Similarities in the glycosylation patterns of D2 dopamine receptors in different bovine brain regions

1987 ◽  
Vol 15 (6) ◽  
pp. 1156-1157
Author(s):  
M. N. LEONARD ◽  
P. G. STRANGE
Keyword(s):  
Dopamine Receptors ◽  
Brain Regions ◽  
Bovine Brain ◽  
D2 Dopamine Receptors

scholarly journals Heterogeneity of D2 dopamine receptors in different brain regions

1987 ◽  
Vol 248 (2) ◽  
pp. 595-602 ◽  
Author(s):  
M N Leonard ◽  
C A Macey ◽  
P G Strange
Keyword(s):  
Caudate Nucleus ◽  
Dopamine Receptors ◽  
Serotonin Receptors ◽  
Brain Regions ◽  
Bovine Brain ◽  
Olfactory Tubercle ◽  
Agonist Binding ◽  
D2 Dopamine Receptors ◽  
Substituted Benzamides ◽  
Nucleotide Regulation

The binding of [3H]spiperone has been examined in membranes derived from different regions of bovine brain. In caudate nucleus, nucleus accumbens, olfactory tubercle and putamen binding is to D2 dopamine and 5HT2 serotonin receptors, whereas in cingulate cortex only serotonin 5HT2 receptor binding can be detected. D2 dopamine receptors were examined in detail in caudate nucleus, olfactory tubercle and putamen using [3H]spiperone binding in the presence of 0.3 microM-mianserin (to block 5HT2 serotonin receptors). No evidence for heterogeneity among D2 dopamine receptors either between brain regions or within a brain region was found from the displacements of [3H]spiperone binding by a range of antagonists, including dibenzazepines and substituted benzamides. Regulation of agonist binding by guanine nucleotides did, however, differ between regions. In caudate nucleus a population of agonist binding sites appeared resistant to guanine nucleotide regulation, whereas this was not the case in olfactory tubercle and putamen.

scholarly journals The glycosylation properties of D2 dopamine receptors from striatal and limbic areas of bovine brain

1988 ◽  
Vol 255 (3) ◽  
pp. 877-883 ◽  
Author(s):  
M N Leonard ◽  
R A Williamson ◽  
P G Strange
Keyword(s):  
Caudate Nucleus ◽  
Dopamine Receptors ◽  
Sodium Cholate ◽  
Bovine Brain ◽  
Affinity Column ◽  
Olfactory Tubercle ◽  
D2 Dopamine Receptors ◽  
Neuraminidase Treatment ◽  
Membrane Bound ◽  
Affinity Interaction

D2 dopamine receptors from bovine brain (caudate nucleus and olfactory tubercle) have been solubilized using sodium cholate/NaCl and their glycoprotein properties studied in terms of their interaction with wheat-germ agglutinin-agarose (WGA-agarose). Under optimal conditions about 65% of the applied D2 dopamine receptors bound to WGA-agarose and could be eluted with N-acetylglucosamine. The ability of receptors to adsorb to the affinity column was shown to be dependent on the cholate and salt concentrations used. Digestion of the membrane bound D2 dopamine receptors with neuraminidase prior to solubilisation reduced the ability of the receptors to bind to WGA-agarose (50% of applied receptors bound) whereas digestion with N-acetylglucosaminidase did not significantly affect binding to WGA-agarose. Digestion with the two enzymes together resulted in a larger decrease in binding to WGA-agarose than was seen with the two enzymes alone (40% of applied receptors bound). Stepwise elution of bound receptors from the WGA-agarose columns using 2.5 mM- and 100-mM-N-acetylglucosamine showed that about 40% of the bound receptors interacted with WGA-agarose in a low-affinity manner, the remainder showing a high-affinity interaction. Neuraminidase treatment reduced the low-affinity population suggesting that the interaction of oligosaccharides bearing sialic acid with WGA-agarose is of lower affinity and that higher-affinity binding is via N-acetylglucosamine. These data are discussed in terms of the heterogeneity of carbohydrate moieties on the D2 dopamine receptors within a brain region. In all the tests applied here, however, receptors from caudate nucleus and olfactory tubercle behaved identically so their glycosylation patterns must be very similar.

