bluuebunny/crossref_metadata_2025 huggingface dataset:
Angiotensin peptides acting at rostral ventrolateral medulla contribute to hypertension of TGR(mREN2)27 rats
Authors: FONTES MARCO A. P. , BALTATU OVIDIU , CALIGIORNE SORDAINI M. , CAMPAGNOLE-SANTOS MARIA J. , GANTEN DETLEV , BADER MICHAEL , SANTOS ROBSON A. S.
Published: December 2017

Fontes, Marco A. P., Ovidiu Baltatu, Sordaini M. Caligiorne, Maria J. Campagnole-Santos, Detlev Ganten, Michael Bader, and Robson A. S. Santos. Angiotensin peptides acting at rostral ventrolateral medulla contribute to hypertension of TGR(mREN2)27 rats. Physiol Genomics 2: 137–142, 2000.—We have previously demonstrated that microinjections of the selective angiotensin-( 1 – 7 ) [ANG-( 1 – 7 )] antagonist, A-779, into the rostral ventrolateral medulla (RVLM) produces a significant fall in mean arterial pressure (MAP) and heart rate (HR) in both anesthetized and conscious rats. In contrast, microinjection of angiotensin II (ANG II) AT1 receptor antagonists did not change MAP in anesthetized rats and produced dose-dependent increases in MAP when microinjected into the RVLM of conscious rats. In the present study, we evaluated whether endogenous ANG-( 1 – 7 ) and ANG II acting at the RVLM contribute to the hypertension of transgenic rats harboring the mouse renin Ren-2 gene, TGR(mREN2)27. Unilateral microinjection of A-779 (0.1 nmol) produced a significant fall in MAP (−25 ± 5 mmHg) and HR (−57 ± 20 beats/min) of awake TGR rats. The hypotensive effect was greater than that observed in Sprague-Dawley (SD) rats (−9 ± 2 mmHg). Microinjection of the AT1 antagonist CV-11974 (0.2 nmol) produced a fall in MAP in TGR rats (−14 ± 4 mmHg), contrasting with the pressor effect observed in SD rats (33 ± 9 mmHg). These results indicate that endogenous ANG-( 1 – 7 ) exerts a significant pressor action in the RVLM, contributing to the hypertension of TGR(mREN2)27 transgenic rats. The role of ANG II at the RVLM seems to be dependent on its endogenous level in this area.
Diet effects on weight gain and body composition in high growth (hg/hg) mice
Authors: CORVA PABLO M. , MEDRANO JUAN F.
Published: December 2017

Corva, Pablo M., and Juan F. Medrano. Diet effects on weight gain and body composition in high growth ( hg/ hg) mice. Physiol Genomics 3: 17–23, 2000.—Nongenetic factors such as nutrition modulate the effects of genes responsible for overgrowth in animals. The goal of this study was to examine the importance of genotype × diet interactions on the effects of a major locus that regulates growth in the mouse. We have examined the phenotype of high growth ( hg), a partially recessive autosomal locus that increases growth rate and mature body size. C57BL/6J (C57) and congenic C57BL/6J- hg/ hg (HG) mice were fed three experimental diets differing in protein and energy content from 3 to 12 wk of age. HG mice grew faster and were, on average, 51% heavier than C57 at 12 wk of age. Feed intake was higher in HG mice but proportional to the increase in body weight. The magnitude of the differences in body size and composition between lines depended on the interaction between genotype and the protein/energy ratio of the diet. In C57, the diets modified the level of fatness without changing adult lean mass. However, in HG the diets differentially affected both linear growth and body composition. In general, HG had higher plasma levels of insulin-like growth factor I at 3 and 12 wk than C57. Plasma insulin did not differ between lines, but leptin was higher for C57 mice fed a high-energy diet. These results show that the effects of hg on growth are modulated by diet composition. Therefore, this mutation could be a valuable model with which to study the genetic and nutritional aspects of overgrowth disorders.
Global expression profiling of yeast treated with an inhibitor of amino acid biosynthesis, sulfometuron methyl
Authors: JIA MELISSA H. , LAROSSA ROBERT A. , LEE JIAN-MING , RAFALSKI ANTONI , DEROSE ELLEN , GONYE GREG , XUE ZHIXIONG
Published: December 2017

Jia, Melissa H., Robert A. LaRossa, Jian-Ming Lee, Antoni Rafalski, Ellen DeRose, Greg Gonye, and Zhixiong Xue. Global expression profiling of yeast treated with an inhibitor of amino acid biosynthesis, sulfometuron methyl. Physiol Genomics 3: 83–92, 2000.—The expression pattern of 1,529 yeast genes in response to sulfometuron methyl (SM) was analyzed by DNA microarray technology. SM, a potent herbicide, inhibits acetolactate synthase, a branched-chain amino acid biosynthetic enzyme. Exposure of yeast cells to 0.2 μg/ml SM resulted in 40% growth inhibition, a Gcn4p-mediated induction of genes involved in amino acid and cofactor biosynthesis, and starvation response. The accumulation of intermediates led to the induction of stress response genes and the repression of genes involved in carbohydrate metabolism, nucleotide biosynthesis, and sulfur assimilation. Extended exposure to SM led to a relaxation of the initial response and induction of sugar transporter and ergosterol biosynthetic genes, as well as repression of histone and lipid metabolic genes. Exposure to 5 μg/ml SM resulted in >98% growth inhibition and stimulated a similar initial expression change, but with no relaxation after extended exposure. Instead, more stress response and DNA damage repair genes become induced, suggesting a serious cellular consequence. Other salient features of metabolic regulation, such as the coordinated expression of cofactor biosynthetic genes with amino acid biosynthetic ones, were evident from our data. A potential link between SM sensitivity and ergosterol metabolism was uncovered by expression profiling and confirmed by genetic analysis.
A novel, sensitive detection system for high-density microarrays using dendrimer technology
Authors: STEARS ROBIN L. , GETTS ROBERT C. , GULLANS STEVEN R.
Published: December 2017

