Response

Wong, Marisa L.; Medrano, Juan F.

doi:10.2144/000112064

Cited by 5 publications

(2 citation statements)

References 2 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Because input amount of amplified cDNA was equivalent across all samples, raw Ct values were inversely proportional to the levels of gene expression (Libus and Storchova, 2006). We chose this approach instead of normalization to housekeeping genes because of the potential for differential expression of such reference transcripts in disease versus control samples (Dheda et al, 2005; Wong and Medrano, 2005). A similar approach has recently been validated in which standardized DNA input amounts for qPCR were used (Bernard et al, 2009, 2011).…”

Section: Methodsmentioning

confidence: 99%

Evidence for Transcriptional Factor Dysregulation in the Dorsal Raphe Nucleus of Patients with Major Depressive Disorder

Kerman¹,

Bernard²,

Bunney³

et al. 2012

Front. Neurosci.

View full text Add to dashboard Cite

Extensive evidence implicates dysfunction in serotonin (5-HT) signaling in the etiology of major depressive disorder (MDD). Dorsal raphe nucleus (DR) is a major source of serotonin in the brain, and previous studies have reported within it alterations in 5-HT-related gene expression, protein levels, receptor binding, and morphological organization in mood disorders. In the present study, we utilized in situ hybridization-guided laser capture microdissection to harvest tissue samples from the middle-caudal subregion of the human DR post-mortem from MDD patients and from psychiatrically normal comparison subjects. Extracted RNA was prepared for gene expression profiling, and subsequent confirmation of select targets with quantitative real-time PCR. Our data indicate expression changes in functional gene families that regulate: (1) cellular stress and energy balance, (2) intracellular signaling and transcriptional regulation, and (3) cell proliferation and connectivity. The greatest changes in expression were observed among transcriptional regulators, including downregulation in the expression of TOB1, EGR1, and NR4A2 and their downstream targets. Previous studies have implicated these gene products in the regulation of functional domains impacted by MDD, including cognitive function, affective regulation, and emotional memory formation. These observations indicate altered function of several transcriptional regulators and their downstream targets, which may lead to the dysregulation of multiple cellular functions that contribute to the pathophysiology of MDD. Future studies will require single cell analyses in the DR to determine potential impact of these changes on its cellular functions and related circuits.

show abstract

Section: Methodsmentioning

confidence: 99%

Evidence for Transcriptional Factor Dysregulation in the Dorsal Raphe Nucleus of Patients with Major Depressive Disorder

Kerman¹,

Bernard²,

Bunney³

et al. 2012

Front. Neurosci.

View full text Add to dashboard Cite

show abstract

“…Nuclear paraspeckle assembly transcript 1 (NEAT1) is one such lncRNA and is over expressed in patients with human immunodeciency virus (HIV-1), 2 which is a serious and fatal worldwide immunodeciency disease. 5,6 So exploiting a rapid, accurate, sensitive and quantitative method for detecting NEAT1 is necessary. The level of NEAT1 expression reects whether the body's antiviral ability is strong or weak.…”

Section: Introductionmentioning

confidence: 99%

A novel label free long non-coding RNA electrochemical biosensor based on green l-cysteine electrodeposition and Au–Rh hollow nanospheres as tags

et al. 2015

View full text Add to dashboard Cite

Nuclear paraspeckle assembly transcript 1 (NEAT1) that reflects whether the body's anti-HIV virus immunity is strong or weak is one of many long non-coding RNAs (lncRNAs). The effective detection of NEAT1 has great scientific significance and clinical value. However, the detection of lncRNA is difficult with existing methods. The quantification of lncRNA by electrochemical methods based on changes in circuit properties such as capacitance, conductance and resistance may solve this problem. In this study, we describe a new label-free strategy based on catalytic signal amplification with single-wall carbon nanotubes wrapped with Au-Rh hollow nanospheres (Au/Rh-HNP@SWCNT). This strategy is useful for detecting the lncRNA NEAT1. Firstly, L-cysteine (L-Cys) was electrodeposited onto a Au electrode and incubated in colloidal Au to fully expose all of the binding sites of the Au particles, which can bind to L-Cys through its thiol-group. It can take advantage of each surface of the Au nanoparticles that is bound to the capture probe, which contains a (GGG) 3 trimer that can bind to the electron mediator hemin. The results indicate that catalysis was noticeably enhanced and that the biosensor provided ultrasensitive detection of the lncRNA NEAT1. The linear calibration of the biosensor ranged from 1 fM mL À1 to 100 nM mL À1 , and the limit of detection was 0.8863 fM mL À1 . This lncRNA biosensor based on Au/Rh-HNP@SWCNT complex signal amplification and an L-Cys Au nano-film exhibited acceptable reproducibility and clear selectivity. This strategy may provide a new alternative for clinical HIV diagnosis through the detection of NEAT1.

show abstract

Validation of reference genes for normalization of real‐time quantitative RT‐PCR data in traumatic brain injury

Cook

Vink

Donkin

et al. 2008

J of Neuroscience Research

View full text Add to dashboard Cite

Careful validation of reference genes used for the normalization of real-time RT-PCR data is required to obtain accurate results regarding gene expression. We evaluated the stability of seven commonly used reference genes in the cerebral cortex and hippocampus of rats 3 days following traumatic brain injury (TBI). HPRT, SDHA, and GUSB were found to be the most stable reference genes in the cerebral cortex, whereas B2MG, TBP, and GAPDH were the most stable in the hippocampus. The use of three reference genes was determined to be the optimal number for accurate normalization of data. To illustrate this point, when our gene of interest, substance P (SP), was normalized against the three most stable reference genes in both brain areas, we found no significant difference between injured and uninjured rats at the 3-day time point. However, when our SP data were normalized to each reference gene individually, SP mRNA level was highly variable depending on the reference gene chosen. The results of the present study highlight the importance of validating reference genes to be used for real-time RT-PCR analysis. The use of the most stable reference genes presented here will allow more accurate normalization of gene expression data in TBI.

show abstract

Response

Cited by 5 publications

References 2 publications

Evidence for Transcriptional Factor Dysregulation in the Dorsal Raphe Nucleus of Patients with Major Depressive Disorder

Evidence for Transcriptional Factor Dysregulation in the Dorsal Raphe Nucleus of Patients with Major Depressive Disorder

A novel label free long non-coding RNA electrochemical biosensor based on green l-cysteine electrodeposition and Au–Rh hollow nanospheres as tags

Validation of reference genes for normalization of real‐time quantitative RT‐PCR data in traumatic brain injury

Contact Info

Product

Resources

About