bluuebunny/crossref_metadata_2025_split huggingface dataset:
Modulation of WNT signalling by inflammatory factor TWEAK, and its consequences on fibroblast-epithelial communication
Authors: Raphael Bella , Matellan Carlos , Manresa Mario 
Published: May 2024

BACKGROUND AND AIMS: A single layer of epithelial cells that are supported by resident mucosal fibroblasts, provides an intestinal barrier against pathogens. In Inflammatory Bowel Disease (IBD), this barrier is disrupted by mechanisms which are poorly understood. Intestinal fibroblasts produce WNT, BMP and Notch mediators that contribute to epithelial homeostasis and an impairment on these signals results in the loss of barrier integrity which has been observed in IBD. In fact, fibroblasts producing these signals may be lost during inflammation. Yet the mediators and mechanism that drive this loss of homeostatic stromal-derived signals are unknown. TWEAK is a member of the tumour necrosis factor superfamily (TNFSF) that was found to be upregulated in IBD biopsies and dysregulated tissue repair and promote chronic inflammation in in vivo murine models. However, whether TWEAK exerts its effects on fibroblasts and how these may contribute to barrier breakdown remains unknown. My central hypothesis is that TWEAK dysregulates the crosstalk between fibroblasts and epithelial cells in the intestinal tract by impairing a homeostatic signalling network. METHODS: To assess the effect of TWEAK on WNT signalling and wound healing in fibroblasts and epithelial cells, the expression of WNT ligands and receptors were assessed. Primary human colonic fibroblasts and human colonic epithelial cells were stimulated with TWEAK at various time points and analysed by qPCR, Western blot and Flow cytometry and scratch wound assay. RESULTS: Both Fibroblasts and epithelial cells displayed abundant Fn14 surface receptor, although epithelial cells at a level lower than that found on colonic fibroblasts. Transcriptional signatures at gene RSPO2 (0.5-fold ± decrease, *p<0.05, n=4), WNT5A (1 fold ± decrease, **p<0.01, n=4) and BMP6 (1 fold ± decrease, ***p<0.001, n=4) and protein level RSPO2 (50% ± decrease respectively, n=5) showed a TWEAK-mediated suppression after prolonged exposure (24-48h), but not in epithelial cells. Moreover, TWEAK delayed fibroblast healing capacity after 24h (50% ± decrease in wound closure, n=4) in a wound-scratch assay. CONCLUSION: TWEAK represses the expression of homeostatic factors in colonic fibroblasts but not on epithelial cells, suggesting this mechanism could be specific to the intestinal stroma. The effect that this impairment of WNT signals in stromal cells may have on their differentiation and communication with the epithelium will be subject for further study. UCD Ad Astra PhD Scholarship. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.
Life in the Slow Lane: The Rate of Protein Synthesis Is Slower in the Protoendothermic Tenrec than the Boreoeutherian Golden-Mantled Ground Squirrel
Authors: Carlson Zachary , Hindle Allyson G , van Breukelen Frank 
Published: May 2024

In order to address mechanisms of how hibernating mammals depress protein synthesis during torpor, we developed primary cell cultures from golden-mantled ground squirrels (Spermophilus [Callospermophilus] lateralis) and tenrecs (Tenrec ecaudatus). During torpor in squirrels, there is an acute uncoupling of translational initiation and elongation at body temperature (Tb) = 18°C as animals enter torpor. As a result, there are robust changes to the polysome profile of squirrels depending on state. Protein synthesis is depressed to 0.23 to 0.5% of active rates during torpor. In the bizarre, Afrotherian Tenrec ecaudatus, metabolism and Tb are remarkably plastic. Tenrecs may be fully active with a Tb of 12 °C or hibernating at a Tb of 28 °C. Moreover, oxygen consumption rates may vary 25 fold in active tenrecs. With this high variability in Tb, one might expect that these tenrecs might not be able to use Tb to control protein synthesis. Liver polysome profiles were confusing- there were no real changes between states or temperature. Rather, all tenrecs had a rather modest polysome profile with little heavy material typical of very active protein synthesis. To address this, we developed liver (do not grow well) and kidney cell (grow very well) cultures from ground squirrels and tenrecs. Our results indicate that the translation rate in tenrec primary cells is ~30 % of the translation rate in golden-mantled ground squirrel (p < 0.05). We interpret these results as tenrecs living ‘Life in the Slow Lane.’ By experiencing these low rates of a process critical to homeostasis, we contend tenrecs are avoiding rapid changes as a consequence of their rapid changes in Tb or metabolism. In effect, ‘driving slowly’ allows a tenrec to avoid major accidents which would imperil the animal. Perhaps not surprisingly, tenrec litters are enormous with as many as 32 confirmed young in a single litter. This research was supported by the Translational Research Institute for Space Health (TRISH). This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.
Sex-Dependent Increases of Hepatic Acylcarnitines by Myocardial Infarction
Authors: Wurm Daniel , Dai Wujing , Gu Haiwei , Chen Qin M. 
Published: May 2024

