Polygenic genetic variation affecting antibody formation underlies hypertensive renal injury in the stroke-prone spontaneously hypertensive rat.

During development of the spontaneously hypertensive rat (SHR), several distinct but closely related lines were generated. Most lines are resistant to hypertensive renal disease. However, the SHR-A3 line (stroke-prone SHR) experiences end-organ injury (EOI) and provides a model of injury susceptibility that can be used to uncover genetic causation. In the present study, we generated a congenic line in which three distinct disease loci in SHR-A3 are concurrently replaced with homologous loci from an injury-resistant SHR line (SHR-B2). Verification that all three loci were homozygously replaced in this triple congenic line [SHR-A3(Trip B2)] while the genetic background of SHR-A3 was fully retained was obtained by whole genome sequencing. Congenic genome substitution was without effect on systolic blood pressure [198.9 ± 3.34 mmHg, mean ± SE, SHR-A3(Trip B2) = 194.7 ± 2.55 mmHg]. Measures of renal injury (albuminuria, histological injury scores, and urinary biomarker levels) were reduced in SHR-A3(Trip B2) animals, even though only 4.5 Mbases of the 2.8 Gbases of the SHR-B2 genome (0.16% of the genome) was transferred into the congenic line. The gene content of the three congenic loci and the functional effects of gene polymorphism within suggest a role of immunoglobulin in EOI pathogenesis. To prove the role of antibodies in EOI in SHR-A3, we generated an SHR-A3 line in which expression from the immunoglobulin heavy chain gene was knocked out (SHR-A3-IGHKO). Animals in the SHR-A3-IGHKO line lack B cells and immunoglobulin, but the hypertensive phenotype is not affected. Renal injury, however, was reduced in this line, confirming a pathogenic role for immunoglobulin in hypertensive EOI in this model of heritable risk.NEW & NOTEWORTHY Here, we used a polygenic animal model of hypertensive renal disease to show that genetic variation affecting antibody formation underlies hypertensive renal disease. We proved the genetic thesis by generating an immunoglobulin knockout in the susceptible animal model.

Adsorption of crystal violet on activated bamboo fiber powder from water: preparation, characterization, kinetics and isotherms.

Biomass-activated carbon has made a great contribution as an adsorbent in the field of dye wastewater treatment. In this study, the response surface method (RSM) based on the Box-Behnken design was used to optimize the preparation process. Bamboo fiber activated carbon (BAC) with a specific surface area of 2892 m2 g-1 and a pore volume of 1.80 cm3 g-1 was prepared. Various characterization methods (SEM, XPS, XRD, and Raman spectroscopy) were used to analyze the micro-structure of BAC. In the microscopic state, the BAC is fibrous and maintains the originally connected pores of the bamboo fiber. After high-temperature activation, the microcrystallinity of BAC decreases, and the degree of graphitization is low, indicating the presence of amorphous carbon. The adsorption capacity of BAC to crystal violet in simulated wastewater was evaluated via an adsorption experiment. Under the following conditions: the dosage of BAC was 0.04 g, the concentration was 600 mg L-1, the adsorption temperature and time were 25 °C and 30 min, respectively, and the as-prepared BAC had a 99.96% removal rate. The adsorption process conformed to the pseudo-second-order kinetic model and Langmuir adsorption isotherm model, indicating that the adsorption process of CV on BAC belonged to monomolecular layer adsorption. The adsorption process occurs spontaneously and is accompanied by heat release, and the maximum adsorption capacity of BAC within a given concentration range could reach 1353.09 mg g-1. SEM-EDS characterization before and after adsorption showed that ion exchange and the presence of oxygen-containing functional groups played an important role in promoting the adsorption process. The results show that BAC considerably affects CV removal, which has great application prospects.

The Assembled Genome of the Stroke-Prone Spontaneously Hypertensive Rat.

