Tyrosine phosphorylation of Kv1.5 is upregulated in intrauterine growth retardation rats with exaggerated pulmonary hypertension.

Intrauterine growth retardation (IUGR) is associated with the development of adult-onset diseases, including pulmonary hypertension. However, the underlying mechanism of the early nutritional insult that results in pulmonary vascular dysfunction later in life is not fully understood. Here, we investigated the role of tyrosine phosphorylation of voltage-gated potassium channel 1.5 (Kv1.5) in this prenatal event that results in exaggerated adult vascular dysfunction. A rat model of chronic hypoxia (2 weeks of hypoxia at 12 weeks old) following IUGR was used to investigate the physiological and structural effect of intrauterine malnutrition on the pulmonary artery by evaluating pulmonary artery systolic pressure and vascular diameter in male rats. Kv1.5 expression and tyrosine phosphorylation in pulmonary artery smooth muscle cells (PASMCs) were determined. We found that IUGR increased mean pulmonary artery pressure and resulted in thicker pulmonary artery smooth muscle layer in 14-week-old rats after 2 weeks of hypoxia, while no difference was observed in normoxia groups. In the PASMCs of IUGR-hypoxia rats, Kv1.5 mRNA and protein expression decreased while that of tyrosine-phosphorylated Kv1.5 significantly increased. These results demonstrate that IUGR leads to exaggerated chronic hypoxia pulmonary arterial hypertension (CH-PAH) in association with decreased Kv1.5 expression in PASMCs. This phenomenon may be mediated by increased tyrosine phosphorylation of Kv1.5 in PASMCs and it provides new insight into the prevention and treatment of IUGR-related CH-PAH.

Identification of quantitative trait loci for mineral elements in grains and grass powder of barley.

Mineral elements in barley (Hordeum vulgare) play an important physiological role in global human health. In this study, quantitative trait loci (QTLs) for concentration of nine mineral elements in barley grain and grass powder were detected in a population of 193 recombinant inbred lines of the barley cross Ziguangmangluoerling x Schooner and the parents. We observed large genetic variation contributing to element concentrations in both grains and grass powder. The mean K, Ca, and Fe concentrations in grass powder were 6.67, 12.00, and 4.58 times that of regenerating barley grains. In grains, 17 QTLs that accounted for 6.36-64.08% of the phenotypic variation in Zn, Mg, Ca, K, Na, Mn, Fe, and P concentrations were identified. In grass powder, seven QTLs were identified; these accounted for 6.03-21.86% of the variation in Ca, Zn, Mg, K, Fe, and Cu concentrations. These QTLs affecting elements in grain and grass powder are so far unreported in barley. To our knowledge, QTLs with pleiotropic effects for three elements were also identified for the first time in barley. The qK1/qMg1/qCa1 region between markers Bmag0211 and GBMS0014 on chromosome 1H was shown to have large additive effects for Mg, Ca, and K concentrations in grains. These additive effects indicated that the high element (Mg, Ca, Zn, Mn, and K) alleles were contributed by Ziguangmangluoerling. These results will further our understanding of the genetic basis of mineral elements and help us develop markers linked with mineral elements for marker-assisted selection breeding in barley.

Isolation and characterization of microsatellite markers for Dendranthema morifolium (Asteraceae) using next-generation sequencing.

Dendranthema morifolium (Asteraceae) is a perennial herbaceous plant native to China. A long history of artificial crossings may have resulted in complex genetic background and decreased genetic diversity. To protect the genetic diversity of D. morifolium and enabling breeding of new D. morifolium cultivars, we developed a set of molecular markers. We used pyrosequencing of an enriched microsatellite library by Roche 454 FLX+ platform, to isolate D. morifolium simple sequence repeats (SSRs). A total of 32,863 raw reads containing 2251 SSRs were obtained. To test the effectiveness of these SSR markers, we designed primers by randomly selecting 100 novel SSRs, and amplified them across 60 cultivars representing five different petal shape groups. Sixteen SSRs were polymorphic with the number of alleles ranging from 6 to 19, and their expected and observed heterozygosities ranging from 0.477 to 0.848, and 0.250 to 0.804, respectively. The polymorphism information content ranged from 0.459 to 0.854 and the inbreeding coefficient ranged from -0.119 to 0.759. An unweighted pair-group method arithmetic average analysis was performed to survey the phylogenetic relationships of these 60 cultivars and five clusters were identified. These markers can be used for investigating genetic relationships and identifying elite alleles through linkage and association analyses.

Study of the expression of GABA(A) receptor in rats during acute lung injury caused by endotoxin.

The objective of the present study was to investigate the role of γ-aminobutyric acid type A receptor (GABA(A)R) in lipopolysaccharide (LPS)-induced acute lung injury (ALI) in rats. Thirty-two male wistar rats were randomly divided into four groups. Rats in the GABA group were pretreated with LPS and GABA, while those in the bicuculline (BIC) group were pretreated with LPS and bicuculline. We assessed the arterial blood gas, dry/wet ratio, and the level of tumor necrosis factor-α (TNF-α), IL-6, malondialdehyde, and superoxide dismutase 6 h after the immunization. Paraffin sections of samples were detected using the steptavidin-peroxidase method. Protein expression was detected using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and western blotting. PaO2 in the LPS group was significantly lower than that in the control rats. Activation of GABA-mediated signaling by GABA increased the expression of GABA(A)R in airway bronchial and alveolar epithelial cells. Blockade of the GABA(A)R by bicuculline limited the expression of this receptor. The GABA group rats had higher levels of tissue TNF-α and IL-6 than in ALI rats and control rats. The BIC group rats demonstrated an opposite expression level compared to the GABA group rats. Our results suggest that the GABA(A)R could aggravate the inflammatory response syndrome and oxidative stress in the lungs and play an essential role in LPS-induced acute lung injury. It provides a novel method to study the incidence and mortality of ALI during the peroperative period.

