bluuebunny/crossref_metadata_2025_split huggingface dataset:
Identification of novel changes in skeletal muscle proteome after sildenafil treatment
Authors: Wu Linda , Garcia Luis , Lipinski Austin , Langlais Paul , Coletta Dawn 
Published: May 2023

Insulin Resistance (IR) is associated with increased matrix proteins, including collagens, fibronectins, and laminins in the endomysium surrounding skeletal muscle. The increased ECM deposition in skeletal muscle IR results in ECM stiffening that decreases muscle integrity and cellular signaling. Recent evidence has shown that Sildenafil, a phosphodiesterase 5a inhibitor, improves insulin sensitivity in humans, and in high fat diet mice, reduces collagen deposition in the matrix surrounding skeletal muscle. In addition, data shows that a short-term treatment of Sildenafil for 7 days increases protein synthesis and decreases muscle fatigue in humans, which may mitigate the effects of IR. However, very few studies have examined the molecular mechanisms and metabolic signaling directly associated with Sildenafil’s influence on skeletal muscle. The aim of our study was to perform an unbiased proteomic analysis on Sildenafil’s effects on skeletal muscle in C2C12 myotubes. We hypothesized that Sildenafil would alter the abundance of proteins in metabolic pathways that are involved in communicating with the ECM. We performed a dose response (n = 3 for each dose; 0.5 μM, 1 μM, 2.5 μM, 5 μM, and 10 μM) and time response (n = 3 for each time; 30 min, 1 hour, 4 hour, 6 hour) curve. Proteomic analyses revealed that the highest number of proteins significantly changing was at 1 μM for 30 min (n = 423). Analysis of the significant proteins revealed 396 and 27 proteins that increased and decreased, respectively. KEGG analysis revealed that 20% of the total changing proteins (n = 83) were in metabolic pathways. The top 10 metabolic protein increasers, represented as a fold change of abundance compared to vehicle, were ASCL3 (1.27 ± 0.08), GT251 (1.12 ± 0.10), ODPA (1.15 ± 0.03), PCKGM (1.08 ± 0.04), PCY1A (1.23 ± 0.04), PGM1 (1.06 ± 0.04), PTG1S (1.21 ± 0.05), PYR1 (1.37 ± 0.09). SAHH (1.19 + 0.07), and THIM (1.13 ± 0.08). Of the 83 metabolic proteins, only 6 were decreasing, P5CR2 (0.78 ± 0.04), ALD1 (0.82 ± 0.11), P4K2A (0.67 ± 0.05), DGAT1 (0.27 ± 0.19), SCLY (0.77 ± 0.15), and PLCD3 (0.61 ± 0.16). The results from this preliminary study conclude that Sildenafil affects proteins involved in metabolic pathways. Further analysis will be performed to confirm the roles of these proteins in relation to ECM signaling/communication in these metabolic pathways. University of Arizona's Research, Innovation & Impact Faculty Seed Grant This is the full abstract presented at the American Physiology Summit 2023 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.
Purinergic receptor activation attenuates pilocarpine-induced water intake
Authors: De Paula Patricia , Gois Pedro A. , Menani José V. 
Published: May 2023

Intraperitoneal (ip) injection of pilocarpine (PILO, muscarinic cholinergic agonist) induces salivation, water intake and changes in body temperature. These effects are due to a direct action on the salivary glands and central nervous system (CNS). ATP (adenosine 5'-triphosphate) is a neurotransmitter in the peripheral and CNS and is responsible for activating purinergic receptors located in various regions including the salivary glands. Preliminary results from our laboratory showed that ATP injected intraperitoneally (ip) reduced PILO-induced salivation, suggesting the participation of peripheral purinergic receptors in salivary secretion. The aim of the present study was to investigate the effects of ip injection of ATP (a natural agonist of purinergic receptors) on water intake and changes in body temperature induced by PILO also administered ip. Male Holtzman rats (300-350 g, n=18) were used. The experimental protocols were approved by the Ethical Committee for Animal Care and Use – CEUA from the Dentistry School of Araraquara, UNESP (Proc. CEUA nº 11/2019). A group of 10 rats received ip injection of PILO (1 mg/kg of body weight) to induce water intake. Ten minutes before, half of the group received an ip injection of ATP (200 mg/kg of body weight) and the other half received saline. Then water was offered in graduated burettes fitted with metal drinking spouts. The results showed that ip injections of ATP (200 mg/kg of body weight) reduced PILO-induced water intake (0.2 ± 0.1 ml/15 min, vs. saline + pilocarpine: 2.2 ± 0.6 ml/15 min, p<0.05). Another group of 8 rats was submitted to a median laparotomy for the placement of body temperature sensors in the abdominal cavity. The results showed that ip injection of PILO did not alter body temperature (0.3 ± 0.1 °C, vs. saline: 0.3 ± 0.1 °C) and the combination of ATP + PILO also did not change body temperature (0.2 ± 0.1 °C). Thus, the results suggest that ip injection of ATP reduced PILO-induced water intake, but PILO alone or combined with ATP did not change body temperature in rats CNPq, PROPE UNESP, FAPESP This is the full abstract presented at the American Physiology Summit 2023 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.
Loss-of-STING function by nitro-fatty acids reduces endometriotic lesion formation in a syngeneic model of endometriosis
Authors: Pitts Mareena , Xu Wei , Garcia-Barrio Minerva , Schopfer Francisco , Thompson Winston , Villacorta Luis 
Published: May 2023

