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

Physical Blocking Neural Tube Closure Affects Radial Intercalation and Neural Crest Midline-directed Migration in Xenopus Dorsal Explants

神经管闭合缺陷(NTDs)是一种严重的先天畸形疾病,在新生儿中有千分之一的发病率.神经管融合前后,多种组织参与形态发生运动.神经管一经融合,神经嵴细胞就会向背侧中线方向产生单极突出并向此方向迁移形成神经管的顶部.与此同时,神经管从腹侧开始发生辐射状切入以实现单层化.在此,我们在非洲爪蟾的移植体中机械阻断神经管的闭合以检测其细胞运动及随后的图式形成.结果显示神经管闭合缺陷的移植体不能形成单层化的神经管,并且神经嵴细胞滞留在侧面区域不能向背侧中线迁移,而对神经前体标记基因的检测显示神经管的背腹图式形成并未受到影响.以上结果表明神经管的融合对于辐射状切入和神经嵴细胞向背侧中线方向的迁移过程是必需的,而对于神经管的沿背腹轴方向的图式形成是非必需的.

Extremely low-frequency electromagnetic field exposure during chronic morphine treatment strengthens downregulation of dopamine D2 receptors in rat dorsal hippocampus after morphine withdrawal

The aim of this study was to investigate the effect of extremely low-frequency electromagnetic field (ELF-EMF) exposure during morphine treatment on dopamine D2 receptor (D2R) density in the rat dorsal hippocampus following withdrawal. Rats were exposed t

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.

A morphologically variant natural population of cyprinids without dorsal fin possibly derived from Mystacoleucus marginatus (Osteichthyes: Teleostes) from the Lancang Jiang River, Yunnan, China

In July 2001, 15 grotesque cyprinid specimens were collected in the Lancangjiang River (of the upper reaches of the Mekong River) in Menglun Town, Xishuanbanna Prefecture, Yunnan Province, PR China. These specimens are characterized by surprising characte

A Dorsal homolog (FcDorsal) in the Chinese shrimp Fenneropenaeus chinensis is responsive to both bacteria and WSSV challenge

Rel/NF kappa B is a family of transcription factors. In the present study, a Rel/NF kappa B family member, Dorsal homolog (FcDorsal) was cloned from the Chinese shrimp Fenneropenaeus chinensis. The full length cDNA of FcDorsal consists of 1627 bp, revealed a 1071 bp open reading frame encoding 357 aa. The predicted molecular weight (MW)of the deduced amino acid sequence of FcDorsal was 39.78 kDa, and its theoretical pl was 8.85. Amino acid sequence analysis showed that FcDorsal contains a Rel homolog domain (RHD) and an IPT/TIG (Ig-like, plexins and transcriptions factors) domain. The signature sequence of dorsal protein existed in the deduced amino acid sequence. Spatial expression profiles showed that FcDorsal had the highest expression level in the hemocytes and lymphoid organ (Oka). The expression profiles in the hemocytes and lymphoid organ were apparently modulated when shrimp were stimulated by bacteria or WSSV. Both Gram-positive (G(+)) bacteria (Micrococcus lysodeikticus) and Gram-negative (G(-)) bacteria (Vibrio anguillarium) injection to shrimp caused the up-regulation of FcDorsal at the transcription level. DsRNA approach was used to study the function of FcDorsal and the data showed that FcDorsal was related to the transcription of Penaeidin 5 in shrimp. The present data provide clues that FcDorsal might play potential important roles in the innate immunity of shrimp. Through comparison of the expression profiles between FcDorsal and another identified Rel/NF kappa B member (FcRelish) in shrimp responsive to WSSV challenge, we speculate that FcDorsal and FcRelish might play different roles in shrimp immunity. (C) 2010 Elsevier Ltd. All rights reserved.

Two new species of Rhagophthalmus Motschulsky (Coleoptera : Rhagophthalmidae) from Yunnan, south-western China, with notes on known, species

Rhagophthalmus fugongensis Li & Liang and Rhagophthalmus lufengensis Li & Ohba are introduced as new to science. The first record of R. tonkineus in China is provided. The male genitalia of R. gibbosulus and R. giganteus are described and illustrated. The female and larva of R. giganteus and the female of R. semisulcatus are illustrated. The distribution of Rhagophthalmus is discussed and a rectification of the relative arrangement of different parts of the male genitalia is provided (the base-piece is on the ventral side of the male genitalia, and the parameres on the dorsal side).

