Splicing Factor Transformer-2β (Tra2β) Regulates the Expression of Regulator of G Protein Signaling 4 (RGS4) Gene and Is Induced by Morphine

Regulator of G protein signaling 4 (RGS4) is a critical modulator of G protein-coupled receptor (GPCR)-mediated signaling and plays important roles in many neural process and diseases. Particularly, drug-induced alteration in RGS4 protein levels is associated with acute and chronic effects of drugs of abuse. However, the precise mechanism underlying the regulation of RGS4 expression is largely unknown. Here, we demonstrated that the expression of RGS4 gene was subject to regulation by alternative splicing of the exon 6. Transformer-2β (Tra2β), an important splicing factor, bound to RGS4 mRNA and increased the relative level of RGS4-1 mRNA isoform by enhancing the inclusion of exon 6. Meanwhile, Tra2β increased the expression of full-length RGS4 protein. In rat brain, Tra2β was co-localized with RGS4 in multiple opioid action-related brain regions. In addition, the acute and chronic morphine treatment induced alteration in the expression level of Tra2β in rat locus coerulus (LC) in parallel to that of RGS4 proteins. It suggests that induction of this splicing factor may contribute to the change of RGS4 level elicited by morphine. Taken together, the results provide the evidence demonstrating the function of Tra2β as a new mediator in opioid-induced signaling pathway via regulating RGS4 expression.     

Heat shock protein 70 interacts with aquaporin-2 and regulates its trafficking

The trafficking of aquaporin-2 (AQP2) involves multiple complex pathways, including regulated, cAMP-, and cGMP-mediated pathways, as well as a constitutive recycling pathway. Although several accessory proteins have been indirectly implicated in AQP2 recycling, the direct protein-protein interactions that regulate this process remain largely unknown. Using yeast two-hybrid screening of a human kidney cDNA library, we have identified the 70-kDa heat shock proteins as AQP2-interacting proteins. Interaction was confirmed by mass spectrometry of proteins pulled down from rat kidney papilla extract using a GST-AQP2 C-terminal fusion protein (GST-A2C) as a bait, by co-immunoprecipitation (IP) assays, and by direct binding assays using purified hsc70 and the GST-A2C. The direct interaction of AQP2 with hsc70 is partially inhibited by ATP, and the Ser-256 residue in the AQP2 C terminus is important for this direct interaction. Vasopressin stimulation in cells enhances the interaction of hsc70 with AQP2 in IP assays, and vasopressin stimulation in vivo induces an increased co-localization of hsc70 and AQP2 on the apical membrane of principal cells in rat kidney collecting ducts. Functional knockdown of hsc70 activity in AQP2 expressing cells results in membrane accumulation of AQP2 and reduced endocytosis of rhodamine-transferrin. Our data also show that AQP2 interacts with hsp70 in multiple in vitro binding assays. Finally, in addition to hsc70 and hsp70, AQP2 interacts with several other key components of the endocytotic machinery in co-IP assays, including clathrin, dynamin, and AP2. To summarize, we have identified the 70-kDa heat shock proteins as a AQP2 interactors and have shown for hsc70 that this interaction is involved in AQP2 trafficking.     

钠-氢交换蛋白 mRNA在人胎儿两个发育阶段中的组织特异性表达

钠－氢交换蛋白（Na^ -H^ exchangers,NHE）至少包含6个不同的亚型,生长因子可激活其表达。目前,对在发育过程中NHE的表达了解甚少。本文利用RT－PCR观察了4种NHE亚型的mRNA在人胎儿的两个不同发育阶段（11周、16周）在不同组织中的表达,以研究它们的发育调控。结果显示,NHE1 mRNA在两种胎龄的多种组织中均有表达,和16周胚胎相比,11周的胚胎的NHE1 mRNA的表达较弱,并且表现出明显的组织差异。据此推测,NHE1的管家（house-keeping）功能可能至少在11周就开始形成,而最迟在16周已基本建立;NHE2和NHE3 mRNA在11周和16周的胚胎组织中的特异性表达呈现相反的变化趋势及组织分布上的重叠,后者与NHE2和NHE3在成人组织中的分布及功能的重叠的特点相吻合;NHE5 mRNA的表达在11周的胚胎组织中比较普遍,而在16周的胚胎组织中则局限在小脑组织中,本研究表明,在人胚胎发育11－16周期间,NHE的组织特异性表达表现出时间依赖性的调控,而在不迟于胚胎发育的第16周,具有“管家功能”的NHE1的基因表达已与成人相似。     

