Tissue-specific mRNA Expression Profiles of Human Nuclear Receptor Subfamilies

Abstract

Pairs of forward and reverse primers and TaqMan probes specific to each human nuclear receptor were prepared. Analysis of the mRNA expression level of each target of 43 nuclear receptors in total RNA from single and pooled specimens of various human organs (liver, kidney, adrenal gland, lung, heart, brain, cerebellum, skeletal muscle, spleen, thymus, thyroid gland, prostate, testis, uterus, placenta, bone marrow, trachea, and salivary gland) was performed by real-time reverse transcription PCR using an ABI PRISM 7700 sequence detector system. The mRNA expression of 33 nuclear receptors (NR1A1, 1A2, 1B1, 1B2, 1B3, 1C1, 1C2, 1C3, 1D1, 1D2, 1F1, 1F2, 1F3, 1H2, 1H3, 1I1, 1I2, 2B1, 2B2, 2B3, 2C1, 2C2, 2F1, 2F2, 3A2, 3B1, 3C1, 3C2, 3C4, 4A1, 4A2, 4A3, and 6A1) was successfully detected in all of the tissues by this method. NR1H4, 2A1, and 3C3 mRNAs were not detectable in the heart, heart, and liver, respectively. NR5A2 mRNA was not detectable in either the brain or cerebellum. NR3A1 mRNA was not detectable in the small intestine, colon, brain, and cerebellum. NR5A1 mRNA was not detectable in the kidney, stomach, small intestine, and colon. NR1I3 mRNA was detected in the liver, kidney, stomach, small intestine, adrenal gland, lung, brain, skeletal muscle, thymus, thyroid gland, prostate, testis, placenta, and trachea. NR2A2 mRNA was detected in the liver, kidney, prostate, testis, uterus, and trachea. NR2E1 mRNA was detected in the adrenal gland, brain, cerebellum, testis, placenta, and bone marrow. NR2E3 mRNA was detected in the adrenal gland, thyroid gland, prostate, testis, uterus, trachea, and salivary gland. This study provides information concerning the tissue distribution of the mRNA expression of 43 human nuclear receptors. The mRNA expression profiles of CYP3A4, CYP3A5 and ABC-transporters are also shown. These results are valuable for establishing a nuclear receptor-mediated screening system for new chemical entities in new drug development.

Purification of full-length human Pregnane and Xenobiotic Receptor: polyclonal antibody preparation for immunological characterization

“…From the earlier reports with RT-PCR and Northern blotting approaches, PXR levels are reported to be relatively higher in liver and intestinal tissues when compared with the other tissues [3,7]. Also, endogenous levels of PXR protein in cultured mammalian cell lines appear to be significantly low.…”

“…Lane1, PXR transfected COS-1 cell extract; Lane 2, sample buffer; Lane 3, Mouse liver nuclear extract. adrenal gland and thyroid gland etc [7]. Due to lack of PXR specific antibodies that could detect low levels of endogenously expressed PXR, these studies mainly relied on either Northern blot or RT-PCR analysis.…”

The Orphan Nuclear Receptor TLX/NR2E1 in Neural Stem Cells and Diseases

“…These studies suggest that the fruit fly and mouse homologues of human TLX are predominantly expressed in NSCs or neuronal precursor cells and are important for the proliferation and self-renewal of NSCs. Human TLX expression is detectable in multiple organs and tissues including the adrenal gland, brain, cerebellum, testis, placenta, and bone marrow [6,15,39]. According to the RNA-seq data from the Illumina Human Body Map 2.0 project (http://www.ensembl.org/info/genome/genebuild/ rnaseq_annotation.html), TLX expression in brain is significantly higher than in other organs and tissues, indicating its evolutionarily conserved function in the CNS.…”

Transcriptional expression analysis of ABC efflux transporters and xenobiotic-metabolizing enzymes in the Chinese rare minnow

“…The PCR reaction mixtures (25 l) consisted of Brilliant II SYBR Green QPCR master mix, 300 nM forward primer and 300 nM reverse primer. The primer pairs used for real-time PCR were shown in Table 1 Comparing the relative expression of the target genes in different organs requires a similar concentration of a reference transcript in all organs, previous studies have evaluated the relative expression of several housekeeping gene (e.g., ˇ-actin, b 2 m, gapdh, hprt, pgk1) in different organs, and chose the housekeeping gene which showed the smallest variation among the tissues studied as the suitable reference gene to compare relative expression levels between different organs (Nishimura et al, 2004(Nishimura et al, , 2009. Therefore, to compare relative expression levels of the ABC genes between different tissues in rare minnow, the expression of six housekeeping genes (ˇ-actin, elongation factor 1-alpha (ef1˛), glyceraldehyde-3-phosphate dehydrogenase (gapdh), TATA box binding protein (tbp), tubulin alpha 1(tub˛1) and 18S rRNA) were assessed in eight rare minnow tissues, and the housekeeping gene which showed the smallest variation among the tissues was selected as the suitable reference gene.…”

“…Comparing the relative expression of the target genes in different organs requires a similar concentration of a reference transcript in all organs, according to previous studies (Nishimura et al, 2004(Nishimura et al, , 2009, the relative expressions of six housekeeping genes (ˇ-actin, ef1˛, gapdh, tbp, tub˛1 and 18S rRNA) were assessed in eight rare minnow tissues studied, the results showed that the expression of ˇ-actin mRNA displayed the smallest variation among the tissues studied, the ratio of highest value to lowest value was 8.47-fold (data showed in Table S1 in the supplementary materials). Therefore, we considered ˇ-actin mRNA to be a suitable endogenous control for measurement of mRNA expression between different tissues.…”