Endometriosis is a painful, chronic inflammatory disease that impacts fertility and the overall well-being in 10% of reproductive age women. Current therapies include hormonal treatment to induce a hypoestrogenic state, causing serious complications from side effects. STING (Stimulator of Interferon Genes) plays an essential role in innate immunity but also exacerbates inflammation in chronic inflammatory diseases. Recently, STING was found aberrantly upregulated in endometriosis patients. Nitro-fatty acids (NO2-FAs) are bioactive lipids that exert anti-inflammatory and protective roles in preclinical models of inflammation and fibrosis, in part via inhibition of STING. We hypothesized that NO2-FAs protect against STING-dependent endometriosis lesion formation and tested its therapeutic potential using a syngeneic transplant model of endometriosis. Mice expressing green fluorescent protein (EGFP) were treated with Pregnant Mare Serum Gonadotrophin (PMSG) to induce decidualization. Decidualized uterine fragments were transplanted into wild-type (WT) female recipients randomized to receive NO2-FA or vehicle. Endometriosis was allowed to develop for 28 days. In vivo imaging of EGFP donor-derived endometrial lesions demonstrates that NO2-FA administration significantly reduces peritoneal endometriotic lesion formation in female recipients. Furthermore, loss-of-STING function, using STING KO female mice as recipients, demonstrates that STING plays a systemic functional role on endometriosis development. Finally, our studies demonstrate that NO2-FAs inhibit cyclic GMP-AMP Synthase (cGAS)-induced type I interferon signaling in bone-marrow derived macrophages (BMDM) and in the peritoneum of WT female mice but not STING KO mice. In summary, we show that STING is a novel, essential factor contributing to the pathogenesis of endometriosis and that NO2-FA administration may serve as a novel non-hormonal therapy against endometriosis by targeting STING. This study is funded by the American Physiological Society William Townshend Predoctoral Fellowship (MP), NIMHD U54-MD007602 (WT), R01-HL162294 (MGB) and R01-HL123333 (LV). This is the full abstract presented at the American Physiology Summit 2023 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.
Transient changes in endothelial and epithelial cell cytoskeleton and morphology upon olfactory receptor signaling
Authors: Nilon Annabella , Scibek Jacob , Curtis Theresa 
Published: May 2023

Olfactory receptors (ORs) are expressed in various non-nasal tissues; however, their physiological roles are largely unknown. ORs are G-protein coupled receptors (GPCRs), and there is a well-established link between different classes of G-proteins and cytoskeletal structure changes affecting cellular morphology that has been unexplored after odorant binding. Thus, the present study was conducted to determine if odorant binding in non-olfactory cells causes cytoskeletal changes that lead to cell morphological changes. To this end, human umbilical vein endothelial cells (HUVECs), which express OR10J5, and human keratinocyte (HaCaT) cells, which express OR2AT4, were used to conduct this study. Using these two different cell lines, it was shown that odorant addition, lyral and Sandalore, respectively, caused a dose-dependent increase in cAMP. Furthermore, HUVECs exposed to lyral exhibited a disassembly of the tubulin-containing microtubules and an increase in cortical actin microfilaments, whereas HaCaT cell exposure to Sandalore exhibited both a disassembly of the tubulin-containing microtubules and cortical actin microfilaments. Even though the endothelial and epithelial cell lines had different cytoskeletal responses to odorant exposure, both exhibited a change in cellular morphology as detected by a decrease in electrical resistance using Electric Cell-substrate Impedance Sensing (ECIS; Applied Biophysics, Inc., Troy, NY). Interestingly, these odorant-induced cytoskeletal and morphological changes occurred rapidly (under 1 hour) but returned to normal after 20 hours, highlighting the complexity of olfactory signaling, which may involve desensitization of the signaling pathway and/or internalization of the odorant receptor. Future experiments will further elucidate the olfactory receptor signaling mechanism in endothelial and epithelial cells so that their physiological role can be fully understood. Funded by Army Research Office STTR #W911NF-19-P-0014 and W911NF-20-C-0014, the SUNY Cortland Undergraduate Research Council, and Research and Sponsored Programs. This is the full abstract presented at the American Physiology Summit 2023 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.
A novel small molecule inhibitor of vascular endothelin signaling and hypertension
Authors: Ezewudo Emmanuella , Menon Sreelakshmi , Zerin Farzana , Moniri Nader , Nguyen Tro , Kashikar Rama , Chougule Mahavir , Rahman Taufiq , Hasan Raquibul 
Published: May 2023

