bluuebunny/crossref_metadata_2025_split huggingface dataset:
Systemic Administration of Serotonin Reduces the Excitability of the Raphé Pallidus‐Brown Adipose Tissue Sympathetic Nerve Pathway
Authors: Mota Clarissa M. D. , Madden Christopher J. , Morrison Shaun F. 
Published: June 2021

Activation of 5‐HT1A receptors in the raphé pallidus (RPa) inhibits the skin cooling‐evoked increases in brown adipose tissue (BAT) sympathetic nerve activity (SNA). Our aim was to determine whether intravenous (iv) 5‐HT would affect BAT SNA and if 5‐HT1A receptors in the RPa play a role in mediating any of the effects of iv 5‐HT. In urethane/α‐chloralose anesthetized, neuromuscularly blocked, ventilated rats, electrical stimulation (100–150 μA, single 1 ms pulses) of the RPa evoked compound potentials in BAT SNA. Iv infusion of 5‐HT (500 μg/h) reduced the amplitude of the RPa‐evoked compound potentials in BAT SNA. Skin cooling‐evoked increases in BAT SNA were also inhibited by the iv infusion of 5‐HT, and by an iv bolus of the 5‐HT1A/7 receptor agonist, 8‐OHDPAT (100 μg in 50 μl). Nanoinjection of the 5‐HT1A receptor antagonist, WAY100635 (10 mM, 100 nl), in the RPa completely prevented the iv 8‐OHDPAT‐induced inhibition of BAT SNA, but not that following iv 5‐HT. In conclusion, iv 5‐HT reduces the excitability of the sympathetic pathway from the RPa to BAT, but this effect is not mediated by activation of 5‐HT1A receptors in the RPa. In contrast, activation of 5‐HT1A receptors in the RPa plays a key role in the inhibition of cold‐evoked BAT SNA by iv administration of 8‐OHDPAT.Support or Funding InformationSupported by São Paulo Research Foundation (FAPESP) grant #2017/24232‐9 (C.M.D.M.) and by NIH R01‐NS091066 (S.F.M.).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Leptin‐induced neuroinflammatory effects in the hypothalamic neurons: role of Sirt1 and FoxO1
Authors: Zheng Hong , Liu Xuefei 
Published: June 2021

Accumulated evidence indicates that central nervous system signaling pathways may contribute to the etiology and pathogenesis of obesity‐induced hypertension. Hypothalamic leptin‐mediated signaling may contribute to the exaggerated sympathoexcitation in obesity. Leptin can influence food intake, energy expenditure and sympathetic nerve activity via hypothalamic pro‐opiomelanocortin (POMC) neurons in the arcuate nucleus (ARCN) of hypothalamus. The aim of the study was to determine the effects of leptin on the neuroinflammation‐mediated high sympathetic nerve activity in obesity. We used hypothalamic neuronal cell mHypoA‐POMC (CLU500) and microglial cell (SIM‐A9) to observe pro‐inflammatory and anti‐inflammatory cytokines levels after 24 hours leptin treatment. We found that leptin incubation increased the levels of pro‐inflammatory cytokines interleukin 6 (IL‐6), interleukin 1β (IL‐1β), and tumor necrosis factor alpha (TNFα) in both POMC and microglial cells. Leptin treatment also reduced the level of lipoxin A4, a potent anti‐inflammatory bioactive lipid formed from arachidonic acid in both cells. Knockdown nutrient‐sensing enzyme sirtuin 1 (Sirt1) by siRNASirt1 significantly attenuated the levels of leptin‐induced pro‐inflammatory cytokines in the POMC and microglial cells. On the contrary, knockdown transcription factor forkhead box protein O1 (FoxO1) by siRNAFoxO1 significantly enhanced leptin‐induced pro‐inflammatory cytokines. Further, in vivo study, we found significant increased protein expressions of IL‐6, IL‐1β and TNFα in the ARCN in 12 weeks high fat induced obese rats. siRNASirt1 reduced leptin‐mediated renal sympathetic nerve activity in obese rats. The study suggests that enhanced leptin‐mediated sympathetic activation in obesity may via hypothalamic neuroinflammatory pathway. We propose Sirt1 and FoxO1 in hypothalamus are key regulators of leptin signaling pathway contributed to the over sympathetic activation in obesity.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Long‐term consequences of acute kidney injury during pregnancy
Authors: Szczepanski Jamie , Spencer Shauna‐Kay , Griffin Ashley , Bowles Teylor , Wallace Kedra 
Published: June 2021

