension
Authors: Muthusamy Abinaya , Gorr Matthew W. , Chinn Amy M. , Insel Paul A. 
Published: June 2021

Pulmonary arterial hypertension (PAH) is a chronic and fatal disease of the lungs. The 3‐year survival rate is 50–60%, as current therapies of PAH have limited effect on mortality. An important feature of PAH is the increased proliferation and contractile activity of pulmonary arterial smooth muscle cells (PASMCs). We cultured patient‐derived PASMCs and performed RNA‐seq on PASMCs from control and PAH lungs. We found that the expression of a G protein‐coupled receptor (GPCR) ADGRG6 (adhesion GPCR G6, or GPR126) is ~10‐fold higher in PAH patient PASMCs compared to control PASMCs. To test its functional activity, we overexpressed ADRGR6 in HEK293 cells, and found that proliferation of the cells decreased by 60% at 48 hours after transfection. Using qPCR, we observed that expression of cell cycle/proliferation markers CDK1, CDK2, and MKI67 was decreased after 48 hours, while expression of apoptosis markers VIM, ITGAM, and PCDH7 was increased. Additional preliminary data using flow cytometry indicate that cells overexpressing ADGRG6 are stalled at the S/G2 cell cycle checkpoint. These data suggest that ADRGRG6 activation could cause reduced proliferation in smooth muscle cells and be a viable target for PAH therapy.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Multidimensional Clustering of Regenerative Alveolar Duct Cells after Murine Pneumonectomy
Authors: Ysasi Alexandra , Bennett Robert , Valenzuela Cristian , Servais Andrew , Wagner Willi , Tsuda Akira , Ackermann Maximilian , Mentzer Steven 
Published: June 2021

Lung regeneration occurs in a variety of adult mammals after surgical removal of one lung (pneumonectomy). Previous studies of murine post‐pneumonectomy compensatory lung growth have identified areas of increased growth in subpleural alveolar ducts. To identify transcriptional patterns in these regenerative “hotspots”, we isolated single cells from alveolar ducts at post‐pneumonectomy days 1, 3 and 7. We used laser microdissection, enzymatic digestion and microfluidic isolation to capture individual cells from alveolar ducts, then performed single‐cell transcriptional analysis using the C1 integrated microfluidic circuit (Fluidigm) and a custom PCR panel designed for lung growth and repair genes. The multi‐dimensional data set was analyzed using visualization software based on the tSNE similarity analysis algorithm (viSNE, Cytobank). The similarity analysis identified six clusters of cells. To obtain a provisional cell type identity for these clusters, previously obtained post‐pneumonectomy cell sorted bulk transcriptional data was projected on the viSNE map using a similarity matrix analysis. The viSNE cluster with the highest overall level of gene expression was genotypically similar to myofibroblasts (64.04% of Acta2+ cells), endothelial progenitor cells (66.67% of sorted CD31+, CD34+cell samples), and type II pneumocytes (83.33% of sorted CD45−, MHC Class II+, phosphine+ cell samples). Another cluster genotypically overlapped primarily with sorted endothelial cells (94.74% of sorted CD31+ cell samples), but also some monocytes (50% of sorted CD11b+ cell samples) and alveolar macrophages (61.54% of sorted CD45+, CD11b−, CD11c+, F4/80+, Gr1− cell samples). One cluster was present only after pneumonectomy (99.23% post‐pneumonectomy, 0.77% un‐operated control), suggesting time point dependent gene activation. The transcriptional pattern of the six clusters was analyzed for genes associated with lung repair, matrix production and angiogenesis. The data demonstrated that multiple cell types (clusters) transcribed genes linked to these basic functions. We conclude that the coordinated gene expression across multiple clusters is likely a response to a shared regenerative microenvironment within the subpleural alveolar ducts.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
N‐Glycan and Heparan Sulphate Components of the Extracellular Matrix are Essential for Laminar Shear Stress Response of the Epithelial Sodium Channel (ENaC)
Authors: Ashley Zoe , Barth Daniel , Baldin Jan Peter , Fronius Martin 
Published: June 2021

