The use of real-time PCR analysis in a gene expression study of Alzheimer’s disease post-mortem brains

Gutala, Ramana; Reddy, P. Hemachandra

doi:10.1016/j.jneumeth.2003.09.005

Cited by 105 publications

(74 citation statements)

References 46 publications

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“…For the neuropeptide, hypocretin, our finding of similar levels between L and V mice (Supplemental Table 1) is consistent with other reports that find no change in expression during lactation [5,7]. The finding of no differences in GAPDH (via either the array or real-time PCR) are consistent with this gene's housekeeping functions [11], although in some paradigms GAPDH expression can be altered [35].…”

Section: Other Comparisons To Previous Studiessupporting

confidence: 91%

Gene array profiling of large hypothalamic CNS regions in lactating and randomly cycling virgin mice

Gammie

Hasen

Awad³

et al. 2005

Molecular Brain Research

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A dramatic example of neuronal and physiological plasticity in adult mammals occurs during the transition from a non-maternal to a maternal, lactating state. In this study we compared gene expression within a large continuous region of the CNS involved in maternal behaviors (hypothalamus, preoptic regions, and nucleus accumbens) between lactating (L) (postpartum Day 7) and randomly cycling virgin (V) outbred mice. Using high density oligonucleotide arrays representing 11,904 genes, two statistical algorithms were used to identify significant differences in gene expression: robust multi array (p < 0.001) (n = 92 genes) and significance analysis of microarrays using a 10% false discover rate (n = 114 genes). 27 common genes were identified as significant using both techniques. A subset of genes (n = 5) were selected and examined by real-time PCR. Our findings were consistent with previous published work. For example, neuropeptide Y (NPY) and proenkephalin were elevated in L mice, whereas POMC was decreased. Increased levels of NPY Y2 receptor and polo-like kinase and decreased levels of endothelin receptor type b in L mice are examples of novel gene expression changes not previously identified. Expression differences occurred in broad classes. Together, our findings provide possible new material on gene expression changes that may support maternal behaviors. The advantages and drawbacks of sampling large CNS regions using arrays are discussed.

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Section: Other Comparisons To Previous Studiessupporting

confidence: 91%

Gene array profiling of large hypothalamic CNS regions in lactating and randomly cycling virgin mice

Gammie

Hasen

Awad³

et al. 2005

Molecular Brain Research

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“…Newer data indicated that β-actin is differentially expressed in the brain specimens of both AD and control subjects [23]. Indeed, the results of recent research in the pathomechanism of AD emphasize the significance of cytoskeletal changes [18].…”

Section: Discussionmentioning

confidence: 99%

“…NFT production is a result of abnormal hyperphosphorylation of the microtubule-associated protein tau and the mitogen-activated protein kinase (MAPK) plays a leading role in this abnormal hyperphosphorylation [27]. The β-actin has a relatively stable expression, but this molecule is considered to be involved in synaptogenesis, neuronal plasticity and clinical conditions like depression and AD, too [23].…”

Section: Introductionmentioning

confidence: 99%

Increase in Alzheimer's related markers preceeds memory disturbances: Studies in vasopressin-deficient Brattleboro rat

Várga

Klausz

Domokos

et al. 2014

Brain Research Bulletin

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Alzheimer's disease (AD) is the most common form of dementia in the elderly. For more effective therapy early diagnostic markers could be beneficial. Therefore we compared one year old rats with adults and examined if changes in possible brain markers of AD preceeded memory decline. We also tested if vasopressin-deficient animals were useful model of AD as vasopressin has well known positive effect on memory and AD patient has decreased vasopressin production.We compared adult (3 month) and old (12 month), normal and vasopressin-deficient Brattleboro rats. To receive a comprehensive picture about their memory we examined their social discrimination, object discrimination and conditioned learning abilities (shuttle box).Amyloid precursor protein (APP), mitogen-activated protein kinase 1 (MAPK1), β-actin and tryptophan 2,3-dioxygenase 2 (TDO2) mRNA levels was measured by quantitative PCR.There was no difference between the memory of adult and aged groups. The vasopressin-deficient rats at both ages showed a weaker performance in the course of social and object discrimination tests and a higher escape failure during the shuttle box experiment. The brain marker mRNAs of the elder animals were higher than the levels of the adults, but the absence of vasopressin had no influence on them.Thus, the one year old rats showed elevated levels of AD-related markers, but memory deficits were observable only in vasopressin deficient animals. Vasopressin does not seem to have pathogenic role in AD. Changes in the studied markers might predict later symptoms, although further studies are required for confirmation.

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“…128,135 These data are consistent with observations that SYP mRNA levels are decreased in the AD hippocampus by qPCR and in situ hybridization. 175,176 Moreover, selective down-regulation of synaptic-related proteins appears to be an early manifestation of AD, as loss of SYP protein correlates with cognitive decline. [177][178][179][180] Quantitative assessment of high-density cDNA microarrays revealed several ESTs are up-regulated in individual NFT-bearing CA1 pyramidal neurons in AD brains compared with normal CA1 neurons in agematched control brains, including the lysosomal hydrolase cathepsin D (CatD or CTSD-Unigene abbreviation).…”

Section: Expression Profile Analysis Of Neurofibrillary Tangles In Admentioning

confidence: 99%

Single cell gene expression profiling in Alzheimer’s disease

et al. 2006

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Summary: Development and implementation of microarray techniques to quantify expression levels of dozens to hundreds to thousands of transcripts simultaneously within select tissue samples from normal control subjects and neurodegenerative diseased brains has enabled scientists to create molecular fingerprints of vulnerable neuronal populations in Alzheimer's disease (AD) and related disorders. A goal is to sample gene expression from homogeneous cell types within a defined region without potential contamination by expression profiles of adjacent neuronal subpopulations and nonneuronal cells. The precise resolution afforded by single cell and population cell RNA analysis in combination with microarrays and real-time quantitative polymerase chain reaction (qPCR)-based analyses allows for relative gene expression level comparisons across cell types under different experimental conditions and disease progression. The ability to analyze single cells is an important distinction from global and regional assessments of mRNA expression and can be applied to optimally prepared tissues from animal models of neurodegeneration as well as postmortem human brain tissues. Gene expression analysis in postmortem AD brain regions including the hippocampal formation and neocortex reveals selectively vulnerable cell types share putative pathogenetic alterations in common classes of transcripts, for example, markers of glutamatergic neurotransmission, synaptic-related markers, protein phosphatases and kinases, and neurotrophins/neurotrophin receptors. Expression profiles of vulnerable regions and neurons may reveal important clues toward the understanding of the molecular pathogenesis of various neurological diseases and aid in identifying rational targets toward pharmacotherapeutic interventions for progressive, late-onset neurodegenerative disorders such as mild cognitive impairment (MCI) and AD.

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The use of real-time PCR analysis in a gene expression study of Alzheimer’s disease post-mortem brains

Cited by 105 publications

References 46 publications

Gene array profiling of large hypothalamic CNS regions in lactating and randomly cycling virgin mice

Gene array profiling of large hypothalamic CNS regions in lactating and randomly cycling virgin mice

Increase in Alzheimer's related markers preceeds memory disturbances: Studies in vasopressin-deficient Brattleboro rat

Single cell gene expression profiling in Alzheimer’s disease

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