BACKGROUND: We report the creation and evaluation of a de novo assembly of the genome of the spontaneously hypertensive rat, the most widely used model of human cardiovascular disease. METHODS: The genome is assembled from long read sequencing (PacBio HiFi and continuous long read data [CLR]) and scaffolded with long-range structural information obtained from Bionano optical maps and proximity ligation sequencing proximity analysis of the genome. The genome assembly was polished with Illumina short reads. Completeness of the assembly was investigated using Benchmarking Universal Single Copy Orthologs analysis. The genome assembly was also evaluated with the rat reference gene set, using NCBI automated protocols. We also generated orthogonal single molecule transcript sequence reads (Iso-Seq) from 8 tissues and used them to validate the coding assembly, to annotate the assembly with RNA transcripts representing unique full length transcript isoforms for each gene and to determine whether divergences between RefSeq sequences and the assembly were attributable to assembly errors or polymorphisms. RESULTS: The assembly analysis indicates that this assembly is comparable in contiguity and completeness to the current rat reference assembly, while the use of HiFi sequencing yields an assembly that is more correct at the single base level. Synteny analysis was performed to uncover the extent of synteny and the presence and distribution of chromosomal rearrangements between the reference and this assembly. CONCLUSION: The resulting genome assembly is reference quality and captures significant structural variation.

Comparative Metabolomic Profiling Reveals Key Secondary Metabolites Associated with High Quality and Nutritional Value in Broad Bean (Vicia faba L.).

High quality and nutritional benefits are ultimately the desirable features that influence the commercial value and market share of broad bean (Vicia faba L.). Different cultivars vary greatly in taste, flavor, and nutrition. However, the molecular basis of these traits remains largely unknown. Here, the grain metabolites of the superior Chinese landrace Cixidabaican (CX) were detected by a widely targeted metabolomics approach and compared with the main cultivar Lingxiyicun (LX) from Japan. The analyses of global metabolic variations revealed a total of 149 differentially abundant metabolites (DAMs) were identified between these two genotypes. Among them, 84 and 65 were up- and down-regulated in CX compared with LX. Most of the DAMs were closely related to healthy eating substances known for their antioxidant and anti-cancer properties, and some others were involved in the taste formation. The KEGG-based classification further revealed that these DAMs were significantly enriched in 21 metabolic pathways, particularly in flavone and flavonol biosynthesis. The differences in key secondary metabolites, including flavonoids, terpenoids, amino acid derivates, and alkaloids, may lead to more nutritional value in a healthy diet and better adaptability for the seed germination of CX. The present results provide important insights into the taste/quality-forming mechanisms and contributes to the conservation and utilization of germplasm resources for breeding broad bean with superior eating quality.

Generation and characterization of coal-based needle coke produced by the co-carbonization of coal liquefaction pitch and anthracene oil.

In this study, we studied the feasibility of preparing high-quality needle coke from coal liquefaction pitch. Nine types of blending pitch (coal liquefaction pitch and anthracene oil mixed with different ratios) were used as raw materials to generate needle coke via the co-carbonization method. Optical microscopy, X-ray diffraction, Raman spectroscopy and scanning electron microscopy were employed to determine the properties (microstructure, distribution of carbon microcrystals, true density and micro-strength) of the needle coke derived by the co-carbonization method. Actually, the co-carbonization of coal liquefaction pitch and anthracene oil was an essential method to control the micro-structure and property of the derived needle coke. Briefly, the needle coke derived by the co-carbonization of coal liquefaction pitch and anthracene oil had a lower content of mosaic structure of 14.17%, ideal carbon crystal content of 82.67%, and true density of 2.296 g cm-3. Briefly, the addition of anthracene oil is a suitable method to adjust the property of coal liquefaction pitch, which is also a good method to produce high-quality needle coke via the co-carbonization of coal liquefaction pitch and anthracene oil. Thus, the use of coal liquefaction pitch and anthracene oil as raw materials to generate high-quality needle coke is a considerable method to realize the clean and high value-added utilization of coal liquefaction pitch.

Preparation and Characterization of Mesocarbon Microbeads by the Co-Polycondensation of High-Temperature Coal Tar Pitch and Coal Pyrolytic Extracts.