Many different diseases and environmental stressors can lead to an increase in oxidative stress. Urban air pollution leads to inhalation of airborne particulate matter in high traffc environments. This has been linked to chronic oxidative stress and increased risk or severity of myocardial infarction (MI). MI is one cause of acute oxidative stress. A key player for cellular defense against oxidative stress is activation of Nrf2, a transcription factor that controls the expression of key antioxidant and cytoprotective genes. Although Nrf2 has been reported to regulate genes for anabolic or lipid metabolism, little is known about the impact of Nrf2 or MI on the biochemical metabolism in the liver. Because the liver is a center for metabolism, metabolomic landscapes were determined in a model of chronic oxidative stress by Nrf2 knockout or acute oxidative stress by induction of MI via left anterior descending coronary artery ligation. 1353 compounds were detected with a quality control coeffcient variation value <20% using LC-MS/MS based non-targeted metabolomics. Differences in the baseline metabolic profiles of male versus female lead to comparison of Nrf2 knockout or MI in a sex specific manner. In male wild type mice, the liver responded to MI by elevating several carnitine derivatives, including linoleoyl carnitine, palmitoyl carnitine, and (11Z, 14Z)-eicosadienoyl carnitine. However, none of these changes were observed in female wild type mice. Either male or female Nrf2 knockout mice failed to respond to MI by elevating carnitines. These acylcarnitine increases were confirmed with a non-targeted lipidomics analysis. The elevated acylcarnitine levels correlate with an increase in carnitine palmitoyl transferase 1, CPT1, the enzyme responsible for the conjugation of carnitine and long-chain fatty acids. After this conjugation, acylcarnitines are transported into the mitochondria via carnitine-acylcarnitine translocase. Once inside the mitochondria, the carnitine moiety is removed via CPT2 and the long-chain fatty acid can be used for beta-oxidation to produce ATP. This data suggest that male mice are responding to MI by altering energy metabolism and increasing carnitines to compensate for energy production. Nrf2 appears to play a role in such metabolic reprogramming. The research programs under the direction of Dr. Qin M. Chen have been supported by NIH R01 GM125212, R01 GM126165, Holsclaw Endowment, the University of Arizona College of Pharmacy start-up fund, and NIEHS T32 ES007091. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.
The effect of baroreflex-induced sympathetic suppression on renal arterial impedance in rats
Authors: Fukuda Yukiko , Kawada Toru , Kataoka Yasuyuki , Peterson Jon , Saku Keita , Alexander Joe , Sunagawa Kenji 
Published: May 2024

