Immunoblot and immunohistochemical comparison of murine monoclonal antibodies specific for the rat D1a and D1b dopamine receptor subtypes

Luedtke, Robert R.; Griffin, Suzy A.; Conroy, Susan S.; Jin, Xialan; Pinto, Aline; Sesack, Susan R.

doi:10.1016/s0165-5728(99)00142-3

Cited by 52 publications

(56 citation statements)

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“…The following antibodies were used to detect the total amount of proteins: rabbit polyclonal anti-GluR1 (1 : 500, Upstate biotechnology, Mundolsheim, France), rabbit polyclonal anti-ERK1/2 (1 : 2000, Upstate biotechnology, Mundolsheim, France), mouse monoclonal anti-D1R (1 : 500), produced as described (Luedtke et al, 1999), rabbit polyclonal anti-adenylate cyclase V/VI (1 : 400, sc-590 (C-17), Santa Cruz Biotechnology, Heidelberg, Germany). Specific anti-Gaolf antibodies were affinity purified rabbit polyclonal antibodies directed against full-length Gaolf and immunoadsorbed on full-length Gas as previously described ).…”

Section: Antibodies and Drugsmentioning

confidence: 99%

“…Tissue microdisks were punched out from the caudateputamen using a stainless steel cylinder (2.2-mm diameter) and stored at À801C before homogenization. Western blotting with antibodies recognizing specifically Gaolf or D1R was performed as described above with the exception that samples for D1R detection were not boiled before loading, but incubated at 371C during 30 min (Luedtke et al, 1999).…”

Section: D1r and Gaolf Immunoblotting And Ac Assaymentioning

confidence: 99%

“…To investigate the consequences of quantitative changes in D1R and Gaolf protein levels on D1R signaling, we used Gnal + /À and Drd1a + /À mice. D1R protein levels were measured by immunoblotting of striatal extracts with an anti-D1R antibody (Luedtke et al, 1999) (Figure 1a and b). D1R protein levels remained unchanged in Gnal + /À ( Figure 1a) in agreement with previous findings using autoradiographic ligand binding assays Drago et al, 1994).…”

Section: Dopamine-stimulated Ac Activity Is Decreased Inmentioning

confidence: 99%

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Quantitative Changes in Gαolf Protein Levels, but not D1 Receptor, Alter Specifically Acute Responses to Psychostimulants

Corvol

Valjent

Pascoli

et al. 2006

Neuropsychopharmacol

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Striatal dopamine D1 receptors (D1R) are coupled to adenylyl cyclase through Gaolf. Although this pathway is involved in important brain functions, the consequences of quantitative alterations of its components are not known. We explored the biochemical and behavioral responses to cocaine and D-amphetamine (D-amph) in mice with heterozygous mutations of genes encoding D1R and Gaolf (Drd1a + /À and Gnal + /À), which express decreased levels of the corresponding proteins in the striatum. Dopamine-stimulated cAMP production in vitro and phosphorylation of AMPA receptor GluR1 subunit in response to D-amph in vivo were decreased in Gnal + /À, but not Drd1a + /À mice. Acute locomotor responses to D1 agonist SKF81259, D-amph and cocaine were altered in Gnal + /À mice, and not in Drd1a + /À mice. This haploinsufficiency showed that Gaolf but not D1R protein levels are limiting for D1R-mediated biochemical and behavioral responses. Gnal + /À mice developed pronounced locomotor sensitization and conditioned locomotor responses after repeated injections of D-amph (2 mg/kg) or cocaine (20 mg/kg). They also developed normal D-amph-conditioned place preference. The D1R/cAMP pathway remained blunted in repeatedly treated Gnal + /À mice. In contrast, D-amph-induced ERK activation was normal in the striatum of these mice, possibly accounting for the normal development of long-lasting behavioral responses to psychostimulants. Our results clearly dissociate biochemical mechanisms involved in acute and delayed behavioral effects of psychostimulants. They identify striatal levels of Gaolf as a key factor for acute responses to psychostimulants and suggest that quantitative alterations of its expression may alter specific responses to drugs of abuse, or possibly other behavioral responses linked to dopamine function.

