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131.
Vandecasteele M M S Royer S Belluscio M Berényi A Diba K Fujisawa S Grosmark A Mao D Mizuseki K Patel J Stark E Sullivan D Watson B Buzsáki G 《Journal of visualized experiments : JoVE》2012,(61):e3568
A major challenge in neuroscience is linking behavior to the collective activity of neural assemblies. Understanding of input-output relationships of neurons and circuits requires methods with the spatial selectivity and temporal resolution appropriate for mechanistic analysis of neural ensembles in the behaving animal, i.e. recording of representatively large samples of isolated single neurons. Ensemble monitoring of neuronal activity has progressed remarkably in the past decade in both small and large-brained animals, including human subjects. Multiple-site recording with silicon-based devices are particularly effective because of their scalability, small volume and geometric design. Here, we describe methods for recording multiple single neurons and local field potential in behaving rodents, using commercially available micro-machined silicon probes with custom-made accessory components. There are two basic options for interfacing silicon probes to preamplifiers: printed circuit boards and flexible cables. Probe supplying companies (http://www.neuronexustech.com/; http://www.sbmicrosystems.com/; http://www.acreo.se/) usually provide the bonding service and deliver probes bonded to printed circuit boards or flexible cables. Here, we describe the implantation of a 4-shank, 32-site probe attached to flexible polyimide cable, and mounted on a movable microdrive. Each step of the probe preparation, microdrive construction and surgery is illustrated so that the end user can easily replicate the process. 相似文献
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133.
Made of more than 40 subunits, the rotenone-sensitive NADH:ubiquinone oxidoreductase (complex I) is the most intricate membrane-bound enzyme of the mitochondrial respiratory chain. In vascular plants, fungi, and animals, at least seven complex I subunits (ND1, -2, -3, -4, -4L, -5, and -6; ND is NADH dehydrogenase) are coded by mitochondrial genes. The role of these highly hydrophobic subunits in the enzyme activity and assembly is still poorly understood. In the unicellular green alga Chlamydomonas reinhardtii, the ND3 and ND4L subunits are encoded in the nuclear genome, and we show here that the corresponding genes, called NUO3 and NUO11, respectively, display features that facilitate their expression and allow the proper import of the corresponding proteins into mitochondria. In particular, both polypeptides show lower hydrophobicity compared to their mitochondrion-encoded counterparts. The expression of the NUO3 and NUO11 genes has been suppressed by RNA interference. We demonstrate that the absence of ND3 or ND4L polypeptides prevents the assembly of the 950-kDa whole complex I and suppresses the enzyme activity. The putative role of hydrophobic ND subunits is discussed in relation to the structure of the complex I enzyme. A model for the assembly pathway of the Chlamydomonas enzyme is proposed. 相似文献
134.
Marko-Varga G Ogiwara A Nishimura T Kawamura T Fujii K Kawakami T Kyono Y Tu HK Anyoji H Kanazawa M Akimoto S Hirano T Tsuboi M Nishio K Hada S Jiang H Fukuoka M Nakata K Nishiwaki Y Kunito H Peers IS Harbron CG South MC Higenbottam T Nyberg F Kudoh S Kato H 《Journal of proteome research》2007,6(8):2925-2935
Personalized medicine allows the selection of treatments best suited to an individual patient and disease phenotype. To implement personalized medicine, effective tests predictive of response to treatment or susceptibility to adverse events are needed, and to develop a personalized medicine test, both high quality samples and reliable data are required. We review key features of state-of-the-art proteomic profiling and introduce further analytic developments to build a proteomic toolkit for use in personalized medicine approaches. The combination of novel analytical approaches in proteomic data generation, alignment and comparison permit translation of identified biomarkers into practical assays. We further propose an expanded statistical analysis to understand the sources of variability between individuals in terms of both protein expression and clinical variables and utilize this understanding in a predictive test. 相似文献
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Background and Aims
Tree diameter, tree height and canopy closure have been described by previous meta-analyses as being important characteristics in roost selection by cavity-roosting bats. However, size and direction of effects for these characteristics varied greatly among studies, also referred to as heterogeneity. Potential sources of heterogeneity have not been investigated in previous meta-analyses, which are explored by correlating additional covariates (moderator variables). We tested whether effect sizes from 34 studies were consistent enough to reject the null hypothesis that trees selected by bats did not significantly differ in their characteristics from randomly selected trees. We also examined whether heterogeneity in tree diameter effect sizes was correlated to moderator variables such as sex, bat species, habitat type, elevation and mean summer temperature.Methods
We used Hedges’ g standardized mean difference as the effect size for the most common characteristics that were encountered in the literature. We estimated heterogeneity indices, potential publication bias, and spatial autocorrelation of our meta-data. We relied upon meta-regression and multi-model inference approaches to evaluate the effects of moderator variables on heterogeneity in tree diameter effect sizes.Results
Tree diameter, tree height, snag density, elevation, and canopy closure were significant characteristics of roost selection by cavity-roosting bats. Size and direction of effects varied greatly among studies with respect to distance to water, tree density, slope, and bark remaining on trunks. Inclusion of mean summer temperature and sex in meta-regressions further explained heterogeneity in tree diameter effect sizes.Conclusions
Regional differences in roost selection for tree diameter were related to mean summer temperature. Large diameter trees play a central role in roost selection by bats, especially in colder regions, where they are likely to provide a warm and stable microclimate for reproductive females. Records of summer temperature fluctuations inside and outside tree cavities that are used by bats should be included in future research. 相似文献138.
