PubMed Health⌕ Search

Biomedical subjects

Jian-hong Luo

Publications and source records attributed to Jian-hong Luo.

12 recordsLinked to original sources
PubMed2006

Ca2+ sparks and Ca2+ glows in superior cervical ganglion neurons.

AIM: Ca2+ release from the endoplasmic reticulum (ER) is an integral component of neuronal Ca2+ signaling. The present study is to investigate properties of local Ca2+ release events in superior cervical ganglion (SCG) neurons. METHODS: Primary cultured SCG neurons were prepared from neonatal rats (P3-P7). Low concentration of caffeine was used to induce Ca2+ release from the ER Ca2+ store, and intracellular Ca2+ was recorded by high-resolution line scan confocal imaging and the Ca2+ indicator Fluo-4. RESULTS: Two populations of local Ca2+ release events with distinct temporal characteristics were evoked by 1.5 mmol/L caffeine near the surface membrane in the soma and the neurites of SCG neurons. Brief events similar to classic Ca2+ sparks lasted a few hundreds of milliseconds, whereas long-lasting events displayed duration up to tens of seconds. Typical somatic and neurite sparks were of 0.3- and 0.52-fold increase in local Fluo-4 fluorescence, respectively. Typical Ca2+ glows were brighter (deltaF/F0 approximately 0.6), but were highly confined in space. The half maximum of full duration of neurite sparks was much longer than those in the soma (685 vs 381 ms). CONCLUSION: Co-existence of Ca2+ sparks and Ca2+ glows in SCG neurons indicates distinctive local regulation of Ca2+ release kinetics. The local Ca2+ signals of variable, site-specific temporal length may bear important implications in encoding a 'memory' of the trigger signal.

Animals↗
PubMed2006

Phorbol-induced surface expression of NR2A subunit homologues in HEK293 cells.

AIM: N-methyl-D-aspartate receptors (NMDAR) are heteromeric complexes primarily assembled from NR1 and NR2 subunits. In normal conditions, NR2 subunits assemble into homodimers in the endoplasmic reticulum (ER). These homodimers remain in the ER until they coassemble with NR1 dimers and are trafficked to the cell surface. However, it still remains unclear whether functional homomeric NMDAR exist in physiological or pathological conditions. METHODS: We transfected GFP-NR2A alone into HEK293 cells, treated the cells with PKC activator 12-myristate-13 acetate (PMA), and then detected surface NR2A subunits with a live cell immunostaining method. We also used a series of NR2A mutants with a partial deletion of its C-terminus to identify the regions that are involved in the PMA-mediated surface expression of NR2A subunits. RESULTS: NR2A subunits were expressed on the cell membrane after incubation with PMA (200 nmol/L, 30 min), although no functional NMDA channels were detected after PMA-induced membrane trafficking. Immunostaining with an ER marker also revealed that NR2A subunits were exported from the ER after PMA treatment. Furthermore, the deletion of amino acids between 1149-1347 or 1354-1464 of NR2A inhibited PMA-induced surface expression of NR2A subunits. CONCLUSION: First, our data suggests that PMA treatment can induce the surface expression of homomeric NR2A subunits. Furthermore, this process is probably mediated by the NR2A C-terminal region between positions 1149 and 1464.

Cell Line↗
PubMed2005

Influence of low dietary histamine on seizure development of chemical kindling induced by pentylenetetrazol in rats.

AIM: To determine the role of dietary low histamine on the seizure development of pentylenetetrazol (PTZ)-induced kindling in rats. METHODS: After 14 d of feeding on a low histamine diet (LH, containing 0.145 mumol/g of histamine), the rats were chemically kindled by repeated intraperitoneal injection of a subconvulsant dose of PTZ (35 mg/kg) once every 48 h, and seizure activity of kindling was recorded for 30 min. Histamine in brain samples was analyzed using a high performance liquid chromatography system with a fluorescence spectrofluorometer. RESULTS: The LH diet induced an increase in seizure response (seizure susceptibility) to the first trial of PTZ, and resulted in facilitation of subsequent PTZ kindling process (seizure development). The histamine levels in the cortex, hippocampus, and hypothalamus of LH-treated rats decreased significantly and these changes correlated well with seizure behavior (r = 0.875, 0.651, and 0.796, respectively). In addition, chronic kindled seizures resulted in a significant increase of the histamine content in the cortex and hypothalamus in the LH-fed groups. CONCLUSION: These findings indicate that the histamine in daily food could influence the brain histaminergic function, and play an important role in regulating seizure susceptibility.

Animals↗
PubMed2005

GM1 stabilizes expression of NMDA receptor subunit 1 in the ischemic hemisphere of MCAo/reperfusion rat.

