Cloning and Tissue Distribution of Novel Splice Variants of the Rat GABABReceptor

Isomoto, Shojiro; Kaibara, Muneshige; Sakurai-Yamashita, Yasuko; Nagayama, Yuji; Uezono, Yasuhito; Yano, Katsusuke; Taniyama, Kohtaro

doi:10.1006/bbrc.1998.9706

Cited by 119 publications

(86 citation statements)

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“…For 13 C/ 15 N isotope labeling, a 13 C/ 15 N buffered minimal dextrose medium (100 mM potassium phosphate (pH 6 or 3), 15 N/YNB, 0.5% (w/v) [ 13 C]glucose, and 0.00004% (w/v) biotin) was used instead of BMG. Cells were then harvested by centrifugation as described above and resuspended in 100 mM potassium phosphate (pH 3), 15 N/YNB, 0.1% (w/v) [ 13 C]glycerol, and 0.00004% (w/v) biotin for 2 h prior to induction. This stage allows a smooth transition between glucose and methanol as the sole carbon source for the cells and was found to reduce cell death.…”

Section: Construction Of Expression Vectors Encoding Ccp1 Ccp2mentioning

confidence: 99%

“…This stage allows a smooth transition between glucose and methanol as the sole carbon source for the cells and was found to reduce cell death. Induction for both 15 N and 13 C/ 15 N isotope labeling was carried out using buffered minimal methanol medium containing 15 N/YNB instead of 14 N/YNB. For 13 C/ 15 N isotope labeling, buffered minimal methanol medium was prepared with [ 13 C]methanol.…”

Section: Construction Of Expression Vectors Encoding Ccp1 Ccp2mentioning

confidence: 99%

“…Sample concentrations were 0.6 mM (CCP1) and 0.7 mM (CCP12 and CCP2). The spectral widths were 5000 Hz in the 1 H dimension and 2000 Hz in the 15 N dimension for all spectra. The CCP1 spectrum was recorded with 64 scans and 512 increments in the 15 N dimension; the CCP2 spectrum was recorded with 8 scans and 128 increments in the 15 N dimension; and the CCP12 spectrum was recorded with 16 scans and 96 increments in the 15 N dimension.…”

Section: -Anilinonaphthalene-8-sulfonic Acid (Ans) Fluorescencementioning

confidence: 99%

“…The spectral widths were 5000 Hz in the 1 H dimension and 2000 Hz in the 15 N dimension for all spectra. The CCP1 spectrum was recorded with 64 scans and 512 increments in the 15 N dimension; the CCP2 spectrum was recorded with 8 scans and 128 increments in the 15 N dimension; and the CCP12 spectrum was recorded with 16 scans and 96 increments in the 15 N dimension. After processing, data matrices were 1024 points in 1 H and 256 points in 15 N. For each spectrum, a sine-bell square window function (with a shift of 90°) was applied in both dimensions.…”

Section: -Anilinonaphthalene-8-sulfonic Acid (Ans) Fluorescencementioning

confidence: 99%

“…Two other variations of the GABA B R1 subunit occur: in the fifth extracellular loop (rat GABA B R1c) and at the C terminus (human GABA B R1d) (13,15). Finally, GABA B R1e is a soluble truncated version of GABA B R1a (16).…”

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confidence: 99%

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Structural Analysis of the Complement Control Protein (CCP) Modules of GABAB Receptor 1a

Blein

Ginham

Uhrı́n

et al. 2004

Journal of Biological Chemistry

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The ␥-aminobutyric acid type B (GABA B ) receptor is a heterodimeric G-protein-coupled receptor. In humans, three splice variants of the GABA B receptor 1 (R1) subunit differ in having one, both, or neither of two putative complement control protein (CCP) modules at the extracellular N terminus, prior to the GABA-binding domain. The in vivo function of these predicted modules remains to be discovered, but a likely association with extracellular matrix proteins is intriguing. The portion of the GABA B R1a variant encompassing both of its CCP module-like sequences has been expressed, as have the sequences corresponding to each individual module. Each putative CCP module exhibits the expected pattern of disulfide formation. However, the second module (CCP2) is more compactly folded than the first, and the three-dimensional structure of this more C-terminal module (expressed alone) was solved on the basis of NMRderived nuclear Overhauser effects. This revealed a strong similarity to previously determined CCP module structures in the regulators of complement activation. The N-terminal module (CCP1) displayed conformational heterogeneity under a wide range of conditions whether expressed alone or together with CCP2. Several lines of evidence indicated the presence of native disorder in CCP1, despite the fact that recombinant CCP1 contributes to binding to the extracellular matrix protein fibulin-2. Thus, we have shown that the two CCP modules of GABA B R1a have strikingly different structural properties, reflecting their different functions.␥-Aminobutyric acid (GABA) 1 is the principal inhibitory neurotransmitter of the vertebrate central nervous system. It is the ligand for both ionotropic GABA type A receptors and metabotropic GABA type B (GABA B ) receptors. GABA B receptors belong to G-protein-coupled receptor class III, which includes metabotropic glutamate receptors, Ca 2ϩ -sensing receptors, and some pheromone and taste receptors (1). Agonist and antagonists of GABA B receptors have been shown to be effective in clinical cases or animal models of nociception, depression, addiction, epilepsy, and cognitive impairment. Selective GABA B receptor ligands could also be useful in the treatment of peripheral nervous system disorders (2, 3).The GABA B receptor is composed of subunits termed GABA B R1 and GABA B R2, both of which are needed for receptor function (4 -7). The two subunits share a similar molecular architecture, common to all class III G-protein-coupled receptors, consisting of a large extracellular N-terminal domain encompassing a ligand-binding site, followed by a transmembrane heptahelical domain and an intracellular C-terminal tail. Coupling to G-proteins is mediated by the intracellular loops connecting the transmembrane helices and the C-terminal region.In class III G-protein-coupled receptors, the extracellular domain of each subunit is proposed to have a dynamic bilobate structure, where the two globular lobes form a "clamshell"-like shape. The current model for the function of these dimers suggest...