Molecular cloning and tissue distribution profiles of the chicken R-spondin1 gene.

Rspo1 belongs to the Rspo family, which is composed of 4 members (Rspo1-4) that share 40 to 60% sequence homology and similar domain organizations, and regulate the WNT signaling pathway via a common mechanism. Rspo1 plays a key role in vertebrate development and is an effective mitogenic factor of gastrointestinal epithelial cells. We report the cloning of chicken Rspo1 and its gene expression distribution among tissues. It contained an open reading frame of 783 bp encoding a protein of 260 amino acids, and its molecular weight was predicted to be 28.80 kDa. Reverse transcription-polymerase chain reaction-based gene expression analysis indicated that chicken Rspo1 was highly expressed in the stomach muscle tissue, but was expressed at low levels in the lung, brain, jejunum, cecum, ileum, spleen, pancreas, kidney, and glandular stomach. These results suggest that Rspo1 plays a major role in muscular immune protection.

Comparative proteomic analysis of the nucleus accumbens during extinction and reinstatement of morphine dependence / Análisis proteómico comparativo del núcleo accumbens durante laextinción y recaída de la dependencia a la morfina

BACKGROUND: The aim of this study was to detect differentially expressed proteins in the nucleus accumbens between the states of extinction and reinstatement of morphine addiction. Numerous studies on the neurobiological mechanisms concerning drug craving and relapse have been reported to date, but data on their relationship with the underlying key molecular mechanisms involved remain limited. METHODS: In this study, 40 male SpragueDawley rats were equally randomized into a saline group and a morphine group. Both groups received drug selfadministration training, after which extinction models were established naturally. The groups were further divided into two subgroups for extinction and reinstatement tests. Cerebral nucleus accumbens masses were measured for total protein extraction. Twodimensional electrophoresis was performed to determine differential protein spots. These differential proteins were then enzymolysed and identified using mass spectrography. RESULTS: The proteins were classified as fatty acidbinding protein, serine/threonine protein phosphatase 2A catalytic subunit beta isoform, serine/threonine protein phosphatase 2A catalytic subunit alpha isoform, serine/threonine protein phosphatase 2A regulatory subunit B² subunit gamma or heat shock protein 90 cochaperone CDC37. CONCLUSION: Significant changes in five proteins were detected between extinction and reinstatement. These proteins are correlated with phosphorylation and the tricarboxylic acid cycle.

ANTECEDENTES: El objetivo de este estudio fue detectar las proteínas diferencialmente expresadas en el núcleo accumbens entre los estados de extinción y recaída de la adicción a la morfina. Hasta la fecha se han reportado numerosos estudios en relación con los mecanismos neurobiológicos del deseo incontenible y recaída en el consumo de drogas, pero los datos sobre su relación con los mecanismos moleculares fundamentales subyacentes implicados, siguen siendo limitados. MÉTODO: En este estudio, 40 ratas machos SpragueDawley fueron por igual asignadas de manera aleatoria a un grupo salino y un grupo de morfina. Ambos grupos recibieron entrenamiento de autoadministración de drogas, después de lo cual se establecieron modelos de extinción de manera natural. A su vez, los grupos fueron luego subdivididos en dos subgrupos para realizar pruebas de extinción y recaída. Se procedió a medir las masas cerebrales del núcleo accumbens para la extracción total de proteína. Se realizó una electroforesis bidimensional para determinar manchas proteicas diferenciales. Estas proteínas diferenciales fueron entonces sometidas a enzimólisis e identificadas mediante espectrografía de masa. RESULTADOS: Las proteínas fueron clasificadas como proteína de unión a ácidos grasos, isoforma beta de la subunidad catalítica serinatreonina proteína fosfatasa 2A, isoforma alfa de la subunidad catalítica serinatreonina proteína fosfatasa 2A, subunidad gamma subunidad B" de la serinatreonina proteína fosfatasa 2A, o la proteína CDC37 cochaperona 90 de choque térmico. CONCLUSIÓN: Se detectaron cambios significativos en cinco proteínas entre la extinción y la recaída. Estas proteínas están correlacionadas con la fosforilación y el ciclo del ácido tricarboxílico.

Comparative proteomic analysis of the nucleus accumbens during extinction and reinstatement of morphine dependence.

BACKGROUND: The aim of this study was to detect differentially expressed proteins in the nucleus accumbens between the states of extinction and reinstatement ofmorphine addiction. Numerous studies on the neurobiological mechanisms concerning drug craving and relapse have been reported to date, but data on their relationship with the underlying key molecular mechanisms involved remain limited. METHODS: In this study, 40 male Sprague-Dawley rats were equally randomized into a saline group and a morphine group. Both groups received drug self-administration training, after which extinction models were established naturally. The groups were further divided into two subgroups for extinction and reinstatement tests. Cerebral nucleus accumbens masses were measured for total protein extraction. Two-dimensional electrophoresis was performed to determine differential protein spots. These differential proteins were then enzymolysed and identified using mass spectrography. RESULTS: The proteins were classified as fatty acid-binding protein, serine/threonine protein phosphatase 2A catalytic subunit beta isoform, serine/threonine protein phosphatase 2A catalytic subunit alpha isoform, serine/threonine protein phosphatase 2A regulatory subunit B2 subunit gamma or heat shock protein 90 co-chaperone CDC37. CONCLUSION: Significant changes in five proteins were detected between extinction and reinstatement. These proteins are correlated with phosphorylation and the tricarboxylic acid cycle.