Mesocarbon microbeads (MCMBs) are a kind of engineering and functional artificial carbon materials generally prepared by the polymerization of polycyclic aromatic hydrocarbons. The physicochemical property of the raw materials plays a key role in the quality of MCMBs. For a detailed analysis of the synergistic effects of the generation of MCMBs, a high-temperature coal tar pitch was used as raw materials, and coal pyrolytic extracts were used as additive to synthesize the MCMBs. The microstructure and morphology of the derived MCMBs were determined by an optical microscope, scanning electron microscope, X-ray diffraction, Raman spectrum, and laser particle size analyzer. In fact, the addition of the coal pyrolytic extracts can adjust the molecular structure of the blending pitch, and the coal pyrolytic extracts can promote the generation of the MCMBs during the co-polycondensation process. The MCMBs obtained by co-polycondensation method have a good degree of sphericity, lower defects in the surface morphology, and a lower charge transfer resistance (Rct) of 4.677 Ω.

mRatBN7.2: familiar and unfamiliar features of a new rat genome reference assembly.

Rat genomic tools have been slower to emerge than for those of humans and mice and have remained less thorough and comprehensive. The arrival of a new and improved rat reference genome, mRatBN7.2, in late 2020 is a welcome event. This assembly, like predecessor rat reference assemblies, is derived from an inbred Brown Norway rat. In this "user" survey we hope to provide other users of this assembly some insight into its characteristics and some assessment of its improvements as well as a few caveats that arise from the unique aspects of this assembly. mRatBN7.2 was generated by the Wellcome Sanger Institute as part of the large Vertebrate Genomes Project. This rat assembly has now joined human, mouse, chicken, and zebrafish in the National Center for Biotechnology Information (NCBI)'s Genome Reference Consortium, which provides ongoing curation of the assembly. Here we examine the technical procedures by which the assembly was created and assess how this assembly constitutes an improvement over its predecessor. We also indicate the technical limitations affecting the assembly, providing illustrations of how these limitations arise and the impact that results for this reference assembly.

Differences in Grain Microstructure and Proteomics of a Broad Bean (Vicia faba L.) Landrace Cixidabaican in China Compared with Lingxiyicun Introduced from Japan.

In response to the germplasm resources' conservation in China, the characters of a superior landrace of broad bean (Vicia faba L.) Cixidabaican (CX) were identified, compared with Lixiyicun (LX) introduced from Japan. The plant morphology and root structure of CX were larger, pods/seeds number and yield per plant were higher, but the size of pods/seeds and single-seed weight were lower than the similar characteristics in LX. The protein content of dry seeds of CX was 4.1% lower than LX, while the amino acids contents showed no difference between the two cultivars. The seed scan electron micrograph showed that the structure of starch granules was similar, while the granules number was lower in CX than LX. iTRAQ-based proteomics showed that 80 differentially abundant proteins (DAPs) were higher, and 45 DAPs were less abundant in the seeds of CX compared to LX, and DAPs were enriched in proteins of carbohydrate and amino acid metabolism. These results verified the importance of the further study of landraces by showing superior traits of CX, which could contribute to the breeding of better-quality varieties.

On-off-on nanosensors of carbon quantum dots derived from coal tar pitch for the detection of Cu2+, Fe3+, and L-ascorbic acid.

Novel nitrogen-doped carbon quantum dots (N-CQDs) were synthesized by a chemical oxidation method using medium-low temperature coal tar pitch as the raw material. Such quantum dots were developed as a highly sensitive fluorescent "on-off-on" switch sensor for the selective and simultaneous sensing of Cu2+ and Fe3+. The as-prepared N-CQDs, which emit blue light, were characterized by TEM images, FT-IR spectra, Raman spectroscopy, XPS analysis, fluorescence spectra, and UV-vis absorption spectra. The results showed that the N-CQDs exhibit outstanding optical properties and high optical stability within the pH range of 4-10, with a quantum yield of approximately 7%. Additionally, the material performed as an "on-off" sensor which can be dramatically extinguished by Cu2+ and Fe3+. A linear relationship between Cu2+ and Fe3+ ion concentration and fluorescence intensity was observed in the range from 0 to 50 µM. The limits of detection of the fluorescent sensor toward Cu2+ and Fe3+ were 0.16 µM and 0.173 µM, respectively. The Fe3+-quenched N-CQDs were restored after adding L-ascorbic acid, due to the redox reaction between Fe3+ and L-ascorbic acid, which resulted in the detachment of Fe3+ from the surface of the N-CQDs. The linear range for the detection of L-ascorbic acid was 0-28 µM. Therefore, the amount of L-ascorbic acid can be measured by using the sensing system consisting of Fe3+ and N-CQDs. In consequence, N-CQDs are considered an important material for the detection of Cu2+ and Fe3+ in water samples or L-ascorbic acid in drugs.

