Journal: Journal of molecular endocrinology

Article Title: Characterization of GPR101 transcript structure and expression patterns

doi: 10.1530/JME-16-0045

Figure Lengend Snippet: A) 1-cell, B) 8-cell, C) 30% epiboly, D) bud, E) 3 somite, F) 20 somite, G) 24hpf, H) 2dpf, I) 5dpf. gpr101 mRNA (blue) is visible from 1-cell to 8-cell stage (A, B), probably representing maternal transcripts, and then reappears during the bud stage (D), approximately 10hpf. Starting at 2dpf (H) a strong and brain-specific staining is seen. Scale bar: 100 µm; hpf: hours post-fertilization; dpf: days post-fertilization.

Article Snippet: The expression levels of human GPR101 were measured by RT-qPCR in a Human Normal cDNA Array containing 48 samples covering all major human normal tissues (HMRT103, Origene, Rockville, MD, USA).

Techniques: Staining

Journal: Journal of molecular endocrinology

Article Title: Characterization of GPR101 transcript structure and expression patterns

doi: 10.1530/JME-16-0045

Figure Lengend Snippet: A) This panel shows the human GPR101 aligned with mouse and rat Gpr101. The track displaying non-human RefSeq genes follows the display conventions for gene prediction tracks. The color shading indicates the level of review the RefSeq record has undergone: predicted (light), provisional (medium), reviewed (dark). The RNAs were aligned against the human genome using blat. A track with the predicted CpG islands is also shown underneath. The number on the left of each island represents the CpG count. B) The mouse Gpr101 mRNA was BLASTed to the human genome to determine the predicted UTRs of GPR101 The alignment predicted a 215 bp 5’UTR located about 2 kb upstream of the start codon and a splicing event in the 3’UTR. C and D) Analysis of the putative human GPR101 promoter region. 5 kb upstream of the predicted GPR101 TSS were aligned to the mouse genome (C). The alignment returned four regions which were further analyzed by MPromDb (orange bar), the UCSC genome browser track annotating TF binding sites assayed by ChIP-seq (grey bars), and Genomatix (yellow bars) (D). The identity of the predicted TFs is shown in Supplementary Figure 1B and 1C. The location of the previously predicted CpG island by CpG Island Searcher (Trivellin et al. 2014) is also shown in panel D (green bar).

Article Snippet: The expression levels of human GPR101 were measured by RT-qPCR in a Human Normal cDNA Array containing 48 samples covering all major human normal tissues (HMRT103, Origene, Rockville, MD, USA).

Techniques: Binding Assay, ChIP-sequencing

Journal: Journal of molecular endocrinology

Article Title: Characterization of GPR101 transcript structure and expression patterns

doi: 10.1530/JME-16-0045

Figure Lengend Snippet: A) Four isoforms were observed in the analyzed samples by both 5’-RACE and RNAseq. All isoforms shared a common 3’UTR, while they differ for the 5’UTR. The coding sequence (CDS) is represented as a dark grey box, the UTRs as a smaller light grey box, and the intron as a black line. B) Relative expression of the four isoforms assessed by RT-qPCR in a pituitary tumor harboring an Xq26.3 microduplication. Isoform 1 is expressed at much higher levels than the other isoforms. C) Isoform 1 expression in different tissues. The relative expression of isoform 1 was determined by RT-qPCR in the following samples: pituitary tumor and lymphocytes of an X-LAG patient, lymphocytes of her unaffected parents, normal pituitary, GH-secreting tumor without Xq26.3 microduplication, HEK293 cells. Isoform 1 expression was detected only in pituitary samples, showing higher expression in the tumor harboring a GPR101 duplication. All values are relative and normalized on ACTB expression.

Article Snippet: The expression levels of human GPR101 were measured by RT-qPCR in a Human Normal cDNA Array containing 48 samples covering all major human normal tissues (HMRT103, Origene, Rockville, MD, USA).

Techniques: Sequencing, Expressing, Quantitative RT-PCR

Journal: Journal of molecular endocrinology

Article Title: Characterization of GPR101 transcript structure and expression patterns

doi: 10.1530/JME-16-0045

Figure Lengend Snippet: A) GPR101 expression in a panel of 48 different human tissues. B) GPR101 expression in a panel of 24 human brain regions. The highest GPR101 expression levels were observed in the nucleus accumbens. All values are relative and were normalized on ACTB expression. The dotted lines represent the average expression in all tissues analyzed in each cDNA panel.

Article Snippet: The expression levels of human GPR101 were measured by RT-qPCR in a Human Normal cDNA Array containing 48 samples covering all major human normal tissues (HMRT103, Origene, Rockville, MD, USA).

