Molecular cloning and characterization of RGA1 encoding a G protein α subunit from rice (Oryza sativa L. IR-36)

A cDNA clone, RGA1, was isolated by using a GPA1 cDNA clone of Arabidopsis thaliana G protein subunit as a probe from a rice (Oryza sativa L. IR-36) seedling cDNA library prepared from roots and leaves. Sequence analysis of genomic clone reveals that the RGA1 gene has 14 exons and 13 introns, and encodes a polypeptide of 380 amino acid residues with a calculated molecular weight of 44.5 kDa. The encoded protein exhibits a considerable degree of amino acid sequence similarity to all the other known G protein subunits. A putative TATA sequence (ATATGA), a potential CAAT box sequence (AGCAATAC), and a cis-acting element, CCACGTGG (ABRE), known to be involved in ABA induction are found in the promoter region. The RGA1 protein contains all the consensus regions of G protein subunits except the cysteine residue near the C-terminus for ADP-ribosylation by pertussis toxin. The RGA1 polypeptide expressed in Escherichia coli was, however, ADP-ribosylated by 10 M [adenylate-³²P] NAD and activated cholera toxin. Southern analysis indicates that there are no other genes similar to the RGA1 gene in the rice genome. Northern analysis reveals that the RGA1 mRNA is 1.85 kb long and expressed in vegetative tissues, including leaves and roots, and that its expression is regulated by light.     

The effect of the loading condition corresponding to functional shoulder activities on trabecular architecture of glenoid

There is little information on bone morphology as it relates to shoulder activities. This study investigated how loads corresponding to functional shoulder activities affect the trabecular architecture of the glenoid. Two different protocols were used. Protocol 1 investigated the material and morphological characteristics of the glenoid by analyzing digitized trabecular bone images obtained from 12 cadaver scapula specimens. Protocol 2 used a finite element analysis (FEA) to compute the principal stress trajectories acting within the glenoid. The principal stresses were derived for five loading conditions, which represent typical functional shoulder activities. The study showed that shoulder activity involved in carrying a light load makes the greatest contribution to the trabecular architecture compared with other shoulder activities considered in this study (p<0.05). With all of the activities considered in this study, the lateral region, particularly in the anterior and posterior portions, showed greater deviation and greater sensitivity to variation under loading conditions than did the other regions (p<0.05). These results suggest that owing to the extra sensitivity of the anterior and posterior parts of the lateral region, these regions may be more informative in the analysis of the trabecular architecture following shoulder musculoskeletal injuries. These results may provide essential design information for shoulder prostheses and contribute to an understanding of morphological changes resulting from injury.     

A post-burst after depolarization is mediated by group i metabotropic glutamate receptor-dependent upregulation of Ca(v)2.3 R-type calcium channels in CA1 pyramidal neurons

Activation of group I metabotropic glutamate receptors (subtypes mGluR1 and mGluR5) regulates neural activity in a variety of ways. In CA1 pyramidal neurons, activation of group I mGluRs eliminates the post-burst afterhyperpolarization (AHP) and produces an afterdepolarization (ADP) in its place. Here we show that upregulation of Ca(v)2.3 R-type calcium channels is responsible for a component of the ADP lasting several hundred milliseconds. This medium-duration ADP is rapidly and reversibly induced by activation of mGluR5 and requires activation of phospholipase C (PLC) and release of calcium from internal stores. Effects of mGluR activation on subthreshold membrane potential changes are negligible but are large following action potential firing. Furthermore, the medium ADP exhibits a biphasic activity dependence consisting of short-term facilitation and longer-term inhibition. These findings suggest that mGluRs may dramatically alter the firing of CA1 pyramidal neurons via a complex, activity-dependent modulation of Ca(v)2.3 R-type channels that are activated during spiking at physiologically relevant rates and patterns.     

园林植物新害虫——女贞粗腿象甲

本文首次报道了发生于贵阳地区女贞属植物上的一种新害虫——女贞粗腿象甲Ochyromera ligustri Warner,简要介绍其分布、寄主植物、形态特征、生物学特性,并提出相应的防治建议。     