Stears, Robin L., Robert C. Getts, and Steven R. Gullans. A novel, sensitive detection system for high-density microarrays using dendrimer technology. Physiol Genomics 3: 93–99, 2000.—To improve signal detection on cDNA microarrays, we adapted a fluorescent oligonucleotide dendrimeric signal amplification system to microarray technology. This signal detection method requires 16-fold less RNA for probe synthesis, does not depend on the incorporation of fluorescent dNTPs into a reverse transcription reaction, generates a high signal-to-background ratio, and can be used to allow for multichannel detection on a single chip. Furthermore, since the dendrimers can be detected individually, it may be possible, by employing dendrimer-binding standards, to calculate the numbers of bound cDNAs can be estimated. These features make the dendrimer signal detection reagent ideal for high-throughput functional genomics research.
Gene expression in hypothalamus and brown adipose tissue of mice divergently selected for heat loss
Authors: ALLAN MARK F. , NIELSEN MERLYN K. , POMP DANIEL
Published: December 2017

Allan, Mark F., Merlyn K. Nielsen, and Daniel Pomp. Gene expression in hypothalamus and brown adipose tissue of mice divergently selected for heat loss. Physiol Genomics 3: 149–156, 2000.—Gene expression was evaluated in mice divergently selected for 16 generations for heat loss, measured by direct calorimetry. The high (MH) heat loss line has ∼50% greater heat loss, ∼35% less body fat, ∼20% greater feed intake, and twofold greater activity levels than the low (ML) heat loss line. At 11 wk, inbred males (developed from MH and ML) were euthanized 3 h after dark for dissection of tissues and extraction of RNA. Differential display PCR (DD-PCR) was used to evaluate transcriptional differences between lines in hypothalamus and brown adipose tissue (BAT). Evaluation was replicated within and across lines, using family pools of mRNA. Two genes were confirmed by competitive RT-PCR and/or Northern analysis to have greater levels of mRNA present in ML relative to MH mice. In both hypothalamus and BAT, the ribosomal protein L3 ( RPL3) gene was expressed at higher levels in ML, whereas an unknown expressed sequence tag (EST) was also found at higher levels in the hypothalamus of ML mice. These results implicate RPL3 in regulation of energy balance and extend the genetic dissection of response to selection to the transcriptional level.
Analysis of mucosal gene expression in inflammatory bowel disease by parallel oligonucleotide arrays
Authors: DIECKGRAEFE B. K. , STENSON W. F. , KORZENIK J. R. , SWANSON P. E. , HARRINGTON C. A.
Published: December 2017

DNA arrays capable of simultaneously measuring expression of thousands of genes in clinical specimens from affected and normal individuals have the potential to provide information about disease pathogenesis not previously possible. Few studies have applied mRNA profiling to diseases involving complex tissues like the intestinal mucosa, reflecting the unique challenges inherent to this type of analysis. We report the analysis of mucosal gene expression in ulcerative colitis (UC) patients and inflamed and noninflamed control specimens. Genes can be used as markers for cell recruitment, activation, and mucosal synthesis of immunoregulatory molecules. Self-organizing maps were applied to cluster and analyze gene expression patterns and were paired with histopathological scores to identify genes associated with increased disease activity. Clustering was achieved on the basis of differences in expression levels across individual specimens. Several inflammatory mediators were identified as likely determinants of characteristic histological features of active UC. These results provide proof of principle for application of functional genomics to larger inflammatory bowel disease populations for gene discovery, to facilitate identification of disease subgroups on the basis of gene expression signatures, and for prediction of disease behavior or optimal therapeutic intervention.
Functional properties of skeletal muscle from transgenic animals with upregulated heat shock protein 70
Authors: NOSEK THOMAS M. , BROTTO MARCO A. P. , ESSIG DAVID A. , MESTRIL RUBEN , CONOVER RICHARD C. , DILLMANN WOLFGANG H. , KOLBECK RALPH C.
Published: December 2017

The influence of inducible heat stress proteins on protecting contracting skeletal muscle against fatigue-induced injury was investigated. A line of transgenic mice overexpressing the inducible form of the 72-kDa heat shock protein (HSP72) in skeletal muscles was used. We examined the relationship between muscle contractility and levels of the constitutive (HSC73) and inducible (HSP72) forms of the 72-kDa heat shock protein in intact, mouse extensor digitorum longus (EDL), soleus (SOL), and the diaphragm (DPH). In all transgenic muscles, HSP72 was expressed at higher levels compared with transgene-negative controls, where HSP72 was below the level of detection. At the same time, HSC73 levels were downregulated in all transgenic muscle types. Shipment-related stress caused an elevation in the levels of HSP72 in all muscles for 1 wk after arrival of the animals. We also found that, although no statistical differences in response to intermittent fatiguing stimulation in the contractile properties of intact transgene-positive muscles compared with their transgene-negative counterparts were observed, the response of intact transgene-positive EDL muscles to caffeine was enhanced. These findings demonstrate that elevated HSP72 does not protect EDL, SOL, or DPH muscles from the effects of intermittent fatiguing stimulation. However, HSP72 may influence the excitation-contraction coupling (ECC) process, either directly or indirectly, in EDL muscle. If the effects on ECC were indirect, then these results would suggest that manipulation of a specific gene might cause functional effects that seem independent of the manipulated gene/protein.
Abrogation of thrombin-induced increase in pulmonary microvascular permeability in PAR-1 knockout mice
Authors: VOGEL STEPHEN M. , GAO XIAOPEI , MEHTA DOLLY , YE RICHARD D. , JOHN THERESA A. , ANDRADE-GORDON PATRICIA , TIRUPPATHI CHINNASWAMY , MALIK ASRAR B.
Published: December 2017