Techniques: Expressing

Journal: Journal of molecular endocrinology

Article Title: Characterization of GPR101 transcript structure and expression patterns

doi: 10.1530/JME-16-0045

Figure Lengend Snippet: A) Preincubation of GPR101 antibody with blocking peptide, resulted in no staining. B) Punctuate GPR101 (green) labeling could not be attributed to somatotrophs (red). C) GPR101 (green) is expressed by LHβ positive cells (red). D–F) High magnification images showing membranous and cytoplasmic distribution of GPR101 in monkey gonadotrophs, excluding nucleus. G–I) GPR101 (green) is expressed by subpopulation of rat somatotrophs (red), as indicated by arrows. Scale bars: A-D: 20 µm; E: 10 µm.

Article Snippet: The expression levels of human GPR101 were measured by RT-qPCR in a Human Normal cDNA Array containing 48 samples covering all major human normal tissues (HMRT103, Origene, Rockville, MD, USA).

Techniques: Blocking Assay, Staining, Labeling

Journal: Journal of molecular endocrinology

Article Title: Characterization of GPR101 transcript structure and expression patterns

doi: 10.1530/JME-16-0045

Figure Lengend Snippet: Panel A shows an H&E stain transverse section of an entire pituitary gland from a 14-week-old fetus at low magnification (50×). The medial (M) and lateral (L) portions of the gland are highlighted. Panel B shows GPR101 staining (brown immunopositivity highlighted) in the lateral portion of a 14-week-old fetal pituitary at higher magnification (400×). Immunopositivity is localized to the cytoplasm and is negative in the nuclei. The different staining intensities for GPR101 at 24 weeks in the lateral (Panel C) and medial (Panel D) portions of the fetal pituitary can be seen (400×). Some positivity for GPR101 in capillaries can also be seen. The increasing GPR101 staining intensity in the lateral portion of the fetal pituitary is illustrated in Panel E at 34 weeks, where very high density is seen (400×). Panels F-I show GPR101 (red) and GH (green) immunofluorescence staining in the anterior pituitary during post-natal life. Almost absent GPR101 expression was seen in a two-year-old female child (F) and in a 21-year-old male (I). Several GPR101 positive cells were present in two males aged 10 (G) and 17 (H) year old. Very little co-localization of GPR101 with GH was seen in Panels G and H. Nuclei (blue) were counterstained with DAPI.

Article Snippet: The expression levels of human GPR101 were measured by RT-qPCR in a Human Normal cDNA Array containing 48 samples covering all major human normal tissues (HMRT103, Origene, Rockville, MD, USA).

Techniques: Staining, Immunofluorescence, Expressing

Journal: Journal of molecular endocrinology

Article Title: Characterization of GPR101 transcript structure and expression patterns

doi: 10.1530/JME-16-0045

Figure Lengend Snippet: A) GPR101 immunoreactive cells and processes were pronounced in accumbens subventricular zone (SVZ) and rhinencephalic differential field (Rh). B) Strong GPR101 immunoreactivity was observed in striatal neuroepithelium and striatal subventricular zone (SVZ) around lateral ventricle (LV) and striatal differentiating field. GPR101 positive cells and cell processes were also observed in the septal area. C) In more posterior sections, GPR101 immunoreactivity was observed throughout the hippocampus and lateral to third ventricle (3V), in the anterior hypothalamus. GPR101 positive cells and processes were also noticed in the amygdaloid differentiating field (A). Scale bar applies to all images: 0.5 mm.

Article Snippet: The expression levels of human GPR101 were measured by RT-qPCR in a Human Normal cDNA Array containing 48 samples covering all major human normal tissues (HMRT103, Origene, Rockville, MD, USA).

Techniques:

Journal: Journal of molecular endocrinology

Article Title: Characterization of GPR101 transcript structure and expression patterns

doi: 10.1530/JME-16-0045

Figure Lengend Snippet: A) and B) WISH staining for gpr101 and gh1 (purple) in 2dpf distinct Zebrafish embryos (ventral view). gpr101 is strongly expressed in the whole brain region (A), while a pituitary-specific staining is visible with the gh1 probe (B). C) and D) Double WISH/immunofluorescence staining for gpr101 (purple) and a pituitary (PRL, green, panel C) and hypothalamic marker [Tyrosine hydroxylase (TH) specific for diencephalic dopaminergic neurons, green, panel D] in a 2dpf Zebrafish embryo (ventral view). gpr101 staining overlaps with the location of pituitary and hypothalamic neurons. The qualitatively different aspect of gpr101 staining in panels C and D is due to the different focal planes selected for PRL and TH. Scale bar: 50 µm applies to all panels.

Article Snippet: The expression levels of human GPR101 were measured by RT-qPCR in a Human Normal cDNA Array containing 48 samples covering all major human normal tissues (HMRT103, Origene, Rockville, MD, USA).

Techniques: Staining, Immunofluorescence, Marker