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The Adhesion GPCR GPR125 is specifically expressed in the choroid plexus and is upregulated following brain injury.

Pickering C, Hägglund M, Szmydynger-Chodobska J, Marques F, Palha JA, Waller L, Chodobski A, Fredriksson R, Lagerström MC, Schiöth HB - BMC Neurosci (2008)

Bottom Line: A single copy of GPR125 was found in many vertebrate genomes.Induction of inflammation by LPS did not change GPR125 expression.However, GPR125 expression was transiently increased (almost 2-fold) at 4 h after traumatic brain injury (TBI) followed by a decrease (approximately 4-fold) from 2 days onwards in the choroid plexus as well as increased expression (2-fold) in the hippocampus that was delayed until 1 day after injury.

View Article: PubMed Central - HTML - PubMed

Affiliation: Uppsala University, Department of Neuroscience, Functional Pharmacology, BMC, Box 593, SE-75124, Uppsala, Sweden. chris.pickering@neuro.uu.se

ABSTRACT

Background: GPR125 belongs to the family of Adhesion G protein-coupled receptors (GPCRs). A single copy of GPR125 was found in many vertebrate genomes. We also identified a Drosophila sequence, DmCG15744, which shares a common ancestor with the entire Group III of Adhesion GPCRs, and also contains Ig, LRR and HBD domains which were observed in mammalian GPR125.

Results: We found specific expression of GPR125 in cells of the choroid plexus using in situ hybridization and protein-specific antibodies and combined in situ/immunohistochemistry co-localization using cytokeratin, a marker specific for epithelial cells. Induction of inflammation by LPS did not change GPR125 expression. However, GPR125 expression was transiently increased (almost 2-fold) at 4 h after traumatic brain injury (TBI) followed by a decrease (approximately 4-fold) from 2 days onwards in the choroid plexus as well as increased expression (2-fold) in the hippocampus that was delayed until 1 day after injury.

Conclusion: These findings suggest that GPR125 plays a functional role in choroidal and hippocampal response to injury.

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Related in: MedlinePlus

In situ hybridization combined with immunohistochemistry on free floating sections using 400 ng of digoxigenin (DIG)-labeled mouse GPR125 antisense probe stained with Fast Red enzyme substrate (A-C), white cell nucleus staining with (1:1000) DAPI (A), green staining with the (1:400) epithelial-cell-specific pan-cytokeratin antibody (A and B) and dark blue (1:500) glial fibrillary acidic protein (GFAP) staining (B and C). A) The z-stack function of the confocal microscope rendered 9 layers together to form this 3-dimensional image illustrating GPR125 expression in the cytoplasm around the DAPI-stained nuclei. Separate images are provided for the 3 channels followed by the merged images in the final panel. B) At higher magnification (63× objective), staining for the cytokeratin protein follows the edge of GPR125-labeled cells which illustrates co-localization. C) To show that GPR125 staining is predominantly in the choroid plexus, GFAP was used to stain ependymal cells which line the walls of the ventricle.
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Figure 4: In situ hybridization combined with immunohistochemistry on free floating sections using 400 ng of digoxigenin (DIG)-labeled mouse GPR125 antisense probe stained with Fast Red enzyme substrate (A-C), white cell nucleus staining with (1:1000) DAPI (A), green staining with the (1:400) epithelial-cell-specific pan-cytokeratin antibody (A and B) and dark blue (1:500) glial fibrillary acidic protein (GFAP) staining (B and C). A) The z-stack function of the confocal microscope rendered 9 layers together to form this 3-dimensional image illustrating GPR125 expression in the cytoplasm around the DAPI-stained nuclei. Separate images are provided for the 3 channels followed by the merged images in the final panel. B) At higher magnification (63× objective), staining for the cytokeratin protein follows the edge of GPR125-labeled cells which illustrates co-localization. C) To show that GPR125 staining is predominantly in the choroid plexus, GFAP was used to stain ependymal cells which line the walls of the ventricle.

Mentions: To prepare the mouse tissue panel, 4 adult, male Sv129 (Alab, Sollentuna, Sweden) were housed in a climate-controlled facility (12 hr light/dark cycle with lights on at 07.00) with constant temperature (22–23°C) and 55% humidity. Animals were given 7 days to acclimatize to the local conditions and had free access to water and R36 food pellets (Labfor Lactamin, Vadstena, Sweden). Between 3 and 6 hours into the light period, animals were decapitated for dissection of brains and various peripheral tissues. Brains were placed in a brain matrix for dissection into areas indicated in Figure 4 while peripheral organs were taken in their entirety. All samples were frozen on a dry ice block before immersion into RNALater solution (Ambion). Following 1 hr of incubation at room temperature to allow complete penetration of the tissue, samples were stored at -80°C until further processing.