We investigated the function of proteinase-activated receptor-1 (PAR-1) in the regulation of pulmonary microvascular permeability in response to thrombin challenge using PAR-1 knockout mice (−/−). Lungs were isolated and perfused with albumin (5 g/100 ml)-Krebs solution at constant flow (2 ml/min). Lung wet weight and pulmonary artery pressure (Ppa) were continuously monitored. We determined the capillary filtration coefficient ( Kfc) and125I-labeled albumin (BSA) permeability-surface area product (PS) to assess changes in pulmonary microvessel permeability to liquid and albumin, respectively. Normal and PAR-1-null lung preparations received in the perfusate: 1) thrombin or 2) selective PAR-1 agonist peptide (TFLLRNPNDK-NH2). In control PAR-1 (+/+) mouse lungs,125I-albumin PS and Kfcwere significantly increased over baseline (by ∼7- and 1.5-fold, respectively) within 20 min of α-thrombin (100 nM) challenge. PAR-1 agonist peptide (5 μM) gave similar results, whereas control peptide (5 μM; FTLLRNPNDK-NH2) was ineffective. At relatively high concentrations, thrombin (500 nM) or PAR-1 agonist peptide (10 μM) also induced increases in Ppaand lung wet weight. All effects of thrombin (100 or 500 nM) or PAR-1 agonist peptide (5 or 10 μM) were prevented in PAR-1-null lung preparations. Baseline measures of microvessel permeability and Ppain the PAR-1-null preparations were indistinguishable from those in normal lungs. Moreover, PAR-1-null preparations gave normal vasoconstrictor response to thromboxane analog, U-46619 (100 nM). The results indicate that the PAR-1 receptor is critical in mediating the permeability-increasing and vasoconstrictor effects of thrombin in pulmonary microvessels.
Multiple blood pressure QTL on rat chromosome 1 defined by Dahl rat congenic strains
Authors: SAAD YASSER , GARRETT MICHAEL R. , RAPP JOHN P.
Published: December 2017

A series of congenic strains were constructed in which segments of chromosome (chr) 1 from Lewis (LEW) rats were introgressed into the Dahl salt-sensitive (S) strain. Three blood pressure quantitative trait loci (QTL) were defined. Two of these (QTL 1a and QTL 1b) were closely linked in the region between 1q31 and 1q35. The third blood pressure QTL (QTL region 2) was close to the centromere between 1p11 and 1q12, which includes the candidate gene Slc9a3 for sodium/hydrogen exchange. The blood pressure QTL 1a and QTL 1b defined here overlap significantly with QTL for disease phenotypes of renal failure, stroke, ventricular mass, and salt susceptibility defined in other rat strains, implying that these disease phenotypes and our blood pressure phenotype have causes in common. QTL 1b also corresponded approximately with a blood pressure QTL described on human chr 15. The QTL region 2 corresponded approximately with blood pressure QTL described on mouse chr 10 and human chr 6.
Melanocortin receptor-mediated mobilization of intracellular free calcium in HEK293 cells
Authors: MOUNTJOY KATHLEEN G. , KONG PHILIP L. , TAYLOR JOHN A. , WILLARD DERRIL H. , WILKISON WILLIAM O.
Published: December 2017

Mouse melanocortin receptors, MC1-R, MC3-R, MC4-R, and MC5-R, when expressed in HEK293 cells and stimulated with either α-melanocyte-stimulating hormone (α-MSH) or desacetyl-α-MSH, mediate increases in intracellular free calcium concentration ([Ca2+]i) with EC50values between 0.3 and 4.3 nM. The increase in [Ca2+]iis cholera toxin sensitive and pertussis toxin insensitive. The mechanism involves calcium mobilization from intracellular stores without a transient rise in inositol trisphosphate. Mouse agouti protein (55 nM) is a competitive antagonist of α-MSH (6-fold) and desacetyl-α-MSH (8-fold), coupling the mMC1-R to increased [Ca2+]i. Agouti protein (55 nM) significantly increased the EC50for α-MSH (3-fold), and 550 nM agouti protein significantly increased the EC50for desacetyl-α-MSH (4-fold), coupling the mMC4-R to a rise in [Ca2+]i. However, agouti protein antagonism of the MC4-R may not be competitive since there was a trend for the maximum response to also increase. There was no significant antagonism of the MC3-R and MC5-R by agouti protein (55 nM). Understanding the physiological relevance of the transduction of a calcium signal by melanocortin peptides may be important for future development of therapeutic targeting of the melanocortin receptors.
Differential gene expression profiling in human brain tumors
Authors: MARKERT JAMES M. , FULLER CATHERINE M. , GILLESPIE G. YANCEY , BUBIEN JAMES K. , McLEAN LEE ANNE , HONG ROBERT L. , LEE KAILIN , GULLANS STEVEN R. , MAPSTONE TIMOTHY B. , BENOS DALE J.
Published: December 2017

