Characterization of acid-sensing ion channels in dorsal horn neurons of rat spinal cord

Acid-sensing ion channels (ASICs) are ligand-gated cation channels activated by extracellular protons. In periphery, they contribute to sensory transmission, including that of nociception and pain. Here we characterized ASIC-like currents in dorsal horn neurons of the rat spinal cord and their functional modulation in pathological conditions. Reverse transcriptase-nested PCR and Western blotting showed that three ASIC isoforms, ASIC1a, ASIC2a, and ASIC2b, are expressed at a high level in dorsal horn neurons. Electrophysiological and pharmacological properties of the proton-gated currents suggest that homomeric ASIC1a and/or heteromeric ASIC1a + 2b channels are responsible for the proton-induced currents in the majority of dorsal horn neurons. Acidification-induced action potentials in these neurons were compatible in a pH-dependent manner with the pH dependence of ASIC-like current. Furthermore, peripheral complete Freund's adjuvant-induced inflammation resulted in increased expression of both ASIC1a and ASIC2a in dorsal horn. These results support the idea that the ASICs of dorsal horn neurons participate in central sensory transmission/modulation under physiological conditions and may play important roles in inflammation-related persistent pain.

Upregulation of acid-sensing ion channel ASIC1a in spinal dorsal horn neurons contributes to inflammatory pain hypersensitivity

Development of chronic pain involves alterations in peripheral nociceptors as well as elevated neuronal activity in multiple regions of the CNS. Previous pharmacological and behavioral studies suggest that peripheral acid-sensing ion channels (ASICs) cont

Differential expression of the Brunol/CELF family genes during Xenopus laevis early development

The BRUNOL/CELF family of RNA-binding proteins plays important roles in post-transcriptional regulation and has been implicated in several developmental processes. In this study, we describe the cloning and expression patterns of five Brunol genes in Xenopus laevis. Among them, only Brunol2 is maternally expressed and the zygotic expression of the other four Brunol genes starts at different developmental stages. During Xenopus development, Brunol1, 4-5 are exclusively expressed in the nervous system including domains in the brain, spinal cord, optic and otic vesicles. Brunol2 and 3 are expressed in both the somatic mesoderm and the nervous system. Brunol2 is also extensively expressed in the lens. In transfected Hela cells, BRUNOL1, 2 and 3 proteins are localized in both the cytoplasm and the nucleus, while BRUNOL4 and 5 are only present in the cytoplasm, indicating their different functions.

人胚神经干细胞移植治疗大鼠脊髓损伤

　目的　探讨人胚神经干细胞( hNSC) 移植治疗脊髓损伤(SCI) 的可行性。方法　分 离、培养和鉴定hNSC;用5 溴22 脱氧尿苷嘧啶(BrdU) 标记hNSC ,并将其移植到14 只T10 半横断 的Wistar 大鼠损伤脊髓内(另外14 只T10 半横断损伤的大鼠作为对照组,仅损伤脊髓内注射 DMEM/ F12 培养液) ,用BrdU 的FITC 免疫荧光染色检测移植细胞的存活和迁徙,用NF2200 、 GFAP 免疫组织化学鉴定移植细胞的分化,BBB 评分评定大鼠功能恢复情况。结果　(1) 获得了大 量的hNSC; (2) 用免疫组织化学可以检测到移植的hNSC 能在体内长时间存活(达2 个月) 并向远 处迁徙,并分化为神经元和胶质细胞; (3) 检测到实验组大鼠BBB 得分明显高于对照组大鼠( P < 0. 01) ,在SCI 后第10 周时实验组和对照组BBB 得分最大差距达到2. 1分。结论　hNSC 移植能促 进SCI 大鼠后肢功能恢复,它是SCI 移植治疗较有价值的细胞资源。

Mechanisms of H+ modulation of glycinergic response in rat sacral dorsal commissural neurons

