Abstract:
:A clonal human embryonic kidney (HEK) 293 cell line was established that stably expressed the rat kappa-opioid receptor (rKOR) with a FLAG epitope at the amino terminus. The Kd for [3H]diprenorphine was 1.1+/-0.2 nM, and the Bmax was 2.6+/-0.4 pmol/mg. Dynorphin A (1-13), U69,593 and naloxone competitively inhibited [3H]diprenorphine binding with Ki values of 2.0, 18 and 18 nM, respectively, in good agreement with previously reported affinities for the unmodified receptor. U69,593 stimulated [35S]GTPgammaS binding in a concentration-dependent manner and caused phosphorylation of mitogen-activated protein (MAP) kinase, indicating that the activated epitope-tagged receptor triggered appropriate signaling pathways. Immunoblot analysis demonstrated that two immunoreactive receptor species with apparent molecular masses of 42 and 52 kDa were expressed. Previous studies indicated that the 42 kDa protein was localized intracellularly and was a precursor of the 52 kDa receptor, which was present at the cell surface. rKOR was extracted from transfected HEK 293 cell membranes with n-dodecyl-beta-D-maltopyranoside. Sequential use of wheat germ agglutinin chromatography, Sephacryl S300 gel filtration chromatography, anti-FLAG immunoaffinity chromatography and SDS/PAGE permitted purification of the 52 kDa receptor. MALDI-TOF mass spectrometry was used to identify peptides derived from rKOR following sequential in-gel digestion with trypsin and cyanogen bromide. Eighteen rKOR peptides were detected, corresponding to 27.1% coverage of the receptor. Precursor-selective MS/MS confirmed the identity of most of these peptides. In addition, we have identified heat shock protein 70 (HSP70) as a rKOR-interacting protein.
journal_name
Brain Resjournal_title
Brain researchauthors
Wannemacher KM,Terskiy A,Bian S,Yadav PN,Li H,Howells RDdoi
10.1016/j.brainres.2008.06.121subject
Has Abstractpub_date
2008-09-16 00:00:00pages
13-26eissn
0006-8993issn
1872-6240pii
S0006-8993(08)01609-0journal_volume
1230pub_type
杂志文章相关文献
BRAIN RESEARCH文献大全abstract::In non-neuronal cultures of cells dissociated from postnatal rat cerebellum astrocytes, identified by the presence of the marker protein glial fibrillary acidic protein (GFAP), displayed two distinct morphological forms. One class was stellate in shape with radially distributed fine processes, while the other class wa...
journal_title:Brain research
pub_type: 杂志文章
doi:10.1016/0165-3806(83)90143-8
更新日期:1983-11-01 00:00:00
abstract::Small iontophoretic application of wheat-germ-agglutinin conjugated to horseradish peroxidase, was used to simultaneously observe, in the rat, the afferent and the efferent connections of the cuneiformis nucleus (Cnf), in the same animal. The results demonstrate that the main Cnf afferent connections originate from: (...
journal_title:Brain research
pub_type: 杂志文章
doi:10.1016/0006-8993(89)90449-6
更新日期:1989-06-19 00:00:00
abstract::A neuron׳s response to a sound can be suppressed by the presentation of a preceding sound. It has been suggested that this suppression is a direct correlate of the psychophysical phenomenon of forward masking, however, forward suppression, as measured in the responses of the auditory nerve, was insufficient to account...
journal_title:Brain research
pub_type: 杂志文章
doi:10.1016/j.brainres.2016.02.043
更新日期:2016-05-15 00:00:00
abstract::The effects of acute ethanol exposure of chick spinal cord neurons were studied in tissue culture, using whole-cell voltage-clamp techniques. Results indicate that ethanol produces a persistent increase in the sensitivity of spinal neurons to GABA and glycine, with no change in input resistance or resting membrane pot...