OBJECTIVEWomen with severe preeclampsia and HELLP syndrome (sPE/HELLP) are more likely to develop acute kidney injury (AKI) compared to women without sPE/HELLP. AKI during pregnancy is associated with rates of maternal mortality and fetal loss that range from 30–60%. Women with a history of sPE/HELLP are more likely to develop hypertension, and have higher incidences of chronic kidney disease, end stage renal failure and cognitive failure compared to women with histories of normal pregnancies. We tested the hypothesis sPE/HELLP leads to brain damage during pregnancy which is exacerbated by AKI.STUDY DESIGNOn gestational day (GD) 12, mini‐osmotic pumps infusing sFlt‐1 and sEng were placed into rats to induce HELLP. A subset of HELLP and NP rats underwent bilateral renal ischemia‐reperfusion surgery for 45 minutes on GD18 to create AKI. After rats had recovered, all rats were placed in metabolic cages for 24hrs. Mean arterial pressure was measured on GD19 followed by Evan's blue infusion to assess blood brain barrier permeability, or tissue collection and euthanization.RESULTSDuring pregnancy HELLP+AKI rats had a significant decrease in urine output compared to NP rats (1.4±0.28 vs. 0.48±0.12mL/hr; p=0.01). MAP was significantly increased in HELLP rats (p<0.0001) and HELLP+AKI rats (p=0.03) compared to NP rats. Kidney weight was also significantly increased in NP+AKI compared to NP rats (p=0.005) and in HELLP+AKI compared to HELLP rats (p=0.003). Urine creatinine levels were significantly lower in all groups of rats compared to NP rats (p=0.01). NP pups were significantly larger compared to HELLP (p=0.006), NP+AKI (p=0.0002) and HELLP+AKI (p=0.0005) pups. Rats with HELLP had increased blood brain barrier permeability in the brainstem/cerebellum compared to NP rats (p=0.04). Similar results were seen in NP+AKI (p=0.04) and HELLP+AKI (p=0.01) rats compared to NP rats. Whole brain water weight was significantly increased in HELLP+AKI rats compared to NP rats (p=0.03).CONCLUSIONThe results from the current study suggest that AKI during pregnancy contributes to oliguria, hypertension, decreased urine creatinine, maternal brain injury and reduced pup size. In the setting of HELLP+AKI brain injury was worsened compared to rats with just AKI or HELLP syndrome. Studies are currently ongoing to examine the progression to chronic kidney disease and cognitive failure in rats with a history of sPE/HELLP with and without AKI during pregnancy.Support or Funding InformationResearch reported in this publication was supported by the National Institute Of General Medical Sciences of the National Institutes of Health under Award Number P20GM121334. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
TRPC6 Inactivation Does Not Protect Against Diabetic Kidney Disease in Streptozotocin (STZ)‐treated Sprague‐Dawley Rats
Authors: Khayyat Naghmeh Hassanzadeh , Kim Eun Young , Dryer Stuart E 
Published: June 2021

Glomerular TRPC6 channels are up‐regulated in rodent models of diabetes mellitus, but the role of TRPC6 in driving progression of experimental diabetic nephropathy is controversial. We recently used CRISPR/Cas9 methods to generate Sprague‐Dawley rats with a deletion in exon 2 of the Trpc6 gene that renders the channels non‐functional [1]. In addition, we observed that animals lacking functional TRPC6 were protected in the chronic puromycin aminonucleoside (PAN) model of acquired FSGS [1]. In the present study, 300‐gram adult Trpc6del/del rats and their wild‐type Trpc6wt/wt littermates were examined in the STZ model of type‐1 diabetes using protocols approved by the University of Houston IACUC committee. In our experiments, diabetes was induced by a single intraperitoneal injection of STZ, while controls received vehicle. Animals were not treated with insulin. STZ‐treated rats with severe hyperglycemia (>450 mg/dl) by 5 days after treatment were used for analysis and were monitored for the next 10 weeks. At 4 and 10 weeks after STZ treatment, compared to vehicle‐treated controls, STZ‐treated rats had severe hyperglycemia (745 ± 144 mg/dl, mean ± SD at 10 weeks). They also had elevated plasma triglycerides and total cholesterol, and failed to gain weight. STZ‐treated rats also exhibited markedly increased urine albumin excretion, increased blood urea nitrogen (BUN), and increased plasma creatinine and decreased creatinine clearance, indicating marked declines in renal function in diabetic rats. Two‐way ANOVA analyses of these datasets revealed no differences between Trpc6wt/wt and Trpc6del/del rats in any of these parameters. There was no time in the course of the experiment where we obtained functional data suggesting a protective effect of TRPC6 inactivation on blood glucose, or kidney function in STZ diabetes. Histological analysis carried out 10 weeks after STZ treatment revealed marked glomerular and tubulointerstitial disease in STZ‐treated rats. Mesangial expansion was the prominent feature in glomeruli of STZ‐treated animals, and glomeruli were less disrupted than we previously observed in the chronic PAN model [1]. Blind semi‐quantitative analysis of glomerular histology revealed a significant effect of STZ but no protective effect of TRPC6 inactivation on glomerular pathology. In summary, TRPC6 inactivation does not prevent declines in renal function and appears to have no protective effect in the STZ model of type‐1 diabetes in Sprague‐Dawley rats. This is in marked contrast to our previous work in chronic PAN nephrosis, where TRPC6 inactivation preserves renal function and histology.Support or Funding InformationSupported by NIH grant R01‐DK104708.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Urinary prorenin/renin and soluble prorenin receptor excretion in Type 1 and Type 2 diabetes
Authors: Ramkumar Nirupama , Stuart Deborah , Kohan Donald 
Published: June 2021