The epithelial sodium channel (ENaC), a mechanosensitive ion channel, is known to be activated by laminar shear stress (LSS) [1]. We hypothesise that the large extracellular loops of ENaC interact with the complex extracellular matrix (ECM) and that the application of LSS deforms both the extracellular matrix (ECM) and the interacting ENaC components resulting in a change in ENaC activity. This study investigated whether or not the N‐glycan and heparan sulphate components of the ECM are specifically involved in the LSS activation of ENaC.Human α, β and γ ENaC subunits were co‐expressed in oocytes surgically harvested from female Xenopus laevis. Two‐electrode voltage‐clamp was used to determine ENaC activity in response to LSS when different components of the ECM were degraded by neuraminidase and heparinase III enzymes. A significant LSS response was observed in ENaC expressing oocytes, compared to native (no ENaC) oocytes; normalized LSS current of 5.5 ± 1.1 compared to 0.06 ± 0.03 (p = 0.003, n =8–14). In ENaC expressing oocytes cleavage of N‐glycan from the ECM with neuraminidase (0.2U/mL for 2h at 21°C) significantly reduced the normalised LSS response from 4.5 ± 0.4 to 1.7 ± 0.4 (p=0.003, n=8). Destruction of heparan sulphate components of the ECM with heparinase III (0.5 U/mL for 30min at 21°C) significantly reduced the LSS response from 5.1 ± 1.2 to 1.2 ± 0.5 (p=0.009, n=12). Data indicates that both removal of N‐glycan and cleavage of heparan sulphate from the ECM reduce the activation of ENaC by LSS, indicating that they are crucial components of the ECM for mechanosensation by ENaC.Support or Funding InformationThis study was supported by the Royal Society Marsden fund.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Exposure of proximal tubule cells to fluid shear stress alters transcription of metabolic pathway enzymes
Authors: Ren Qidong , Eshbach Megan L. , Rittenhouse Natalie L. , Long Kimberly R. , Rbaibi Youssef , Locker Joseph , Poholek Amanda C. , Weisz Ora A. 
Published: June 2021

Cells lining the proximal tubule (PT) of the kidney are specialized for efficient recovery of ions, glucose, and proteins from the glomerular filtrate. Despite their high glucose levels, PT cells in vivo utilize gluconeogenic rather than glycolytic pathways to maintain the robust metabolic activity needed to carry out their functions. In contrast, the reliance of PT cell cultures on glycolytic metabolism has been a significant limitation in studying the regulation of PT function in vitro. We discovered recently that opossum kidney (OK) cells cultured under continuous fluid shear stress (FSS) develop morphological and functional characteristics that more closely resemble PT cells in vivo compared with cells cultured under static conditions. We performed RNA Seq on OK cells maintained under static conditions or exposed to FSS for up to 96 h. Principal component analysis confirmed significant time‐ and FSS‐dependent changes in gene expression. Consistent with enzyme activity profiles described in the PT, we observed an increase in transcripts encoding enzymes in the gluconeogenic pathway in OK cells cultured under FSS, and a concomitant reduction of transcripts encoding glycolytic enzymes. Current studies are underway to extend and validate our observations using complementary approaches. Our data provide further support that OK cell culture under FSS provides an improved model of the PT.Support or Funding InformationTsinghua MD Scholars ProgramNIH R01 DK101484This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Persistent disruption of lateral junctional complexes and actin cytoskeleton in parotid salivary glands following radiation treatment
Authors: Wong Wen Yu , Pier Maricela , Limesand Kirsten H. 
Published: June 2021