Expression of nonstructural rotavirus protein NSP4 mimics Ca2+ homeostasis changes induced by rotavirus infection in cultured cells 下载免费PDF全文
Díaz Y Chemello ME Peña F Aristimuño OC Zambrano JL Rojas H Bartoli F Salazar L Chwetzoff S Sapin C Trugnan G Michelangeli F Ruiz MC 《Journal of virology》2008,82(22):11331-11343
Rotavirus infection modifies Ca2+ homeostasis, provoking an increase in Ca2+ permeation, the cytoplasmic Ca2+ concentration ([Ca2+]cyto), and total Ca2+ pools and a decrease in Ca2+ response to agonists. A glycosylated viral protein(s), NSP4 and/or VP7, may be responsible for these effects. HT29 or Cos-7 cells were infected by the SA11 clone 28 strain, in which VP7 is not glycosylated, or transiently transfected with plasmids coding for NSP4-enhanced green fluorescent protein (EGFP) or NSP4. The permeability of the plasma membrane to Ca2+ and the amount of Ca2+ sequestered in the endoplasmic reticulum released by carbachol or ATP were measured in fura-2-loaded cells at the single-cell level under a fluorescence microscope or in cell suspensions in a fluorimeter. Total cell Ca2+ pools were evaluated as 45Ca2+ uptake. Infection with SA11 clone 28 induced an increase in Ca2+ permeability and 45Ca2+ uptake similar to that found with the normally glycosylated SA11 strain. These effects were inhibited by tunicamycin, indicating that inhibition of glycosylation of a viral protein other than VP7 affects the changes of Ca2+ homeostasis induced by infection. Expression of NSP4-EGFP or NSP4 in transfected cells induced the same changes observed with rotavirus infection, whereas the expression of EGFP or EGFP-VP4 showed the behavior of uninfected and untransfected cells. Increased 45Ca2+ uptake was also observed in cells expressing NSP4-EGFP or NSP4, as evidenced in rotavirus infection. These results indicate that glycosylated NSP4 is primarily responsible for altering the Ca2+ homeostasis of infected cells through an initial increase of cell membrane permeability to Ca2+. 相似文献
139.
Absence of Mate Choice and Postcopulatory Benefits in a Species with Extreme Sexual Size Dimorphism 下载免费PDF全文
Marie‐Claire Chelini Eileen A. Hebets 《Ethology : formerly Zeitschrift fur Tierpsychologie》2016,122(2):95-104
Most hypotheses related to the evolution of female‐biased extreme sexual size dimorphism (SSD) attribute the differences in the size of each sex to selection for reproduction, either through selection for increased female fecundity or selection for male increased mobility and faster development. Very few studies, however, have tested for direct fitness benefits associated with the latter – small male size. Mecaphesa celer is a crab spider with extreme SSD, whose males are less than half the size of females and often weigh 10 times less. Here, we test the hypotheses that larger size in females and smaller size in males are sexually selected through differential pre‐ and postcopulatory reproductive benefits. To do so, we tested the following predictions: matings between small males and large females are more likely to occur due to mate choice; females mated to small males are less likely to accept second copulation attempts; and matings between small males and large females will result in larger clutches of longer‐lived offspring. Following staged mating trials in the laboratory, we found no support for any of our predictions, suggesting that SSD in M. celer may not be driven by pre‐ or post‐reproductive fitness benefits to small males. 相似文献
140.
Andrew Wu Philip J. Budge John Williams Marie R. Griffin Kathryn M. Edwards Monika Johnson Yuwei Zhu Stella Hartinger Hector Verastegui Ana I. Gil Claudio F. Lanata Carlos G. Grijalva 《PloS one》2015,10(6)