journal_title:Brain research
pub_type: 杂志文章
doi:10.1016/0006-8993(88)90098-4
更新日期:1988-07-12 00:00:00
abstract::Morphological changes in neocortical somatostatin- and neuropeptide-Y-immunoreactive cells in senile dementia of the Alzheimer type (SDAT) were studied using light-microscopic immunohistochemical methods. The density of somatostatin-immunoreactive cells in the neocortex did not decrease in cases of SDAT compared with ...
journal_title:Brain research
pub_type: 杂志文章
doi:10.1016/0006-8993(86)91099-1
更新日期:1986-04-02 00:00:00
abstract::The stapedius muscle activity associated with vocalization was analyzed in young and adult roosters. Our results show that remarkable differences in the behavior of vocalization-related stapedius muscle activity exist between these two ages. Unlike young roosters, electrical stimulation in the midbrain of adult cocks ...
journal_title:Brain research
pub_type: 杂志文章
doi:10.1016/0006-8993(90)90823-t
更新日期:1990-10-08 00:00:00
abstract::Basic fibroblast growth factor (FGF-2) isoforms of the adrenal medulla are differentially expressed during rat postnatal development. While the 18 and 23 kDa isoforms continuously rise towards the adult expression level, the 21 kDa isoform displays a peak expression at postnatal day 28. The peak expression of the 21 k...
journal_title:Brain research
pub_type: 杂志文章
doi:10.1016/s0006-8993(97)00341-7
更新日期:1997-05-23 00:00:00
abstract::Axon-to-myelin transfer of lipids precursors have been studied in the rabbit optic system by intraocular injection of [32P]orthophosphate, [14C]glycerol and [3H]glycerol. Choline and ethanolamine phosphoglycerides and myelin showed increasing [32P]-radioactivity between 7 and 21 days following injection, while [3H]- a...
journal_title:Brain research
pub_type: 杂志文章
doi:10.1016/0006-8993(83)90136-1
更新日期:1983-06-20 00:00:00
abstract::The intent of this study was to investigate neurochemical and behavioural effects of aversive stimulation and the impact of auditory background noise. Using in vivo microdialysis, hippocampal acetylcholine was extracted and subjected to HPLC analysis while male Wistar rats were exposed to aversive stimulation similar ...
journal_title:Brain research
pub_type: 杂志文章
doi:10.1016/s0006-8993(00)02842-0
更新日期:2000-11-03 00:00:00
abstract::The amygdala has long been implicated in conditioned fear. The mesencephalic dopaminergic system provides a rich innervation to the amygdala [J.H. Fallon, P. Ciofi, Distribution of monoamines within the amygdala, in: J.P. Aggleton (Ed.), The Amygdala: Neurobiological Aspects of Emotion, Memory and Mental Dysfunction, ...
journal_title:Brain research
pub_type: 杂志文章
doi:10.1016/s0006-8993(99)01291-3
更新日期:1999-05-08 00:00:00
abstract::The stimulated (40 Hz) release of enkephalin from the myenteric plexus can be modulated by multiple types of opiate receptor. The direction of the modulation is not fixed but is bimodal. Both an inhibition and an enhancement of evoked release is observed depending on the concentration of opioid agonist that is used. E...
journal_title:Brain research
pub_type: 杂志文章
doi:10.1016/0006-8993(89)91594-1
更新日期:1989-12-11 00:00:00
abstract::Effects of pre-treatment of synaptic membranes with beta-adrenoceptor agonists and cholera toxin on [3H]clonidine and [3H]yohimbine binding were examined in rat cerebral cortex. Pre-incubation of cerebral cortical membranes with isoproterenol (10 or 200 microM) or dobutamine (1, 10 or 200 microM) at 37 degrees C for 4...
journal_title:Brain research
pub_type: 杂志文章
doi:10.1016/0006-8993(85)90531-1
更新日期:1985-03-11 00:00:00
abstract::Systemic administration of mesenchymal stem cells (MSCs) following cerebral infarction exerts functional improvements. Previous research has suggested potential therapeutic mechanisms that promote neuroprotection and synaptogenesis. These include secretion of neurotrophic factors, remodeling of neural circuits, restor...
