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High UV Excitation Intensity Induces Photoconversion of DAPI During Wide-Field Microscopy 高紫外激发强度诱导DAPI在宽视场显微镜下的光转换
Pub Date : 2014-04-30 DOI: 10.26443/msurj.v9i1.158
C. Brown
Background: Multi-color fluorescence microscopy is dependent on the spectral specificity of the dyes and probes used for localization. One of the most commonly used fluorescent DNA dyes is DAPI, which is usually excited by UV light to emit in the blue visible light range. Herein, we describe a pattern of decreasing DAPI fluorescence upon extended UV exposure, closely followed by an increase in emission maxima in the green range. Methods: UV-induced photo-conversion of DAPI to green-fluorescing photoproducts was studied on Chinese hamster ovary cells using wide-field fluorescence microscopy, at different UV exposure times and intensities. Imaging was done in repetitive cycles, by alternating between a DAPI and FITC filter cube and following this with a 1 second UV excitation time. The effect of differing UV light intensities on the photo- conversion process was not previously described in the literature. Results: Upon image analysis from a large sample of cells, the rate of photo-conversion was shown to be dependent on both the duration of UV excitation and the intensity of the UV light source. Both the process of DAPI depletion and photo-product growth showed biphasic exponential patterns of change. Furthermore, the level of DAPI fluorescence intensity was found to be negatively correlated with the green fluorescence of the photo-product. Limitations: This study did not examine the effect of differing mounting media or a variation in DAPI concentration on the rate of DAPI photo-conversion. Also, the exact light dosage to the system was not measured from the 100W Hg bulb. Photo-bleaching of green fluorescence in cells not stained with DAPI was not measured to control for bleaching of endogenous cell molecules. Conclusion: Based on our findings, a set of recommendations was formulated in order to help reduce the effects of UV-induced DAPI photo-conversion.
背景:多色荧光显微镜依赖于用于定位的染料和探针的光谱特异性。最常用的荧光DNA染料之一是DAPI,它通常被紫外光激发,在蓝色可见光范围内发射。在这里,我们描述了一种模式,减少DAPI荧光在延长紫外线照射,紧随其后的是在绿色范围内的发射最大值的增加。方法:利用宽视场荧光显微镜研究了不同紫外照射时间和强度下,DAPI在中国仓鼠卵巢细胞中转化为绿色荧光产物。通过在DAPI和FITC滤光片立方体之间交替进行重复循环成像,然后进行1秒的紫外线激发时间。不同紫外光强度对光转化过程的影响在以前的文献中没有描述过。结果:通过对大量细胞样本的图像分析,显示出光转换速率取决于紫外线激发的持续时间和紫外线光源的强度。DAPI耗竭和光产物生长过程均呈双相指数型变化。此外,DAPI的荧光强度水平与光产物的绿色荧光呈负相关。局限性:本研究没有检查不同的安装介质或DAPI浓度变化对DAPI光转化率的影响。此外,系统的确切光剂量不是从100W Hg灯泡测量的。未检测DAPI染色细胞中绿色荧光的光漂白,以控制内源性细胞分子的漂白。结论:基于我们的研究结果,提出了一套建议,以帮助减少紫外线诱导的DAPI光转化的影响。
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引用次数: 0
Microstimulation to the Middle Temporal Area and its Effect on the Generation of Microsaccades 中颞区微刺激及其对微跳的影响
Pub Date : 2014-04-30 DOI: 10.26443/msurj.v9i1.157
Haider Riaz, A. Golzar
It has been repeatedly shown that neural activity in different brain structures can be correlated with perceptual and cognitive functions using electrical microstimulation. Currently, microstimulation is the only method that can demonstrate causal links between neural activity and specific cognitive functions. This study investigates the effects of microstimulation to the MT area of the visual cortex on the production of microsaccades for several seconds. Microsaccades are a type of fixational eye movement characterized by their quickness and low amplitude. The preliminary findings in this paper suggest that microstimulation to the MT area causes an increase in frequency and peak velocity of microsaccades. However, a more in-depth analysis to establish a correlation between the two was unsuccessful. These results suggest further investigation into the effects of microstimulation on microsaccades – using more sophisticated and reliable data collecting and analyzing techniques – is necessary.
