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Myriam Forneris

Publications and source records attributed to Myriam Forneris.

3 recordsLinked to original sources

Age-dependent decrease in the ghrelin gene expression in the human adrenal cortex: a real-time PCR study.

Numerous lines of evidence indicate that ghrelin, an endogenous ligand of the growth hormone-secretagogue receptor, is expressed in the human and rat adrenal cortex. In this study, we examined whether ghrelin gene expression undergoes changes in the human adrenal cortex during aging. Semi-quantitative real-time reverse transcription-polymerase chain reaction demonstrated a highly significant negative correlation between ghrelin mRNA and age in adrenal cortex of 27 patients (aged from 33 to 82 years), who underwent unilateral adrenalectomy/nephrectomy for kidney cancer. No significant differences in the level of adrenal ghrelin expression were observed between males and females. Since it has been previously shown that ghrelin exerts a marked growth-stimulating action on cultured adrenocortical cells, we hypothesize that the down-regulation of ghrelin gene transcription in adrenals could be associated with the reported decrease in adrenal DNA synthesis and mitogenic activity during aging.

Adrenal Cortex↗

Similar sequence-free amplification of human glyceraldehyde-3-phosphate dehydrogenase for real time RT-PCR applications.

One of the major applications of real time polymerase chain reaction (PCR) is relative quantification, where the expression of a target gene is determined as a ratio to a stably expressed reference gene, the so-called housekeeping gene. Glyceraldehyde-3-phosphate dehydrogenase (GAPD) is a glycolytic enzyme, which is active in all mammalian tissues and is frequently used as housekeeping gene in expression studies. The functional locus maps to human chromosome 12p13, but several GAPD-related sequences, including processed pseudogenes, GenBank homologous sequences and computationally predicted sequences are present along the human genome. Due to the high level of GAPD-related sequences it is very important to avoid genomic DNA amplification when GAPD is used as endogenous control in mRNA quantification. We have outlined a GAPD couple of primers that avoid any genomic DNA amplification for real time reverse transcription PCR applications by SYBR-Green Dye. These new designed primers are an useful and chip alternative to probe technologies, and can carry out specific and reproducible data in mRNA expression studies.

Base Sequence↗

Adrenomedullin and vascular endothelium growth factor genes are overexpressed in the regenerating rat adrenal cortex, and AM and VEGF reciprocally enhance their mRNA expression in cultured rat adrenocortical cells.

Adrenomedullin (AM) is an endogenous regulatory peptide, which exerts growth promoting and neoangiogenic actions in several normal and neoplastic tissues. Evidence has been provided that AM and the pro-angiogenic peptide vascular endothelium growth factor (VEGF) are expressed in the adrenal gland, and we investigated whether their gene transcription is modified in a model of rapid rat adrenal growth, namely regeneration after enucleation and contralateral adrenalectomy. Real-time polymerase chain reaction (PCR) showed that AM and VEGF mRNAs were significantly increased in regenerating adrenals with respect to the intact adrenocortical tissue of sham-operated control rats. Subsequent studies were carried out on dispersed rat adrenocortical cells cultured in vitro for 72 h. Conventional PCR demonstrated that cultured cells expressed AM and VEGF mRNAs. Moreover, real-time PCR showed that 24 h exposure to 10-8 M AM or 50 ng/ml VEGF significantly raised the expression of VEGF and AM, respectively, without affecting that of their own mRNA, suggesting the occurence of autocrine-paracrine up-regulatory mechanisms. Taken together, our findings allow us to conceive that the coordinate up-regulation of AM and VEGF expression may favor cell proliferation and neoangiogenesis, thereby leading to a rapid restoration of morphology and function of regenerating adrenals.

Adrenal Cortex↗