84 resultados para actin filament
Resumo:
A novel technique for high-power extracavity pulse compression with a nonlinear solid material is demonstrated. Before spectral broadening by self-phase modulation in the solid material, a short filament generated in argon is used as a spatial filter, which works for a uniform spectrum broadening over the spatial profile. Compensated by chirped mirrors, a 15-fs pulse is generated from a 32-fs input laser pulse. A total transmission larger than 80% after the solid material is achieved.
Resumo:
Temperature controlled filamentation is experimentally demonstrated in a temperature gradient gas-filled tube. The proper position of the tube is heated by a furnace and two ends of the tube are cooled by air. The experimental results show that multiple filaments are shrunken into a single filament or no filament only by increasing the temperature at the beginning of the filament. This technique offers another degree of freedom of controlling the filamentation and opens a new way for intense monocycle pulse generation through gradient temperature in a noble gas.
Resumo:
Conical emission (CE) has been investigated experimentally by laser pulses with different pulse durations and spectral bandwidths. The results show that the overall CE curve will shift as the varying of spectral bandwidth of pump laser pulse. But for pump laser pulses which have same spectral bandwidth but different pulse duration, the CE angles will be same at the spectral region close to the pump wavelength while will be different at the spectral region far away from the pump wavelength. We have also fitted the measured CE angles with X-wave model. The calculated curves and the measured CE curves match reasonably well. The best fits indicate that the group velocity of the filament pulse may be greatly controlled by controlling the spectral bandwidth of pump laser pulse. (C) 2008 Elsevier B.V. All rights reserved.
Formation of X-waves at fundamental and harmonics by infrared femtosecond pulse filamentation in air
Resumo:
We experimentally observe the formation of X-waves at fundamental, third harmonic, and fifth harmonic wavelengths by infrared (central wavelength at similar to 1500 nm) femtosecond laser pulse filamentation in air. By fitting the angularly resolved spectra of the fundamental and harmonic waves using X-wave relations, we confirm that all the X-waves have nearly the same group velocity, indicating that they are locked in space and time during their propagation in filament.
Resumo:
The authors report the measurement of the angularly resolved spectrum of the third harmonic generated in a femtosecond filament in air and its evolution with increasing pump power. Pumped by a focused infrared ultrashort pulse with a carrier wavelength of 1270 nm, a pulse duration of similar to 20 fs, and pulse energy up to 487 mu J, the generated third harmonic is composed of an on-axis emission and a conical ring emission. When the pump power is sufficiently high, angularly resolved spectra with significant X-like feature could be observed, indicating the formation of nonlinear X wave at third harmonic. (c) 2008 American Institute of Physics.
Resumo:
A novel technique of controlling the evolution of the filamentation was experimentally demonstrated in an argon gas-filled tube. The entrance of the filament was heated by a furnace and the other end was cooled with air, which resulted in the temperature gradient distribution along the tube. The experimental results show that multiple filaments are merged into a single filament and then no filament by only increasing the temperature at the entrance of the filament. Also, the filament can appear and disappear after increasing the local temperature and input pulse energy in turn. This technique offers another degree of freedom to control the filamentation and opens a new way for multi-mJ level monocycle pulse generation through filamentation in the noble gas.
Resumo:
We investigate the evolution of filamentation in air by using a longitudinal diffraction method and a plasma fluorescence imaging technique. The diameter of a single filament in which the intensity is clamped increases as the energy of the pump light pulse increases, until multiple filaments appear. (c) 2006 Optical Society of America.
Resumo:
The authors report the investigation of filament and supercontinuum generation by focusing a femtosecond laser beam into water doped with silver nanoparticles. The silver nanoparticles enhance the nonlinear optical response of water, leading to broadening of supercontinuum spectra in self-focused femtosecond filaments. During the propagation of the supercontinuum light in the filament, the silver nanoparticles preferentially scatter the short-wavelength light near the plasmon resonant wavelength peak, followed by the scattering of the long-wavelength light. Thus, a side view of the filament shows a full-color spectrum in the visible range, which is herein called "rainbow filament." (c) 2007 American Institute of Physics.
Resumo:
Two-cycle optical pulses with duration of 5 fs and energy of 0.7 mJ have been generated at 1 kHz by compressing the 38 fs laser pulses from a carrier-envelope phase (CEP) controlled Ti:sapphire laser system through a cascade filamentation compression technique. A simple and effective method is developed to suppress multiple filament formation and stabilize a single filament by inserting a soft aperture with an appropriate diameter into the driving laser beam prior to focusing, resulting in an excellent compressed beam quality. The good beam quality and potentially higher peak power make this ultrashort laser pulse source a significant tool for high-field physics applications. (C) 2007 Optical Society of America.