Gene expression profiling of three human temporal lobe brain tissue samples (normal) and four primary glioblastoma multiforme (GBM) tumors using oligonucleotide microarrays was done. Moreover, confirmation of altered expression was performed by whole cell patch clamp, immunohistochemical staining, and RT-PCR. Our results identified several ion and solute transport-related genes, such as N-methyl-d-aspartate (NMDA) receptors, α-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA)-2 receptors, GABAA receptor subunits α3, β1, β2, and β3, the glutamate transporter, the glutamate/aspartate transporter II, the potassium channel KV2.1, hKVβ3, and the sodium/proton exchanger 1 (NHE-1), that are all downregulated in the tumors compared with the normal tissues. In contrast, aquaporin-1, possibly aquaporins-3 and -5, and GLUT-3 message appeared upregulated in the tumors. Our results also confirmed previous work showing that osteopontin, nicotinamide N-methyltransferase, murine double minute 2 (MDM2), and epithelin (granulin) are upregulated in GBMs. We also demonstrate for the first time that the cytokine and p53 binding protein, macrophage migration inhibitory factor (MIF), appears upregulated in GBMs. These results indicate that the modulation of ion and solute transport genes and heretofore unsuspected cytokines (i.e., MIF) may have profound implications for brain tumor cell biology and thus may identify potential useful therapeutic targets in GBMs.
Artificial selection for intrinsic aerobic endurance running capacity in rats
Authors: KOCH LAUREN GERARD , BRITTON STEVEN L.
Published: December 2017

Artificial selection for intrinsic aerobic endurance running capacity was started using genetically heterogeneous N:NIH stock of rats as a founder population ( n = 168). Selection for low and high capacity was based upon distance run to exhaustion on a motorized treadmill using a velocity-ramped running protocol. The starting velocity was 10 m/min and was increased by 1 m/min every 2 min (slope was constant at 15°). At each generation, within-family selection was practiced using 13 families for both the low and high lines. A rotational breeding paradigm maintained the coefficient of inbreeding at less than 1% per generation. On average the founder population ran to exhaustion in 355 ± 11 m. Six generations of selection produced lines that differed in running capacity by 171%, with most of the change occurring in the high line. At generation 6 the low line ran 310 ± 8 m and the high line 839 ± 21 m at exhaustion. Selection for running capacity produced changes in body weight as a correlated trait. By generation 6, the low-line females were 20% heavier than the high-line females, and the low-line males were 16% heavier than the high-line males.
Senescence-related changes in gene expression in muscle: similarities and differences between mice and men
Authors: WELLE STEPHEN , BROOKS ANDREW , THORNTON CHARLES A.
Published: December 2017

A microarray study of the effect of senescence in mice on gene expression in muscle has been published recently. The present analysis was done to evaluate the extent to which the age-related differences in gene expression in murine muscle are also evident in human muscle. RNA extracted from muscle of young (21–24 yr) and old men (66–77 yr) was studied both by serial analysis of gene expression (SAGE) and by oligonucleotide microarrays. SAGE tags were detected for 61 genes homologous to genes reported to be differentially expressed in young and old murine muscle. The microarray had probe sets for 70 homologous genes. For 17 genes, there was evidence for a similar age-related change in expression in muscles of mice and men. For 32 other genes, there was evidence that the effect of age on the level of expression is not the same in mice and men. There was no evidence that older human muscle has increased expression of the stress response genes that are increased in old murine muscle.
Cloning and functional characterization of an uncoupling protein homolog in hummingbirds
Authors: VIANNA CLAUDIA R. , HAGEN THILO , ZHANG CHEN-YU , BACHMAN ERIC , BOSS OLIVIER , GEREBEN BALAZS , MORISCOT ANSELMO S. , LOWELL BRADFORD B. , BICUDO JOSÉ EDUARDO P. W. , BIANCO ANTONIO C.
Published: December 2017

The cDNA of an uncoupling protein (UCP) homolog has been cloned from the swallow-tailed hummingbird, Eupetomena macroura. The hummingbird uncoupling protein (HmUCP) cDNA was amplified from pectoral muscle (flight muscle) using RT-PCR and primers for conserved domains of various known UCP homologs. The rapid amplification of cDNA ends (RACE) method was used to complete the cloning of the 5′ and 3′ ends of the open reading frame. The HmUCP coding region contains 915 nucleotides, and the deduced protein sequence consists of 304 amino acids, being ∼72, 70, and 55% identical to human UCP3, UCP2, and UCP1, respectively. The uncoupling activity of this novel protein was characterized in yeast. In this expression system, the 12CA5-tagged HmUCP fusion protein was detected by Western blot in the enriched mitochondrial fraction. Similarly to rat UCP1, HmUCP decreased the mitochondrial membrane potential as measured in whole yeast by uptake of the fluorescent potential-sensitive dye 3′,3-dihexyloxacarbocyanine iodide. The HmUCP mRNA is primarily expressed in skeletal muscle, but high levels can also be detected in heart and liver, as assessed by Northern blot analysis. Lowering the room’s temperature to 12–14°C triggered the cycle torpor/rewarming, typical of hummingbirds. Both in the pectoral muscle and heart, HmUCP mRNA levels were 1.5- to 3.4-fold higher during torpor. In conclusion, this is the first report of an UCP homolog in birds. The data indicate that HmUCP has the potential to function as an UCP and could play a thermogenic role during rewarming.
Time course of LPS-induced gene expression in a mouse model of genitourinary inflammation
Authors: SABAN MARCIA R. , HELLMICH HELEN , NGUYEN NGOC-BICH , WINSTON JOHN , HAMMOND TIMOTHY G. , SABAN RICARDO
Published: December 2017

