Abstract：
:Analysis of gene expression in the brain is a valuable tool to study the function of the brain under normal and pathological conditions. Although there are many techniques used to measure gene expression the validity of any such experiment is directly related to the quality of the RNA in the samples. The most readily available source of human brain tissue is post-mortem and while frozen tissue is sometimes available, most archived tissue is fixed and paraffin-embedded. The use of fixed tissue for expression analysis introduces variables, which must be considered in the experimental design. In addition, factors associated with clinical variability of the patient and with tissue procurement can affect RNA transcript levels. In order to illustrate the effects of two common tissue fixatives, formalin and ethanol, on the quality of RNA for expression analysis we compare RNA extracted from these fixed tissues to the gold standard, flash-frozen tissue. We describe RNA extraction from fixed tissue and ways to assess the quality or intactness of the RNA using reverse transcription combined with polymerase chain reaction amplification. An advantage of using archived tissue is the ease with which single cells or subpopulations of cells can be obtained by laser microdissection. The successful isolation of RNA from microdissected cells is also presented. From our results and a review of the literature we conclude that RNA from fixed tissues is a viable source of RNA for expression analysis which should enable new experimental approaches and discoveries as long as attention is given to variables that can affect RNA at all levels of analysis.
journal_name
Neurochem Resjournal_title
Neurochemical researchauthors
Van Deerlin VM,Gill LH,Nelson PTdoi
10.1023/a:1020996519419keywords:
subject
Has Abstractpub_date
2002-10-01 00:00:00pages
993-1003issue
10eissn
0364-3190issn
1573-6903journal_volume
27pub_type
杂志文章abstract：:Cyclin-dependent kinase 5 (cdk5) is a proline-directed serine/threonine kinase that is activated mostly by association with its activators, p35 and p39. Initially projected as a neuron-specific kinase, cdk5 is expressed ubiquitously and its kinase activity solely depends on the presence of its activators, which are al...
journal_title：Neurochemical research
pub_type： 杂志文章,评审
doi：10.1007/s11064-008-9885-4
更新日期：2009-06-01 00:00:00
abstract：:Axonal regeneration across the site of spinal cord lesion is often aborted in adult mammalian species. The use of DNA vaccine to nullify the inhibitory molecules has been shown to be effective in promoting axonal regeneration in injured spinal cord. The possible molecular mechanisms, however, remain to be elucidated. ...
journal_title：Neurochemical research
pub_type： 杂志文章
doi：10.1007/s11064-009-9951-6
更新日期：2009-09-01 00:00:00
abstract：:Crayfish glutamic acid decarboxylase (GAD), like the homologous enzymes from other species, is inhibited by carbonyl-trapping agents (e.g. aminooxyacetic acid; AOAA) and sulfhydryl reagents (e.g. 5,5'-dithiobis-(2-nitrobenzoic acid); DTNB). It also is inhibited by the product GABA, many anions (e.g. SCN- and Cl-), and...
journal_title：Neurochemical research
pub_type： 杂志文章
doi：10.1007/BF00964526
更新日期：1984-07-01 00:00:00
abstract：:Sandhoff disease is an incurable neurodegenerative disorder caused by mutations in the lysosomal hydrolase β-hexosaminidase. Deficiency in this enzyme leads to excessive accumulation of ganglioside GM2 and its asialo derivative, GA2, in brain and visceral tissues. Small molecule inhibitors of ceramide-specific glucosy...
journal_title：Neurochemical research
pub_type： 杂志文章
doi：10.1007/s11064-013-0992-5
更新日期：2013-04-01 00:00:00
abstract：:Brains of Alzheimer disease patients in early stages of dementia contain an increased 24(S)-hydroxycholesterol (cerebrosterol)/cholesterol ratio when compared to controls. In this study, effects of amyloid beta peptides and of racemic 24-hydroxycholesterol were evaluated in vitro on undepleted or cholesterol-depleted ...
