神经元型一氧化氮合酶在血管性痴呆大鼠海马中的表达

目的 探讨神经元型一氧化氮合酶(nNOS)在血管性痴呆(VD)大鼠海马中的表达。方法 将60只大鼠随机分为：对照组、VD12h组、VD1d组、VD3d组、VD7d组。采用反复夹闭双侧颈总动脉方法建立VD大鼠模型，用HE染色观察各组大鼠海马CA1区神经元的数目；应用免疫组化染色和Western印迹方法检测nNOS在大鼠海马中的表达。结果 VD12h组、VD1d组、VD3d组、VD7d组大鼠海马CA1区神经元数均明显下降。nNOS在对照组大鼠海马CA1区中弱表达．在VD12h组表达增强．VD1d组进一步增强，VD3d和7d组表达逐渐减弱。结论 nNOS可能参与缺血早期海马神经元的损害，是VD的发病机制之一。     

Different effects of eNOS and nNOS inhibition on transient forebrain ischemia

To clarify the functions of nitric oxide (NO) induced by either neuronal NO synthase (nNOS) or endothelial NO synthase (eNOS) after transient cerebral ischemia, we investigated the effects of L-N(5)-(1-iminoethyl)ornithine (L-NIO), a relatively selective eNOS inhibitor, and 7-nitroindazole (7-NI), a relatively selective nNOS inhibitor, on hippocampal dysfunction caused by cerebral ischemia. We measured mean arterial blood pressure (MABP), hippocampal blood flow, direct current (DC) potential, CA1 population spike (PS) and extracellular concentrations of glutamate from rat hippocampus after transient forebrain ischemia, which was induced by four-vessel occlusion for 10 min. L-NIO (20 mg/kg) and 7-NI (25 mg/kg) were administered intraperitoneally 20 min before ischemia. L-NIO, but not 7-NI, increased MABP before, during and after ischemia, compared with the vehicle group. 7-NI, but not L-NIO, reduced the amplitude of anoxic depolarization induced by ischemia. 7-NI recovered the PS amplitude in part 60 min after ischemia. 7-NI, but not L-NIO, reduced the ischemia-induced levels of glutamate. These results indicate that nNOS inhibition with 7-NI improves, at least in part, hippocampal dysfunction after ischemia, while eNOS inhibition with L-NIO worsens it.     

17Beta-estradiol stimulates regeneration of sciatic nerve in female mice

In ovariectomized mice with and without estrogen replacement, regeneration of the sciatic nerve after crush injury was studied. Functional recovery, quantified with sciatic functional index was significantly accelerated in estrogen-treated mice throughout the regeneration. On semi-thin sections of sciatic nerves in estrogen-treated mice we registered a greater total number of regenerating nerve fibers at the first week, and a higher mean axonal area at the third week of regeneration. Our results demonstrated that estrogen treatment enhances regeneration of the sciatic nerve.     

17 Beta-estradiol suppresses AMPA-induced increases in regional cerebral O2 consumption

We tested the hypothesis that 17 beta-estradiol would reduce the cerebral O2 consumption response resulting from glutamate receptor stimulation by alpha amino-3-hydroxy-5-methyl-isoxazole-4-propionate (AMPA). Fourteen ovariectomized rats were separated into 17 beta-estradiol (0.5 mg 21 day release pellet) and control (placebo pellet) groups to determine cerebral blood flow (14C-iodoantipyrine) and O2 consumption (microspectrophotometry). After topical cortical stimulation with 10(-3) M and 10(-4) M AMPA, cerebral blood flow increased significantly in both groups in a concentration-dependent manner. Cerebral O2 extraction was not significantly different in any region of the 17 beta-estradiol treated group. In the placebo treated group, the O2 extraction in the saline treated cortex and in the 10(-3) M AMPA treated cortex was significantly higher when compared to the 10(-4) M AMPA treated cortex. Cerebral O2 consumption in the control group increased by 20%, from 5.2 +/- 0.6 to 6.1 +/- 0.7, with 10(-4) M AMPA and significantly increased by 64% to 8.5 +/- 0.8 ml O2 min-1 100 g-1 with 10(-3) M AMPA. The 17 beta-estradiol group demonstrated no statistically significant difference in O2 consumption between the saline treated and AMPA treated cortex. Thus, 17 beta-estradiol reduced the effects of AMPA in increasing cerebral O2 consumption.     

17Beta-estradiol suppresses AMPA-induced increases in regional cerebral O2 consumption

Abstract

We tested the hypothesis that 17beta-estradiol would reduce the cerebral O₂ consumption response resulting from glutamate receptor stimulation by alpha amino-3-hydroxy-5-methyl-isoxazole-4-propionate (AMPA). Fourteen ovariectomized rats were separated into 17beta-estradiol (0.5 mg 21 day release pellet) and control (placebo pellet) groups to determine cerebral blood flow (¹⁴C-iodoantipyrine) and O₂ consumption (microspectrophotometry). After topical cortical stimulation with 10^-3 M and 10-4 M AMPA, cerebral blood flow increased significantly in both groups in a concentration-dependent manner. Cerebral O₂ extraction was not significantly different in any region of the 17beta-estradiol treated group. In the placebo treated group, the O₂ extraction in the saline treated cortex and in the 10^-3 M AMPA treated cortex was significanly higher when compared to the 10^-4 M AMPA treated cortex. Cerebral O₂ consumption in the control group increased by 20%, from 5.2 ± 0.6 to 6.1 ± 0.7, with 10^-4 M AMPA and significantly increased by 64% to 8.5 ± 0.8 ml O₂ min^-1 100 g^-1 with 10^-3 M AMPA. The 17beta-estradiol group demonstrated no statistically significant difference in O₂ consumption between the saline treated and AMPA treated cortex. Thus, 17beta-estradiol reduced the effects of AMPA in increasing cerebral O₂ consumption.     