In this study, self-organizing map (SOM) gene cluster techniques are applied to the analysis of cDNA microarray analysis of gene expression changes occurring in the early stages of genitourinary inflammation. We determined the time course of lipopolysaccharide (LPS)-induced gene expression in experimental cystitis. Mice were euthanized 0.5, 1, 4, and 24 h after LPS instillation into the urinary bladder, and gene expression was determined using four replicate Atlas mouse cDNA expression arrays containing 588 known genes at each time point. SOM gene cluster analysis, performed without preconditions, identified functionally significant gene clusters based on the kinetics of change in gene expression. Genes were classified as follows: 1) expressed at time 0; 2) early genes (peak expression between 0.5 and 1 h); and 3) late genes (peak expression between 4 and 24 h). One gene cluster maintained a constant level of expression during the entire time period studied. In contrast, LPS treatment downregulated the expression of some genes expressed at time 0, in a cluster including transcription factors, protooncogenes, apoptosis-related proteins (cysteine protease), intracellular kinases, and growth factors. Gene upregulation in response to LPS was observed as early as 0.5 h in a cluster including the interleukin-6 (IL-6) receptor, α- and β-nerve growth factor (α- and β-NGF), vascular endothelial growth factor receptor-1 (VEGF R1), C-C chemokine receptor, and P-selectin. Another tight cluster of genes with marked expression at 1 h after LPS and insignificant expression at all other time points studied included the protooncogenes c-Fos, Fos-B, Fra-2, Jun-B, Jun-D, and Egr-1. Almost all interleukin genes were upregulated as early as 1 h after stimulation with LPS. Nuclear factor-κB (NF-κB) pathway genes collected in a single cluster with a peak expression 4 h after LPS stimulation. In contrast, most of the interleukin receptors and chemokine receptors presented a late peak of expression 24 h after LPS coinciding with the peak of neutrophil infiltration into the bladder wall. Selected cDNA microarray observations were confirmed by RNase protection assay. In conclusion, the cDNA array experimental approach provided a global profile of gene expression changes in bladder tissue after stimulation with LPS. SOM techniques identified functionally significant gene clusters, providing a powerful technical basis for future analysis of mechanisms of bladder inflammation.
Monitoring expression of genes involved in drug metabolism and toxicology using DNA microarrays
Authors: GERHOLD DAVID , LU MEIQING , XU JIAN , AUSTIN CHRISTOPHER , CASKEY C. THOMAS , RUSHMORE THOMAS
Published: December 2017

Oligonucleotide DNA microarrays were investigated for utility in measuring global expression profiles of drug metabolism genes. This study was performed to investigate the feasibility of using microarray technology to minimize the long, expensive process of testing drug candidates for safety in animals. In an evaluation of hybridization specificity, microarray technology from Affymetrix distinguished genes up to a threshold of ∼90% DNA identity. Oligonucleotides representing human cytochrome P-450 gene CYP3A5 showed heterologous hybridization to CYP3A4 and CYP3A7 RNAs. These genes could be clearly distinguished by selecting a subset of oligonucleotides that hybridized selectively to CYP3A5. Further validation of the technology was performed by measuring gene expression profiles in livers of rats treated with vehicle, 3-methylcholanthrene (3MC), phenobarbital, dexamethasone, or clofibrate and by confirming data for six genes using quantitative RT-PCR. Responses of drug metabolism genes, including CYPs, epoxide hydrolases ( EHs), UDP-glucuronosyl transferases ( UGTs), glutathione sulfotransferases ( GSTs), sulfotransferases ( STs), drug transporter genes, and peroxisomal genes, to these well-studied compounds agreed well with, and extended, published observations. Additional gene regulatory responses were noted that characterize metabolic effects or stress responses to these compounds. Thus microarray technology can provide a facile overview of gene expression responses relevant to drug metabolism and toxicology.
Genetic loci determining bone density in mice with diet-induced atherosclerosis
Authors: DRAKE THOMAS A. , SCHADT ERIC , HANNANI KAMBIZ , KABO J. MICHAEL , KRASS KELLY , COLINAYO VERONICA , GREASER LLOYD E. , GOLDIN JONATHAN , LUSIS ALDONS J.
Published: December 2017

This study investigates the phenotypic and genetic relationships among bone-density-related traits and those of adipose tissue and plasma lipids in mice with diet-induced atherosclerosis. Sixteen-month-old female F2 progeny of a C57BL/6J and DBA/2J intercross, which had received an atherogenic diet for 4 mo, were examined for multiple measures of femoral bone mass, density, and biomechanical properties using both computerized tomographic and radiographic methods. In addition, body weight and length, adipose tissue mass, plasma lipids and insulin, and aortic fatty lesions were assessed. Bone mass was inversely correlated with extent of atherosclerosis and with a prooxidant lipid profile and directly correlated with body weight, length, and, most strongly, adipose tissue mass. Quantitative trait locus (QTL) analysis, using composite interval mapping (CIM) and multi-trait analysis, identified six loci with multi-trait CIM LOD scores > 5. Three of these coincided with loci linked with adipose tissue and plasma high-density lipoprotein. Application of statistical tests for distinguishing close linkage vs. pleiotropy supported the presence of a potential pleiotropic effect of two of the loci on these traits. This study shows that bone mass in older female mice with atherosclerosis has multiple genetic determinants and provides phenotypic and genetic evidence linking the regulation of bone density with adipose tissue and plasma lipids.
Ureteric bud derivatives express angiotensinogen and AT1 receptors
Authors: PRIETO MINOLFA , DIPP SUSANA , MELEG-SMITH SUZANNE , EL-DAHR SAMIR S.
Published: December 2017