Objective: We examined the effect of baroreflex on renal arterial impedance (ZRA) to gain insights into renal hemodynamics and local autoregulation. Methods: This study involved male Wistar-Kyoto rats (n = 6). We measured renal arterial pressure (PRA) and renal arterial flow (FRA). For high-resolution renal arterial impedance (ZRA) over the frequency range of 0.1-70 Hz, random pacing was employed. We isolated the bilateral carotid sinus baroreceptor regions and utilized a servo pump to control carotid sinus pressure (CSP) either at 60 mmHg or 140 mmHg. We controlled mean arterial pressure (AP) at 100 mmHg (AP-fix) or allowed to vary in response to changes in CSP (AP-variable). A three‐element Windkessel (3-WK) model was used to quantify ZRA (R1: proximal resistance, R2: distal resistance, and C: renal compliance). Results: With AP-fix, an increase of CSP from 60 mmHg to 140 mmHg did not change mean PRA (99.85 ± 0.16 vs. 98.00 ± 1.22 mmHg, P = ns) or mean FRA (8.73 ± 0.41 vs. 8.10 ± 0.38 min/mL, Δ = −7%, P = ns). This rise of CSP did not significantly affect R1 (0.74 ± 0.11 vs. 0.61 ± 0.12 mmHg·min/mL, P = ns) or R2 (8.13 ± 1.08 vs. 8.79 ± 0.85 mmHg·min/mL, P = ns), but it did increase C by 20% (4.05 ± 0.47 vs. 4.88 ± 0.35 (×10−5) mL/mmHg, P < 0.05). With AP-variable, elevating CSP from 60 mmHg to 140 mmHg significantly lowered mean PRA by 38% (105.42 ± 2.84 vs. 65.15 ± 4.87 mmHg, P < 0.01), but did not change mean FRA (9.00 ± 0.48 vs. 7.92 ± 0.66 min/mL, Δ = −12%, P = ns). Changes in CSP did not affect R1 (0.77 ± 0.12 vs. 1.23 ± 0.24 mmHg·min/mL, P = ns), but they did reduce R2 by 41% (8.01 ± 1.08 vs. 4.76 ± 0.79 mmHg·min/mL, P < 0.01) and increase C almost five-fold (3.79 ± 0.36 vs. 22.16 ± 6.91 (×10−5) mL/mmHg, P < 0.05). Conclusion: The minor change in FRA (−12%) despite a 38% drop in PRA suggests the presence of local autoregulation to buffer changes in renal blood flow. The primary effect of renal hemodynamic regulation on ZRA was found in R2. When AP was allowed to vary in response to changes in CSP, increasing CSP, the baroreflex-induced sympathetic suppression decreased R2. However, when AP was held constant at 100 mmHg, the reduction in R2 was not observed, even with baroreflex activation resulting from changes in CSP. Hence, the local renal autoregulation may counteract the baroreflex, maintaining renal blood flow unchanged. Such a mechanism serves to protect the glomeruli from exogenously induced high mean AP. NTT Research, Inc. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.
Glycoprotein VI Platelet Receptor Blockade Attenuates Progression of Established Abdominal Aortic Aneurysms in Murine Models
Authors: Brunner Seth , Benson Tyler , Spuzzillo Anthony , Wadding-Lee Caris , Coughlin Taylor , Cameron Scott , Owens Phillip 
Published: May 2024

Objective: Abdominal aortic aneurysms (AAA) frequently feature the formation of a nonocclusive intraluminal thrombus (ILT) along the site of aortic dilation. Platelets are known to maintain hemostasis and propagate thrombosis through several redundant activation mechanisms, yet the role of platelet activation in the pathogenesis of AAA associated ILT is still poorly understood. Thus, we sought to investigate how platelet activation via the glycoprotein VI receptor pathway impacts the pathogenesis of AAA. Methods and Results: RNA sequencing was performed on age-matched control human aortic tissue, AAA aortic tissue, and AAA ILT. Platelet transcripts comprised 25% of significantly enriched genes in the comparison of AAA thrombus to AAA tissue and control aortic wall, with GPVI increased by 9.1 Log2-FC (P = 3.21E−08). Circulating GPVI was significantly elevated in platelets isolated from AAA patients versus age-matched controls. Soluble GPVI (sGPVI—indicator of platelet activation), was significantly increased in the plasma of patients with fast-growing AAAs compared to slow-growing AAAs and healthy control subjects. To determine if GPVI blockade was protective from AAA progression, mice underwent either the angiotensin II (AngII) infusion or the topical elastase models of aneurysm formation. Following model initiation, aortic diameter was monitored by weekly ultrasounds. After two weeks, mice which developed an aneurysm (defined as an aortic diameter of >1.2mm) were randomized into control (IgG—50μg) or treatment (JAQ1—50μg, monoclonal GPVI antibody) groups. As expected, platelets isolated from JAQ1 treated mice did not activate in response to the GPVI specific agonist convulxin. Furthermore, JAQ1 treatment attenuated aneurysm progression, increased type I collagen deposition, attenuated macrophage accumulation, and increased survival in both murine AAA models versus IgG control treated mice. Conclusions: Platelets play a critical role in AAA pathophysiology and specific blockade of the GPVI-mediated activation pathway attenuates progression of established aneurysms in two independent murine models. Thus, our work demonstrates that specific targeting of GPVI to blunt platelet activation in AAA could be a potential therapeutic for a pathology with no current pharmacological treatments. R01HL147171-04 and R01HL158801-02. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.
The SNX17-Retriever endosomal recycling pathway is a key regulator of synaptic function and plasticity in hippocampal neurons
Authors: Chavis Garrett , Rivero-Rios Pilar , Tsukahara Takao , Uygun Tunahan , Iwase Shigeki , Weisman Lois , Sutton Michael 
Published: May 2024

