1.Response of P2X receptors in CA1 region of rat hippocampus to ATP and suramin
Journal of Third Military Medical University 2003;0(16):-
Objective To investigate the effects of ATP, suramin, ivermectin (IVM) and low pH value on pyramidal cells of rat hippocampus. Methods Pyramidal cells were rapidly dissociated from the hippocampus of 7-day postnatal rats by mechanical and enzymatic methods. Effects of ATP, suramin, IVM and low pH value on P2X receptor of neurons were studied by the technique of whole cell patch clamp. Results The transmembrane current was affected by ATP and suramin. There were different effects of IVM and low pH value on ATP induced current on different neurons. Conclusion There is extensive expression of P2X receptor on hippocampus pyramidal cells of rats. The expression of P2X receptors subunits are different among neurons.
2.Effect of hypoxia on NR1 subunit of NMDA receptor in rat cortex and hippocampus
Xigui WU ; Yandong ZHAO ; Huaizhen RUAN
Journal of Third Military Medical University 2003;0(18):-
Objective To observe the expression and phosphorylation of N-methyl-D-aspartate (NMDA) receptor NR1 subunit in rat cortex and hippocampus under high altitude hypoxia. Methods The adult SD rats were exposed to hypobaric hypoxia imitating 5 500-meter high altitude for 8 h daily for 3, 7, 14, 21 d. Immunohistochemical staining, Western blotting and co-immunoprecipitation were used to detect the expression and phosphorylation of NR1 in rat cortex and hippocampus. Results Both immunohistochemistry and Western blotting showed NR1 expression in rat cortex and hippocampus was increased under hypoxia in a time-dependent manner. The tyrosine phosphorylation of hypoxia groups was increased, and reached the peak on day 14 after hypoxia, then decreased, still higher than that of control groups till day 21 (P
3.Rapid inhibition of ATP-induced currents by dexamethasone in rat dorsal root ganglion neurons
Xiaohong LIU ; Junwei ZENG ; Huaizhen RUAN
Medical Journal of Chinese People's Liberation Army 1983;0(05):-
Objective Glucocorticoid hormone may nongenomically affect cell functions in addition to its classic effects on gene expression. The purpose of present study was to explore whether dexamethasone, a synthetical glucocorticoid hormone, has a rapid nongenomic effect on ATP-induced currents in rat dorsal root ganglion (DRG) neurons and the related signal transduction pathway. Methods The effects of dexamethasone on ATP-induced currents were studied on cultured DRG neurons using patch clamp technique. Results Three types of currents (transient, sustained and biphasic) were evoked by ATP (100 ?mol/L) in cultured DRG neurons. When DRG neurons were pretreated with dexamethasone (0.01-10?mol/L) for 30s, inhibition of the transient current and the transient component of the biphasic current evoked by ATP in DRG neurons was observed. The inhibitory effect of dexamethasone was dose-dependent. However, dexamethasone did not seem to affect the sustained current and the sustained component of the biphasic current induced by ATP. The inhibitory effect of dexamethasone on ATP-induced currents was blocked by glucocorticoid receptor antagonist RU38486 (10?mol/L) and protein kinase A inhibitor H-89 (10?mol/L), but not by G protein inhibitor GDP-?-S (0.2mmol/L) and protein kinase C inhibitor chelerythrine chloride (10?mol/L). Conclusions Dexamethasone can selectively inhibit the transient current mediated by P2X3 receptors in DRG neurons. The inhibitory effect of dexamethasone might be mediated by glucocorticoid receptor through activating PKA signal pathway. These results suggest that glucocorticoid hormone might participate in the control of pain by modulating the actions of extracellular ATP in sensory neurons.
4.Expression change of P2X_3 receptor in spinal dorsal horn and dorsal root ganglion in inflammatory pain rats
Lu DU ; Jinhai ZHANG ; Huaizhen RUAN
Journal of Third Military Medical University 1984;0(02):-
Objective To investigate the expression change of P2X3 receptor in spinal dorsal horn and dorsal root ganglion(DRG) in rats with experimental inflammatory pain.Methods Totally 30 SD rats received a subcutaneous injection of carrageenan in the right paw to establish inflammatory pain model,and then divided randomly and equally into 5 groups,that is 6,12,24,72 and 168 h after injection.Another 6 rats undergoing a subcutaneous injection of normal saline in the right paw served as control.Pain threshold was assayed by thermal radiation and von Frey filament in 2,4,6,12,24,72 and 168 h after injection.Expression of P2X3 receptor in spinal dorsal horn and DRG of every group was measured by immunohistochemical staining(IHC).Results After injection of carrageenan,thermal hyperalgesia and mechanical allodynia appeared in the rats.The thermal withdrawl latency(TWL) and 50% paw withdrawl threshold(PWT) were decreased obviously from 2 h,descended to the lowest at 12 h,then returned to the normal level after 72 h.The expression of P2X3 receptor in spinal dorsal horn was increased obviously from 24 h,while that at DRG was increased obviously from 12 h.Both of the expressions lasted for 72 h,and returned to the normal level at 168 h after injection.Conclusion Thermal hyperalgesia and mechanical allodynia appear in the rats after carrageenan injection,and the P2X3 receptor is activated and up-regulated in the DRG and spinal cord,suggesting that P2X3 receptor may play an important role in the incidence and development of inflammatory pain.