Trace metals dramatically boost oxygen electrocatalysis of N-doped coal-derived carbon for zinc-air batteries.

The commercialization of metal-air batteries requires efficient, low-cost, and stable bifunctional electrocatalysts for reversible electrocatalysis of the oxygen reduction reaction (ORR) and the oxygen evolution reaction (OER). The modification of natural coal by heteroatoms such as N and S, or metal oxide species, has been demonstrated to form very promising electrocatalysts for the ORR and OER. However, it remains elusive and underexplored as to how the impurity elements in coal may impact the electrocatalytic properties of coal-derived catalysts. Herein, we explore the influence of the presence of various trace metals that are notable impurities in coal, including Al, Si, Ca, K, Fe, Mg, Co, Mn, Ni, and Cu, on the electrochemical performance of the prepared catalysts. The constructed Zn-air batteries are further shown to be able to power green LED lights for more than 80 h. The charge-discharge polarization curves exhibited excellent and durable rechargeability over 500 (ca. 84 h) continuous cycles. The promotional effect of the trace elements is believed to accrue from a combination of electronic structure modification of the active sites, enhancement of the active site density, and formation of a conductive 3-dimensional hierarchical network of carbon nanotubes.

Natural genetic variation in Stim1 creates stroke in the spontaneously hypertensive rat.

Similar to humans, the risk of cerebrovascular disease in stroke-prone spontaneously hypertensive rats (SHR-A3/SHRSP) arises from naturally occurring genetic variation. In the present study, we show the involvement of genetic variation affecting the store-operated calcium signaling gene, Stim1, in the pathogenesis of stroke in SHR. Stim1 is a key lymphocyte activation signaling molecule and contains functional variation in SHR-A3 that diverges from stroke-resistant SHR-B2. We created a SHR-A3 congenic line in which Stim1 was substituted with the corresponding genomic segment from SHR-B2. Compared with SHR-A3 rats, Stim1 congenic SHR-A3 (SHR-A3(Stim1-B2)) have reduced cerebrovascular disease in response to salt loading including lower neurological deficit scores and cerebral edema. Microbleeds and major hemorrhages occurred in over half of SHR-A3 rats. These lesions were absent in SHR-A3(Stim1-B2) rats. Loss of Stim1 function in mice and humans is associated with antibody-mediated autoimmunity due to defects in T lymphocyte helper function to B cells. We investigated autoantibody formation using a high-density protein array to detect the presence of IgG and IgM autoantibodies in SHR-A3. Autoantibodies to key cerebrovascular stress proteins were detected that were reduced in the congenic line.

Stim1 Polymorphism Disrupts Immune Signaling and Creates Renal Injury in Hypertension.

Background Spontaneously hypertensive rats of the stroke-prone line (SHR-A3) develop hypertensive renal disease as a result of naturally occurring genetic variation. Our prior work identified a single-nucleotide polymorphism unique to SHR-A3 that results in truncation of the carboxy terminus of STIM1. The SHR-B2 line, which is also hypertensive but resists hypertensive renal injury, expresses the wild-type STIM1. STIM1 plays a central role in lymphocyte calcium signaling that directs immune effector responses. Here we show that major defects in lymphocyte function affecting calcium signaling, nuclear factor of activated T cells activation, cytokine production, proliferation, apoptosis, and regulatory T-cell development are present in SHR-A3 and attributable to STIM1. Methods and Results To assess the role of Stim1 variation in susceptibility to hypertensive renal injury, we created a Stim1 congenic line, SHR-A3(Stim1-B2), and STIM1 function was rescued in SHR-A3. We found that Stim1 gene rescue restores disturbed lymphocyte function in SHR-A3. Hypertensive renal injury was compared in SHR-A3 and the SHR-A3(Stim1-B2) congenic line. Histologically assessed renal injury was markedly reduced in SHR-A3(Stim1-B2), as were renal injury biomarker levels measured in urine. Stim1 deficiency has been linked to the emergence of antibody-mediated autoimmunity. Renal glomerular immunoglobulin deposition was greater in SHR-A3 than SHR-B2 and was reduced by Stim1 congenic substitution. Serum anti-double-stranded DNA antibody titers in SHR-A3 were elevated compared with SHR-B2 and were reduced in SHR-A3(Stim1-B2). Conclusions Stim1 deficiency in lymphocyte function originating from Stim1 truncation in SHR-A3 combines with hypertension to create end organ disease and may do so as a result of antibody formation.