Xerostomia and hyposalivation are debilitating side effects for patients treated with ionizing radiation for head and neck cancer. Despite technological advances, collateral damage to the salivary glands remains a significant problem for patients and severely diminishes their quality of life. During the wound healing process, restoration of junctional contacts is necessary to maintain polarity, structural integrity, and orientation cues for secretion. However, little is known about whether these structural molecules are impacted following radiation damage and more importantly following tissue restoration. Here, we evaluated changes in adherens junctions and cytoskeletal regulators in an injury model where mice were irradiated with 5 Grey (5Gy) and a restoration model where mice injected post‐radiation with insulin like growth factor 1 (IGF1) are capable of restoring salivary function. Using co‐immunoprecipitation, we determined that there is a decrease in β‐catenin bound to E‐cadherin following damage that is restored to untreated levels with IGF1. Via its adaptor proteins, β‐catenin links the cadherins to the cytoskeleton and part of this regulation is mediated through Rho‐associated coiled‐coil kinase (ROCK) signaling. In our model, F‐actin organization is also disrupted and there is an induction of ROCK activity. These findings illustrate that radiation induces a disruption of the cadherin‐catenin complex, alters F‐actin organization, and induces ROCK signaling at early and chronic stages where salivary hyposalivation is observed. Understanding the role of these components and its regulation may be important in tissue restoration where a coordinated and polarized epithelium are necessary for functionality.Support or Funding InformationThis work was supported in part by NIH R01 DE023534 received by Kirsten Limesand and stipend support for Wen Yu Wong was provided by the Cancer Biology Training Grant, T32CA009213‐38This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Early life stress induces vascular expression of pro‐oxidant, proinflammatory genes in adulthood in an HDAC9‐dependent manner
Authors: Pettway Yasminye D. , Ho Dao H. , McPherson Kasi , Seifert Michael E. , Pollock Jennifer S. 
Published: June 2021

Early life stress (ELS) has been correlated with an increased risk of developing cardiovascular disease (CVD) in adulthood; however, mechanisms linking ELS and CVD risk are unknown. Toll‐like receptor (TLR) activation, specifically TLR4, has been implicated as a key link between inflammation and CVD. TLR4 signaling can activated NADPH oxidases (NOXs) that produce superoxide to increase CVD risk. Further, multiple studies have found that NF‐kB activation, an important downstream step in the TLR4 signaling pathway, requires the activation of certain NOX isoforms, including NOX2 and NOX4. We utilized a mouse model of maternal separation with early weaning (MSEW) to investigate the effect of ELS on the expression and activation of genes encoding these pro‐oxidant, proinflammatory proteins. MSEW mice were separated 4hr/day during postnatal day (PD)2–5, 8 hr/day during PD6‐16, and weaned at PD17. Normally‐reared control mice were left undisturbed and weaned at PD21. Experiments were conducted on 3–6 month old male mice. We have previously shown that MSEW induces NOX‐dependent superoxide production and vascular endothelial dysfunction. We also found increased gene expression of NOX2, NOX4, and TLR4 in the aorta of MSEW mice. Recent immunohistochemical studies have also revealed increased aortic TLR4 protein expression in MSEW mice (n=5/group). Due to similar patterns of expression of genes involved in the TLR4 signaling pathway, we further hypothesized that a common upstream transcription factor could be a driver of these ELS‐induced gene changes. We focused this study on histone deacetylases (HDACs), as numerous studies have implicated HDACs in the modulation of proinflammatory responses and the progression of a variety of CVDs. Interestingly, we found that MSEW induced aortic HDAC9 protein expression, exclusive of other HDAC isoforms. A 3‐hr ex vivo pretreatment with trichostatin A (TSA), a pan‐inhibitor of HDACs, improved vascular endothelial function in MSEW mice (n=5/group, p=0.047). Further, ex vivo TSA treatment led to a trend of decreased expression of NOX2, NOX4, and TLR4 genes (n=4–8/group). These results suggest that ELS leads to HDAC9‐dependent pro‐oxidative and proinflammatory gene expression that increase CVD risk in adulthood. They also further emphasize the role of TLR4 signaling on superoxide production and vascular endothelial function. Future studies aim to clarify the role of TLR4 signaling in these phenotypes, as well as the effect of increased HDAC9 expression on the transcriptional regulation of TLR4 and other genes that could confer CVD risk in adulthood.Support or Funding InformationThis research was funded by PO1HL69999 and the UAB PREP Program.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.