骨形态发生蛋白9诱导成骨相关TGFβⅡ型受体的鉴定分析

前期研究发现骨形态发生蛋白9(bone morphogenetic protein 9,BMP9)具有较强的诱导骨形成的能力,但目前对其诱导骨形成相关的分子机制了解甚少．采用重组腺病毒的方法成功构建显性负性突变的转化生长因子(TGF)βⅡ型受体的腺病毒,将其与BMP9共同导入靶细胞,通过体外细胞实验和体内动物实验,初步鉴定分析与BMP9诱导成骨密切相关的TGFβⅡ型受体．体外细胞实验发现：三种显性负性突变的TGFβⅡ型受体,即dnBMPRⅡ、dnActRⅡ和dnActRⅡB在体外能抑制由BMP9诱导的碱性磷酸酶(alkaline phosphatase,ALP)表达和钙盐沉积,并可抑制BMP9诱导的Smad信号途径的激活．动物实验也显示,dnBMPRⅡ、dnActRⅡ和dnActRⅡB可抑制BMP9诱导的异位成骨,提示相应的野生型TGFβⅡ型受体即BMPRⅡ、ActRⅡ和ActRⅡB很可能与BMP9诱导成骨有关．而靶细胞中均只表达BMPRⅡ和ActRⅡ,并不表达ActRⅡB.随后,采用RNA干扰(RNA interference,RNAi)对BMPRⅡ和ActRⅡ进行基因沉默,结果发现BMPRⅡ和ActRⅡ的表达受到抑制后,由BMP9诱导的荧光素酶活性和ALP活性也相应受到抑制．因此,BMPRⅡ和ActRⅡ是与BMP9诱导成骨分化相关的TGFβⅡ型受体．     

短时高温胁迫对瓜实蝇生长发育及繁殖的影响

为明确瓜实蝇对短时高温胁迫的耐受性。利用人工气候箱模拟短时高温胁迫,测定了不同高温处理(34、36、38、40、42、44、46、48℃)12 h,对不同发育阶段瓜实蝇的存活率和生长发育的影响。结果表明短时高温显著影响瓜实蝇的存活,随温度升高,瓜实蝇各虫态的存活率逐渐降低;高温处理12 h后瓜实蝇卵、幼虫、蛹、雌成虫、雄成虫的致死中温度LT 50分别为35.48、37.55、41.85、43.62、43.32℃;34~42℃短时高温胁迫对瓜实蝇各虫态的发育历期无明显影响,44℃时其发育历期均显著增长;46℃、48℃处理下各虫态死亡率较高,不能正常发育;随着处理温度的升高,雌成虫产卵前期不断增长,单雌产卵量呈下降趋势,成虫寿命不断缩短,后代雌性比增大。44℃及以上的短时高温胁迫不利于瓜实蝇的生长发育,40℃及以上的短时高温胁迫不利于瓜实蝇的繁殖,雌性瓜实蝇对短时高温的胁迫的适应性强于雄性,随着处理温度的升高,后代雌性比例增大。     

骨形态发生蛋白9通过JNKs激酶途径调控间充质干细胞成骨分化

前期研究发现骨形态发生蛋白9(bone morphogenetic protein 9,BMP9)除了通过经典Smad途径外,也可通过丝裂原活化蛋白激酶(mitogen activated protein kinases,MAPKs)中的p38激酶途径调控间充质干细胞成骨分化．本研究继续探讨MAPKs的重要成员c-Jun氨基末端激酶(c-Jun N-terminal kinases,JNKs)对于BMP9诱导间充质干细胞成骨分化的调控作用．利用BMP9重组腺病毒感染间充质干细胞,通过体外细胞实验和体内动物实验,初步分析BMP9是否可通过JNKs激酶途径调控间充质干细胞成骨分化．结果表明：BMP9可通过促进JNKs激酶磷酸化而导致其活化;JNKs抑制剂SP600125可抑制由BMP9诱导的间充质干细胞的碱性磷酸酶(alkaline phosphatase,ALP)活性、骨桥蛋白(osteocpontin,OPN)和骨钙素(osteocalcin,OCN)表达以及钙盐沉积;利用抑制剂SP600125抑制JNKs激酶活性后,BMP9诱导Runx2的表达和转录活性,以及Smad经典途径的激活也相应受到抑制;RNA干扰导致JNKs基因沉默同样也可抑制BMP9诱导的间充质干细胞成骨分化以及裸鼠皮下异位成骨．因此,BMP9可通过活化JNKs激酶途径,从而调控间充质干细胞成骨分化．     

骨形态发生蛋白9通过p38激酶途径调控间充质干细胞C3H10T1/2成骨分化

前期研究发现骨形态发生蛋白9(bone morphogenetic protein 9,BMP9)具有较强的诱导间充质干细胞成骨分化的能力.为进一步揭示其诱导和调控间充质干细胞成骨分化的机理,利用BMP9重组腺病毒感染间充质干细胞C3H10T1/2,通过体外细胞实验和体内动物实验,初步分析BMP9是否可通过p38激酶途径调控间充质干细胞成骨分化.结果发现,BMP9可以通过促进p38激酶磷酸化而导致其活化,p38抑制剂SB203580可抑制由BMP9诱导的C3H10T1/2细胞的碱性磷酸酶(alkalinephosphatase,ALP)活性、骨桥蛋白(osteopontin,OPN)表达和钙盐沉积,而且利用抑制剂SB203580抑制p38激酶活性后,BMP9诱导的Smad经典途径的激活也相应受到抑制,RNA干扰导致p38基因沉默同样也可抑制BMP9诱导的ALP活性、OPN表达、钙盐沉积以及裸鼠皮下异位成骨.因此,BMP9可通过活化p38激酶途径调控间充质干细胞C3H10T1/2成骨分化.