The measurement of adenosine and estrogen receptor expression in rat brains following ovariectomy using quantitative PCR analysis

Rose’Meyer, Roselyn Barbara; Mellick, Albert S.; Garnham, Bronwyn Gaye; Harrison, Glenn; Massa, Helen Maureen; Griffiths, Lyn R.

doi:10.1016/s1385-299x(02)00219-2

Cited by 89 publications

(74 citation statements)

References 22 publications

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“…These results were confirmed by absolute mRNA quantification using standard curves from serial dilution of purified plasmids (data not shown). The ratio between the two methods varied between 0.8 and 1.8 according to the subunits, in agreement with previous comparative studies (23). This variability is attributed to distinct primer efficiency and, overall, cumulative errors introduced by the additional steps of the absolute mRNA quantification (spectrophotometry, calculations of molecular weight) (24).…”

Section: Discussionsupporting

confidence: 89%

Ubiquitous and Kidney-Specific Subunits of Vacuolar H+-ATPase Are Differentially Expressed during Nephrogenesis

Jouret¹,

Auzanneau²,

Debaix³

et al. 2005

Journal of the American Society of Nephrology

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The vacuolar H؉ -ATPase (V-ATPase) is a ubiquitous multisubunit pump that is responsible for acidification of intracellular organelles. In the kidney, a particular form of V-ATPase, made of specific subunits isoforms, has been located at the plasma membrane of intercalated cells (IC). Mutations in genes encoding IC-specific subunits cause infant distal renal tubular acidosis (dRTA), suggesting that the segmental distribution of these subunits is acquired at birth or during early infancy. However, the comparative ontogeny of the IC-specific versus the ubiquitous subunits of V-ATPase and the mechanisms involved in their segmental expression remain unknown. Real-time reverse transcription-PCR, in situ hybridization, immunoblotting, immunostaining, and subcellular fractionation analyses characterized the expression and distribution of V-ATPase subunits, transcription factors, and differentiation markers during mouse nephrogenesis. Ubiquitous A, E1, B2, G1, and C1 subunits showed an early (embryonic day 13.5 [E13.5]) and stable expression throughout nephrogenesis, followed by a slight increase around birth. The developmental pattern of a1 was bimodal, with early induction, gradual decrease during organogenesis, and neonatal increase. These patterns contrasted with the later (from E15.5) and progressive expression of IC-specific a4, B1, G3, and C2 subunits, after the induction of the forkhead transcription factor Foxi1. From E15.5, Foxi1 mRNA was detected in IC, where it co-distributed with B1 in late nephrogenesis. Immunostaining showed that the distribution of ubiquitous E1 and B2 was acquired from E15.5, whereas a4 was located in IC during late nephrogenesis. Subcellular fractionation showed that in both fetal and mature (cortex and medulla) kidneys, E1 and a4 were located in endosomes. These data demonstrate a differential expression and a coordinate regulation of IC-specific versus ubiquitous V-ATPase subunits during nephrogenesis. They provide new insights into the complex regulation of V-ATPase subunits, the maturation of IC along the nephron, and the pathophysiology of hereditary dRTA. T he vacuolar-type proton ATPase (V-ATPase) is a ubiquitous multisubunit pump that is essential for the acidification of intracellular organelles by coupling ATP hydrolysis to transmembrane proton transport (1). The VATPase has been located in a variety of intracellular compartments, including clathrin-coated vesicles, endosomes, and lysosomes. Acidification of these intracellular organelles is a prerequisite for a number of important intracellular processes, such as ligand-receptor dissociation, receptor recycling and ligand degradation, storage, or intracellular targeting (2). In addition to its intracellular distribution, V-ATPase has been located in the plasma membrane of specialized epithelial cells of the kidney and epididymis, as well as in the ruffled border membrane of osteoclasts, where it functions in urinary or semen acidification and in bone resorption, respectively (3). J Am Soc NephrolThe structure of V-ATPase inc...

