Abstract：
:Changes in expression of phenotypic markers characterizing the cells comprising intact neurospheres are difficult to determine easily and accurately. The problem is compounded by the diversity of cell populations and phenotypes involved, and consequent non-uniform protein expression across the neurosphere wall, or around the circumference. Therefore, interpreting the effects of phenotype modifying conditions has been a complex and demanding task. Here, we report a novel direct method for measuring protein expression in immunofluoerescently labeled intact neurospheres by densitometric image analysis of optical cross-sections, obtained by scanning laser confocal microscopy. To demonstrate our methodology, hollow human neurospheres were exposed to basic fibroblast growth factor (FGF2), which reportedly induces neuronal commitment in monolayer cultures of neuroprogenitor cells derived from neurospheres. We determined that this treatment downregulated nestin and vimentin, protein markers accepted to indicate an immature, uncommitted phenotype. Neuron specific enolase was only marginally affected. Our strategy allows quantitation of changes in expression of marker proteins that is comparable to Western blot analysis. In addition to discriminating heterogeneity in protein expression, suitable optics may allow the resolution down to single cell level. We propose that this novel strategy, with or without confocal microscopy, may be applied to other biological systems. Analysis of protein expression by the cells comprising tubular or cylindrical cellular structures, or approximately spherical cell aggregates, can be performed efficiently using a small sample size.
journal_name
J Neurosci Methodsjournal_title
Journal of neuroscience methodsauthors
Moeller ML,Dimitrijevich SDdoi
10.1016/j.jneumeth.2004.04.014subject
Has Abstractpub_date
2004-10-15 00:00:00pages
43-50issue
1eissn
0165-0270issn
1872-678Xpii
S0165027004001621journal_volume
139pub_type
杂志文章abstract：BACKGROUND:Tree shrews are proposed as an alternative animal model to nonhuman primates due to their close affinity to primates. Neuroimaging techniques are widely used to study brain functions and structures of humans and animals. However, tree shrews are rarely applied in neuroimaging field partly due to the lack of ...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/j.jneumeth.2017.09.006
更新日期：2018-01-01 00:00:00
abstract：:We present a simple and efficient method for expressing cDNAs in Purkinje neurons (PNs) present in heterogeneous mouse cerebellar cultures. The method combines the transfection of freshly dissociated cerebellar cells via nucleofection with the use of novel expression plasmids containing a fragment of the L7 (Pcp2) gen...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/j.jneumeth.2011.06.006
更新日期：2011-09-15 00:00:00
abstract：:The reproducibility of motor unit twitches obtained using spike-triggered averaging (STA) was examined in the human first dorsal interosseus. For each motor unit (30 total) a series of STA twitches was derived using a 30 s averaging window. Within each averaging window, eight independent measures characterizing motor ...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/s0165-0270(03)00077-3
更新日期：2003-06-30 00:00:00
abstract：:The physical basis for electrical stimulation of excitable tissue, as used by electrophysiological researchers and clinicians in functional electrical stimulation, is presented with emphasis on the fundamental mechanisms of charge injection at the electrode/tissue interface. Faradaic and non-Faradaic charge transfer m...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章,评审
doi：10.1016/j.jneumeth.2004.10.020
更新日期：2005-02-15 00:00:00
abstract：BACKGROUND:Functional magnetic resonance imaging is well suited to link neural population activation with movement parameters of complex natural arm movements. However, currently existing MR-compatible arm tracking devices are not constructed to measure arm joint movement parameters of unrestricted movements. Therefore...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/j.jneumeth.2017.01.015
更新日期：2017-03-15 00:00:00
abstract：:In vitro and in vivo models of experimental glioma are useful tools to gain a better understanding of glioblastoma (GBM) and to investigate novel treatment strategies. However, the majority of preclinical models focus on treating solid intracranial tumours, despite surgical resection being the mainstay in the standard...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/j.jneumeth.2017.04.019
更新日期：2017-06-01 00:00:00
abstract：:The design of a simple and inexpensive battery powered lamp controller for microscope lamps commonly used in patch clamp and other biophysical studies is described. The circuit linearizes the normally exponential voltage-intensity curve of standard 12-V lamps by using an operational amplifier to provide feedback to th...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/0165-0270(91)90098-k