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Section: Antibodies and Drugsmentioning

confidence: 99%

Section: D1r and Gaolf Immunoblotting And Ac Assaymentioning

confidence: 99%

Section: Dopamine-stimulated Ac Activity Is Decreased Inmentioning

confidence: 99%

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Quantitative Changes in Gαolf Protein Levels, but not D1 Receptor, Alter Specifically Acute Responses to Psychostimulants

Corvol

Valjent

Pascoli

et al. 2006

Neuropsychopharmacol

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“…3D), which indicated no translocation of D5R-YFP by the drug pretreatment. 3.4 D1R and D5R co-expressed in the same cell kept their distinct trafficking properties Dopamine D1 receptors can not only form the homodimer [16,17] , but also form the heterodimer with other GPCRs [17,18] . The heterodimerized GPCRs can result in the mutual modulation of their function and/or the redistribution of their subcellular locations [17][18][19][20] .…”

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confidence: 99%

神经细胞中多巴胺d1和d5受体不同亚细胞分布和胞内转运的特性

Jin

2009

Neurosci. Bull.

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Objective To explore the possible differential trafficking properties of the dopamine D1-like receptor subtypes, D1 receptor and D5 receptor. Methods To visualize distributions of dopamine D1-like receptor subtypes at subcellular level, the yellow and cyan variants of green fluorescent protein (GFP) were used to tag D1 and D5 receptors. After transfection with the tagged dopamine receptors, the neuroblastoma cells NG108-15 were treated with D1 agonist SKF38393 or acetylcholine (ACh). Then we observed the subcellular distributions of the tagged receptors under the confocal microscopy and tried to determine trafficking properties by comparing their distribution patterns before and after the drug treatment. Results In resting conditions, D1 receptors located in the plasma membrane of NG108-15 cells, while D5 receptors located in both plasma membrane and cytosol. With the pre-treatment of SKF38393, the subcellular distribution of D1 receptors was changed. The yellow particle-like fluorescence of tagged D1 receptors appeared in the cytosol, indicating that D1 receptors were internalized into cytosol from the cell surface. Same situation also occurred in ACh pre-treatment. In contrast, the subcellular distribution of D5 receptors was not changed after SKF38393 or ACh treatment, indicating that D5R was not translocated to cell surface. Interestingly, when D1 and D5 receptors were co-expressed in the same cell, both kept their distinct subcellular distribution patterns and the trafficking properties. Conclusion Our present study reveals that in NG108-15 nerve cells, dopamine D1 and D5 receptors exhibit differential subcellular distribution patterns, and only D1 receptor has a marked trafficking response to the drug stimulation. We further discuss the potential role of the differential trafficking properties of D1-like receptors in complex modulation of DA signaling.

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“…Dopamine receptor D1 is more prevalent than the D5 receptor and is expressed in the brain as well as in various other tissues. In the brain the D5 receptor gene is expressed at much lower levels than the D1 receptor gene and also shows a more restricted expression profile (Ciliax et al, 2000;Luedtke et al, 1999). Dopamine D5 receptors are found in the cerebral cortex, thalamus, diagonal band area, striatum, substantia nigra, and hippocampus (Luedtke et al, 1999).…”

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Conditional inactivation of the dopamine receptor 5 gene: Flanking the Drd5 gene with loxP sites

Heyer

Xiao

Bugaj-Gaweda

et al. 2002

Genesis

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Immunoblot and immunohistochemical comparison of murine monoclonal antibodies specific for the rat D1a and D1b dopamine receptor subtypes

Cited by 52 publications

References 40 publications

Quantitative Changes in Gαolf Protein Levels, but not D1 Receptor, Alter Specifically Acute Responses to Psychostimulants

Quantitative Changes in Gαolf Protein Levels, but not D1 Receptor, Alter Specifically Acute Responses to Psychostimulants

神经细胞中多巴胺d1和d5受体不同亚细胞分布和胞内转运的特性

Conditional inactivation of the dopamine receptor 5 gene: Flanking the Drd5 gene with loxP sites

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