已经多次证明,不同脑结构的神经活动可以通过电微刺激与感知和认知功能相关联。目前,微刺激是唯一可以证明神经活动和特定认知功能之间因果关系的方法。本研究探讨了对视觉皮层MT区进行数秒微刺激对微跳产生的影响。微扫视是一种以快速、低振幅为特征的眼球注视运动。初步结果表明,微刺激可引起微跳频的增加和峰值速度的增加。然而,更深入的分析,以建立两者之间的相关性是失败的。这些结果表明,有必要使用更复杂、更可靠的数据收集和分析技术,进一步研究微刺激对微眼动的影响。
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引用次数: 0
Parrotfish Body Size As An Indicator of Diurnal Fish Species Richness On Fringing Coral Reefs in Barbados 巴巴多斯边缘珊瑚礁上鹦嘴鱼体大小作为鱼类物种日丰富度的指标
Pub Date : 2014-04-30 DOI: 10.26443/msurj.v9i1.155
A. Eger, Catherine Pigeon-Dubeau, Lauriane Sibileau
Background: Coral reefs around the world are host to some of the most condensed and varied ecosystems. However, over the past decades, their biodiversity has alarmingly decreased. A rapid and reliable way of assessing their status is urgently needed to monitor and prevent their decline. The purpose of this study is to assess whether or not family Scaridae (common name: parrotfish) body size can be used as an index to evaluate the diurnal fish diversity on coral reefs. Methods: We selected 6 accessible reefs on the West coast of Barbados and measured the size of parrotfish we encountered, as well as the number of fish species present on the reef; this data was then plotted and statistically analyzed to establish a possible correlation. Results: Our results show that reef fish species richness is strongly correlated to both the ratio of large to small parrotfish and the average parrotfish size. It is however the large to small ratio that exhibited the strongest relationship. Further analysis revealed that the population size of large parrotfish correlates with reef biodiversity with a Pearson’s r coefficient of 0.97. Conclusion: This relationship could be due to the fact that large parrotfish (greater than or equal to 20 cm) have increased grazing rates compared to smaller ones, this increased grazing promotes coral polyp recruitment, thereby benefiting the diversity observed on coral reefs. Further research is needed to elucidate whether these results can be extended to other areas of the Caribbean and provide conservation efforts with an easy tool to survey and protect coral reef ecosystems.
背景:世界各地的珊瑚礁是一些最浓缩和最多样化的生态系统的宿主。然而,在过去的几十年里,它们的生物多样性已经惊人地减少。目前迫切需要一种快速可靠的方法来评估它们的状况,以监测和防止它们的减少。本研究的目的是评估是否可以用鹦嘴鱼科(俗称:鹦嘴鱼)的体型作为评价珊瑚礁鱼类昼夜多样性的指标。方法:我们在巴巴多斯西海岸选择了6个可到达的珊瑚礁,测量了我们遇到的鹦嘴鱼的大小,以及珊瑚礁上存在的鱼类种类数量;然后对这些数据进行绘制和统计分析,以确定可能的相关性。结果:珊瑚鱼物种丰富度与大、小鹦嘴鱼的比例和鹦嘴鱼的平均尺寸密切相关。然而,大与小的比例表现出最强的关系。进一步分析表明,大型鹦嘴鱼种群规模与珊瑚礁生物多样性的相关系数为0.97。结论:这种关系可能是由于大型鹦嘴鱼(大于或等于20厘米)比较小的鹦嘴鱼增加了放牧率,这种增加的放牧促进了珊瑚虫的补充,从而有利于在珊瑚礁上观察到的多样性。需要进一步的研究来阐明这些结果是否可以推广到加勒比的其他地区,并为保护工作提供一个简单的工具来调查和保护珊瑚礁生态系统。
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引用次数: 0
Development and Characterization of a Directional Gamma-ray Detector 定向伽玛射线探测器的研制与表征
Pub Date : 2014-04-30 DOI: 10.26443/msurj.v9i1.156
Felix Cormier, Marcel P. Georgin, Stephen Koelbl, R. Oda
Background: This work characterizes the first generation of detectors from the Hanna Laboratory to implement Silicon Photomultipliers and a heptagonal scintillator conguration. The purpose of the device is to determine the angle at which a radioactive source is located. Methods: The development of the detector consisted of three phases: construction(September 2012- December 2012), simulation and characterization (April 2013). The experimental portion of the work consisted of placing a 137 Cs source at an arbitrary location, measuring the count rates in each scintillator panel and analysing the results. Results: The detector’s function was validated by confirming the inverse square law with a radioactive source moving away from the detector. Furthermore, with a x2 summation method of analysis the angular position of a source was determined with an accuracy