Inactivation of the renin-angiotensin system interferes with the morphogenesis of the renal medulla. Thus ureteric bud (UB) derivatives may be a target for angiotensin production and action. To begin to test this hypothesis, we examined the cellular expression of angiotensinogen (Ao) and AT1 receptor proteins during rat metanephrogenesis by immunohistochemistry. In addition, we tested whether UB-derived cells in culture express the Ao and AT1 proteins. On embryonic day E15, Ao and AT1 are expressed in the UB branches and stromal mesenchyme. S-shaped bodies, including the vascular cleft, express AT1 but not Ao. The metanephric mesenchyme and pretubular aggregates are Ao negative and AT1 negative. Expression of Ao and AT1 in UB branches and ampullae is also observed on E16. However, UB expression of Ao is transient and is no longer detectable in the developing distal nephron beyond E17. On E17, both Ao and AT1 are expressed in capillary loop glomeruli and proximal tubules, whereas UB branches express AT1 only. By E18, the majority of Ao immunoreactivity is clustered in terminally differentiated proximal tubules, whereas AT1 receptors are expressed in both proximal and distal nephron segments. The specificity of Ao and AT1 staining was documented by the elimination/attenuation of immunoreactivity after preadsorption of the primary antibodies with their respective antigens. Consistent with the in vivo findings, the AT1 protein is abundantly expressed in cellular lysates of mouse UB ( E11.5) and IMCD3 (adult) cells. Moreover, AT1 receptors in UB and IMCD3 cells are functional, since angiotensin II treatment elicits the tyrosine phosphorylation of the mitogen-activated protein kinases, ERK1/2. To our knowledge, this is the first demonstration of Ao and AT1 protein expression in the developing distal nephron. Angiotensin II may have a paracrine role in the ontogeny of the collecting system.
Ren1dand Ren2 cooperate to preserve homeostasis: evidence from mice expressing GFP in place of Ren1d
Authors: PENTZ ELLEN S. , LOPEZ MARIA LUISA S. SEQUEIRA , KIM HYUNG-SUK , CARRETERO OSCAR , SMITHIES OLIVER , GOMEZ R. ARIEL
Published: December 2017

To distinguish the contributions of Ren1dand Ren2 to kidney development and blood pressure homeostasis, we placed green fluorescent protein (GFP) under control of the Ren1drenin locus by homologous recombination in mice. Homozygous Ren1d-GFP animals make GFP mRNA in place of Ren1dmRNA in the kidney and maintain Ren2 synthesis in the juxtaglomerular (JG) cells. GFP expression provides an accurate marker of Ren1dexpression during development. Kidneys from homozygous animals are histologically normal, although with fewer secretory granules in the JG cells. Blood pressure and circulating renin are reduced in Ren1d-GFP homozygotes. Acute administration of losartan decreases blood pressure further, suggesting a role for Ren2 protein in blood pressure homeostasis. These studies demonstrate that, in the absence of Ren1d, Ren2 preserves normal kidney development and prevents severe hypotension. Chronic losartan treatment results in compensation via recruitment of both Ren1d- and Ren2-expressing cells along the preglomerular vessels. This response is achieved by metaplastic transformation of arteriolar smooth muscle cells, a major mechanism to control renin bioavailability and blood pressure homeostasis.
Changes in the mitochondrial proteome from mouse hearts deficient in creatine kinase
Authors: KERNEC FLORENCE , ÜNLÜ MUSTAFA , LABEIKOVSKY WLADIMIR , MINDEN JONATHAN S. , KORETSKY ALAN P.
Published: December 2017

Creatine kinase (CK) is an abundant enzyme, important for maintenance of high-energy phosphate homeostasis in many tissues including heart. Double-knockout CK (DbKO-CK) mice missing both the muscle (MM) and sarcomeric mitochondrial (ScMit) isoforms of CK have recently been studied. Despite a large change in skeletal muscle function in DbKO-CK mice, there is little functional change in the heart. To investigate whether there are specific changes in cardiac mitochondrial proteins associated with the loss of MM- and ScMit-CK isoforms, we have used difference gel electrophoresis (DIGE) to compare mitochondrial proteins from wild-type and DbKO-CK mice. Mass spectrometry fingerprinting was used to identify 40 spots as known mitochondrial proteins. We have discovered that the loss of MM- and ScMit-CK isoforms did not cause large scale changes in heart mitochondrial proteins. The loss of ScMit-CK was readily detected in the DbKO-CK samples. We have also detected a large decrease in the precursor form of aconitase. Furthermore, two mitochondrial protein differences have been found in the parent mouse strains of the DbKO-CK mice.
KGF alters gene expression in human airway epithelia: potential regulation of the inflammatory response
Authors: PRINCE LAWRENCE S. , KARP PHILIP H. , MONINGER THOMAS O. , WELSH MICHAEL J.
Published: December 2017

Keratinocyte growth factor (KGF) regulates several functions in adult and developing lung epithelia; it causes proliferation, stimulates secretion of fluid and electrolytes, enhances repair, and may minimize injury. To gain insight into the molecular processes influenced by KGF, we applied KGF to primary cultures of well-differentiated human airway epithelia and used microarray hybridization to assess the abundance of gene transcripts. Of 7,069 genes tested, KGF changed expression levels of 910. Earlier studies showed that KGF causes epithelial proliferation, and as expected, treatment altered expression of numerous genes involved in cell proliferation. We found that KGF stimulated transepithelial Cl−transport, but the number of cystic fibrosis (CF) transmembrane conductance regulator (CFTR) transcripts fell. Although transcripts for ClC-1 and ClC-7 Cl−channels increased, KGF failed to augment transepithelial Cl−transport in CF epithelia, suggesting that KGF-stimulated Cl−transport in differentiated airway epithelia depends on the CFTR Cl−channel. Interestingly, KGF decreased transcripts for many interferon (IFN)-induced genes. IFN causes trafficking of Stat dimers to the nucleus, where they activate transcription of IFN-induced genes. We found that KGF prevented the IFN-stimulated trafficking of Stat1 from the cytosol to the nucleus, suggesting a molecular mechanism for KGF-mediated suppression of the IFN-signaling pathway. These results suggest that in addition to stimulating proliferation and repair of damaged airway epithelia, KGF stimulates Cl−transport and may dampen the response of epithelial cells to inflammatory mediators.
Rat mitochondrial ATP synthase ATP5G3: cloning and upregulation in pancreas after chronic ethanol feeding
Authors: LI HA-SHENG , ZHANG JI-YING , THOMPSON BRYAN S. , DENG XIAO-YING , FORD MICHAEL E. , WOOD PAUL G. , STOLZ DONNA B. , EAGON PATRICIA K. , WHITCOMB DAVID C.
Published: December 2017

