Real-time PCR analysis of leptin and leptin receptor expression in the rat prostate, and effects of leptin on prostatic acid phosphatase release

Abstract

Abstract.We have recently demonstrated the expression of leptin and leptin receptor (Ob-R) isoforms a, b, c, e and f in the rat seminal vesicles and prostate. The aim of the present study was to provide a semiquantitative real-time PCR estimation of leptin/Ob-R isoform mRNA expression in the seminal vesicles and individual components of rat prostate, and to ascertain the in vitro effects of leptin on prostate acid phosphatase release. The highest expression of the leptin and Ob-R genes was in the seminal vesicles and lateral prostate lobe, respectively. Of the various isoforms, Ob-Rb displayed the highest and Ob-Re the lowest expression. Leptin (10 -8 and 10 -6 M) enhanced acid phosphate release from seminal vesicles, and (10 -6 M) decreased it from the coagulating lobe. Taken together, our findings support the contention that leptin may be involved in the autocrine-paracrine functional regulation of rat seminal vesicles and prostate. The physiological relevance of the marked heterogeneity of the different prostate lobes in both their leptin/Ob-R expression and functional response to leptin remains to be addressed.

The Expression of Leptin and Its Receptor During Tumorigenesis of Diffuse Gliomas such as Astrocytoma and Oligodendroglioma - Grade II, III and IV (NOS)

“…The real-time PCR analysis of expression of leptin and its receptor has been carried out so far only in rat prostrate, which revealed the amplification curves of leptin cDNA alone (Malendowicz et al, 2006). In another study on pituitary adenomas, intact Ob-Rb was documented in 89.6% of the cases by RT-PCR (Jin et al, 1999).…”

Steroidogenic acute regulatory protein gene expression, steroid-hormone secretion and proliferative activity of adrenocortical cells in the presence of proteasome inhibitors: In vivo studies on the regenerating rat adrenal cortex

“…Total RNA was extracted from the frozen regenerating adrenals and intact gland of sham-operated animals, and reverse transcribed to cDNA (18)(19)(20)(21). Semiquantitative real-time PCR was performed as previously detailed (22)(23)(24) in a Roche LightCycler 2.0 with software version 4.0, using the following primers: i) StAR sense (684-703), 5'-CCTGAGCAAAGCGG TGTCAT-3' and antisense (850-870), 5'-GCAAGTGGCTGG CGAACTCTA-3' (amplifon, 187 bp; accession number, NM-03155); and ii) HPRT (hypoxanthine guanine phosphoribosyl transferase 1) sense (391-412), 5'-CAGTCAACGGGGG ACATAAAAG-3' and antisense (515-536), 5'-ATTTTGG GGCTGTACTGCTTGA-3' (amplicon, 146 bp; accession number, NM-01258).…”

Neuromedin-U stimulates enucleation-induced adrenocortical regeneration in the rat

“…Total RNA was extracted from the frozen specimens, and reverse transcribed to cDNA (26)(27)(28)(29). PCR was carried out, as previously detailed (30)(31)(32)(33), in a Roche LightCycler 2.0, using the following primers: NMUR1 sense (481-500), 5'-GCCATCTGGGTCTTCGCTAT-3' and antisense (797-816), 5'-CACCTGTCTGCGTTCCCTAT-3' (336 bp; accession number, AF242873). The PCR program was: denaturation step at 94˚C for 10 min, followed by 35 cycles of three amplification steps (94˚C for 30 sec, annealing at 55˚C for 60 sec and extension at 68˚C for 120 sec).…”