journal_title:Brain research
pub_type: 杂志文章
doi:10.1016/j.brainres.2018.05.033
更新日期:2018-09-15 00:00:00
abstract::This study investigated the relationship between central cholinergic neurons and insulin secretion in bilateral adrenalectomized fed rats. Neostigmine (a cholinesterase inhibitor, 5 x 10(-8) mol) administered into the third cerebral ventricle produced significant increases in hepatic venous plasma insulin and glucose ...
journal_title:Brain research
pub_type: 杂志文章
doi:10.1016/0006-8993(89)91003-2
更新日期:1989-07-24 00:00:00
abstract::An interesting feature of the kindling phenomenon relates to the finding that kindling established in one region of the brain may reduce the number of stimulations required to establish the phenomenon in a second region. It has been proposed that this 'transfer' phenomenon reflects the extent to which seizures arising...
journal_title:Brain research
pub_type: 杂志文章
doi:10.1016/s0006-8993(97)00779-8
更新日期:1997-10-10 00:00:00
abstract::The role of glial cells in nitric oxide production in the cerebellum of conscious rats was investigated with a glial selective metabolic inhibitor, fluorocitrate. The levels of nitric oxide metabolites (nitrite plus nitrate) in the dialysate following in vivo microdialysis progressively increased to more than 2-fold t...
journal_title:Brain research
pub_type: 杂志文章
doi:10.1016/s0006-8993(97)00372-7
更新日期:1997-07-11 00:00:00
abstract::Accumulating evidence suggests that the neuropeptide substance P (SP) and its principal receptor neurokinin 1 (NK1) play a specific role in the behavioral response to opioids and stress that may help to initiate and maintain addictive behavior. In animal models, the NK1 receptor is required for opioids to produce thei...
journal_title:Brain research
pub_type: 杂志文章,评审
doi:10.1016/j.brainres.2009.11.014
更新日期:2010-02-16 00:00:00
abstract::In order to characterize the neurons responsible for ACTH release and the suppression produced by adrenocortical steroids, we have studied the pharmacological sensitivity of neurons in the region containing the highest concentration of corticotropin releasing factor (CRF) in the brain and probably responsible for most...
journal_title:Brain research
pub_type: 杂志文章
doi:10.1016/0006-8993(83)90368-2
更新日期:1983-08-01 00:00:00
abstract::Finger representation on the first somatosensory cortex was mapped in conscious monkeys by single-unit recording. While representation was somatotopic in area 3, this was not the case in area 2, where the representation of different fingers overlapped. Our results indicate that area 2 is not a pure replica of the rece...
journal_title:Brain research
pub_type: 杂志文章
doi:10.1016/0006-8993(80)91139-7
更新日期:1980-09-22 00:00:00
abstract::Axonal projections of medial brain stem areas rich in serotonin-containing neurons were identified in layers I and II of cat medullary dorsal horn using EM autoradiography. Following [3H]amino acid injections into the brain stem, labeled axonal endings were found throughout layers I and II but were most numerous in la...
journal_title:Brain research
pub_type: 杂志文章
doi:10.1016/0006-8993(81)90729-0
更新日期:1981-01-26 00:00:00
abstract::The effects of target and non-target cells on the growth and function of intrastriatal grafts of mesencephalic dopamine neurons have been studied in rats with unilateral 6-hydroxydopamine-induced lesions of the nigrostriatal dopamine pathway. Cell suspensions of ventral mesencephalon from 14-15-day-old rat fetuses (ri...
journal_title:Brain research
pub_type: 杂志文章
doi:10.1016/0165-3806(86)90174-4
更新日期:1986-01-01 00:00:00
abstract::Physical exercise can induce brain plasticity and reduce the cognitive decline observed in type 1 diabetes mellitus (T1DM). We investigated the effects of physical exercise to prevent or reverse spatial memory deficits produced by diabetes and some biochemical and immunohistochemical changes in hippocampal astrocytes ...