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Section: Discussionsupporting

confidence: 89%

Ubiquitous and Kidney-Specific Subunits of Vacuolar H+-ATPase Are Differentially Expressed during Nephrogenesis

Jouret¹,

Auzanneau²,

Debaix³

et al. 2005

Journal of the American Society of Nephrology

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“…Competition assays can be used to compare expression levels of the same gene in different samples, while the absolute Q-PCR technologies can use standard curves to estimate the specific amount of a specific target (Rose' Meyer et al, 2003). Determination of cycle threshold (CT) in real time thermocyclers uses a similar approach to that used in semiquantitative analysis in conventional multiplex PCR, for which values are calculated based on an endogenous standard which is usually a housekeeping gene such as the Molecular markers and prostate cancer 197 β-2M gene.…”

Section: Discussionmentioning

confidence: 99%

“…Therefore, a comparative analysis is an adequate and consistent procedure to investigate gene expression levels and is not dependent upon absolute levels of expression (Rose' Meyer et al, 2003).…”

Section: Discussionmentioning

confidence: 99%

Differential expression of the KLK2 and KLK3 genes in peripheral blood and tissues of patients with prostate cancer

et al. 2006

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We used the multiplex semi-quantitative reverse-transcriptase PCR (RT-PCR) to investigate kallikrein 2 and 3 (KLK2 and KLK3) mRNA levels in prostate tissue from 42 prostate cancer patients, 33 of whom were also assessed for peripheral blood KLK2 expression by qualitative semi-nested RT-PCR. We found that KLK2 was an important tissue biomarker for distinguishing between prostate cancer patients and those with benign prostatic hyperplasia, particularly when KLK2 expression was > 60% of that of the β2-microglobulin constitutive gene. Patients with an average relative expression value ≥ 0.6 (cutoff value) had an eleven-fold higher chance of having prostate cancer. When one or two tissues samples were evaluated for KLK2 expression using the cutoff value the estimated chance of having prostate cancer was increased by seven times for one positive sample and 45 times for two positive samples. There was no significant correlation between KLK3 gene expression and prostate cancer diagnosis. Logistic regression for blood and tissue KLK2 expression successfully detected 92% of the prostate cancer cases. The detection of KLK2 in blood showed a sensitivity of 59% and a specificity of 82%. This study indicates that the KLK2 gene may be a useful molecular marker for the diagnosis of prostate cancer and that analysis of KLK2 expression in blood and tissues could provide a novel approach for the clinical investigation of this type of cancer.

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“…Therefore, estrogen might directly modulate the neuroprotection by caffeine through an action on the A 2A R, although there is no information currently on such an interaction. It is reported (Rose'Meyer et al, 2003) that A 2A R expression is significantly decreased after OVX. However, we found no difference in striatal A 2A R binding densities between OVX mice treated with estrogen and placebo (data not shown).…”

Section: Mechanism Of Interaction Between Caffeine and Estrogenmentioning

confidence: 94%

Estrogen Prevents Neuroprotection by Caffeine in the Mouse 1-Methyl-4-Phenyl-1,2,3,6-Tetrahydropyridine Model of Parkinson's Disease

Xu¹,

Xu²,

Brown-Jermyn³

et al. 2006

J. Neurosci.

127

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The measurement of adenosine and estrogen receptor expression in rat brains following ovariectomy using quantitative PCR analysis

Cited by 89 publications

References 22 publications

Ubiquitous and Kidney-Specific Subunits of Vacuolar H+-ATPase Are Differentially Expressed during Nephrogenesis

Ubiquitous and Kidney-Specific Subunits of Vacuolar H+-ATPase Are Differentially Expressed during Nephrogenesis

Differential expression of the KLK2 and KLK3 genes in peripheral blood and tissues of patients with prostate cancer

Estrogen Prevents Neuroprotection by Caffeine in the Mouse 1-Methyl-4-Phenyl-1,2,3,6-Tetrahydropyridine Model of Parkinson's Disease

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