The Adhesion GPCR GPR125 is specifically expressed in the choroid plexus and is upregulated following brain injury.

Pickering C, Hägglund M, Szmydynger-Chodobska J, Marques F, Palha JA, Waller L, Chodobski A, Fredriksson R, Lagerström MC, Schiöth HB - BMC Neurosci (2008)

In situ hybridization combined with immunohistochemistry on free floating sections using 400 ng of digoxigenin (DIG)-labeled mouse GPR125 antisense probe stained with Fast Red enzyme substrate (A-C), white cell nucleus staining with (1:1000) DAPI (A), green staining with the (1:400) epithelial-cell-specific pan-cytokeratin antibody (A and B) and dark blue (1:500) glial fibrillary acidic protein (GFAP) staining (B and C). A) The z-stack function of the confocal microscope rendered 9 layers together to form this 3-dimensional image illustrating GPR125 expression in the cytoplasm around the DAPI-stained nuclei. Separate images are provided for the 3 channels followed by the merged images in the final panel. B) At higher magnification (63× objective), staining for the cytokeratin protein follows the edge of GPR125-labeled cells which illustrates co-localization. C) To show that GPR125 staining is predominantly in the choroid plexus, GFAP was used to stain ependymal cells which line the walls of the ventricle.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2571103&req=5

Figure 4: In situ hybridization combined with immunohistochemistry on free floating sections using 400 ng of digoxigenin (DIG)-labeled mouse GPR125 antisense probe stained with Fast Red enzyme substrate (A-C), white cell nucleus staining with (1:1000) DAPI (A), green staining with the (1:400) epithelial-cell-specific pan-cytokeratin antibody (A and B) and dark blue (1:500) glial fibrillary acidic protein (GFAP) staining (B and C). A) The z-stack function of the confocal microscope rendered 9 layers together to form this 3-dimensional image illustrating GPR125 expression in the cytoplasm around the DAPI-stained nuclei. Separate images are provided for the 3 channels followed by the merged images in the final panel. B) At higher magnification (63× objective), staining for the cytokeratin protein follows the edge of GPR125-labeled cells which illustrates co-localization. C) To show that GPR125 staining is predominantly in the choroid plexus, GFAP was used to stain ependymal cells which line the walls of the ventricle.
Mentions: To prepare the mouse tissue panel, 4 adult, male Sv129 (Alab, Sollentuna, Sweden) were housed in a climate-controlled facility (12 hr light/dark cycle with lights on at 07.00) with constant temperature (22–23°C) and 55% humidity. Animals were given 7 days to acclimatize to the local conditions and had free access to water and R36 food pellets (Labfor Lactamin, Vadstena, Sweden). Between 3 and 6 hours into the light period, animals were decapitated for dissection of brains and various peripheral tissues. Brains were placed in a brain matrix for dissection into areas indicated in Figure 4 while peripheral organs were taken in their entirety. All samples were frozen on a dry ice block before immersion into RNALater solution (Ambion). Following 1 hr of incubation at room temperature to allow complete penetration of the tissue, samples were stored at -80°C until further processing.

Bottom Line: A single copy of GPR125 was found in many vertebrate genomes.Induction of inflammation by LPS did not change GPR125 expression.However, GPR125 expression was transiently increased (almost 2-fold) at 4 h after traumatic brain injury (TBI) followed by a decrease (approximately 4-fold) from 2 days onwards in the choroid plexus as well as increased expression (2-fold) in the hippocampus that was delayed until 1 day after injury.

View Article: PubMed Central - HTML - PubMed

Affiliation: Uppsala University, Department of Neuroscience, Functional Pharmacology, BMC, Box 593, SE-75124, Uppsala, Sweden. chris.pickering@neuro.uu.se

ABSTRACT

Background: GPR125 belongs to the family of Adhesion G protein-coupled receptors (GPCRs). A single copy of GPR125 was found in many vertebrate genomes. We also identified a Drosophila sequence, DmCG15744, which shares a common ancestor with the entire Group III of Adhesion GPCRs, and also contains Ig, LRR and HBD domains which were observed in mammalian GPR125.

Results: We found specific expression of GPR125 in cells of the choroid plexus using in situ hybridization and protein-specific antibodies and combined in situ/immunohistochemistry co-localization using cytokeratin, a marker specific for epithelial cells. Induction of inflammation by LPS did not change GPR125 expression. However, GPR125 expression was transiently increased (almost 2-fold) at 4 h after traumatic brain injury (TBI) followed by a decrease (approximately 4-fold) from 2 days onwards in the choroid plexus as well as increased expression (2-fold) in the hippocampus that was delayed until 1 day after injury.

Conclusion: These findings suggest that GPR125 plays a functional role in choroidal and hippocampal response to injury.

Show MeSH
Related in: MedlinePlus