Resumo:
水稻Angrp-1基因编码富含甘氨酸蛋白质。氨基酸序列分析表明该蛋白在氨基端可能存在一个信号肽序列,其后为富含甘氨酸序列。亲水性指数显示AnGRP-1蛋白包含两个区域。第一个区域由氨基端的1到27氨基酸残基组成。为强疏水性区。该肽段的氨基端为带正电荷号肽特征。与水稻Osgrp-1和菜豆grp1.8基因编码的信号肽进行同源性比较,发现这三种信号肽具有alafLvLLNIg同源序列。AnGRP-1蛋白的第二区域由第28到183氨基酸残基组成。该区域富含甘氨酸,具轻度亲水性,并且含有15个Gly-Tyr-Gly基序,这些基序中的酷西安酸残基之间相互作用,有可能形成分子间或分子内的连接。 利用水稻原生质体瞬间表达系统,比较Angrp-1基因启动子283bp和-1020bp片段,以及Ubiquitin启动子、Actin启动子和35S启动子的活性。结果表明,在水稻原生质体瞬间表达中,Angrp-1基因的两个启动子片段活性较低,接近本底。三个对照的组成型启动子中,以Ubiquitin启动子的活性最强,Actin启动子次之, 35S启动子最弱。根据以上结果推测水稻Angrp-1基因的表达可能具有组织或发育特异异表达和特性。 为研究Angrp-1基因的稳定表达与调控特性,将Angrp-1基因的启动子缺失片段1020bp、941bp、568bp和182bp分别与GUS基因融合,得到pGRP6-121、pGRP7-121、pGRP8-121和pGRP9-121等4个克隆,进行烟草转化,获得转基因植株。GUS活性测定表明在-1020到-568之间,随缺失增多,启动子活性下降。当缺失至-182时,182bp的启动子片段活性高于568bp片段。因此,Angrp-1基因5'端-1020到-568之间存在正调控序列,在-568至-182之间存在负调控序列。对pGRP6-121的烟草转基因植株进行GUS组织染色和活性测发现,GUS基因在维管组织优先表达。在根、茎、叶三种器官中,叶和茎的表达量最高。营养生长期的表达量高于生殖生长期。用2283bp和1020bp的启动子片段分别与GUS基因融合,转化水稻,转化体的GUS组织学染色表明Gus基因具有维管束优先表达特性。因此,Angrp-1基因的表达具有较明显的组织、器官和发育特异性。 水稻AnGRP-1蛋白的免疫组织定位分析表明,Angrp-1基因产物主要定位于水稻的维管组织。利用免疫金方法对AnGRP-1蛋白进行超微结构定位,发现金颗粒主要分布于水稻细胞的细胞壁中。因此,可基本确定AnGRP-1蛋白为细胞壁蛋白。在细胞内的内质网和高尔基体上面或附近也有金颗粒的分布,表明至少有部分AnGRP-1蛋白经过内质网-高尔基体-质膜途径,最终达到细胞壁。 为研究Angrp-1基因编码的信号肽功能,将其信号肽与GUS的氨基端融合,分别置于35S启动子和Angrp-1基因1020bp启动子片段控制之下,得到克隆p35s-SP-GUS-121和pGRP6-SP-GUS-121,分别以pBI121和pGRP6-121作对照,进行烟草基因转化。转化体的GUS活性分析表明,在35S启动子控制下,加与不加信号肽对GUS活性影响不大。但是,在Angrp-1基因的1020bp启动子片段作用下,信号肽与GUS融合后的转化体中其本不表现GUS活性。利用免疫金方法进行的超微结构定位表明,在p35S-SP-GUS-121和pGRP6-SP-GUS-121转化的烟草植株中,均可在细胞壁中检测室GUS产物的存在,因此AnGRP-1的信号肽能引导GUS从细胞质分泌细胞壁中。 以Angrp-1基因5'端的-568到+1作探针,发现Angrp-1基因以在水稻窄叶青和京系17两个亲本中存在多态性,并进一步将Angrp-1基因定位于水稻第10号染色体上。
Resumo:
在高等植物有性生殖过程中,花粉管作为运输精子到胚珠的载体,它的生长具有高度极性化,并且要依赖于微丝。由于花粉管本身所具的特性,它已经成为研究细胞相互识别、胞内和胞外信号的模式系统。本文为了研究微丝在白杄(Picea meyeri Rehd. et Wils.)花粉管生长中的作用,我们应用不同浓度的微丝聚合抑制剂Latrunculin B (LATB) 处理花粉管,并通过激光共聚焦显微镜观察微丝聚合状态的动态变化。结果发现,在低浓度下的LATB能使花粉管中的微丝严重解聚,且抑制其顶端生长。 我们进一步利用蛋白质组学的手段,分析了白杄花粉管微丝解聚后蛋白质的表达图谱。通过双向电泳分离出500个左右考马斯亮兰染色的蛋白质斑点,经过软件分析发现,其中大部分蛋白质的表达量未发生变化,而只有110个蛋白斑点有较大变化。将这些蛋白斑点从胶上切下酶解后用于质谱鉴定,最终鉴定出35个蛋白,其中有18个为上调蛋白,17个下调蛋白。根据其主要功能,通常可分为碳水化合物代谢、胁迫反应、信号和细胞扩展等几类。我们发现由于微丝解聚引起的能量代谢水平降低,可能与依赖于信号传导的微丝重组过程相关。此外,当LATB浓度增加到50 nM时,与细胞壁多糖合成相关的两个蛋白,如reversibly glycosylated polypeptide和type IIIa membrane protein cp-wap13几乎不表达,这说明当微丝聚合完全被抑制后,依赖于微丝的分泌系统也受到影响,从而引起相应蛋白质变化,最终导致细胞壁成分合成的减少。细胞骨架蛋白actin的下调,进一步说明微丝在花粉管生长过程中起着提供或支持的一种机制,也就是能调节信号介导的花粉管生长,并使其在特定的时期到达特定的部位,从而完成植物的受精作用。