OBJECTIVE: To determine the protective effect of monosialoganglionside (GM1) and evaluate the influence of GM1 on expression of N-methyl-D-aspartate receptor subunit 1 (NMDAR1) in Sprague-Dawley (SD) rats with focal cerebral ischemia-reperfusion (I/R). METHODS: Left middle cerebral artery (MCA) was occluded by an intraluminal suture for 1 h and the brain was reperfused for 72 h in SD rats when infarct volume was measured, GM1 (10 mg/kg) was given ip (intraperitoneally) at 5 min (group A), 1 h (group B) and 2 h (group C) after MCA occlusion (MCAo). Expression of NMDAR1 was detected by Western blot at various time after reperfusion (4 h, 6 h, 24 h, 48 h and 72 h) in ischemic hemispheres of the rats with or without GM1 administered. RESULTS: (1) Adjusted relative infarct volumes of groups A and B were significantly smaller than that of group C and the control group (P<0.01 and P<0.05, respectively). (2) Expression level of NMDAR1 was temporally high at 6 h after reperfusion, and dipped below the normal level at 72 h after reperfusion. GM1 at 5 min after MCAo significantly suppressed the expression of NMDAR1 at 6 h after reperfusion (P<0.05 vs the control). At 72 h after reperfusion, the NMDAR1 expression level of rats treated with GM1 administered (at 5 min or 2 h after MCAo) was significantly higher than that of the control (P<0.05). CONCLUSION: GM1 can time-dependently reduce infarct volume in rats with focal cerebral I/R partly through stabilizing the expression of NMDAR1.

Animals↗
PubMed2004

[Effects of pentylenetetrazol at subconvulsant and convulsant dose on NMDA receptor subunits in rats].

OBJECTIVE: To investigate the expression of N-methyl-D-asparate (NMDA) receptor subunit proteins after administration of different doses of pentylenetetrazol (PTZ). METHODS: After ip injection of a subconvulsant (35 mg/kg) and convulsant (50 mg/kg) dose of PTZ, the rats were decapitated at different time points. The levels of cortical NR1 NR2A and NR2B subunit proteins were detected by immunoblotting. RESULT: 35 mg/kg PTZ and 50 mg/kg PTZ elicited different behavioral changes (P<0.001). The NR2A subunit in the cortex significantly increased 1 h after PTZ injection (P<0.05). For the 50 mg/kg group, both the NR2A and NR2B subunits proteins increased at 1 h in the cortex and then decreased; the protein levels returned to normal after 48 h. However, NR1 subunit had no changes. CONCLUSION: The NR2 subunit is involved in PTZ-induced seizure.

Animals↗
PubMed2004

[Role of area postrema of medulla in regulation of rat cardiovascular activity].

OBJECTIVE: To explore the role of area postrema (AP) of medulla in control of cardiovascular functions in rat. METHODS: (1) Sprague Dawley rats were anaesthetized with urethane and pentobarbital and the AP was stimulated by electrical stimulus with intensity of 0.1 mA and frequencies ranged 10 approximate, equals 80 Hz. (2) Excitatory amino acid L-glutamate (L- Glu, 0.1 approximate, equals 0.5 mol/L) was microinjected into AP in urethane anaesthetized rats and the changes of mean arterial pressure (MAP) and heart rate (HR) were recorded. RESULT: (1) When the frequencies of 10 Hz, 20 Hz and 40 Hz were used, the electrical stimulation of AP caused decrease of MAP and HR (P<0.001),while the electrical stimulation with the frequencies of 60 Hz and 80 Hz caused an increase of MAP (P<0.05) but a decrease of HR (P<0.001). (2) Microinjection of L-Glu at 0.1 mol/L had no effect on MAP and HR (P>0.05), but it decreased MAP and HR at 0.15 mol/L (P<0.001, P<0.05). The MAP was increased (P<0.001) but HR (P<0.05) was decreased at the concentrations of 0.2 mol/L and 0.5 mol/L, respectively. CONCLUSION: Alterations of MAP and HR induced by electrical or chemical stimulation on AP of medulla are related to the frequency of electrical stimulation or concentration of L-Glu.

Animals↗
PubMed2003

[Correlation between basic expression level of NMDA receptor NR1 subunit protein in hippocampus and learning ability in rats].

OBJECTIVE: To investigate the relationship between basic expression level of NMDA receptor NR1 subunit protein in hippocampus and learning ability of rats. METHODS: Using a novel-object recognition model and Morris water maze,the novel-object recognition ability and spatial memory of SD rats were ranked, and grouped as the high (top 20 %) and the low (bottom 20%), respectively. NR1 subunit protein levels in hippocampus were measured by quantitative immunoblotting with NR1 subunit specific antibody. RESULT: The level of NR1 subunit protein in hippocampus in the high novel-object recognition ability group was 60% (P<0.01), higher than that in the low one, and in the high spatial memory group it was 45.4 % (P<0.05), higher than that in the low one, respectively. CONCLUSION: The basic expression level of NR1 subunit protein in hippocampus is related to novel-object recognition ability and spatial memory of rats.

Animals↗
PubMed2003

[Surface expression and co-localization of NMDA receptor and AMPA receptor on dendritic tree of hippocampal neurons in culture].