Individuals with chronic excessive alcohol ingestion are put at the risk of acute and chronic pancreatitis. Underlying molecular mechanisms are unknown. Differential gene expression in the pancreas was profiled using mRNA differential display by comparison between control and ethanol-consuming rats. Male Wistar rats were fed with diets containing 6.7% (vol/vol) ethanol for 4 wk. A cDNA tag that was overexpressed in the pancreas of rats fed ethanol was isolated. A 723-bp cDNA was cloned from a rat pancreatic cDNA library, which encodes a novel rat mitochondrial ATP synthase subunit 9, isoform 3 (ATP5G3), which is homologous to a human ATP5G3 gene. Real-time PCR demonstrated that all three nuclear gene isoforms (ATP5G1, ATP5G2, and ATP5G3) were consistently upregulated in the pancreas of alcohol-consuming rats, parallel with mitochondrial injury. The cellular response to mitochondrial damage and metabolic stress may reflect an adaptive process for mitochondrial repair in pancreatic acinar cells during chronic ethanol ingestion.
NewDyscalcloci for myocardial cell necrosis and calcification (dystrophic cardiac calcinosis) in mice
Authors: IVANDIC BORIS T. , UTZ H. FRIEDRICH , KACZMAREK PIOTR M. , AHERRAHROU ZOUHAIR , AXTNER SUSANNE B. , KLEPSCH CAROLA , LUSIS ALDONS J. , KATUS HUGO A.
Published: December 2017

Dystrophic cardiac calcinosis (DCC) occurs among certain inbred strains of mice and involves necrosis and subsequent calcification as response of myocardial tissue to injury. Using a complete linkage map approach, we investigated the genetics of DCC in an F2intercross of resistant C57BL/6J and susceptible C3H/HeJ inbred strains and identified previously a major predisposing quantitative trait locus (QTL), Dyscalc1, on proximal chromosome 7. Analysis of inheritance suggested, however, that DCC is influenced by additional modifier QTL, which have as yet not been mapped. Here, we report the identification by composite interval mapping of the DCC loci Dyscalc2, Dyscalc3, and Dyscalc4 on chromosomes 4, 12 and 14, respectively. Together, the four Dyscalc loci explained 47% of the phenotypic variance of DCC, which was induced by a high-fat diet. Additive epistasis between Dyscalc1 and Dyscalc2 enhanced DCC. Examining recombinant inbred strains, we propose a 10-cM interval containing Dyscalc1 and discuss potential candidate genes.
Identification of genomic regions controlling plasma FSH concentrations in Meishan-White Composite boars
Authors: ROHRER G. A. , WISE T. H. , LUNSTRA D. D. , FORD J. J.
Published: December 2017

The Chinese Meishan (ME) breed of pig is unique for many reproductive traits. Compared with Western breeds of swine, ME females reach puberty earlier, ovulate more ova per estrus, and have greater uterine capacity, while intact males (boars) have smaller testes and extremely elevated plasma levels of pituitary-derived glycoprotein hormones. In an effort to identify the genetic mechanisms controlling the elevated plasma levels of pituitary-derived glycoprotein hormones [in particular, follicle-stimulating hormone (FSH)] and to determine whether some of these genetic factors are also responsible for differences in other phenotypes, we scanned the entire genome for regions that affected plasma FSH in boars from a Meishan-White Composite (equal contributions of Chester White, Landrace, Large White, and Yorkshire) resource population. Initially, the entire genome of 121 boars was scanned for regions that potentially influenced plasma FSH. The most significant genomic regions were further studied in a total of 436 boars. Three genomic regions located on chromosomes 3, 10, and X apparently possess genes that significantly affect FSH level, and one region provided suggestive evidence for the presence of FSH-controlling genes located on chromosome 8. The region on the X chromosome also affected testes size. Similar genomic regions to those identified on chromosomes 3, 8, and 10 in this study have been identified to affect ovulation rate in female litter mates, supporting the hypothesis that plasma FSH in pubertal boars and ovulation rate in females is controlled by a similar set of genes.
Oxygen-dependent expression of hypoxia-inducible factor-1α in renal medullary cells of rats
Authors: ZOU AI-PING , YANG ZHI-ZHANG , LI PIN-LAN , COWLEY ALLEN W.
Published: December 2017