Germ-line genetic variation in the immunoglobulin heavy chain creates stroke susceptibility in the spontaneously hypertensive rat.

The risk of cerebrovascular disease in stroke-prone spontaneously hypertensive rats (SHR-A3/SHRSP) arises from naturally occurring genetic variation. In the present study we show the involvement of SHR genetic variation that affects antibody formation and function in the pathogenesis of stroke. We have tested the involvement in susceptibility to stroke of genetic variation in IgH, the gene encoding the immunoglobulin heavy chain by congenic substitution. This gene contains functional natural variation in SHR-A3 that diverges from stroke-resistant SHR-B2. We created a SHR-A3 congenic line in which the IgH gene was substituted with the corresponding haplotype from SHR-B2. Compared with SHR-A3 rats, congenic substitution of the IgH locus [SHR-A3(IgH-B2)] markedly reduced cerebrovascular disease. Given the role in antibody formation of the IgH gene, we investigated the presence of IgG and IgM autoantibodies and their targets using a high-density protein array containing ~20,000 recombinant proteins. High titers of autoantibodies to key cerebrovascular stress proteins were detected, including FABP4, HSP70, and Wnt signaling proteins. Serum levels of these autoantibodies were reduced in the SHR-A3(IgH-B2) congenic line.

Genotypic differences in leaf secondary metabolism, plant hormones and yield under alone and combined stress of drought and salinity in cotton genotypes.

Drought and salinity stress highly affect the plant growth and production around the world. Secondary metabolites play a main role in adaptation to the environment and in overcoming stress conditions. In order to investigate the effect of drought and salinity, alone or in combination, on secondary metabolism-related enzyme activities, plant hormones and yield parameters, a greenhouse pot experiment was conducted using two cotton genotypes Zhongmian 23 (salt tolerant) and Zhongmian 41 (salt sensitive). Results showed that single and combined drought and salinity stresses caused remarkable decrease in plant height, bolls and lint yield in the order as follows: D + S > salinity > drought, and Zhongmian 41 > Zhongmian 23. Lower H2 O2 and superoxide but higher proline content and secondary metabolism-related enzyme activities were observed in Zhongmian 23 under drought and salinity, both alone and combined, compared with control in Zhongmian 41. Our findings suggest that controlling reactive oxygen species (ROS) levels and increasing activities of secondary metabolism-related enzymes in Zhongmian 23 might be an effective mechanism to reduce the negative effects of drought and salinity stress. However, cinnamyl alcohol dehydrogenase (CAD), and shikimate dehydrogenase (SKDH) activities were markedly decreased in Zhongmian 41 under salinity stress alone as compared with control. Meanwhile, Zhongmian 23 had higher expression levels of genes related to secondary metabolism (c.f. phenylalanine ammonia-lyase, PAL; polyphenol oxidase, PPO and CAD) under the three stresses compared to Zhongmian 41. The content of flavonoids and phenols were significantly enhanced under drought and D + S, with higher accumulation in Zhongmian 23. Phenols content in Zhongmian 23 remained unchanged under salinity as relative to control, but were significantly reduced in Zhongmian 41. In addition, callose content, chitinase activities and abscisic acid (ABA) and Indole-3-acetic acid (IAA) were more induced in Zhongmian 23 under drought, salinity and D + S, than in Zhongmian 41. Our results suggest that high tolerance to D + S stress in Zhongmian 23 is closely related to elevated callose, chitinase, flavonoids and phenols contents and higher secondary metabolism-related enzyme activities and their transcript levels.

Susceptibility to Hypertensive Renal Disease in the Spontaneously Hypertensive Rat Is Influenced by 2 Loci Affecting Blood Pressure and Immunoglobulin Repertoire.