BackgroundActivation of the intra‐renal, rather than systemic, renin angiotensin system (RAS) has been suggested to play a role in the development of diabetic kidney disease. In this study, we examined plasma and urinary prorenin/renin levels and urinary soluble prorenin receptor (sPRR) levels in mice with early Type 1 and Type 2 diabetes.MethodsMale Akita (Type 1 model bearing a mutated Ins2 gene) and db/db (Type 2 model bearing a mutated LepR gene) mice were studied from 10–18 weeks age and metabolic balance studies conducted at 4 week intervals on a normal salt diet.ResultsDiabetic mice were hyperglycemic (Akita: 479 ± 31, db/db: 314 ± 60, control: 152 ± 12 mg/dL), polyuric (Akita: 22.2 ± 3.9, db/db: 10.9 ± 2.3, controls 1.6 ± 0.1 ml/day) and polydipsic (Akita: 24.6 ± 3.4, db/db: 11.5 ± 2.4, controls: 4.2 ± 0.5 ml/day) at 10 weeks which continued until 18 weeks. Compared to controls, Akita mice had lower body weight (23.4 ± 0.6 vs controls: 29.2 ± 0.3 grams) while db/db mice had higher body weight (51.9 ± 0.3 g/day) at 10 weeks which continued until 18 weeks. Compared to controls, urinary angiotensinogen and total prorenin/renin levels were increased in Akita mice at 10 weeks and remained elevated at 14 and 18 weeks of age (Table 1); these increases were proportionally greater than changes in urinary albumin excretion. Urinary sPRR excretion was markedly increased in Akita mice at 14 and 18 weeks. In contrast, db/db mice had higher urinary albumin excretion, urinary angiotensinogen and urinary prorenin/renin excretion relative to controls at all 3 time points but undetectable urinary sPRR excretion until 18 weeks. Plasma renin concentration was similar between the controls and diabetic mice at all time points. There was mild glomerular hypertrophy at 18 weeks of age in db/db but not Akita mice.ConclusionThere is early activation of the intra‐renal but not systemic RAS in Type 1 and 2 diabetes. However, the pattern of intra‐renal RAS component activation appears to differ between Type 1 and 2 diabetes.Support or Funding InformationUniversity of Utah Internal Medicine Academic Excellence Seed GrantThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Sensory Receptors Are Found in a Non‐sensory Organ: The Bladder
Authors: Smith Tilmira A. , Berkowitz Dan E. , Pluznick Jennifer L. 
Published: June 2021