of 10˚ and a precision of 12˚. With a normalisation method of analysis the angular position of a source was found with an accuracy of 2˚ and a corresponding precision of 2˚. Limitations: The quality of the electronics handling the signal from the silicon photomultipliers limited our resolution. Occasional double counts occur when a large amount of energy is imparted to the scintillator. Furthermore, the custom-built circuitry lowered the signal-to-noise ratio such that large distances were not feasible due to electronic noise constraints. Finally, simulation data analysis showed that the break of one circuit only had a small effect on the x2 method of analysis. Conclusions: In conclusion, the design of the detector and the analysis techniques were shown to be suitable for short range angular resolution of a gamma-ray source. Both distance trials and a simulation of the detector prototype confirmed the validity of our design and of the analysis methods used. These promising results at short distances motivate further work in electronic circuit design to improve the range while maintaining both accuracy and precision.
背景:这项工作的特点是第一代探测器从汉纳实验室实现硅光电倍增管和七方体闪烁体配置。该装置的目的是确定放射源所在的角度。方法:探测器的研制分为构建(2012年9月- 2012年12月)、仿真和表征(2013年4月)三个阶段。这项工作的实验部分包括在任意位置放置一个137cs光源,测量每个闪烁体面板的计数率并分析结果。结果:当辐射源远离探测器时,通过确认平方反比定律验证了探测器的功能。利用x2求和分析法确定了光源的角位置,精度为10˚,精度为12˚。用归一化分析方法确定了光源的角位置,其精度为2˚,相应的精度为2˚。限制:处理硅光电倍增管信号的电子设备的质量限制了我们的分辨率。当大量的能量被传递给闪烁体时,偶尔会发生重复计数。此外,定制电路降低了信噪比,因此由于电子噪声的限制,长距离传输是不可行的。最后,仿真数据分析表明,其中一条电路的断路对x2分析方法的影响很小。结论:探测器的设计和分析技术适用于伽玛射线源的短距离角分辨。距离试验和探测器原型的模拟都证实了我们的设计和所使用的分析方法的有效性。这些在短距离上的有希望的结果激励了电子电路设计的进一步工作,以提高范围,同时保持准确性和精度。
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引用次数: 0
Neuroplasticity and Post-Synaptic Rebound-Induced Spiking at Purkinje Cell-Deep Cerebellar Nuclei Synapses 浦肯野细胞-小脑深部核突触的神经可塑性和突触后反弹诱导的尖峰
Pub Date : 2014-04-30 DOI: 10.26443/msurj.v9i1.160
Jenna Hotton
Background: Within the cerebellum white matter are located four pairs of nuclei, collectively known as the deep cerebellar nuclei (DCN) (1). In the cerebellum, signal integration from pre-cerebellar structures via excitatory parallel fibers and climbing fibers in the cerebellar cortex occurs in GABAergic Purkinje cells (PC) (2). The main target of these PC cells is the DCN (2) and approximately 85% of GABAergic input on the DCN is from PCs (3). Furthermore, PCs outnumber DCN neurons (26:1) (2). Therefore, despite receiving substantial inhibition from Purkinje cells, DCN neurons are still active at rest showing regular spiking or spontaneous bursts (4). DCN neurons fire spontaneously at approximately 10-50 Hz (5). Given this unique anatomy of PC-DCN synapses, characterization of this synaptic circuit is important in understanding the overall role of the DCN in the brain. Methods: The findings of 28 studies, including a few reviews, are reported in this paper. Studies selected focused principally on characterization of DCN circuitry properties and the role these properties have in the functioning of the DCN. Most studies employed in vivo and/or in vitro cellular recordings in rodents, among other models. Studies ranged from 1984 to 2013. Summary: This review outlines current findings on the forms of plasticity found in the DCN, the function of the DCN and the connections between the DCN and other brain regions. In short, neurons in the DCN demonstrate both synaptic and non-synaptic plasticity. Cerebellar involvement in motor activity has been extensively studied therefore, not surprisingly; DCN neurons form connections with the motor cortex but also the prefrontal cortex. PC input on the DCN influences spike rate and timing through fluctuations in PC synchrony, and rebound depolarization.