5.he roles of hypoxia and glutamic acid on NMDA receptor channel of hypothalamic neuron in rats
Penghui CHEN ; Huaizhen RUAN ; Xigui WU
Journal of Third Military Medical University 2001;23(4):429-431
Objective To study the effects of hypoxia and glutamic acid on the kinetic properties of NMDA receptor channel of the hypothalamic neurons in rats. Methods Cell-attached mode patch clamp technique was employed to record the single channel current of the NMDA receptor. Results The open probability of NMDA receptor channel was increased after acute hypoxia compared with that of normal state, the open time τ1,τ2 was changed from (0.33±0.10)ms,(4.36±0.26)ms to (0.93±0.22)ms,(7.64±0.72)ms, and the close time τ1,τ2 was from (18.03±3.50)ms,(171.50±19.10)ms to (3.42±1.02)ms,(19.39±3.07)ms. The mean open probability was changed from 0.12±0.05 in normal state to 0.66±0.36 in hypoxia state. Furthermore, glutamic acid can increase open time and open probability of NMDA receptor channel,decrease close time. Conclusion The excitability and the open probability of NMDA receptor channel of hypothalamic neurons increased by hypoxia is related to glutamate.
6.Dorsal root ganglion P2X_3 receptor expression and electrophysiological characteristics in rats with neuropathic pain
Xiaohong LIU ; Junwei ZENG ; Jinhai ZHANG ; Huaizhen RUAN
Journal of Third Military Medical University 1983;0(04):-
Objective To investigate the P2X3 receptor expression and electrophysiological characteristics in dorsal root ganglion (DRG) in rat with neuropathic pain caused by chronic constriction injury of sciatic nerve (CCI) Methods P2X3 receptor expressions in L4,L5 and L6 DRG following CCI were observed by using a polyclonal antibody to label the P2X3 receptor ATP-activated currents in corresponding DRG neurons following CCI were observed by using electrophysiological technique Results A significant increase in P2X3 immunoreactivity was observed in the ipsilateral (injured) L4,L5 and L6 DRG and spinal cord on 7,14 d after CCI In small diameter neurons,a significant increase in the number of cells exhibiting a transient current to ATP was observed on 7,14 d after CCI Moreover,amplitude of these currents was increased Conclusion After CCI,the expression and function of P2X3 receptor in corresponding DRG are increased
7.Effect of altitude hypoxia on somatostatin and ?-aminobutyric acid content of rat hypothalamus
Huaizhen RUAN ; Xiaotang FAN ; Jinhai ZHANG ; Bibo YUAN ; Xigui WU
Chinese Journal of Pathophysiology 1986;0(01):-
AIM:To observe the effect of altitude hypoxia on ?-aminobutyric (GABA) content and prepro-somatostatin mRNA (PPS-mRNA) in the rat hypothalamus. METHODS: Using altitude hypoxia model,in situ hybridization and amino acid analyzer, the number of PPS-mRNA and GABA content in rat hypothalamus was determined. RESULTS:After altitude hypoxia, the contents of GABA in hypothalamus and the number of PPS-mRNA neurons in periventricular nucleus (PeVN), paraventricular nucleus (PaVN) and arcuate nucleus (ArcN) increased significiantly. Bicuculline, a GABA receptor antagonits, could enhance PPS-mRNA expression evoked by altitude hypoxia, but had no effect on GABA content. CONCLUSION: Altitude hypoxia can induce neurotransmitter imbalance of hypothalamus.
8.Effect of altitude hypoxia on glutamate, aspartate and NOS in the rat hypothalamus
Huaizhen RUAN ; Xiaotang FAN ; Jinhai ZHANG ; Bibo YUAN ; Xigui WU
Chinese Journal of Pathophysiology 2000;0(11):-
AIM: To observe the effect of altitude hypoxia on glutamate, aspartate and nitric oxide synthase (NOS) in the rat hypothalamus. METHODS: Using altitude hypoxia model, amino acid analysis system and the NADPH-d histochemistry, we determined the content of glutamate, aspartate and the number of NADPH-d neurons in the rat hypothalamus. RESULTS: After altitude hypoxia, the contents of glutamate, aspartate in the hypothalamus of rats were increased significantly, densely and deeply stained NADPH-d neurons were seen in hypothalamic paraventricular nucleus (PVN)and supraoptic nucleus(SON). If rat were pretreated with the NMDA receptor blockers Ketamine (ip,40 mg/Kg)or AP-V(i.c.v, 10 ?g) , the number of NADPH-d neurons in the rat hypothalamic PVN and SON was markedly less than that in corresponding altitude hypoxia group. CONCLUSION: NMDA receptor may take part in the expression of hypothalamic NOS induced by altitude hypoxia.
9.Relationship between NO changes and LTP production in rat hippocampus in vitro
Zhian HU ; Jun LUO ; Haidi LI ; Huaizhen RUAN ; Xicheng LI ;
Journal of Third Military Medical University 2003;0(20):-
Objective To observe the time course of the changes in nitric oxide (NO) content and the activity of NO synthase (NOS) during hippocampal long term potentiation (LTP) in vitro . Methods The production and maintenance of LTP were detected by using extracelluar electrophysiological recording. NO content and NOS activity were determined by biochemical reaction. The expression of NOS mRNA was detected by in situ hybridization. Results Conditioning stimulation for 10 min induced LTP production and significant increases in NO content, NOS activity and expression of NOS mRNA. However, at 60 min after conditioning stimulation, LTP remained stably but NO content and NOS activity returned to the pre conditioning stimulation level. Moreover, the NOS mRNA was overexpressed at the early stage of LTP production. Conclusion Significant NO changes may occur at the early stage of LTP formation.
10.The experiences of project pattern in practical course of neurobiology
Penghui CHEN ; Huaizhen RUAN ; Wenqin CAI ; Xigui WU
Chinese Journal of Medical Education Research 2005;0(05):-
Neurobiology is to study the structure,function,development and regeneration of the nervous system at molecular,cellular and whole level.The objective of neurobiological teaching is to explore the importont and difficult points in teaching' the present article attempts to design a project pattern in practical course of neurobiology,which is suited to learning mode of medical college students.