Resumo:
花粉管是种子植物受精过程中的雄性生殖单位载体,具有典型的顶端极性生长特点,因此成为近年来研究植物细胞间相互识别、胞内外信号传递的模式系统。裸子植物花粉管与被子植物花粉管相比具有萌发时间长、生长缓慢等特点。一氧化氮(NO)作为一个重要的胞内信号分子,参与调节植物生长发育和多种生理过程,但是,有关NO对花粉管生长的作用机制目前尚不清楚。本研究以裸子植物白皮松(Pinus bungeana)花粉为材料,应用不同浓度的NO释放剂SNAP和SNP, NO清除剂cPTIO和哺乳动物一氧化氮合酶抑制剂L-NNA处理,对白皮松花粉萌发和花粉管伸长进行了细胞学研究,从而为进一步揭示NO调控裸子植物花粉管生长机理提供参考。 本论文首先研究了NO释放剂、清除剂及NOS抑制剂对白皮松花粉萌发和花粉管生长的影响。结果显示,SNAP和SNP能够促进白皮松花粉萌发和花粉管伸长,并具有浓度效用,但对其花粉管的形态特征无明显影响;cPTIO和L-NNA能够抑制白皮松花粉萌发和花粉管生长,并且具有浓度效应,同时还可使花粉管顶端膨大呈球形,并丧失花粉管的极性生长。运用NO特异探针DAF-2DA标记显示,SNAP和SNP处理可促进花粉管胞内NO产生;经cPTIO和L-NNA处理后,花粉管内荧光强度比对照花粉管明显减弱。上述结果表明,NO参与裸子植物花粉管极性生长,并且适量NO可以促进花粉管的生长。 用显微注射技术将Ca2+特异探针注射入白皮松花粉管,以检测白皮松花粉管胞内Ca2+浓度梯度。结果显示,SNAP和SNP处理后,NO荧光增加的同时,花粉管顶端Ca2+浓度梯度增加。与此相反,cPTIO和L-NNA处理后,NO荧光降低的同时,花粉管顶端Ca2+浓度梯度也相应降低。应用非损伤微测技术测定花粉管胞外Ca2+内流,结果显示,SNAP和SNP促进胞外Ca2+内流,而cPTIO和L-NNA则抑制胞外Ca2+内流。据此,我们推测,在白皮松花粉管中,NO可能通过调节胞外Ca2+内流来调节胞内Ca2+浓度梯度,然而,我们不能排除在NO信号传递过程中,胞内Ca2+库可能影响胞内Ca2+浓度梯度。 通过微丝特异探针标记的白皮松花粉管显示,SNAP和SNP可使花粉管顶端细微丝束解聚,而cPTIO和L-NNA处理后,花粉管中微丝聚合,尤其在花粉管顶端形成粗的微丝束,并一直延伸到花粉管的最顶端。结合上述Ca2+结果,我们认为,经NO处理后,白皮松花粉管微丝骨架的动态变化,可能是通过Ca2+浓度来进行调控的。通过FM4-64探针标记显示,正常生长白皮松花粉管胞吞作用主要集中在顶端和亚顶端,并且形成倒“V”形的分布模式,SNAP与SNP处理后荧光分布模式与对照花粉管类似,但达到饱和所用时间相对较短;而L-NNA处理后顶端膨大丧失极性的花粉管,荧光分布在膨大花粉管靠近质膜的区域,未见倒“V”形分布模式;经L-NNA处理后,顶端没有膨大的花粉管,荧光几乎均匀分布于整个花粉管,并且L-NNA处理后达到饱和的时间较对照长。上述结果表明,适量NO能够促进白皮松花粉管胞吞。 免疫抗体标记技术分析发现,对照花粉管的酯化果胶质和AGPs都集中在顶端,酸性果胶质分布在侧壁,胼胝质均匀分布在整个花粉管壁上。L-NNA处理后的花粉管,在其顶端出现酸性果胶和酯化果胶,以及有胼胝质积累,而AGPs则分布在花粉管的基部。运用傅里叶红外光谱技术(Fourier Transform Infared Spectroscopy, FTIR)技术分析白皮松花粉管顶端细胞壁成分,结果显示,SNAP处理后花粉管顶端酯化果胶增加而酸性果胶降低;与之相反,经L-NNA处理后的花粉管,其顶端酯化果胶降低而酸性果胶增加。 综上所述,在白皮松花粉管中,NO促进胞外Ca2+内流,从而维持胞内Ca2+浓度梯度,进而影响花粉管顶端微丝骨架的组装,促进囊泡运输,使花粉管顶端酯化果胶累积,最终促进花粉管的正常生长。
Resumo:
The origin of cytoskeleton and the origin of relevant intracellular transportation system are big problems for understanding the emergence of eukaryotic cells. The present article summarized relevant information of evidences and molecular traces on the origin of actin, tubulin, the chaperonin system for folding them, myosins, kinesins, axonemal dyneins and cytoplasmic