Many ionotropic receptors are modulated by extracellular H+. So far, few studies have directly addressed the role of such modulation at synapses. In the present study, we investigated the effects of changes in extracellular pH on glycinergic miniature inhibitory postsynaptic currents (mIPSCs) as well as glycine-evoked currents (I-Gly) in mechanically dissociated spinal neurons with native synaptic boutons preserved. H+ modulated both the mIPSCs and I-Gly, biphasically, although it activated an amiloride-sensitive inward current by itself. Decreasing extracellular pH reversibly inhibited the amplitude of the mIPSCs and I-Gly, while increasing external pH reversibly potentiated these parameters. Blockade of acid-sensing ion channels (ASICs) with amiloride, the selective antagonist of ASICs, or decreasing intracellular pH did not alter the modulatory effect of H+ on either mIPSCs or I-Gly, H+ shifted the EC50 of the glycine concentration-response curve from 49.3 +/- 5.7 muM at external pH 7.4 to 131.5 +/- 8.1 muM at pH 5.5, without altering the Cl- selectivity of the glycine receptor (GlyR), the Hill coefficient and the maximal I-Gly, suggesting a competitive inhibition of I-Gly by H+. Both Zn2+ and H+ inhibited I-Gly. However, H+ induced no further inhibition of I-Gly in the presence of a saturating concentration of Zn2+. In addition, H+ significantly affected the kinetics of glycinergic mIPSCs and I-Gly. It is proposed that H+ and/or Zn2+ compete with glycine binding and inhibit the amplitude of glycinergic mIPSCs and I-Gly. Moreover, binding of H+ induces a global conformational change in GlyR, which closes the GlyR Cl- channel and results in the acceleration of the seeming desensitization of IGly as well as speeding up the decay time constant of glycinergic mIPSCs. However, the deprotonation rate is faster than the unbinding rate of glycine from the GlyR, leading to reactivation of the undesensitized GlyR after washout of agonist and the appearance of a rebound I-Gly. H+ also modulated the glycine cotransmitter, GABA-activated current (I-GABA). Taken together, the results support a 'conformational coupling' model for H+ modulation of the GlyR and suggest that W may act as a novel modulator for inhibitory neurotransmission in the mammalian spinal cord.

Murine MyaK, a member of a family of yeast YAK1-related genes, is highly expressed in hormonally modulated epithelia in the reproductive system and in the embryonic central nervous system

We have cloned a mouse homologue (designated Myak) of the yeast protein kinase YAK1. The 1210 aa open reading frame contains a putative protein kinase domain, nuclear localization sequences and PEST sequences. Myak appears to be a member of a growing family of YAK1-related genes that include Drosophila and human Minibrain as well as a recently identified rat gene ANPK that encode a steroid hormone receptor interacting protein. RNA blot analysis revealed that Myak is expressed at low levels ubiquitously but at high levels in reproductive tissues, including testis, epididymis, ovary, uterus, and mammary gland, as well as in brain and kidney. In situ hybridization analysis on selected tissues revealed that Myak is particularly abundant in the hormonally modulated epithelia of the epididymis, mammary gland, and uterus, in round spermatids in the testis, and in the corpora lutea in the ovary, Myak is also highly expressed in the aqueduct of the adult brain and in the brain and spinal cord of day 12.5 embryos, Mol. Reprod. Dev. 55:372-378, 2000. (C) 2000 Wiley-Liss, Inc.

Developmental toxicity and alteration of gene expression in zebrafish embryos exposed to PFOS

Perfluorooetanesulfonate (PFOS) is a persistent organic pollutant, the potential toxicity of which is causing great concern. In the present study, we employed zebrafish embryos to investigate the developmental toxicity of this compound. Four-hour post-fertilization (hpf) zebrafish embryos were exposed to 0.1, 0.5, 1, 3 and 5 mg/L PFOS. Hatching was delayed and hatching rates as well as larval survivorship, were significantly reduced after the embryos were exposed to 1, 3 and 5 mg/L PFOS until 132 hpf. The fry displayed gross developmental malformations, including epiboly deformities, hypopigmentation, yolk sac edema, tail and heart malformations and spinal curvature upon exposure to PFOS concentrations of I mg/L or greater. Growth (body length) was significantly reduced in the 3 and 5 mg/L PFOS-treated groups. To test whether developmental malformation was mediated via apoptosis, flow cytometry analysis of DNA content, acridine orange staining and TUNEL assay was used. These techniques indicated that more apoptotic cells were present in the PFOS-treated embryos than in the control embryos. Certain genes related to cell apoptosis, p53 and Bax, were both significantly up-regulated upon exposure to all the concentrations tested. In addition, we investigated the effects of PFOS on marker genes related to early thyroid development (hhex and pax8) and genes regulating the balance of androgens and estrogens (cyp19a and cyp19b). For thyroid development, the expression of hhex was significantly up-regulated at all concentrations tested, whereas pax8 expression was significantly up-regulated only upon exposure to lower concentrations of PFOS (0.1, 0.5, 1 mg/L). The expression of cyp19a and of cyp19b was significantly down-regulated at all exposure concentrations. The overall results indicated that zebrafish embryos constitute a reliable model for testing the developmental toxicity of PFOS, and the gene expression patterns in the embryos were able to reveal some potential mechanisms of developmental toxicity. (C) 2008 Elsevier Inc. All rights reserved.