背景:在小脑白质中有四对核,统称为小脑深部核(DCN)(1)。在小脑中,通过小脑皮层的兴奋性平行纤维和攀爬纤维,来自小脑前结构的信号整合发生在gaba能浦肯野细胞(PC)中(2)。这些PC细胞的主要目标是DCN (2), DCN上大约85%的gaba能输入来自PC(3)。此外,PC的数量超过DCN神经元(26:1)(2)。尽管受到浦肯野细胞的大量抑制,DCN神经元在休息时仍然活跃,表现出规律的峰突或自发爆发(4)。DCN神经元自发放电的频率约为10-50 Hz(5)。鉴于PC-DCN突触的独特解剖结构,表征这种突触回路对于理解DCN在大脑中的整体作用非常重要。方法:对28篇相关研究的结果进行回顾性分析。所选择的研究主要集中在DCN电路特性的表征以及这些特性在DCN功能中的作用。大多数研究采用啮齿类动物和其他模型的体内和/或体外细胞记录。研究时间从1984年到2013年。摘要:本文概述了DCN可塑性的形式、DCN的功能以及DCN与大脑其他区域之间的联系的最新发现。简而言之,DCN中的神经元表现出突触和非突触的可塑性。因此,小脑参与运动活动已被广泛研究,这并不奇怪;DCN神经元与运动皮层以及前额皮质形成连接。在DCN上的PC输入通过PC同步的波动和反弹去极化影响尖峰速率和定时。
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引用次数: 0
Studying a poxvirus gene capture model through recombination and reactivation 痘病毒基因重组再激活捕获模型的研究
Pub Date : 2013-03-31 DOI: 10.26443/msurj.v8i1.106
K. Johnson, D. Evans
Introduction: Vaccinia poxvirus (VACV) is a double stranded DNA virus that replicates in the cytoplasm of infected cells. Some VACV genes resemble homologs of host genes and appear to have been captured from the cell; however, since poxviruses are confined to the cytoplasm, researchers are unclear as to how these viruses acquire this homology (1). If a cellular mRNA was accidentally reverse transcribed into cDNA, which could occur during retrovirus co-infection, a poxvirus might be able to incorporate this sequence into its own genome through rare non-homologous (homology-independent) recombination. Methods: We modeled this process using two different recombination systems and substituted a DNA encoding mycophenolic acid (MPA), a selectable marker, for the hypothetical non-homologous host cDNA. We prepared DNA constructs containing this marker along with 20 base pairs homologous to the 5’ and 3’ flanking regions of the VACV-encoded NotI restriction site. A construct without this flanking homology was also prepared. The “passive” recombination system used a helper poxvirus to reactivate VACV DNA; in contrast, VACV infected BSC40 cells were transfected with the construct in the “active” recombination system. Results: The “passive” recombination system generated 105 PFU/mL of reactivated VACV; however, no recombinants containing the selectable marker were detected. The “active recombination” method generated 106 PFU/mL of total VACV and approximately 10 PFU/mL of recombinant virus for both homology containing and non-homology containing constructs. Discussions: We were unable to determine the recombination frequency of the “passive system” because recombinant virus was not detected. Based on virus titers determined from plaque assays, we approximated the recombination frequency of the “active system” to be ≤ 10-5. We are currently cloning and sequencing viruses resulting from non-homologous recombination to determine where the MPA marker is located. Preliminary analysis of these types of clones (data not shown in this paper) suggests that the transfected DNAs are being incorporated into a diversity of sites, some located near the boundary of the VACV genome where the right terminal inverted repeat begins. In summary, our findings suggest that the recombination frequencies in both methods are very low and better methods of selection are needed to observe these rare events. Future studies of recombinant clones are needed to gain a better understanding of this non- homologous gene capture process.