OBJECTIVE: To investigate the developmental profiles on surface expression and co-localization of NMDA receptor clusters and AMPA receptor clusters on dendrite in cultured hippocampal neurons of rats. METHODS: Green fluorescent protein tagged GluR2 subunit (GFP-GluR2) and FLAG tagged NR2B subunit (FLAG-NR2B) were transfected into cultured hippocampal neurons at 5 days in vitro (DIV5). FLAG-NR2B containing NMDA receptor clusters and GFP-GluR2 containing AMPA receptor clusters expressed on membrane surface were then labeled in living neurons using anti FLAG mAb/Cy3-conjugated anti-mouse antibody and anti-GFP pAb/Alex488-conjugated anti-rabbit antibody. RESULT: The numbers of receptor cluster per 100 microm dendrite in the neurons at DIV7 and DIV19 were 39.7+/-5.0 and 64.7+/-6.1 (P<0.01) for NR2B-NMDAR, 59.1+/-3.3 and 99.7+/-6.4 (P<0.01) for GluR2-AMPAR, and 29.9+/-4.5 and 37.5+/-2.5(P<0.05) for the co-localized, respectively. At DIV7 and DIV19, 75.4% and 57.9% NR2B-NMDAR clusters were co-localized with GluR2-AMPAR; and 50.6% and 37.6% GluR2-AMPAR clusters were co-localized with NR2B-NMDAR, respectively. CONCLUSION: The density of NR2B-NMDAR containing and GluR2-AMPAR containing receptor clusters increases during development of hippocampal neurons in culture. Although the co-localized clusters are increased as well in an unit length of dendrite, the extent to which the two receptor clusters are co-localized decreases. These data imply a possible change in the partnership of AMPA receptor subtype and NMDA receptor subtype at newly formed synapses during development.

Animals↗
PubMed2003

[Surface expression of NMDA receptors composed of NR1 subunit and NR2A subunit mutants with partially deleted C-terminus in HEK293 cells].

OBJECTIVE: To examine the potential function of NMDA receptor NR2A subunit C-terminus in assembling and surface expression of the receptor in HEK293 cells. METHODS: Five vectors GFP- NR2ADeltaC1- DeltaC5 were constructed for expressing N-terminally GFP-tagged NR2A with C-terminal deletion at different regions by using conventional techniques of molecular cloning. The deleted region for NR2ADeltaC1-Delta C5 was 897L-1017S, 1024D-1142P, 1149D-1347G, 1354S-1464V, and 897L-1464V. These plasmids were transfected alone or co-transfected with NR1-1a into HEK293 cells. The surface NMDA receptors were immuno-stained using rabbit antibody against GFP and Cy3 conjugated secondary antibody in living cells. RESULT: The vectors GFP-NR2ADeltaC1-DeltaC5 were generated and all of them expressed GFP fluorescence in the transfected cells. Surface NMDA receptors were detected by immuno-labeling with anti-GFP in the cells co-transfected by NR1-1a and any one of GFP-NR2ADeltaC1-DeltaC5. However, no surface expression of NR2A proteins was found in the transfected cells with any one of these plasmids alone. CONCLUSION: Within the region downstream from the 897L of NR2A subunit, neither a particular domain directly interacted with ER retention domain in NR1-1a C1 cassette, nor that determining ER retention of NR2A subunit itself has been found, indicating that more complicated mechanisms might exist in which the subunit assembling and targeting to plasma membrane of NMDA receptors undergo.

Cell Line↗
PubMed2003

[Construction of expression vectors for C-terminally deleted NR2B subunit mutants and their application in the study of assembling of NMDA receptors].

OBJECTIVE: To investigate the role of NR2B subunit C-terminus in assembling and surface expression of NMDA receptor subtype composed of NR1-1a/NR2B subunits. METHODS: Eight vectors NR2BDelta1-Delta8) expressing GFP-tagged NR2B subunit mutants with various deletion in the carboxyl-terminal region were generated by conventional molecular cloning techniques. Each of these vectors was transfected alone, or co-transfected with NR1-1a into HEK293 cells. NR1-1a/GFP-NR2B receptors on membrane surface of the living transfected cells were immuno-stained using rabbit antibody against GFP followed by Cy3 conjugated secondary antibody. RESULTS: The eight vectors NR2BDelta1-Delta8 were successfully constructed. No surface labeling of GFP-tagged NMDA receptor was found for those transfected cells with NR1-1a, GFP-NR2B, and GFP-NR2BDelta1-Delta8 alone. GFP-tagged NMDA receptors were immuno-stained by anti-GFP for those cells co-transfected by NR1-1a and GFP-NR2B or GFP-NR2BDelta1-Delta6, which were mutants with partially deleted c-terminus at different region. However, positive stained was not found for those cells co-transfected by NR1-1a and GFP-NR2BDelta 7 (lack of most C-terminus and with PDZ binding motif fused with TM4) or GFP-NR2BDelta8 (lack of whole C-terminus). CONCLUSIONS: The formation of NR1-1a/NR2B sub-type NMDA receptor requires co-expression and assembling of NR1-1a and NR2B subunits. Shield or inhibition of ER retention motif within C1 cassette of NR1-1a subunit by NR2B subunit when assembling is not dependent on any particular region in NR2B C-terminus.

Cell Line↗