High blood pressure exerts its deleterious effects on health largely through acceleration of end-organ diseases. Among these, progressive loss of renal function is particularly important, not only for the direct consequences of kidney damage but also because loss of renal function is associated with amplification of other adverse cardiovascular outcomes. Genetic susceptibility to hypertension and associated end-organ disease is non-Mendelian in both humans and in a rodent model, the spontaneously hypertensive rat (SHR). Here, we report that hypertensive end-organ disease in the inbred SHR-A3 line is attributable to genetic variation in the immunoglobulin heavy chain on chromosome 6. This variation coexists with variation in a 10 Mb block on chromosome 17 that contains genetic variation in 2 genes involved in immunoglobulin Fc receptor signaling. Substitution of these genomic regions into the SHR-A3 genome from the closely related, but injury-resistant, SHR-B2 line normalizes both biomarker and histological measures of renal injury. Our findings indicate that genetic variation leads to a contribution by immune mechanisms hypertensive end-organ injury and that, in this rat model, disease is influenced by differences in germ line antibody repertoire.

Red blood cell ß-adrenergic receptors contribute to diet-induced energy expenditure by increasing O2 supply.

Diet-induced obesity (DIO) represents the major cause for the current obesity epidemic, but the mechanism underlying DIO is unclear. ß-Adrenergic receptors (ß-ARs) play a major role in sympathetic nervous system-mediated (SNS-mediated) diet-induced energy expenditure (EE). Rbc express abundant ß-ARs; however, a potential role for rbc in DIO remains untested. Here, we demonstrated that high-fat, high-caloric diet (HFD) feeding increased both EE and blood O2 content, and the HFD-induced increases in blood O2 level and in body weight gain were negatively correlated. Deficiency of ß-ARs in rbc reduced glycolysis and ATP levels, diminished HFD-induced increases in both blood O2 content and EE, and resulted in DIO. Importantly, specific activation of cAMP signaling in rbc promoted HFD-induced EE and reduced HFD-induced tissue hypoxia independent of obesity. Both HFD and pharmacological activation cAMP signaling in rbc led to increased glycolysis and ATP levels. These results identify a previously unknown role for rbc ß-ARs in mediating the SNS action on HFD-induced EE by increasing O2 supply, and they demonstrate that HFD-induced EE is limited by blood O2 availability and can be augenmented by increased O2 supply.

Mycophenolate mofetil prevents cerebrovascular injury in stroke-prone spontaneously hypertensive rats.

Stroke-prone spontaneously hypertensive rats (SHR-A3) develop strokes and progressive kidney disease as a result of naturally occurring genetic variations. We recently identified genetic variants in immune signaling pathways that contribute to end-organ injury. The present study was designed to test the hypothesis that a dysregulated immune response promotes stroke susceptibility. We salt-loaded 20 wk old male SHR-A3 rats and treated them with the immunosuppressant mycophenolate mofetil (MMF, 25 mg/kg/day po) (n = 8) or vehicle (saline) (n = 9) for 8 wk. Blood pressure (BP) was measured weekly by telemetry. Compared with vehicle-treated controls, MMF-treated SHR-A3 rats had improved survival and lower neurological deficit scores (1.44 vs. 0.125; P < 0.02). Gross morphology of the brain revealed cerebral edema in 8 of 9, and microbleeds and hemorrhages in 5 of 9 vehicle-treated rats. These lesions were absent in MMF-treated rats. Brain CD68 expression, indicating macrophage/microglial activation, was upregulated in vehicle-treated rats with microbleeds and hemorrhages but was undetectable in the brains of MMF-treated rats. MMF also prevented renal injury in SHR-A3 rats, evidenced by reduced proteinuria (albumin:creatinine) from 7.52 to 1.05 mg/mg (P < 0.03) and lower tubulointerstitial injury scores (2.46 vs. 1.43; P < 0.01). Salt loading resulted in a progressive increase in BP, which was blunted in rats receiving MMF. Our findings provide evidence that abnormal immune activation predisposes to cerebrovascular and renal injury in stroke-prone SHR-A3 rats.

Defective Store-Operated Calcium Entry Causes Partial Nephrogenic Diabetes Insipidus.