简介:痘痘病毒(VACV)是一种双链DNA病毒,在感染细胞的细胞质中复制。有些疫苗病毒基因类似于宿主基因的同源物,似乎是从细胞中捕获的;然而,由于痘病毒局限于细胞质,研究人员尚不清楚这些病毒是如何获得这种同源性的(1)。如果细胞mRNA在逆转录病毒共感染过程中意外地逆转录为cDNA,痘病毒可能通过罕见的非同源(同源无关)重组将该序列整合到其自身基因组中。方法:我们使用两种不同的重组系统模拟了这一过程,并将一个编码霉酚酸(MPA)的DNA(一个可选择的标记)替换为假设的非同源宿主cDNA。我们制备了包含该标记的DNA构建体以及与vacv编码的NotI酶切位点的5 '和3 '侧区同源的20个碱基对。也制备了一个没有这种侧翼同源的结构体。“被动”重组系统使用辅助痘病毒来重新激活VACV DNA;相比之下,在“活性”重组系统中转染了感染了VACV的BSC40细胞。结果:“被动”重组系统产生105 PFU/mL的再激活VACV;然而,没有检测到含有可选择标记的重组。“活性重组”方法产生了106 PFU/mL的总VACV和大约10 PFU/mL的重组病毒,包含同源性和非同源性。讨论:我们无法确定“被动系统”的重组频率,因为没有检测到重组病毒。根据从空斑测定中确定的病毒滴度,我们估计“活性系统”的重组频率≤10-5。我们目前正在克隆和测序由非同源重组产生的病毒,以确定MPA标记的位置。对这些克隆类型的初步分析(数据未在论文中显示)表明,转染的dna被整合到多种位点,其中一些位于靠近VACV基因组边界的右末端倒置重复开始的地方。总之,我们的研究结果表明,两种方法的重组频率都很低,需要更好的选择方法来观察这些罕见的事件。为了更好地了解这种非同源基因捕获过程,需要对重组克隆进行进一步的研究。
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引用次数: 0
Stress reactivity during evaluation by the opposite sex: comparison of responses induced by different psychosocial stress tests 异性评价时的应激反应:不同心理社会应激测试引起的反应的比较
Pub Date : 2013-03-31 DOI: 10.26443/msurj.v8i1.105
L. Goff, N. Ali, J. Pruessner
It is becoming increasingly difficult for researchers to continue their high rates of publication when funding budgets are running tighter than ever. It is therefore in a researcher’s best interest to utilize more economical tests whenever possible. This project aims to compare various stress tests in order to determine whether the new, cost-efficient Maastricht Acute Stress Test (MAST) activates a physiological and subjective stress response with the same effectiveness as pre-existing, more resource-intensive tests. This study demonstrated that the MAST produces a response similar to that of the previously predominant Trier Social Stress Test (TSST). Meanwhile, database data shows that the purely physiological Cold Pressor Task (CPT) lags behind in terms of response elicited. These findings may allow for a more cost-efficient yet highly effective stress task to become available to researchers.
在资金预算比以往任何时候都紧张的情况下,研究人员要继续保持高发表率变得越来越困难。因此,尽可能使用更经济的测试符合研究人员的最大利益。本项目旨在比较各种压力测试,以确定新的、具有成本效益的马斯特里赫特急性压力测试(MAST)是否能激活生理和主观压力反应,其效果与现有的、资源更密集的测试相同。这项研究表明,MAST产生的反应类似于以前占主导地位的特里尔社会压力测试(TSST)。同时,数据库数据显示,纯生理冷压任务(CPT)在引发的反应方面滞后。这些发现可能为研究人员提供一种更经济高效的压力任务。
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引用次数: 4
Operational transformation in cooperative software systems 协同软件系统中的操作转换
Pub Date : 2013-03-31 DOI: 10.26443/msurj.v8i1.113
C. Leung
Modern cooperative software systems involve multiple concurrent users undertaking a common task in a real-time distributed environment, such as editing a shared text document. Maintaining data consistency, transaction causality, and replication convergence in such an environment, while providing fast client responsiveness, is a substantial challenge for classical distributed computing techniques. Operational transformation (OT) is a class of concurrency algorithms and data models that supports these functionalities, which has drawn significant research attention in the past decade. In this review, we discuss the basic components of operational transformation models, the algorithms involved, and their actual implementations in real-world networked systems. We compare several existing OT control algorithms, the transformation functions and properties supported by each of the algorithms, and the trade-offs that are made with respect to each one. The data and operational models used in OT are well suited for high- latency environments such as the Internet, making them more frequently used in modern web services. Although many different OT control algorithms exist, choosing the most effective one often depends on the particular operations that an application must support.