Augmented endothelin-1 (ET-1) signaling causes profound vasoconstriction, smooth muscle proliferation, endothelial dysfunction, oxidative stress, and vascular inflammation and sensitivity, which are often present in resistant hypertension, salt-sensitive hypertension and hypertension among African Americans. Here, we sought to examine the role of indole-3 acetic acid (IAA) as a novel inhibitor of vascular ET-1 signaling and hypertension. IAA, popularly known as auxin, is the most potent plant growth hormone, which is also produced in the human body via tryptophan metabolism by gut microbiota. Although the role of IAA as a potent plant growth hormone has been characterized, little is known about the biological activity of this compound in humans. Importantly, studies on the role of IAA as a regulator of arterial contractility, vascular inflammation and hypertension remain completely unexplored. Our data demonstrates that IAA inhibits ET-1-evoked vasoconstriction in resistance mesenteric arteries, ROS production in human aortic SMCs, expression of adhesion molecule VCAM-1 on EC surface, leukocyte adhesion, and ECE-1 activity. IAA dose-dependently inhibits the activity of a purified human endothelin converting enzyme-1 (ECE-1) that converts the precursor big ET to its active form ET-1 and is upregulated in hypertension. Consistent with these findings, we show that treatment of Dahl salt-sensitive (DSS) hypertensive rats, a model of augmented ET-1 signaling, with an oral extended-release, in situ gel-based formulation of IAA (IAA-oragel) reduces blood pressure (BP) elevation, suggesting a key role for IAA in BP regulation. Taken together, this study presents a novel inquiry into regulation of ET-1 function by IAA and could identify IAA as the first dual action inhibitor of the ET system. This is the full abstract presented at the American Physiology Summit 2023 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.
A Western Diet Modifies the Activation Profile of Circulating Platelets
Authors: Corken Adam , Wahl Elizabeth , Sikes James , Thakali Keshari 
Published: May 2023

Background: While it is known that obesity increases the risk for cardiovascular disease (CVD), there has been no clear investigation into the effect an obesogenic diet exerts onto circulating platelets. As platelets are the critical actors in the formation of occlusive clots that manifest as a myocardial infarction or stroke, it is pertinent to establish any potential link between obesity and platelet activity. Methods: To examine the effect of a Western diet on baseline platelet activity, male C57BL6/J mice were fed either a control (10% fat, 7% sucrose, 0.3% salt Research Diets Inc. D12450J) or Western (40% fat, 30% sucrose, 8% salt Research Diets Inc. D06111701) diet ad libitum for 20 weeks beginning at the time of weaning. At 12, 16 and 20 weeks after beginning diet administration whole blood was collected via the retro-orbital sinus. Blood was used to generate platelet rich plasma (PRP) which was stimulated with collagen (20mg/ml) prior to staining for flow cytometry analysis. Flow cytometry staining utilized antibodies to recognize the platelet receptors GPIbα, P-Selectin and the activated form of integrin αIIbβ3 (JON/A). Additionally, body composition was likewise determined at these time points using Echo-MRI while bodyweight was measured weekly. Results: Western diet fed mice gained slightly more weight than control mice and had a higher percentage of body fat (measured by EchoMRI). Surprisingly, the two groups differed in GPIbα and activated αIIbβ3 expression on circulating platelets at all three observed time points. Specifically, platelets from control-fed mice displayed significantly higher GPIbα expression throughout the study while Western diet fed animals had higher levels of activated αIIbβ3. There was no difference in platelet expression of P-Selectin between the groups at any time point. Conclusions: Platelet activation is not a binary process where platelets display a definitive “active” or “inactive” phenotype. Though terminal platelet activation elicits de-granulation and an increase in P-Selectin expression, the spectrum of platelet stimulation preceding this is marked by variable incidences of integrin activation and GPIbα shedding. Here we demonstrate proof of principle that dietary factors, such as the consumption of a Western diet, can facilitate early changes to the stimulation pattern of circulating platelets which is then maintained long-term. While the consequence of this altered platelet stimulation profile is unknown with regards to the pathways of hemostasis and thrombosis, further investigations of downstream signaling and amplification pathways are needed. Funding: Arkansas Children's Research Institute/Arkansas Bioscience Institute Postgraduate Fellowship: AWD-037176 This is the full abstract presented at the American Physiology Summit 2023 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 role of estradiol on sensory nerve-mediated dilation in postmenopausal women
Authors: Del Vecchio Angelica , Welti Laura , McGinty Shane , Schwab Allyson , Haigh Katherine , Wong Brett , Wenner Megan 
Published: May 2023