Developmental expression of CagMdkb during gibel carp embryogenesis

Midkine (Mdk) genes have been revealed to have different expression patterns in vertebrates and therefore, additional studies on Mdk expression patterns are required in more species. In this study, CagMdkb has been cloned and characterized from a SMART cDNA library of 10-somite stage embryos of Carassius auratus gibelio. Its full length cDNA is 1091 bp and encodes a sequence of 147 amino acids, which shows 97.3% identity to zebrafish Mdkb on the amino acid level. RT-PCR analysis reveals that CagMdkb is first transcribed in gastrula embryos and maintains a relatively stable expression level during subsequent embryogenesis. Western blot analysis reveals a 19 kDa maternal CagMdkb protein band and the zygotic CagMdkb protein is expressed from gastrula stage. At around 10 somite stage, the 19 kDa CagMdkb is processed to another protein band of about 17 kDa, which might be the secreted form with the 21-residue signal peptide removed. With immunofluorescence analysis, maternal CagMdkb protein was found to be localized in each blastamere cell of early embryos. The zygotic CagMdkb positive fluorescence signal was detected from a pair of large neurons at 18-somite stage. At the later stages, CagMdkb protein was also extended to numerous small neurons in the forebrain, midbrain and hindbrain, as well as to nerve fibers in the spinal cord. Co-localization with 3A10 antibody revealed CagMdkb immunoreactivity on developing Mauthner neurons, a member of reticulospinal neurons. In addition, ectopic expression of CagMdkb in early embryos of gibel carp and zebrafish suppressed head formation and CagMdkb function was found to depend on secretory activity. All these findings indicate that CagMdkb plays an important role in neural development during gibel carp embryogenesis and there is functional conservation of Mdkb in fish head formation.

Design and path planning for a remote-brained service robot

This article introduced an effective design method of robot called remote-brain, which is made the brain and body separated. It leaves the brain in the mother environment, by which we mean the environment in which the brain's software is developed, and talks with its body by wireless links. It also presents a real robot TUT06-B based on this method which has human-machine interaction, vision systems, manipulator etc. Then it discussed the path planning method for the robot based on ant colony algorithm in details, especially the Ant-cycle model. And it also analyzed the parameter of the algorithm which can affect the convergence. Finally, it gives the program flow chat of this algorithm.

System size dependence of associated yields in hadron-triggered jets STAR Collaboration

We present results on the system size dependence of high transverse momentum di-hadron correlations at root s(NN) = 200 GeV as measured by STAR at RHIC. Measurements in d + Au, Cu + Cu and Au + Au collisions reveal similar jet-like near-side correlation yields (correlations at small angular separation Delta phi similar to 0, Delta eta similar to 0) for all systems and centralities. Previous measurements have shown Chat the away-side (Delta phi similar to pi) yield is suppressed in heavy-ion collisions. We present measurements of the away-side Suppression as a function of transverse momentum and centrality in Cu + Cu and Au + Au collisions. The suppression is found to be similar in Cu + Cu and An + An collisions at a similar number of participants. The results are compared to theoretical calculations based on the patron quenching model and the modified fragmentation model. The observed differences between data and theory indicate that the correlated yields presented here will further constrain dynamic energy loss models and provide information about the dynamic density profile in heavy-ion collisions. (C) 2009 Elsevier B.V. All rights reserved.

文冠果壳苷对侧脑室注射Aβ_(1-42)致小鼠学习记忆障碍的改善作用

目的研究文冠果壳苷对侧脑室注射β肽淀粉样蛋白1-4(2Aβ1-42)小鼠学习记忆障碍的改善作用,并初步探讨了其作用机制。方法通过避暗和水迷宫实验检测了小鼠的学习记忆能力;使用分光光度计检测小鼠大脑组织中乙酰胆碱酯酶(AchE)和胆碱乙酰转移酶(ChAT)活性的变化。结果小鼠侧脑室注射凝聚态Aβ1-42显著降低了小鼠被动回避学习记忆能力及空间学习记忆能力(P<0.01),并且小鼠大脑AchE和ChAT活性显著降低(P均<0.05);而文冠果壳苷能够显著减少避暗错误次数,延长潜伏期(分别P<0.05、P<0.01);剂量依赖性地缩短水迷宫实验中小鼠到达安全台的游泳时间(分别P<0.05、P<0.01),不同程度地抑制Aβ1-42模型小鼠的AchE和ChAT活性的降低(P<0.05和<0.01)。结论文冠果壳苷对侧脑室注射Aβ1-42小鼠学习记忆障碍具有显著的改善作用,其作用机制可能与保护中枢胆碱能神经细胞,改善中枢胆碱能神经系统功能等有关。