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Section: Construction Of Expression Vectors Encoding Ccp1 Ccp2mentioning

confidence: 99%

Section: Construction Of Expression Vectors Encoding Ccp1 Ccp2mentioning

confidence: 99%

Section: -Anilinonaphthalene-8-sulfonic Acid (Ans) Fluorescencementioning

confidence: 99%

Section: -Anilinonaphthalene-8-sulfonic Acid (Ans) Fluorescencementioning

confidence: 99%

mentioning

confidence: 99%

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Structural Analysis of the Complement Control Protein (CCP) Modules of GABAB Receptor 1a

Blein

Ginham

Uhrı́n

et al. 2004

Journal of Biological Chemistry

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Differential expression of ?-aminobutyric acid type B receptor-1a and -1b mRNA variants in GABA and non-GABAergic neurons of the rat brain

et al. 2000

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GABA_BR1 receptor protein expression in human mesial temporal cortex: Changes in temporal lobe epilepsy

Muñoz

Arellano

DeFelipe

2002

J of Comparative Neurology

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Immunocytochemistry was used to examine gamma-aminobutyric acid beta (GABA)(B)R1a-b protein expression in the human hippocampal formation (including dentate gyrus, hippocampus proper, subicular complex, and entorhinal cortex) and perirhinal cortex. Overall, GABA(B)R1a-b immunostaining was intense and widespread but showed differential areal and laminar distributions of labeled cells. GABA(B)R1a-b-immunoreactive (-ir) neurons were found in the three main layers of the dentate gyrus, the most intense labeling being present in the polymorphic layer, whereas the granule cells were moderately immunoreactive. Except for slight variations, similar distribution patterns of GABA(B)R1a-b immunostaining were found along the different subfields of the Ammon's horn (CA1-CA4). The highest density of GABA(B)R1a-b-ir neurons was localized in the stratum pyramidale, where virtually every pyramidal cell was intensely immunoreactive, including the proximal part of the apical dendrites. Within the subicular complex, a more intense GABA(B)R1a-b immunostaining was found in the subiculum than in the presubiculum or parasubiculum, especially in the pyramidal and polymorphic cell layers. In the entorhinal cortex, distribution of GABA(B)R1a-b immunoreactivity was localized mainly in both pyramidal and nonpyramidal cells of layers II, III, and VI and in the superficial part of layer V, with layers I, IV, and deep layer V being less intensely stained. In the perirhinal cortex, the most intense GABA(B)R1a-b immunoreactivity was located in the deep part of layer III and in layer V and was mainly confined to medium-sized and large pyramidal cells. Thus, the differential expression, but widespread distribution, of GABA(B)R1a-b protein found in the present study suggests the involvement of GABA(B) receptors in many circuits of the human hippocampal formation and adjacent cortical structures. Interestingly, the hippocampal formation of epileptic patients (n = 8) with hippocampal sclerosis showed similar intensity of GABA(B)R1a-b immunostaining in the surviving neurons located within or adjacent to those regions presenting neuronal loss than in the controls. However, surviving neurons in the granule cell layer of the dentate gyrus displayed a significant reduction in immunostaining in 7 of 8 patients. Therefore, alterations in inhibitory synaptic transmission through GABA(B) receptors appears to affect differentially certain hippocampal circuits in a population of epileptic patients. This reduction in GABA(B)R1a-b expression could contribute to the pathophysiology of temporal lobe epilepsy.

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Cloning and Tissue Distribution of Novel Splice Variants of the Rat GABABReceptor

Cited by 119 publications

References 0 publications

Structural Analysis of the Complement Control Protein (CCP) Modules of GABAB Receptor 1a

Structural Analysis of the Complement Control Protein (CCP) Modules of GABAB Receptor 1a

Differential expression of ?-aminobutyric acid type B receptor-1a and -1b mRNA variants in GABA and non-GABAergic neurons of the rat brain

GABA_BR1 receptor protein expression in human mesial temporal cortex: Changes in temporal lobe epilepsy

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Cloning and Tissue Distribution of Novel Splice Variants of the Rat GABABReceptor

Cited by 119 publications

References 0 publications

Structural Analysis of the Complement Control Protein (CCP) Modules of GABAB Receptor 1a

Structural Analysis of the Complement Control Protein (CCP) Modules of GABAB Receptor 1a

Differential expression of ?-aminobutyric acid type B receptor-1a and -1b mRNA variants in GABA and non-GABAergic neurons of the rat brain

GABABR1 receptor protein expression in human mesial temporal cortex: Changes in temporal lobe epilepsy

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GABA_BR1 receptor protein expression in human mesial temporal cortex: Changes in temporal lobe epilepsy