BACKGROUND: Local heating of the skin elicits a biphasic vasodilatory response. The initial peak of this response is largely mediated by sensory nerve mechanisms. Receptors for estradiol (E2), the primary form of estrogen, have been found on peripheral sensory neurons indicating E2 impacts the nociceptive response. In animal models, E2 may decrease pain tolerance or act as an analgesic but it is currently unclear how E2 affects sensory nerve-mediated responses in women, particularly in E2-deficient postmenopausal women. PURPOSE: We tested the hypothesis that E2 augments sensory nerve-mediated vasodilation during local heating in E2-deficient postmenopausal women. METHODS: We measured cutaneous vasodilation to local heating (42°C) via laser Doppler flowmetry during microdialysis perfusions of lactated Ringer’s (control) and in a subset (n=4), L-NAME (non-selective nitric oxide synthase inhibition; 15mM) in 10 healthy postmenopausal women (55±2 yrs) at baseline (BL) and after 7 days E2 administration using a transdermal patch (E2; 0.1 mg/day, Vivelle-Dot). Cutaneous vascular conductance (CVC) was calculated as SkBF/MAP normalized to maximal vasodilation achieved by perfusion of sodium nitroprusside (28mM) and heating to 43°C. RESULTS: All women were less than 10 years post menopause (average 5±3 years), normotensive (SBP: 123±12 mmHg; DBP 77±5 mmHg) and not obese (BMI: 24±3 kg/m2). Sensory nerve-mediated dilation was unaffected by E2 administration (E2: 56±8 vs BL: 55±13 %CVCmax; p=0.72). Inhibition of eNOS with L-NAME did not blunt sensory nerve-mediated dilation and was not impacted by E2 (E2: 71±18 vs BL: 56±33 %CVCmax; p=0.55). In a subset (n=5) serum E2 concentration increased after E2 administration (E2: 80.42±11.90 vs BL: 25.74±4.42; p=0.0005) CONCLUSIONS: These preliminary findings suggest E2 administration does not impact sensory nerve-mediated dilation in postmenopausal women. Furthermore, sensory nerve-mediated dilation does not appear to be mediated by nitric oxide. Additional research is needed to further understand the impact of sex hormones and aging on sensory nerve-mediated dilation in women. Research supported by AHA award 16SDG30700015 and NIH Grant P20 GM113125 This is the full abstract presented at the American Physiology Summit 2023 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.
Polycystin 1 ciliary localization is controlled by mechanosensitive signaling that involves secretion of ATP
Authors: Gresko Nikolay , Caplan Michael 
Published: May 2023

Autosomal dominant polycystic kidney disease (ADPKD) is caused by mutations in the genes, that encode Polycystin1(PC1) and Polycystin 2 (PC2), respectively. PC1 is a 460kD multi-spanning membrane protein that undergoes multiple proteolytic cleavages. At least two of those cleavages release fragments of the PC1 C-terminus. A population of the PC1 protein localizes to the primary cilium where, together with PC2, it is thought to participate in cellular mechano-sensation. PC1 and PC2 can form a cation-permeable heterotetrameric channel that may regulate Ca+2 influx into the cilia. Recent studies demonstrate that PC1 can function as an atypical GPCR and that it can interact with Wnt ligands.We report that activation of the PC1 receptor function regulates its ciliary localization in cultured LLC-PK1 cells and show that this effect can be modified by ligand binding or by mechanical stimuli. The ciliary quantity of the full length PC1 is dramatically reduced by exposure to the PC1-stimulatory ligand Wnt9b. Furthermore, we show that a protein construct that corresponds to a constitutively active form of the PC1 receptor is absent from the primary cilia whereas a protein construct that corresponds to a constitutively inactive form of PC1 is prominently localized to the primary cilia. Inhibition of β-arrestin with barbadin alters the distribution of the constitutively active PC1 construct, resulting in its accumulation in the cilium. Taken together, these data suggest that the ciliary localization of PC1 is governed by its receptor activity through a mechanism that resembles that of GPCR receptor desensitization. We find that mechanical stimuli also lead to removal of PC1 from the cilium. This effect is due to mechanical stimulus-induced release of ATP into the medium. Enzymatic hydrolysis of extracellular ATP prevents the effects of mechanical stimuli on PC1 ciliary localization, whereas addition of exogenous ATP to the medium is sufficient to induce departure of PC1 from the cilium in the absence of mechanical stimuli. We show that both Wnt9b and mechanical stimuli exert their effects on PC1 ciliary localization through the β-arrestin dependent cascade. These findings illuminate an exciting and largely untapped aspect of PC1 biology. NIH RO1 DK072612 and DOD PR191158 This is the full abstract presented at the American Physiology Summit 2023 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.