Recent research has found that “sensory” receptors (including olfactory receptors (ORs), taste receptors (TRs), and opsins) play functional roles throughout the body. However, the expression of these receptors has not yet been carefully examined in the bladder. In this study, we utilized customized Taqman low density gene array cards to perform quantitative PCR and identify novel sensory receptors in mouse bladder tissue. We extracted RNA from murine bladder (2 male, 2 female), and then screened for the presence of sensory receptors using custom Taqman arrays which included probesets for 43 ORs, 35 bitter TRs, 3 TRs for umami/sweet, 5 opsins, and 7 G‐proteins and/or accessory proteins associated with sensory signaling pathways, as well as a Gapdh control. Using these arrays, we found the expression of 30 sensory transcripts in all male and female bladder samples, with 11 receptors being relatively well‐expressed (Ct values<~30). The 11 highly expressed receptors included 2 opsins (Opn1sw, Opn3), 3 ORs (Olfr558, Olfr99, Olfr78), 5 TRs (Tas1r3, Tas2r143, Tas2r135, Tas2r108, Tas2r126), and the G protein for olfaction (Gnal). We did not note any sex differences, and the top hit (Opn1sw) was the same for all four arrays. Notably, two of the ORs found in the bladder (Olfr78, Olfr558) are known to respond to bacterial metabolites, which may be relevant in the setting of cystitis/urinary tract infections (UTIs). In the future, determining the role of these novel bladder sensory receptors has the potential to enhance our understanding of bladder function, and may lead to novel treatments for conditions such as cystitis/UTIs.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 gene expression in adrenal glands of mice lacking the circadian clock protein PER1
Authors: Barral Dominique , Douma Lauren , Masten Sarah , Cheng Kit‐Yan , Gumz Michelle 
Published: June 2021

The circadian clock regulates a variety of physiological responses throughout a 24‐hour period. At a molecular level, the clock is governed by four core transcription factors, including PER1 and CRY, which are found in every cell in the human body. All reported circadian clock gene KO mice have a blood pressure (BP) phenotype. Aldosterone, a mineralocorticoid hormone and product of the renin‐angiotensin‐aldosterone system (RAAS), is secreted by the adrenal cortex and leads to increased sodium reabsorption by the kidneys in response to low BP or low blood volume. Previously we reported that 129/sv mice lacking the circadian clock gene Per1 had reduced BP and reduced plasma aldosterone levels associated with decreased expression of HSD3B1, a key enzyme in the steroid hormone synthesis pathway. Conversely, we found that male C57BL/6 Per1 KO mice have higher BP than control mice and develop non‐dipping hypertension (a lack of dip in BP during the rest period) on a high salt diet plus mineralocorticoid treatment. Non‐dipping hypertension has been associated with an increased risk for cardiovascular disease in humans. Interestingly, female Per1 KO mice do not develop non‐dipping hypertension on a high salt diet plus mineralocorticoid treatment. The goal of this study was to determine if expression of genes involved in the aldosterone synthesis pathway is altered in C57BL/6 Per1 KO mice. These deviances could explain the BP phenotype observed at the physiological level in male mice. We obtained adrenal glands from male and female WT and Per1 KO mice at noon (midpoint of the mouse rest phase), the same time at which serum was collected for aldosterone measurements. RNA was isolated from these tissues, converted to cDNA, and mRNA levels of genes involved in circadian rhythm and aldosterone synthesis were measured by quantitative real time PCR (qPCR). Preliminary data show that Cry1 is significantly down‐regulated in female Per1 KO mice by about 50% versus WT (n=5/6 per group, p<0.05). Expression of aldosterone synthesis genes CYP11B and HSD3B1 did not appear to be significantly altered in the adrenal tissue of male or female Per1 KO mice compared to WT. In contrast to what was observed with 129/sv Per1 KO, plasma aldosterone levels did not differ between C57BL/6 Per1 KO vs. WT. The observed difference in Cry1 expression suggests that RAAS and the circadian clock may work together in the maintenance of homeostatic BP. In the future we expect to perform similar experiments using male and female WT and Per1 KO mice adrenal tissue collected at a midnight time point in order to further investigate the communication occurring at the molecular level between RAAS and the circadian clock.Support or Funding InformationNIH/NIDDK 1R01DK109570; The Gatorade Trust through the UF Department of MedicineThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Effect of Angiotensin II on Chronic Renal Disease: Participation of NADPH Oxidase Enzymes, Endoplasmic Reticulum Stress and Tubular Na+ Transporters
Authors: Lins Bruna Bezerra , Casare Fernando Augusto Malavazzi , de Souza Maria Oliveira 
Published: June 2021