The insertion and removal of integral membrane proteins is a key mechanism for the regulation of synaptic function. Once surface proteins are internalized, they can either be routed to lysosomes for degradation or can be recycled back to the cell surface. Two recycling protein complexes - the SNX27-Retromer complex and the recently discovered SNX17-Retriever complex - are the main drivers in recycling internalized cargoes in non-neuronal cells. In neurons, the SNX27-Retromer pathway has been shown to regulate surface expression of AMPA-type glutamate receptors (AMPARs) during long-term potentiation (LTP), but almost nothing is known regarding the role of the SNX17-Retriever pathway at synapses. Here, using cultured hippocampal neurons, we demonstrate that the SNX17-Retriever pathway localizes to both synaptic and extra-synaptic sites and that RNAi-mediated disruption of this pathway leads to the functional and structural loss of excitatory synapses. Moreover, we find that synaptic stimulation leads to the recruitment of SNX17 to synaptic sites in a manner that depends on Ca2+ influx through NMDARs, activation of CaMKII, and Retriever complex binding. Knockdown of either SNX17 or the Retriever subunit DSCR3 prevents structural plasticity of dendritic spines during LTP. Finally, this loss of structural plasticity is associated with altered traffcking of the cell adhesion molecule β1 Integrin, which is a SNX17 cargo in non-neuronal cells and as we show, in neurons as well. Additionally, SNX17-Retriever is shown to not play a role in AMPAR recycling. Preliminary studies suggest that SNX27 and SNX17 sort distinct cargos in neurons, suggesting that these two recycling pathways each play important, but unique, roles at synaptic sites. National Institute of Neurological Disorders and Stroke (NINDS) of the National Institutes of Health (NIH) under award numbers R01-NS129198 to L.S. Weisman and M.A. Sutton, R01-NS099340 and R01- NS064015 to L.S. Weisman, R01-NS097498 to M.A. Sutton, and R21-NS125449, and R01-NS116008 to S. Iwase, by the National Institute of Mental Health (NIMH) of the NIH under award number R21-MH127485 to S. Iwase. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.
Normal Skeletal Muscle Growth and Function Require Insulin Receptors Expressed by Cholinergic Motor Neurons
Authors: Hyun Uisu , Kim Youngjun , Sohn Jong-Woo 
Published: May 2024

It is well known that insulin stimulates skeletal muscle growth. However, it is currently unknown how insulin does so since it was previously shown that insulin receptors expressed by skeletal muscles are dispensable in this regard. In this study, we focused on insulin receptors expressed by the cholinergic motor neurons and generated conditional knockout mice which lack insulin receptors specifically in the cholinergic neurons. We found that insulin activates cholinergic motor neurons and that insulin receptors expressed by the cholinergic neurons are required for normal growth of skeletal muscle. We also noted that the conditional knockout mice have underdeveloped neuromuscular junction and compromised skeletal muscle function. Together, our study suggests that insulin stimulates and activates cholinergic motor neurons to promote normal skeletal muscle growth and function. This work was supported by the National Research Foundation of Korea (NRF-2022R1A2C3005613 to Jong-Woo Sohn) funded by the Korean Ministry of Science and ICT. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.
Cyclic strain promotes MCP-1 expression in Dahl Salt Sensitive (SS) rat vascular smooth muscle cells (VSMCs) through the TGF-β pathway
Authors: Feng Conrad , Xing Dongqi , Ying Wei-zhong , Guan Zhengrong , Sanders Paul 
Published: May 2024

