Real-time RT-PCR for the Detection of Beta-adrenoceptor Messenger RNAs in Small Human Endomyocardial Biopsies

Moniotte, Stéphane; Vaerman, Jean‐Pierre; Kockx, Mark; Larrouy, Dominique; Langin, Dominique; Noirhomme, Philippe; Balligand, Jean‐Luc

doi:10.1006/jmcc.2001.1475

Cited by 74 publications

(50 citation statements)

References 25 publications

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“…To determine whether expression of the -AR was the same in the three cell lines mRNA levels of 1-AR, 2-ARand3-AR was determines by RT-PCR as described (13). The data in Fig.…”

Section: Resultsmentioning

confidence: 99%

Role of β-adrenergic receptors in the anti-obesity and anti-diabetic effects of zinc-α2-glycoprotien (ZAG)

Russell

Tisdale

2012

Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biolog

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Objectives:The goal of the current study is to determine whether the -adrenoreceptor (-AR) plays a role in the anti-obesity and anti-diabetic effects of zinc-2-glycoprotein (ZAG). Material and Methods:This has been investigated in CHO-K1 cells transfected with the human 1-, 2-, 3-AR and in ob/ob mice. Cyclic AMP assays were carried out along with binding studies. Ob/ob mice were treated with ZAG and glucose transportation and insulin were examined in the presence or absence of propranonol.Results: ZAG bound to the 3-AR with higher affinity (Kd 46+1nM) than the 2-AR (Kd 71+3nM) while there was no binding to the 1-AR, and this correlated with the increases in cyclic AMP in CHO-K1 cells transfected with the various -AR and treated with ZAG. Treatment of ob/ob mice with ZAG increased protein expression of 3-AR in gastrocnemius muscle, and in white and brown adipose tissue, but had no effect on expression of 1-and 2-AR. A reduction of body weight was seen and urinary glucose excretion, increase in body temperature, reduction in maximal plasma glucose and insulin levels in the oral glucose tolerance test, and stimulation of glucose transport into skeletal muscle and adipose tissue, was completely attenuated by the non-specific -AR antagonist propranolol. Conclusion:The results suggest that the effects of ZAG on body weight and insulin sensitivity in ob/ob mice are manifested through a -3AR, or possibly a 2-AR.

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Section: Resultsmentioning

confidence: 99%

Role of β-adrenergic receptors in the anti-obesity and anti-diabetic effects of zinc-α2-glycoprotien (ZAG)

Russell

Tisdale

2012

Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biolog

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“…While early studies into mRNA detection in rats (Muzzin et al 1991) and humans (Berkowitz et al 1995) Northern blotting or ribonuclease (RNAse) protection assays, this has meanwhile almost fully been superseded by real-time polymerase chain reaction (RT-PCR) techniques. The following studies give examples of primer pairs used for the measurement of β 3 -adrenoceptor mRNA in mice (Lefrere et al 2002;Monjo et al 2005), rats (Alemzadeh and Tushaus 2004;Hatakeyama et al 2004) and humans (Moniotte et al 2001;Nomiya and Yamaguchi 2003). Based upon the sequence information of the β 3 -adrenoceptor, it also appears possible to develop antisense or small interfering RNA (siRNA) approaches, but to the best of our knowledge, neither has been applied to this target until now.…”

Section: Detection Of β 3 -Adrenoceptorsmentioning

confidence: 99%

Tools to study β3-adrenoceptors

Vrydag

Michel

2007

Naunyn-Schmied Arch Pharmacol

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β 3 -adrenoceptors mediate some of the effects of catecholamines on tissues such as blood vessels or the urinary bladder and are putative targets for the treatment of diseases such as the overactive bladder syndrome. Progress in the understanding of the presence, function, and regulation of β 3 -adrenoceptors has been hampered by a lack of highly specific tools. "Classical" is not selective for β 3 -adrenoceptors, at least in humans, and may actually be a partial agonist. Radioligands, which are suitable either for the selective labeling of β 3 -adrenoceptors or for the nonselective labeling of all β-adrenoceptor subtypes, are also missing. β 3 -and β 1 /β 2 double knockout mice have been reported, but their usefulness for extrapolations in humans is questionable based upon major differences between humans and rodents with regard to the ligand recognition and expression profiles of β 3 -adrenoceptors. While the common availability of more selective agonists and antagonists at the β 3 -adrenoceptor is urgently awaited, the limitations of the currently available tools need to be considered in studies of β 3 -adrenoceptor for the time being.

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“…Quantification of ABL transcripts as an internal control for the amount and quality of cDNA was performed for all samples, using the forward primer 5 0 -GCCGCTCGTTGGAACTCCAAGG-3 0 and reverse primer 5 0 -TGACTGGCGTGATGTAGTTTGCTT-3 0 (Tchirkov et al, 1998). Several studies have confirmed that the ABL gene is an appropriate reference in quantitative real-time PCR studies of both pathological and normal cells (Buonamici et al, 2002;Moniotte et al, 2001). During initial optimisation of PCR conditions, amplified products were analysed using agarose gels to ensure correct product size.…”

Section: Quantitative Real-time Rt -Pcr For Htert Mrnamentioning

confidence: 99%

Clinical implications of quantitative real-time RT–PCR analysis of hTERT gene expression in human gliomas

Tchirkov

Rolhion

Jl³

et al. 2003

Br J Cancer

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The presence of telomerase activity in a glioma may be a predictor of its malignant potential. Activation of telomerase is regulated at the transcriptional level of the human telomerase reverse transcriptase (hTERT). Here, we evaluated whether the amount of hTERT mRNA provides a molecular marker of glioma malignancy that would have clinical utility. We used a real-time RT -PCR to assess the number of hTERT transcripts in primary tumour samples derived from 70 glioma patients. Results were standardised by quantifying the number of ABL transcripts as internal control and expressed as hTERT/ABL ratio. The percentage of patients with detectable hTERT mRNA markedly increased with enhanced malignancy: low-grade gliomas expressed hTERT in one out of 14 cases (7.1%), anaplastic gliomas in four out of 13 cases (30.8%) and glioblastoma multiforme (GBM) tumours in 30 out of 43 cases (69.8%). The mean hTERT/ABL ratio was significantly higher in GBMs than in non-GBMs. Subdividing hTERT/ABL ratios as low (p25%) and high (425%), we found that the overall survival among hTERT-positive GBMs was significantly worse in high hTERT expressors than in low hTERT expressors (P ¼ 0.0082). We conclude that the amount of hTERT mRNA may represent a diagnostic and prognostic indicator for GBM patients.

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Real-time RT-PCR for the Detection of Beta-adrenoceptor Messenger RNAs in Small Human Endomyocardial Biopsies

Cited by 74 publications

References 25 publications

Role of β-adrenergic receptors in the anti-obesity and anti-diabetic effects of zinc-α2-glycoprotien (ZAG)

Role of β-adrenergic receptors in the anti-obesity and anti-diabetic effects of zinc-α2-glycoprotien (ZAG)

Tools to study β3-adrenoceptors

Clinical implications of quantitative real-time RT–PCR analysis of hTERT gene expression in human gliomas

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