Store-operated calcium entry (SOCE) is the mechanism by which extracellular signals elicit prolonged intracellular calcium elevation to drive changes in fundamental cellular processes. Here, we investigated the role of SOCE in the regulation of renal water reabsorption, using the inbred rat strain SHR-A3 as an animal model with disrupted SOCE. We found that SHR-A3, but not SHR-B2, have a novel truncating mutation in the gene encoding stromal interaction molecule 1 (STIM1), the endoplasmic reticulum calcium (Ca(2+)) sensor that triggers SOCE. Balance studies revealed increased urine volume, hypertonic plasma, polydipsia, and impaired urinary concentrating ability accompanied by elevated circulating arginine vasopressin (AVP) levels in SHR-A3 compared with SHR-B2. Isolated, split-open collecting ducts (CD) from SHR-A3 displayed decreased basal intracellular Ca(2+) levels and a major defect in SOCE. Consequently, AVP failed to induce the sustained intracellular Ca(2+) mobilization that requires SOCE in CD cells from SHR-A3. This effect decreased the abundance of aquaporin 2 and enhanced its intracellular retention, suggesting impaired sensitivity of the CD to AVP in SHR-A3. Stim1 knockdown in cultured mpkCCDc14 cells reduced SOCE and basal intracellular Ca(2+) levels and prevented AVP-induced translocation of aquaporin 2, further suggesting the effects in SHR-A3 result from the expression of truncated STIM1. Overall, these results identify a novel mechanism of nephrogenic diabetes insipidus and uncover a role of SOCE in renal water handling.

Hypertensive renal injury is associated with gene variation affecting immune signaling.

BACKGROUND: The spontaneously hypertensive rat (SHR) strain exists in lines that contrast strongly in susceptibility to renal injury in hypertension. These inbred lines share common ancestry, and only 13% of their genomes arise from different ancestors. METHODS AND RESULTS: We used next gen sequencing to detect natural allelic variation in 5 genes of the immunoreceptor signaling pathway (IgH, Dok3, Src, Syk, and JunD) that arise from different ancestors in the injury-prone SHR-A3 and the resistant SHR-B2 lines. We created an intercross between these lines, and in the F2 progeny, we observed that the inheritance of haplotype blocks containing the SHR-A3 alleles of these 5 genes correlated with increased albuminuria and histological measures of renal injury. To test whether accumulated genetic variation in this pathway may create a therapeutic target in hypertensive renal injury, rats of both lines were treated with the immunosuppressant mycophenolate mofetil (MMF). MMF reduced proteinuria (albumin to creatinine ratio) from 6.6 to 1.2 mg/mg (P<0.001) in SHR-A3. Glomerular injury scores were reduced in MMF-treated SHR-A3 from 1.6 to 1.4 (P<0.002). Tubulo-interstitial injury was reduced in MMF-treated SHR-A3 from 2.62 to 2.0 (P=0.001). MMF treatment also reduced renal fibrosis in SHR-A3 (3.9 versus 2.0; P<0.001). CONCLUSIONS: Polygenic susceptibility to renal injury in hypertension arises in association with genetic variation in genes that participate in immune responses and is dramatically improved by reduction of immune system activity.

Glutamate mediates the function of melanocortin receptor 4 on Sim1 neurons in body weight regulation.

The melanocortin receptor 4 (MC4R) is a well-established mediator of body weight homeostasis. However, the neurotransmitter(s) that mediate MC4R function remain largely unknown; as a result, little is known about the second-order neurons of the MC4R neural pathway. Single-minded 1 (Sim1)-expressing brain regions, which include the paraventricular nucleus of hypothalamus (PVH), represent key brain sites that mediate melanocortin action. We conditionally restored MC4R expression in Sim1 neurons in the background of Mc4r-null mice. The restoration dramatically reduced obesity in Mc4r-null mice. The anti-obesity effect was completely reversed by selective disruption of glutamate release from those same Sim1 neurons. The reversal was caused by lower energy expenditure and hyperphagia. Corroboratively, selective disruption of glutamate release from adult PVH neurons led to rapid obesity development via reduced energy expenditure and hyperphagia. Thus, this study establishes glutamate as the primary neurotransmitter that mediates MC4Rs on Sim1 neurons in body weight regulation.