SRp38基因研究进展

SR蛋白在前体mRNA可变剪接调控中发挥重要作用。可变剪接调节因子SRp38作为一种新近发现的具有神经及生殖组织特异性的SR蛋白,有典型的SR蛋白结构特征并能够调控GluR-B、TRK-C以及NCAML1等基因的可变剪接,但与其他SR蛋白不一致的是,SRp38可以在一定条件下(有丝分裂M期,热休克)抑制前体mRNA剪接,从而防止错误剪接的出现。SRp38的RRM结构域可以识别特殊的RNA序列并跟U1snRNP结合,而其RS结构域则参与调控前体mRNA剪接。SRp38的磷酸化状态可以影响其调控功能的发挥,在有丝分裂M期及热休克时,该蛋白质均呈去磷酸化状态。SRp38在爪蟾胚胎神经发育过程中发挥作用并且可以同TLS(translocation liposarcoma)蛋白相互作用,提示其可能通过调节前体mRNA可变剪接在神经系统的发育分化以及在肿瘤的发生中扮演角色。     

中国紫金牛属圆齿组果实微形态特征研究

对国产紫金牛科紫金牛属圆齿组(Ardisia Sect. Crispardisia)16种植物的果实表面进行了扫描电镜观察。经观察果实为球形,表面多无毛,稀有柔毛。多数种具腺点,极少数无腺点或不明显,腺点疣状、粒状或横棒状,腺点常与果实表面突起具对应关系。果实表面纹饰大致可分为3种:基本平滑、多皱、波状网纹。某些种不同部位的纹饰差异较为明显。本研究结果对种间鉴别具有重要意义,并且不支持将珍珠伞A. maculosa归并入钮子果A. polysticta中。     

Morphogenesis of Pistillate Flowers of Cercidiphyllum japonicum(Cercidiphyllaceae)

Floral morphogenesis and the development of Cercidiphyllum japonicum Sieb.et Zucc.were observed by scanning electronmicroscopy(SEM).The results showed that the pistillate inflorescences were congested spikes with the flowers arrangedopposite.Great differences between the so-called"bract"and the vegetative leaf were observed both in morphogenesis andmorphology.In morphogenesis,the"bract"primordium is crescent-shaped,truncated at the apex and not conduplicate,has no stipule primordium at the base but does have some inconspicuous teeth in the margin that are not glandular.Theleaf primordium is triangular,cycloidal at the apex,conduplicate,has two stipule primordia at the base,has one gland-toothat the apex occurring at first and some gland-teeth in the margin that occur later.In morphology,the"bract"is also differentto the vegetative leaf in some characteristics that were also illustrated in the present paper.Based on the hypothesis thatthe bract is more similar to the vegetative leaf than the tepal,we considered that the so-called"bract"of C.japonicum mightbe the tepal of the pistillate flower in morphological nature.Therefore,each pistillate flower contains a tepal and a carpel.We did not find any trace of other floral organs in the morphogenesis of the pistillate flower.Therefore we consideredthat the unicarpellate status of extant Cercidiphyllum might be to highly reduce and advance characteristics that make theextant Cercidiphyllum isolated from both fossil Cercidiphyllum-like plants and its extant affinities.     