更新日期：1991-10-01 00:00:00
abstract：BACKGROUND:Emotion recognition plays a key role in multimedia. To enhance the sensation of reality, smell has been incorporated into multimedia systems because it can directly stimulate memories and trigger strong emotions. NEW METHOD:For the recognition of olfactory-induced emotions, this study explored a combination...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/j.jneumeth.2020.108599
更新日期：2020-01-21 00:00:00
abstract：:The measurement of amino acid neurotransmitters by high-performance liquid chromatography (HPLC) has emerged as a reliable and sensitive method. This paper describes a method which employs electrochemical (EC) detection of amino acid derivatives formed by a reaction with o-phthalaldehyde (OPA) in the presence of sulph...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/0165-0270(94)00132-z
更新日期：1995-03-01 00:00:00
abstract：:Amyloid-beta (Abeta) deposits are one of the hallmarks of the neuropathological degeneration observed in Alzheimer's disease (AD) and Abeta concentrations have been reported to vary in different brain regions of AD patients. Abeta is produced by the sequential cleavage of amyloid precursor protein (APP) by beta-secret...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/j.jneumeth.2006.05.001
更新日期：2006-10-30 00:00:00
abstract：BACKGROUND:Currently the problem of incorporating priori information into an independent component analysis (ICA) model is often solved under the framework of constrained ICA, which utilizes the priori information as a reference signal to form a constraint condition and then introduce it into classical ICA. However, it...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/j.jneumeth.2017.03.018
更新日期：2017-05-01 00:00:00
abstract：:In this study, a correlation matrix based hierarchical clustering (CMBHC) method is introduced to extract multiple correlation patterns from resting-state functional magnetic resonance imaging (fMRI) data. It was applied to spontaneous fMRI signals acquired from anesthetized rats, and the results were then compared wi...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/j.jneumeth.2012.08.016
更新日期：2012-10-15 00:00:00
abstract：BACKGROUND:With advances in cell capture, gene expression can now be studied in neuronal subtypes and single cells; however, studying epigenetic mechanisms that underlie these changes presents challenges. Moreover, chromatin immunoprecipitation (ChIP) protocols optimized for low cell number do not adequately address te...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/j.jneumeth.2016.02.006
更新日期：2016-04-01 00:00:00
abstract：:Following bilateral injection of manganese (Mn) into the rat's motor cortex, electrical stimulation of the cortex is shown to increase the transport, uptake and accumulation of Mn in the corticospinal tract (CST), as assessed by manganese-enhanced magnetic resonance imaging (MEI). T(1)-weighted gradient echo images we...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/j.jneumeth.2006.02.001
更新日期：2006-09-30 00:00:00
abstract：BACKGROUND:Certain inherited progressive neurodegenerative disorders, such as spinocerebellar ataxia (SCA), affect neurons in large areas of the central nervous system (CNS). The selective expression of disease-causing and therapeutic genes in susceptible regions and cell types is critical for the generation of animal ...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/j.jneumeth.2013.12.004
更新日期：2014-02-15 00:00:00
abstract：:The Porsolt forced swim test (FST) is one of the most widely used behavioral tests in the evaluation of the antidepressant effects of drugs. It is based on the fact that these drugs reduce the depression-related behaviors of learned helplessness. The model has been modified for use in mice. In contrast to rats, mice a...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/j.jneumeth.2005.04.010
更新日期：2005-11-30 00:00:00
abstract：BACKGROUND:The trigeminal subnucleus caudalis (Vc) plays a critical role in transmission and modulation of nociceptive afferent inputs, and exhibits a similar layer construction to the spinal dorsal horn. However, afferent inputs enter the brainstem and project to a separately located nucleus. It has previously been di...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/j.jneumeth.2017.05.019
更新日期：2017-07-15 00:00:00
abstract：:Preferential exploration of novel locations and objects by rodents has been used to test the effects of various manipulations on object recognition memory. However, manual scoring is time-consuming, requires extensive training, and is subject to inter-observer variability. Since rodents explore primarily by sniffing, ...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/j.jneumeth.2007.11.002
更新日期：2008-03-15 00:00:00
abstract：:This report describes technical adaptations of a traumatic brain injury (TBI) model-largely inspired by Marmarou-in order to monitor microdialysis data and PtiO2 (brain tissue oxygen) before, during and after injury. We particularly focalize on our model requirements which allows us to re-create some drastic pathologi...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/j.jneumeth.2004.04.037