scholarly journals Antisera specific for D2 dopamine receptors

1993 ◽  
Vol 289 (3) ◽  
pp. 789-794 ◽  
Author(s):  
P L Chazot ◽  
A J Doherty ◽  
P G Strange
Keyword(s):  
Dopamine Receptors ◽  
Cho Cells ◽  
Receptor Gene ◽  
Bovine Brain ◽  
Extracellular Loop ◽  
Intracellular Loop ◽  
D2 Dopamine Receptors ◽  
Western Blots ◽  
Recombinant Cho Cells ◽  
Third Intracellular Loop

Antisera have been raised against two peptides from the sequence of D2 dopamine receptors: peptide 1 from the predicted second extracellular loop and peptide 2 from the predicted third intracellular loop. The antisera recognize specifically a 95 kDa band in Western blots of several bovine brain regions, which corresponds to the denatured D2 dopamine receptor, whereas in recombinant CHO cells expressing D2 dopamine receptors a 80 kDa band is seen. The antisera immunoprecipitate 10-20% of the D2 dopamine receptors from soluble preparations of bovine brain. The antisera recognize D2 dopamine receptors in immunofluorescence analyses of recombinant CHO cells bearing the receptor gene. The antisera directed against the third intracellular loop, but not those against the second extracellular loop, will interfere with the coupling of D2 dopamine receptors and G-proteins in bovine brain preparations.

Purification of D2-dopamine receptors from bovine brain

1986 ◽  
Vol 14 (6) ◽  
pp. 1138-1139 ◽  
Author(s):  
SIMON WORRALL ◽  
RICHARD A. WILLIAMSON ◽  
PHILIP G. STRANGE
Keyword(s):  
Dopamine Receptors ◽  
Bovine Brain ◽  
D2 Dopamine Receptors

Partial purification of D2 dopamine receptors from bovine brain

1987 ◽  
Vol 15 (6) ◽  
pp. 1155-1156 ◽  
Author(s):  
RICHARD A. WILLIAMSON ◽  
SIMON WORRALL ◽  
PHILIP G. STRANGE
Keyword(s):  
Dopamine Receptors ◽  
Bovine Brain ◽  
Partial Purification ◽  
D2 Dopamine Receptors

scholarly journals Importance of thiol groups in ligand binding to D2 dopamine receptors from brain and anterior pituitary gland

1992 ◽  
Vol 281 (2) ◽  
pp. 377-380 ◽  
Author(s):  
P L Chazot ◽  
P G Strange
Keyword(s):  
Ligand Binding ◽  
Pituitary Gland ◽  
Dopamine Receptors ◽  
Dopamine Receptor ◽  
Thiol Group ◽  
Brain Regions ◽  
Subsequent Treatment ◽  
D2 Dopamine Receptor ◽  
Receptor Binding Site ◽  
D2 Dopamine Receptors

The effects of the thiol group reagent 5,5′-dithiobis-(2-nitrobenzoic acid) (DTNB) on D2 dopamine receptors have been examined in three brain regions (caudate nucleus, putamen and olfactory tubercle), and in the anterior and neurointermediate lobes of the pituitary gland. Whereas the receptors in brain were insensitive to DTNB, a dose-dependent inhibition of [3H]spiperone binding to D2 receptors in both lobes of the pituitary gland was observed. The effects in the pituitary could be reversed by subsequent treatment with dithiothreitol and could be prevented by prior occupancy of the receptor binding site with a ligand. The effect of DTNB was to decrease the number of ligand-binding sites without altering the affinity of those remaining. The results show that modification of a thiol group of the D2 dopamine receptor in the two lobes of the pituitary gland tested here significantly affects ligand binding. There is therefore a difference between the D2 dopamine receptor populations in brain and pituitary in their sensitivity to modification by DTNB, and this may imply the existence of different receptor isoforms in the two issues.

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