深圳马峦山华南紫萁群落及其物种多样性特征研究

对深圳马峦山地区华南紫萁占优势的植物群落进行研究,结果表明：华南紫萁群落中共有维管植物48科69属82种,种类组成具有明显的南亚热带性质;年龄结构显示群落的主要优势种属于稳定型种群,群落总体处于稳定状态;群落的频度指数规律为A＞B＜C＞D＞E,与RaunKiaer频度定律、海南岛山地雨林优势种群的频度规律等不相符合;华南紫萁种群树高和个体数百分比显示,种群高度0.3~0.7 m植株占比例较大,种群处于旺盛发展期;物种多样性指数为SP＝16.42,SW＝4.56,均匀度指数为E＝0.33,E′＝0.83,群落物种多样性指数和均匀度指数均较高,接近典型南亚热带常绿阔叶林顶极群落类型。本研究揭示了华南紫萁野外生存状态,为该种的保护和利用提供了基础资料。     

黄三七(毛茛科)茎次生木质部导管穿孔板的研究

利用扫描电子显微镜对东亚特有植物黄三七（Souliea vaginata（Maxim.） Franch.）茎的次生木质部离析材料进行了观察,结果表明,黄三七茎次生木质部中的导管分子端壁上具网状穿孔板（麻黄式穿孔板）、梯状穿孔板、网状-梯状混合穿孔板、网状-梯状-单穿孔混合型穿孔板、梯状-单穿孔混合型穿孔板及单穿孔板,同时也观察到了端壁多穿孔板和侧壁穿孔板,并对不同类型穿孔板中纹孔膜的残留也进行了观察。其中,网状穿孔板、各种过渡类型的穿孔板均为毛茛科植物中首次报道。根据观察结果,对导管分子穿孔板的演化及黄三七属植物的系统位置进行了分析。     

黄嘌呤氧化还原酶的结构、功能和作用

黄嘌呤氧化还原酶(XOR)参与人体内的嘌呤代谢,并且是这一代谢过程的限速酶。其终产物是活性氧(包括OH·、H2O2和O2-)和尿酸。这两种产物参与体内多种生理活动。从XOR基因的结构、XOR蛋白的分子结构和基本功能、控制XOR活性的多个环节以及XOR的两种催化产物活性氧和尿酸在生理和病理情况下的功能及机制进行了总结,以期对XOR的发现、研究历史及现状和有待解决的问题有一个系统的了解。     

The Anticonvulsant and Neuroprotective Effects of Oxysophocarpine on Pilocarpine-Induced Convulsions in Adult Male Mice

Epilepsy is one of the prevalent and major neurological disorders, and approximately one-third of the individuals with epilepsy experience seizures that do not respond well to available medications. We investigated whether oxysophocarpine (OSC) had anticonvulsant and neuroprotective property in the pilocarpine (PILO)-treated mice. Thirty minutes prior to the PILO injection, the mice were administrated with OSC (20, 40, and 80 mg/kg) once. Seizures and electroencephalography (EEG) were observed, and then the mice were killed for Nissl and Fluoro-jade B (FJB) staining. The oxidative stress was measured at 24 h after convulsion. Western blot analysis was used to examine the expressions of the Bax, Bcl-2, and Caspase-3. In this study, we found that pretreatment with OSC (40, 80 mg/kg) significantly delayed the onset of the first convulsion and status epilepticus (SE) and reduced the incidence of SE and mortality. Analysis of EEG recordings revealed that OSC (40, 80 mg/kg) significantly reduced epileptiform discharges. Furthermore, Nissl and FJB staining showed that OSC (40, 80 mg/kg) attenuated the neuronal cell loss and degeneration in hippocampus. In addition, OSC (40, 80 mg/kg) attenuated the changes in the levels of Malondialdehyde (MDA) and strengthened glutathione peroxidase and catalase activity in the hippocampus. Western blot analysis showed that OSC (40, 80 mg/kg) significantly decreased the expressions of Bax, Caspase-3 and increased the expression of Bcl-2. Collectively, the findings of this study indicated that OSC exerted anticonvulsant and neuroprotective effects on PILO-treated mice. The beneficial effects should encourage further studies to investigate OSC as an adjuvant in epilepsy, both to prevent seizures and to protect neurons in brain.