更新日期：2004-12-30 00:00:00
abstract：BACKGROUND:Manual annotation of seizures and interictal-ictal-injury continuum (IIIC) patterns in continuous EEG (cEEG) recorded from critically ill patients is a time-intensive process for clinicians and researchers. In this study, we evaluated the accuracy and efficiency of an automated clustering method to accelerat...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/j.jneumeth.2020.108966
更新日期：2020-10-22 00:00:00
abstract：:The silanization of glass, particularly Pyrex, was studied using reaction conditions that might be applied in the fabrication of ion-sensitive microelectrodes of the liquid-membrane type. The efficacy was tested by measuring the hydrophobicity (contact angle) or electrical resistivity of the treated surface. Aminosila...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/0165-0270(83)90037-7
更新日期：1983-07-01 00:00:00
abstract：:Although the classical Golgi method is a powerful means for structural analysis of the brain, it is generally considered to be an unpredictable technique making anatomists wary of using it. Often, even when successful staining has occurred, deposits of silver chromate crystals on the surface of the tissue obscure exam...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/0165-0270(95)00160-3
更新日期：1996-05-01 00:00:00
abstract：:An understanding of how the brain processes information requires knowledge of its underlying wiring diagrams, as well as insights into the relationship between circuit architecture and physiological function. Notably, rabies virus based single-cell genetic manipulations that can facilitate an experimental link between...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章,评审
doi：10.1016/j.jneumeth.2019.108365
更新日期：2019-09-01 00:00:00
abstract：BACKGROUND:Neuroevolution comprises the use of evolutionary computation to define the architecture and/or to train artificial neural networks (ANNs). This strategy has been employed to investigate the behavior of rats in the elevated plus-maze, which is a widely used tool for studying anxiety in mice and rats. NEW MET...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/j.jneumeth.2016.06.010
更新日期：2016-09-01 00:00:00
abstract：BACKGROUND:Over the past decade, pattern decoding techniques have granted neuroscientists improved anatomical specificity in mapping neural representations associated with function and cognition. Dynamical patterns are of particular interest, as evidenced by the proliferation and success of frequency domain methods tha...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/j.jneumeth.2018.06.019
更新日期：2018-10-01 00:00:00
abstract：:Electrical potentials recorded extracellularly from the sinus gland of the isopod, Oniscus asellus, were stored on video tape with the aid of a digital-audio (DA) processor and a video cassette recorder (VCR). The DA processor transforms the analog signal to digital pulses of equal amplitude and converts these pulses ...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/0165-0270(85)90058-5
更新日期：1985-10-01 00:00:00
abstract：BACKGROUND:The use of wearable technology is an emerging field of research in movement disorders. This paper introduces a clinical study to evaluate the feasibility, clinical correlation and reliability of using a system based in smartwatches to quantify tremor in essential tremor (ET) patients and check its acceptance...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/j.jneumeth.2018.02.015
更新日期：2018-06-01 00:00:00
abstract：:A flow rate monitor system for constant perfusion of media for maintaining physiological preparations, particularly in vitro brain slices, is described. The system consists of 3 components: the counter with a drip rate meter and alarm; a drop sensor; and a remote DC power supply. It provides accurate and immediate rea...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/0165-0270(88)90024-6
更新日期：1988-02-01 00:00:00
abstract：BACKGROUND:This manuscript describes a new, multidimensional and data-driven approach to identify outlying datapoints from a first-level fMRI dataset. NEW METHOD:Using three different indicators of data corruption (the fast variance component of DVARS [Δ%D-var], scan-to-scan total displacement [STS], and each scan's o...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/j.jneumeth.2019.02.008
更新日期：2019-04-15 00:00:00
abstract：:This article is part of a study on cortical microcircuitry in which flash evoked potentials (FEPs) were recorded from the anesthetized rat visual cortex. The FEPs were subjected to current source density analysis (CSD). One of the limitations of the CSD method is the need for recording sites outside the sampling volum...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/0165-0270(88)90056-8
更新日期：1988-06-01 00:00:00