journal_title：Neurochemical research
pub_type： 杂志文章
doi：10.1007/s11064-007-9443-5
更新日期：2008-03-01 00:00:00
abstract：:There is increasing evidence that proteins normally involved in the cell cycle play a role in the regulation of neuronal apoptotic death following various insults. However, it is not clear if the same mechanisms regulate cell death of oligodendrocytes as well. In this study, we investigated the mechanism of ceramide-i...
journal_title：Neurochemical research
pub_type： 杂志文章
doi：10.1023/b:nere.0000021236.32785.37
更新日期：2004-05-01 00:00:00
abstract：:Charge microheterogeneity of myelin basic protein is known to affect its conformation and function. Here, the citrullinated myelin basic protein charge isomer, component-8, was shown to be more susceptible to stromelysin-1 cleavage than myelin basic protein component-1. Since levels of component-8 are increased in mul...
journal_title：Neurochemical research
pub_type： 杂志文章
doi：10.1007/s11064-006-9116-9
更新日期：2006-08-01 00:00:00
abstract：:Exposure to Hg2+ below 10 micrometer destroys synaptosomal membrane-associated sialidase of bovine brain in situ. Inhibition by Cu2+ occurs only at relatively higher concentrations, and is demonstrable after the synaptosomal membrane preparation has been presaturated with Cu2+ . Pb2+ does not inhibit enzymatic activit...
journal_title：Neurochemical research
pub_type： 杂志文章
doi：10.1007/BF00964363
更新日期：1978-02-01 00:00:00
abstract：:One of the many pharmacological targets of ethanol is the GABA inhibitory system, and chronic ethanol (CE) is known to alter the polypeptide levels of the GABA(A )receptor subunits in rat brain regions. In the present study, we investigated the regulation of the tyrosine kinase phosphorylation of the GABA(A) receptor ...
journal_title：Neurochemical research
pub_type： 杂志文章
doi：10.1007/s11064-007-9288-y
更新日期：2007-07-01 00:00:00
abstract：:The release of preloaded D-[3H]aspartate, an unmetabolizable analogue of L-glutamate, was studied in superfused hippocampal slices from 7-day-old and 3-month-old (adult) mice under various cell-damaging conditions, including hypoxia, hypoglycemia, ischemia, oxidative stress and the presence of free radicals and metabo...
journal_title：Neurochemical research
pub_type： 杂志文章
doi：10.1023/a:1021052725921
更新日期：1999-08-01 00:00:00
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journal_title：Neurochemical research
pub_type： 杂志文章
doi：10.1023/a:1021608514646
更新日期：2003-01-01 00:00:00
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journal_title：Neurochemical research
pub_type： 杂志文章
doi：10.1007/s11064-011-0649-1
更新日期：2012-03-01 00:00:00
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journal_title：Neurochemical research
pub_type： 杂志文章
doi：10.1023/a:1022434209787
更新日期：1998-06-01 00:00:00
abstract：:Primary malignant brain cancer, one of the most deadly diseases, has a high rate of recurrence after treatment. Studies in the past several years have led to the hypothesis that the root of the recurrence may be brain tumor stem cells (BTSCs), stem-like subpopulation of cells that are responsible for propagating the t...
journal_title：Neurochemical research
pub_type： 杂志文章,评审
doi：10.1007/s11064-009-0079-5
更新日期：2009-12-01 00:00:00
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journal_title：Neurochemical research
pub_type： 杂志文章
doi：10.1007/s11064-006-9149-0
更新日期：2006-10-01 00:00:00
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journal_title：Neurochemical research
pub_type： 杂志文章
doi：10.1007/s11064-004-6879-8
更新日期：2004-11-01 00:00:00
abstract：:Myelinating cells of both the peripheral and central nervous systems (CNSs) undergo dramatic cytoskeletal reorganization in order to differentiate and produce myelin. Myelinating oligodendrocytes in the CNS show a periodic actin pattern, demonstrating tight regulation of actin. Furthermore, recent data demonstrate tha...