dyneins. On this basis the authors proposed a series of works, which should be done in the future, and indicated the ways for reaching the targets. These targets are mainly: 1) the reconstruction of evolutionary path from MreB protein of archaeal ancestor of eukaryotic cells to typical actin; 2) the finding of the MreB or MreB-related proteins in crenarchaea and using them to examine J. A. Lake's hypothesis on the origin of eukaryote from "eocytes" (crenarchaea); 3) the examinations of the existence and distribution of cytoskeleton made of MreB-related protein within coccoid archaea, especially in amoeboid archaeon Thermoplasm acidophilum; 4) using Thermoplasma as a model of archaeal ancestor of eukaryotic cells; 5) the searching for the homolog of ancestral dynein in present-day living archaea. During the writing of this article, Margulis' famous spirochaete hypothesis on the origin of flagella and cilia was unexpectedly involved and analyzed from aspects of tubulins, dyneins and spirochaetes. Actually, spirochaete cannot be reasonably assumed as the ectosymbiotic ancestor of eukaryotic flagella and cilia, since their swing depends upon large amount of bacterial flagella beneath the flexible outer wall, but not depends upon their intracellular tubules and the assumed dyneins. In this case, if they had "evolved" into cilia and lost their bacterial flagella, they would immediately become immobile! In fact, tubulin and dynein-like proteins have not been found in any spirochaete.
Resumo:
TRIM5α(tripartite motif protein 5-alpha)蛋白是恒河猴体内一种非常重要的限制因子,能抑制人免疫缺陷病毒(HIV-1,human immunodeficiency virus type 1)、马感染性贫血病毒(EIAV, equine infectious anemia virus)和猫免疫缺陷病毒(FIV, feline immunodeficiencyvirus)等逆转录病毒的复制.恒河猴TRIM5α的组织分布以及在受到外界刺激时TRIM5α mRNA表达量的变化研究还未见报道.本研究从中国恒河猴的各组织中提取总RNA,以β-actin基因作为内参照,通过半定量RT-PCR检测各组织中TRIMSα mRNA的表达.选择HIV-GFP-VSVG假病毒感染外周血单核细胞(peripheral blood mononuclear cell,PBMC),非特异性刺激剂--佛波脂(Phorbol myfismte acetate,PMA)+离子霉素(ionomycin,Ion)及CD28抗体+CD49d抗体分别共刺激恒河猴PBMC,研究不同刺激对恒河猴TRIM5α mRNA表达水平的影响.结果表明:TRIM5α mRNA表达于所研究的恒河猴21种组织中,免疫系统和泌尿生殖系统组织中表达量最高,而神经系统组织,如大脑、脊髓中表达较少,其他组织中未见明显的表达差异;HIV-GFP-VSVG感染和用PMA+Ion、CD28抗体+CD49d抗体分别共刺激PBMC能促进PBMC中TRIM5α mRNA的转录水平的上调.