现代协同软件系统涉及多个并发用户在实时分布式环境中执行共同任务,例如编辑共享文本文档。在这样的环境中维护数据一致性、事务因果关系和复制收敛,同时提供快速的客户机响应,这对传统的分布式计算技术来说是一个重大挑战。操作转换(OT)是一类支持这些功能的并发算法和数据模型,在过去十年中引起了大量的研究关注。在这篇综述中,我们讨论了操作转换模型的基本组成部分,所涉及的算法,以及它们在现实世界网络系统中的实际实现。我们比较了几种现有的OT控制算法,每种算法支持的转换函数和属性,以及相对于每种算法所做的权衡。OT中使用的数据和操作模型非常适合Internet等高延迟环境,这使得它们在现代web服务中得到更频繁的使用。尽管存在许多不同的OT控制算法,但选择最有效的算法通常取决于应用程序必须支持的特定操作。
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引用次数: 0
Autosomal Recessive Spastic Ataxia of Charlevoix-Saguenay (ARSACS): a once obscure neurodegenerative disease with increasing significance for neurological research 常染色体隐性痉挛性共济失调(ARSACS):一种曾经不为人知的神经退行性疾病,在神经学研究中越来越重要
Pub Date : 2013-03-31 DOI: 10.26443/msurj.v8i1.114
Xinlu Li
Background: Autosomal Recessive Spastic Ataxia of Charlevoix-Saguenay (ARSACS) is a rare cerebellar ataxia occurring in the Charlevoix-Saguenay population in Quebec with high incidence as a result of founder effects. Following the discovery of the gene responsible for the disease, many other patient groups have been identified worldwide and the characterization of the gene product, sacsin, has unveiled similarities between the pathogenic mechanism of ARSACS and those of other major neurodegenerative disease. Summary: The core symptoms of ARSACS consist of a triad of early-onset cerebellar ataxia, peripheral neuropathy and spasticity, which is accounted by degeneration of Purkinje neurons. The gene responsible for the disease is located on chromosome 13q11 and encodes for the chaperone sacsin. Drp-1, a GTPase crucial for regulating mitochondrial fission/fusion dynamics, has been identified as a potential substrate of sacsin, suggesting a link between the pathogenic mechanisms of ARSACS and prevalent neurodegenerative diseases such as Alzheimer’s, Parkinson’s and Huntington’s diseases.