IntroductionChronic kidney disease (CKD) associated with hypertension is a serious disease of high incidence in the health system, and its progression may be associated with renal disorders induced by angiotensin II (Ang II). Thus, the objective of this study was to investigate the mechanisms by which Ang II participates in renal injury associated with CKD, considering the contribution of the NADPH oxidase enzymatic complex to the generation of reactive oxygen species (ROS) and the associated factors with of endoplasmic reticulum (ER) stress, as well as the changes in the tubular sodium (Na+) transporters expression.MethodsMale Wistar rats weighting between 150 and 200 g were treated with Ang II (200 ng/kg/min, by osmotic mini pump, model 2006, ALZET) for 42 days and/or losartan (10 mg/kg/day per gavage) for 14 days and organized into four groups: Control; Treated with Ang II; Treated with losartan and co‐treated with Ang II / losartan. During treatment, the tail blood pressure was evaluated. After treatment, the total renal tissue was used for gene expression analyzes by real‐time polymerase chain reaction (qPCR) and protein expression by western blotting. Immunohistochemistry and immunofluorescence staining were evaluated in the kidney cortex. Statistical analysis was assessed by one‐way ANOVA followed by the Bonferroni's test. Significance was assumed at p<0.05.ResultsThe results indicate that in Ang II chronic treatment induced significant increase in mRNA expression for Nox2 (p<0.01 vs CTL), Nox4 (p<0.01 vs CTL), p22phox (p<0.01 vs CTL), p47phox (p<0.001 vs CTL), Atf4 (p<0.05 vs CTL) and eIF2α (p<0.05 vs CTL), and protein expression of NOX2 (p<0.01 vs CTL), eIF2α (p<0.05 vs CTL) and PERK (p<0.001 vs CTL). Ang II also induced increased protein expression of Na+ exchangers NHE1 (p<0.01 vs CTL) and NHE3 (p<0.05 vs CTL), co‐transporters NKCC2 (p<0.001 vs CTL) and NCC (p<0.01 vs CTL), also Na+ epithelial channel ENaC α isoform in the medulla (p<0.001 vs CTL) and β isoform in cortex (p<0.01 vs CTL). Furthermore, the all stimulatory effects of Ang II were decreased by co‐treatment with losartan.Conclusionour results suggest that the chronic infusion of Ang II was able to stimulate the expression of mRNA to factors associated with ER stress and oxidative stress which may be related with increased expression of NHE1, NHE3, NKCC2, NCC and ENaC (α and β subunits).Support or Funding InformationCNPq (140141/2016‐2), CAPES, FAPESP.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Role of neutral amino acid transporter LAT4 in mouse epithelia
Authors: Rajendran Anuradha , Poncet Nadège , Oparija Lalita , Herzog Brigitte , Verrey François 
Published: June 2021

Amino acids (AAs) are not only building blocks of proteins, but also play key roles in energy metabolism and as (precursors of) neurotransmitters. To maintain amino acid homeostasis, small intestine (SI) and kidney play a major role by absorbing amino acids from food and preventing their loss in the urine, respectively. Given their zwitterionic nature, neutral AAs require transport proteins to cross cell membranes. LAT4 (Slc43a2), a facilitated diffusion uniporter specific for essential neutral amino acids (Leu, Ile, Val, Met and Phe) was first identified by Bodoy et al., 2005 and then shown by us to be highly expressed in the basolateral membranes of SI and kidney tubule epithelia. LAT4 global knockout caused a postnatal malnutrition‐like condition leading to death within 10 days, which was hypothesized to be due to a severe defect in intestinal absorption of AAs (Guetg et al. 2015). To test this hypothesis, we generated an intestine‐specific LAT4 knockout mouse model using Cre‐LoxP system. Intestine specific LAT4 knockout pups were viable and did not show any significant changes in body weight gain as compared to their control littermates even up to 2 months after birth. Quantitative PCR, western blot and immunofluorescence data showed a nearly 95% efficient knockout of LAT4 throughout the SI (n=4). However, these mice displayed no visible phenotype. Amino acid transport along the SI was then assessed by gavaging an AA mix containing radiolabelled LAT4 substrate 3H‐Leu. Blood samples were collected from the tail vein 10′ before and 2′,5′, 10′, 20′ and 60′ after gavage. Mice were then euthanized, SI divided into 4 equal parts and the intestinal tissue and its contents collected separately. Measuring the distribution of 3H‐Leu using a β‐counter revealed an accumulation of Leu in the initial segment of the SI of knockout mice (n=5, P<0.001 by ANOVA), accompanied by a delayed bolus movement. Thus, we show that LAT4 knockout in SI is not the sole or major reason for the lethal phenotype of LAT4 global knockout pups, although it could have contributed to the lethality. In parallel, we have generated a doxycycline‐inducible kidney tubule‐specific LAT4 knockout mouse model to assess the effects of LAT4 deletion in kidney, both in young and adult animals. Results obtained with this new model will allow us to determine whether the urinary loss of amino acids may have strongly contributed to the lethal phenotype of LAT4 global knockout mice or whether it was caused by the defect of LAT4 in other tissues.Support or Funding InformationThis project is funded by a grant of the Swiss National Foundation (SNF) to FV.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.