journal_title:Brain research
pub_type: 杂志文章
doi:10.1016/j.brainres.2016.10.024
更新日期:2017-01-15 00:00:00
abstract::There was a distinct sex-related difference in the anterior pituitary level of substance P-like immunoreactivity (SP-LI) in adult rats; the anterior pituitaries of male rats contained a higher concentration of SP-LI than those of females. Neonatal castration (day 1) resulted in a marked decrease of the SP-LI level in ...
journal_title:Brain research
pub_type: 杂志文章
doi:10.1016/0006-8993(83)90863-6
更新日期:1983-08-29 00:00:00
abstract::In order to investigate the interplay between visuo-spatial attention and central attention, we varied the relative probability (25% vs. 75%) of the responses to lateralized targets in an attentional blink paradigm. When the first target was associated with a less probable response, we observed a larger attentional bl...
journal_title:Brain research
pub_type: 杂志文章
doi:10.1016/j.brainres.2007.09.004
更新日期:2007-12-14 00:00:00
abstract::gamma-L-Glutamyl-L-aspartic acid (gamma-LGLA) has been isolated from Datura stramonium; its structure has been determined and it was then synthesized. In male Swiss mice intraperitoneal administration of the natural peptide (125 nmol/kg) or of the synthetic peptide (25-2500 nmol/kg) 24 h after acquisition of a Y-maze ...
journal_title:Brain research
pub_type: 杂志文章
doi:10.1016/0006-8993(88)90878-5
更新日期:1988-04-19 00:00:00
abstract::Using the teased fiber recording method, we have compared pathophysiological properties of afferent axons injured in the infraorbital nerve (ION) vs the sciatic nerve in rats. Both myelinated and unmyelinated axons ending in ION neuromas produced much less ongoing discharge than those ending in sciatic nerve neuromas....
journal_title:Brain research
pub_type: 杂志文章
doi:10.1016/0006-8993(92)90753-v
更新日期:1992-05-01 00:00:00
abstract::Studies of first (L1) and second (L2) language representation in the brain have not identified the timing and locations of neural regions involved in L1 and L2 processing. Magnetoencephalography offers high spatial and temporal resolution and can be employed to disentangle subtle timing and neural control differences ...
journal_title:Brain research
pub_type: 杂志文章
doi:10.1016/j.brainres.2012.05.054
更新日期:2012-07-27 00:00:00
abstract::The effect of MK-801 on striatal dopamine (DA) release was investigated by using an in vivo microdialysis technique in the freely moving rat. Systemic injection of MK-801 (0.25, 0.5, 1, 2 mg/kg, i.p.) reduced the extracellular level of DA significantly and produced no change in the level of 3,4-dihydroxyphenylacetic a...
journal_title:Brain research
pub_type: 杂志文章
doi:10.1016/0006-8993(90)90197-j
更新日期:1990-09-24 00:00:00
abstract::A subset of olfactory receptor neurons of the Caribbean spiny lobster Panulirus argus possesses receptors for L-glutamate that can mediate both excitatory and inhibitory responses (P.C. Daniel, M.F. Burgess, C.D. Derby, Responses of olfactory receptor neurons in the spiny lobster to binary mixtures are predictable usi...
journal_title:Brain research
pub_type: 杂志文章
doi:10.1016/s0006-8993(97)00816-0
更新日期:1997-10-17 00:00:00
abstract::The distribution of cortical neurons which project to the spinal cord was investigated in the developing and adult rat using the retrograde transport of horseradish peroxidase (HRP). Following injections of HRP into the cervical spinal cord of a postnatal day 4 rat, retrogradely labeled neurons are located in layer Vb...
journal_title:Brain research
pub_type: 杂志文章
doi:10.1016/0165-3806(84)90161-5
更新日期:1984-04-01 00:00:00