背景:常染色体隐性痉挛性共济失调(ARSACS)是一种罕见的小脑性共济失调,发生在魁北克的Charlevoix-Saguenay人群中,由于创始人效应,发病率很高。随着该疾病致病基因的发现,世界范围内也发现了许多其他患者群体,基因产物sacin的特征揭示了ARSACS与其他主要神经退行性疾病的致病机制之间的相似性。摘要:ARSACS的核心症状包括早发性小脑共济失调、周围神经病变和痉挛三联征,这与浦肯野神经元变性有关。导致这种疾病的基因位于染色体13q11上,编码伴侣蛋白sacsin。Drp-1是一种对调节线粒体裂变/融合动力学至关重要的GTPase,已被确定为sacin的潜在底物,这表明ARSACS的致病机制与阿尔茨海默病、帕金森病和亨廷顿病等流行的神经退行性疾病之间存在联系。
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引用次数: 1
Selecting for multicellularity in the unicellular alga Chlamydomonas reinhardtii 单细胞藻莱茵衣藻的多细胞选择
Pub Date : 2013-03-31 DOI: 10.26443/msurj.v8i1.108
Tyler Moulton, Dr. Graham Bell
Background: Researchers have recently begun experimentally exploring the origins of multicellularity (4-6). Their studies have found that the transition to a multicellular state may have been surprisingly simple, considering its profound implications for the history of life (3). This study experimentally selected for multicellularity in the unicellular biflagellated alga Chlamydomonas reinhardtii. This organism is especially interesting because it is basal to the Volvocaceae—a family of biflagellates whose evolutionary transition from unicellular organisms to complex forms have been meticulously characterized (2). The present study aimed to recreate the early stages of this transition, starting from incomplete cytokinesis after cell division. Methods: The procedure was modeled loosely on the experiment performed by Ratcliff et al. (4) in which the authors successfully selected for multicellular Saccharomyces Cerevisiae—unicellular baker’s yeast. Three experimental replicates and one control for nine strains of C. reinhardtii were cultured in round-bottom vials in shaking incubators. Prior to each transfer (every 3-4 days), each culture was slowly mixed, and selection lines were then gently and briefly centrifuged. This applied a selection pressure which rendered heavier (clustered) cells more fit. The very bottom ~2% of the tubes’ contents was transferred, and cell cultures were examined for multicellularity. Results: Six of nine lines of C. reinhardtii demonstrated an increased frequency of C. reinhardtii existing in a two- to four-celled state (the paired cell state accounted for 88% of these clusters)—consistent with the first step toward multicellularity as outlined by Kirk (2). A close study of cell division in the line which exhibited the strongest shift towards the multicellular phenotype suggests that true multicellularity began to evolve in this experiment. A multicellular phenotype did not become fixed in any population. Conclusion: Our findings suggest that an artificial selection pressure is capable of inducing the evolution of multicellularity. Expanding upon this study could help us understand the mechanisms underlying the evolution of multicellularity. Limitations: Sporadic data, possibly the result of difficulties in the procedure, prevented us from rigorously examining the effect of selection through time, limiting our ability to describe the evolutionary response. In addition, the study of individual cells, due to its time-consuming nature, was limited to one replicate of one line exhibiting a pronounced multicellular response. Thorough replication would be required before drawing a strong conclusion from this assay.
背景:研究人员最近开始实验探索多细胞生物的起源(4-6)。他们的研究发现,考虑到其对生命史的深远影响,向多细胞状态的转变可能非常简单(3)。本研究实验选择了单细胞双鞭毛藻莱茵衣藻的多细胞性。这种生物特别有趣,因为它是volvocaceae的基础,volvocaceae是一种双鞭毛虫,其从单细胞生物到复杂形式的进化转变已经被细致地描述过(2)。目前的研究旨在重建这种转变的早期阶段,从细胞分裂后不完全的细胞质分裂开始。方法:该过程松散地模仿Ratcliff等人(4)进行的实验,其中作者成功地选择了多细胞酿酒酵母-单细胞面包酵母。对9株莱茵哈德梭菌在摇瓶中摇瓶培养3个实验重复和1个对照。在每次转移之前(每3-4天),将每种培养物缓慢混合,然后将选品系轻轻短暂离心。这施加了一种选择压力,使更重的(聚集的)细胞更适合。将试管中最底部约2%的内容物转移,并检测细胞培养的多细胞性。结果:9个莱茵哈特瓢虫系中有6个显示出莱茵哈特瓢虫以2到4细胞状态存在的频率增加(配对细胞状态占这些集群的88%)-与Kirk概述的多细胞化的第一步一致(2)。对向多细胞表型转变最强的细胞系进行的细胞分裂的密切研究表明,真正的多细胞化开始在该实验中进化。多细胞表型并没有在任何群体中固定下来。结论:我们的研究结果表明,人工选择压力能够诱导多细胞生物的进化。扩展这项研究可以帮助我们理解多细胞进化的机制。局限性:零星的数据,可能是过程中困难的结果,使我们无法严格检查选择随时间的影响,限制了我们描述进化反应的能力。此外,对单个细胞的研究,由于其耗时的性质,被限制在一个复制的一个系表现出明显的多细胞反应。在从该试验中得出强有力的结论之前,需要进行彻底的复制。
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引用次数: 4
期刊
McGill Science undergraduate research journal : MSURJ
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