Hypoxia-inducible factor-1α (HIF-1α) is a transcription factor that regulates the oxygen-dependent expression of a number of genes. This transcription factor may contribute to the abundant expression of many genes in renal medullary cells that function normally under hypoxic conditions. The present study was designed to determine the characteristics of HIF-1α cDNA cloned from the rat kidney and the expression profile of HIF-1α in different kidney regions and to explore the mechanism activating or regulating HIF-1α expression in renal medullary cells. A 3,718-bp HIF-1α cDNA from the rat kidney was first cloned and sequenced using RT-PCR and TA cloning technique. It was found that 823 amino acids deduced from this renal HIF-1α cDNA had 99%, 96%, and 90% identity with rat, mouse, or human HIF-1α deposited in GenBank, respectively. The 3′-untranslated region of HIF-1α mRNA from the rat kidney contained seven AUUUA instability elements, five of which were found to be conserved among rat, mouse, and human HIF-1α. Northern blot analyses demonstrated a corticomedullary gradient of HIF-1α mRNA expression in the kidney, with the greatest abundance in the renal inner medulla. Western blot analyses also detected a higher HIF-1α protein level in the nuclear extracts from the renal medulla than the renal cortex. A classic loop diuretic, furosemide (10 mg/kg ip), markedly increased renal medullary Po2levels from 22.5 to 52.2 mmHg, which was accompanied by a significant reduction of HIF-1α transcripts in renal medullary tissue. In in vitro experiments, low Po2, but not elevated osmolarity, was found to significantly increase HIF-1α mRNA in renal medullary interstitial cells and inner medullary collecting duct cells. These results indicate that HIF-1α is more abundantly expressed in the renal medulla compared with the renal cortex. Increased abundance of HIF-1α mRNA in the renal medulla may represent an adaptive response of renal medullary cells to low Po2.
Gene expression following acute morphine administration
Authors: LOGUINOV A. V. , ANDERSON L. M. , CROSBY G. J. , YUKHANANOV R. Y.
Published: December 2017

The long-term response to neurotropic drugs depends on drug-induced neuroplasticity and underlying changes in gene expression. However, alterations in neuronal gene expression can be observed even following single injection. To investigate the extent of these changes, gene expression in the medial striatum and lumbar part of the spinal cord was monitored by cDNA microarray following single injection of morphine. Using robust and resistant linear regression (MM-estimator) with simultaneous prediction confidence intervals, we detected differentially expressed genes. By combining the results with cluster analysis, we have found that a single morphine injection alters expression of two major groups of genes, for proteins involved in mitochondrial respiration and for cytoskeleton-related proteins. RNAs for these proteins were mostly downregulated both in the medial striatum and in lumbar part of the spinal cord. These transitory changes were prevented by coadministration of the opioid antagonist naloxone. Data indicate that microarray analysis by itself is useful in describing the effect of well-known substances on the nervous system and provides sufficient information to propose a potentially novel pathway mediating its activity.
Bowing before Dual Gods: How Structured Flexibility Sustains Organizational Hybridity
Authors: Smith Wendy K. , Besharov Marya L.
Published: December 2017

Organizations increasingly grapple with hybridity—the combination of identities, forms, logics, or other core elements that would conventionally not go together. Drawing on in-depth longitudinal data from the first ten years of a successful social enterprise—Digital Divide Data, founded in Cambodia—we induce an empirically grounded model of sustaining hybridity over time through structured flexibility: the interaction of stable organizational features and adaptive enactment processes. We identify two stable features—paradoxical frames, involving leaders’ cognitive understandings of the two sides of a hybrid as both contradictory and interdependent, and guardrails, consisting of formal structures, leadership expertise, and stakeholder relationships associated with each side—that together facilitate ongoing adaptation in the meanings and practices of dual elements, sustaining both elements over time. Our structured flexibility model reorients research away from focusing on either stable or adaptive approaches to sustaining hybridity toward understanding their interaction, with implications for scholarship on hybridity, duality, and adaptation more broadly.
More and Less Effective Updating: The Role of Trajectory Management in Making Sense Again
Authors: Christianson Marlys K.
Published: December 2017

This study examines how updating—the process of revising provisional sensemaking to incorporate new cues—occurs within teams during unexpected events. I compare how 19 teams of emergency department staff managed the same unexpected event (a broken piece of equipment) in a medical simulation scenario. Using a microethnographic approach to analyze video recordings of these teams, I conduct a fine-grained examination of how updating takes place and find considerable variation in its effectiveness across teams. I show that the effectiveness of updating depends not only on how teams remake sense but also on how they engage in trajectory management, balancing the work of updating with their ongoing work (in this case, patient care). Trajectory management practices related to monitoring cues and managing engaging tasks facilitated effective updating and allowed teams to detect and identify the problem caused by the broken piece of equipment and correct it before it led to serious consequences. More-effective teams monitor and rapidly interpret cues, confirming them with others and evaluating changes over time; they then investigate cues, develop plausible explanations, and quickly test them, monitoring cues for feedback. Less-effective teams fail to monitor and confirm cues with others, overlook or misinterpret cues, and delay investigating cues and developing plausible explanations; they also delay testing explanations, often being sidetracked by patient care tasks.
Train Them to Retain Them: Work Readiness and the Retention of First-time Women Workers in India
Authors: Ranganathan Aruna
Published: December 2017

To explore when and why workplace training facilitates the retention of first-time workers from historically underrepresented groups in formal employment, I combine ethnographic fieldwork at an Indian garment factory employing first-time women workers, personnel data over a two-year period, and survey data from a sample of new hires. I find that training is effective at preventing first-time women workers from dropping out soon after they are hired when it is conducted by trainers with many years of experience. Rather than focusing only on job-specific skills, training conducted by experienced trainers includes the basic work-readiness skills of self-presentation, interpersonal communication, work–life separation, and self-reliance needed to survive at work. I find that first-time women workers quasi-randomly assigned to experienced trainers had about a 20-percent greater probability of being retained after three months, and these workers reported that they felt more ready for work than those assigned to less-experienced trainers. My results imply that for the majority of workers from historically underrepresented groups who are entering the workplace for the first time, training is important to foster their retention, and organizations that focus on both the attributes of the people delivering that training and its content have a greater chance of keeping these workers for the long term.