Prior studies have suggested a role for TGF-β in the generation of hypertension in SS rats. We have shown that production of MCP-1 (CCL-2) and TGF-β1 increase in the vasculature of hypertensive SS rats. This study aims to explore novel mechanisms of hypertension development. We hypothesize that TGF-β1 regulated the production of MCP-1 by VSMCs in the setting of cyclic strain. Primary cultures of SS aortic VSMCs in early (<6) passages were grown in standard fashion in Dulbecco’s Modified Eagle’s Medium supplemented with 10% fetal bovine serum. Cultured cells were identified as smooth muscle cells by their characteristic morphology and positive immunostaining for α-smooth muscle actin. In the first study, VSMCs were treated with recombinant active TGF-β1 (1 ng/mL and 10 ng/mL) for 6 h, then harvested for MCP-1 mRNA quantification by SYBR green qPCR. Medium was also collected to determine MCP-1 levels using ELISA. TGF-β1 increased MCP-1 mRNA expression by 2.0- and 3.1-fold in VSMC incubated in medium containing 1 ng/mL and 10 ng/mL of TGF-β1, respectively. In a second series of studies, VSMCs were seeded onto 6-well BioFlex plates. Some cells were pre-treated with 10 μmol/mL of an ALK4/5 inhibitor. Cyclic strain, an in vitro model of hypertension, was performed using the FlexCell 6000 and a standardized regimen (8 h, 15% elongation @ 1 Hz). Cells and medium were then harvested for mRNA analyses and determination of MCP-1 levels. When compared with expression by VSMCs maintained under static condition, cyclic strain increased (p<0.05) MCP-1 mRNA expression 4.2-fold and TGF-β1 mRNA 1.93-fold. Cyclic strain also increased (p<0.01) MCP-1 protein in the medium (static condition vs cyclic strain, 112.6 ± 7.1 pg/mL vs 222.5 ± 6.5 pg/mL, n=6). Pre-treatment of VSMC with the ALK4/5 inhibitor reduced mRNA expression of MCP-1 mediated by cyclic strain (Cyclic strain vs Cyclic strain + AKL4/5 inhibitor, 4.5 ± 1.06 vs 2.88 ± 0.47, n=6, P < 0.01). Thus, cyclic strain promotes MCP-1 production by VSMCs through the TGF-β pathway. The data support the need for further study of this interaction as well as a potential role in arterial hypertension in this rodent model. Department of Veterans Affairs Basic Sciences R&D (BSRD) Service (1 I01 BX005640) National Institutes of Health (P30 DK079337). This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.
RBX1-CRLs Promote Perinatal Heart Development by Regulating Hippo-YAP Signaling
Authors: Zambrano-Carrasco Josue , Wang Wenjuan , Ayala Juan , Sibley John , Carruth Jessica , Jiao Kai , Li Jie 
Published: May 2024

Background: During development, the heart is the first organ to form, ensuring the pumping of blood through the body. Defects in cardiac morphogenesis can lead to congenital heart diseases (CHD) or predispose the heart to cardiomyopathies in adult life. The molecular mechanisms underlying this process remain incompletely understood. Cullin-RING E3 ubiquitin ligases (CRLs) are multi-subunit complexes critical to maintaining protein homeostasis by targeting diverse protein substrates for ubiquitination and subsequent degradation. In CRLs, RBX1 serves as the catalytic subunit that promotes the transfer of ubiquitin to the target protein. Germline ablation of RBX1 in worms, flies, and mice caused embryonic lethality. Deletion of RBX1 in zebrafish resulted in cardiac malformation and dysfunction, which was rescued by endothelial, but not myocardial, RBX1 overexpression. However, the cardiomyocyte-autonomous role of RBX1 in the heart in vertebrates is unknown. Therefore, we hypothesize that cardiomyocyte RBX1 is indispensable for cardiac development. Methods and Results: We generated a cardiomyocyte-specific RBX1 knockout (RBX1CKO) mouse model via αMHCCre. RBX1-deficient mouse embryos were initially morphologically indistinguishable from the littermate controls at E14.5, but began to show peripheral edema and hemorrhage at E16.5, and eventually died around E18.5. Histological analysis of RBX1CKO hearts revealed severe myocardial hypoplasia and bi-ventricular non-compaction by E14.5. Bulk RNA sequencing and proteomic analyses of RBX1CKO hearts identified perturbations in glycolytic to oxidative transition and dysregulation of sarcomeric proteins. EdU incorporation assay showed a significant reduction in cardiomyocyte proliferation in E14.5 RBX1-deficient hearts, accompanied by the impaired nuclear translocation of YAP, a co-transcriptional factor essential for cardiomyocyte proliferation and maturation. Mechanistically, RBX1 deficiency destabilized multiple CRLs and resulted in the accumulation of Hippo kinases and adaptor proteins such as SAV1, LATS1/2, and MOB1, leading to the elevated phosphorylation and inactivation of YAP. We further demonstrated that loss of RBX1 inhibited the degradation of the Hippo adaptor MOB1, likely due to suppressed ubiquitination. Conclusion: Our study identifies RBX1 as an essential regulator of ventricular maturation and found that RBX1-mediated proteolysis of Hippo kinases serves as a critical checkpoint to ensure YAP activation and, hence, cardiomyocyte proliferation and maturation during cardiac development. AHA Predoctoral Fellowship, American Heart Association, 23PRE1012641. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.