journal_title：Neurochemical research
pub_type： 杂志文章,评审
doi：10.1007/s11064-019-02753-0
更新日期：2020-03-01 00:00:00
abstract：:Emerging data suggests that mitochondrial dysfunction is prominently involved in Alzheimer disease (AD) progression. Sirtuin-3 (Sirt3) is a member of the sirtuin family of nicotinamide adenine dinucleotide dependent deacetylases that regulates a variety of mitochondrial functions and suppresses mitochondria-related ph...
journal_title：Neurochemical research
pub_type： 杂志文章
doi：10.1007/s11064-015-1630-1
更新日期：2015-08-01 00:00:00
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journal_title：Neurochemical research
pub_type： 杂志文章
doi：10.1007/s11064-007-9508-5
更新日期：2008-02-01 00:00:00
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journal_title：Neurochemical research
pub_type： 杂志文章
doi：10.1007/s11064-012-0915-x
更新日期：2013-02-01 00:00:00
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journal_title：Neurochemical research
pub_type： 杂志文章
doi：10.1007/BF00971589
更新日期：1988-08-01 00:00:00
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journal_title：Neurochemical research
pub_type： 杂志文章
doi：10.1007/s11064-010-0203-6
更新日期：2010-09-01 00:00:00
abstract：:The K-stimulated release of [3H]ACh from rat midbrain slices prelabeled by incubation with [3H]choline was dependent on extracellular Ca. Phenobarbital inhibited the K-stimulated [3H]ACh release and the IC50 was equal to that found for K-stimulated endogenous ACh release. These results support the suggestion that barb...
journal_title：Neurochemical research
pub_type： 杂志文章
doi：10.1007/BF00964710
更新日期：1980-07-01 00:00:00
abstract：:Impaired glucose transport across brain tissue barriers causes infantile seizures, developmental delay and acquired microcephaly. Since the first report in 1991 (De Vivo et al, NEJM, 1991) 17 patients have been identified with the glucose transporter protein syndrome (GTPS). The diagnostic feature of the syndrome is a...
journal_title：Neurochemical research
pub_type： 杂志文章
doi：10.1023/a:1022544131826
更新日期：1999-04-01 00:00:00
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journal_title：Neurochemical research
pub_type： 杂志文章
doi：10.1007/s11064-019-02839-9
更新日期：2019-08-01 00:00:00
abstract：:Passive avoidance (PA) conditioning is a fear motivated task able to initiate a cascade of altered gene expression within the hippocampus, a structure critical to learning and memory. We have previously shown that neurofibromin (NF1) and amyloid precursor protein (APP), two genes implicated in cognitive function, are ...
journal_title：Neurochemical research
pub_type： 杂志文章
doi：10.1007/s11064-012-0949-0
更新日期：2013-03-01 00:00:00
abstract：:Studies on primary astrocytes cultured in vitro have shown that process formation involves changes in cytoskeletal proteins and release of tension on the substratum. Actin filament reorganization has previously been found to be the major cytoskeletal change occurring during process formation. These changes are relativ...
journal_title：Neurochemical research
pub_type： 杂志文章
doi：10.1023/a:1022413718447
更新日期：1998-03-01 00:00:00
abstract：:Estrogen action is necessary for evidencing the stimulatory action of mifepristone and naloxone on prolactin (PRL) secretion during late pregnancy. Our aim is to determine the mechanism mediating this facilitator action of estrogens. To investigate the hypothalamic mechanisms involved in estrogen actions in PRL secret...
journal_title：Neurochemical research
pub_type： 杂志文章
doi：10.1007/s11064-016-2067-x
更新日期：2016-12-01 00:00:00
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journal_title：Neurochemical research
pub_type： 杂志文章,评审
doi：10.1007/BF00966762
更新日期：1993-09-01 00:00:00
abstract：:A double ligand enzyme-linked immunosorbent assay (ELISA) has been developed to detect antibodies against brain tissue antigens in the sera of patients with neurological diseases. The sera were tested on human white matter homogenate. The technique consists of successive incubations with the human serum to be tested, ...
journal_title：Neurochemical research
pub_type： 杂志文章
doi：10.1007/BF00964651
更新日期：1985-04-01 00:00:00