基因工程化分泌神经营养因子一3的鼠胚神经干细胞实验研究

：探索以Lentivirus为载体，构建同时表达绿色荧光蛋白(GFP)和神经营养因子一3(NT一3)的基因工程化鼠胚神经于细胞(NSC)的可行性。方法：体外分离培养鼠胚NSC，用同时携带NT一3和GFP的lentivirus转染构建工程化NSC；用荧光显微镜、鼠胚背根神经结培养(Dorsal Root Ganglion，DRG)、Westem blot等方法检测基因工程NSC 的转基因表达。结果：荧光显微镜观察到几乎100％的工程化NSC表达GFP：DRG培养和Westem blot检测到基因工程化NSC能高效分泌NT一3蛋白。结论：以IJentivirus为载体，构建同时携带并稳定表达GFP和N‘r_3的基因工程化鼠胚NSC是可行的，可为脊髓损伤基础研究提供有价值的细胞资源。

Lentivirus介导表达多基因的人胚基因工程神经干细胞的实验研究

目的:探索以Lentivirus为载体,构建同时携带并表达多基因的基因工程人胚神经干细胞(hum an neu鄄ral stem cell,hNSC)的可行性,为脊髓损伤治疗的研究提供材料。方法:培养和鉴定hNSC;用携带绿色荧光蛋白(green fluorescence protein,GFP)和神经营养因子-3(neurotrophic factor-3,NT-3)的Lentivirus转染hNSC;用荧光显微镜观察、鼠胚背根神经结培养(dorsal root ganglion,DRG)和Slot blot等方法检测基因工程hNSC的多基因表达情况。结果:培养获得了大量的hNSC;荧光显微镜观察到几乎100%的hNSC表达GFP;基因工程hNSC的培养液能促使大鼠DRG旺盛生长;Slot blot检测到基因工程hNSC能高效分泌NT-3蛋白。结论:以Lentivirus为载体能构建同时携带并稳定表达多基因的基因工程hNSC,为脊髓损伤治疗的基础研究及进一步临床应用提供了有价值的细胞资源。

A review of the cyprinoid fish genus Barbodes Bleeker, 1859, from Yunnan, China, with descriptions of two new species

Fishes of the genus Barbodes in Yunnan have been reviewed and 2 new species are described on the basis of specimens deposited in Kunming institute of Zoology, Chinese Academy of Sciences. Counts and measurements follow Chu and Chen (1989). Barbodes heterostomus is distinguished by its terminal mouth, with gape being horizontal in the male, and acclivitous in the female, last unbranched dorsal ray smooth with upper 1/3 articulated; dorsal fin origin anterior to pelvic fin origin; no dark lateral band on sides of body; gill rakers 13-19; lateral line scales 24-29; longest caudal ray length about 2 times that of shortest. It is distributed in Longchuanjiang and Dayingjiang (upper Irrawaddy). Barbodes baoshanensis is distinguished by its smooth last unbranched dorsal ray with upper 1/3-1/2 articulated; dorsal fin origin anterior to pelvic fin origin; sides of body with a dark longitudinal band; gill rakers 13-14; lateral line scales 23-28. It occurs in Nujiang (upper Salween) and Longchuanjiang. A key to the species of Barbodes in Yunnan is provided.

Taxonomic review of the genus Sinocrossocheilus Wu (Teleostei : Cyprinidae), with a description of four new species

All Sinocrossocheilus species, except S. microstomatus, are reviewed. Four new species, S. labiata, S. papillolabra, S. nigrovittata, and S. longibulla, are described. The genus Sinocrossocheilus differs from other genera of Cyprinidae by the last simple dorsal fin ray being unserrated and unossified, the last unbranched anal fin ray being unserrated and unossified, the 5-branched anal fin rays, the mouth gap being inferior, the rostral cap covering the lower jaw and connecting directly with the lower lip, a row of fleshy lobes on the lower jaw, and a cloudy black spot above the pectoral fin. Sinocrossocheilus labiata is small and has 22 predorsal scales; S. longibulla has a very large air bladder; S. papillolabra possesses a well-developed ventral fin and a wide band covered by fleshy papillae on the lower lip; and S. nigrovittata possesses black longitudinal stripes along the lateral line. Crossocheilus bamaensis and Crossocheilus liuchengensis are transferred to the genus Sinocrossocheilus. Sinocrossocheilus species are endemic to the central and eastern Yunnan-Guizhou Plateau of China, where river systems are anfractuous, including seasonal rivers, cave rivers, underground rivers, and streamlets between mountains. These separated rivers probably provide conditions for the allopatric speciation of the Sinocrossocheilus.

A new species of the genus Tor from Yunnan, China (Teleostei : Cyprinidae)

A new species of the subfamily Barbinae, Tor yingjiangensis, is recognized from Yunnan province, China. It can be distinguished from other Tor species by the following combination of characters: last simple dorsal-fin ray osseous and non-serrated; no forward directed predorsal procumbent spine; head length considerably longer than body depth; no tubercles on the snout or sides of the face; 18-20 gill rakers on the outside of first gill arch; 24-26 lateral line scales; median lobe of lower lip short, its posterior margin truncate, not extending to the vertical across the inner corners of the mouth; and the condition of the lower lip consistent in individuals of different sizes. The new species has been misidentified previously as Tor putitora (Hamilton, F. 1822. An account of the fishes found in the River Ganges and its Branches. Edinburgh & London. 405 pp), which occurs in the Ganges and Indus River basins. The new species can easily be distinguished from T. putitora by having 3-3.5 (vs. 2.5) scales from lateral line to pelvic-fin origin, shorter caudal peduncle length (13.0% vs. 17.2% of standard length), lesser body depth (26.4% vs. 24.0% of standard length) and longer caudal peduncle depth (12.0% vs. 10.9% of standard length), no longitudinal stripe present along side of body, and eyes visible in ventral view of head.