Melatonin regulates neuronal plasticity in the hippocampus

The influence of melatonin on hippocampal evoked potentials initiated by low- and high-frequency electrical stimulations and by two pulses applied in rapid succession was investigated. In confirmation of our previous studies, melatonin attenuated the population spike triggered by low-frequency stimulation (0.03 Hz). High-frequency stimulation (HFS; 100 Hz for 1 sec, three times every 10 sec), which in control slices permanently facilitated neuronal excitability (347% +/- 32%), was also able to amplify the melatonin-depressed potential (467.8% +/- 59.6%). Because melatonin is a hydrophobic molecule, it was dissolved and applied in ethanol. Ethanol (0.4%) by itself reduced the magnitude of HFS-induced potentiation (233.5% +/- 16.8%). The slices stimulated with two pulses separated with a delay longer than 15 msec demonstrated a facilitation of the response to the second stimuli (paired-pulse facilitation; PPF). The influence of melatonin (100 microM) on PPF was biphasic: Shortly after addition of melatonin, PPF was briefly (5-10 min) reversed to paired-pulse inhibition (PPI), which gradually returned to a stable PPF. Ethanol (0.4%) applied without melatonin exerted only a marginal, facilitatory effect on PPF. The delay between two successively applied pulses, shorter than 13 msec, resulted in attenuation of the response to the second stimuli (PPI). Melatonin (100 microM) reversed the attenuation of the second potential within 15-20 min following its application. Ethanol applied by itself at the concentration of 0.4% temporarily (5-10 min), but significantly, depressed the second potential. These results demonstrate the ability of melatonin to modulate specific forms of plasticity in hippocampal pyramidal neurons.     

17Beta-estradiol blocks NMDA-induced increases in regional cerebral O(2) consumption

We tested the hypothesis that 17beta-estradiol would reduce the cerebral O(2) consumption response to stimulation of N-methyl-D-aspartate (NMDA) receptors. We determined NMDA receptor density in 10 ovariectomized Wistar female rats equally divided into a control group and 17beta-estradiol (500 microg/21 days) treated group. An autoradiographic assay using 125I-MK-801, an NMDA antagonist, was used to measure specific binding to NMDA receptors. Another 14 ovariectomized rats were separated into 17beta-estradiol and control groups to determine cerebral blood flow (14C-iodoantipyrine) and O(2) consumption (microspectrophotometry). 17Beta-estradiol caused a 20% decrease in specific binding to cortical NMDA receptors. After topical cortical stimulation with 10(-3)M and 10(-4)M NMDA, blood flow increased significantly in control from 73+/-5 in the saline treated cortex to 110+/-8 ml/min/100 g with 10(-3)M NMDA. In contrast, there was no significant change in blood flow in the 17beta-estradiol treated animals. Cerebral O(2) extraction increased significantly in the 10(-3)M NMDA treated cortex in both groups. Cerebral O(2) consumption in the control group significantly increased by 53%, from 3.7+/-0.2 to 5.7+/-0.5 with 10(-4)M NMDA and 72% to 6.4+/-2.4 ml O(2)/min/100 g with 10(-3)M NMDA. The 17beta-estradiol group demonstrated no significant difference between the saline treated and NMDA treated cortex. Thus, 17beta-estradiol blocked the effects of NMDA on cerebral O(2) consumption and this was associated with a slightly decreased number of NMDA receptors.     

Mannotriose regulates learning and memory signal transduction in the hippocampus

Rehmannia is a commonly used Chinese herb, which improves ieaming and memory. However, the crucial components of the signal transduction pathway associated with this effect remain elusive. Pri- mary hippocampal neurons were cultured in vitro, insulted with high-concentration （1 × 10-4 mol/L） cor- ticosterone, and treated with 1 × 104 mol/L mannotriose. Thiazolyl blue tetrazolium bromide assay and western blot analysis showed that hippocampal neuron survival rates and protein levels of glucocorti- cold receptor, serum and glucocorticoid-regulated protein kinase, and brain-derived neurotrophic factor were all dramatically decreased after high-concentration corticosterone-induced injury. This effect was reversed by mannotriose, to a similar level as RU38486 and donepezil. Our findings indicate that mannotriose could protect hippocampal neurons from high-concentration corticosterone-induced injury. The mechanism by which this occurred was associated with levels of glucocorticoid receptor protein, serum and glucocorticoid-regulated protein kinase, and brain-derived neurotrophic factor.     

Estrogen regulates bcl-x expression in rat hippocampus

In this study, we examined whether experimental alterations of circulating estrogen levels are associated with changes in the expression of bcl-x, an inhibitor of apoptosis. We report that bcl-x mRNA expression in rat hippocampus significantly decreases after reduction of estrogen levels resulting from ovariectomy. Exposure of ovariectomized rats to 17beta-estradiol for either 5 or 28 days restored bcl-x mRNA expression to levels at or above those observed in sham-ovariectomized control animals. These data demonstrate that physiological levels of estrogen regulate hippocampal expression of bcl-x, an important modulator of neuronal apoptosis. Estrogen-mediated regulation of bcl-x may be relevant to the maintenance of neuronal viability and may contribute to the mechanism of estrogen neuroprotection.     

delta 9-THC and 17-beta-estradiol in hippocampus

Electrophysiological field potentials recorded from in vitro hippocampal slice preparations show dose-dependent differences in response to 17-beta-estradiol (E2) and delta-9-tetrahydrocannabinol (THC) added to the incubation medium. Using a wide range of doses (1 pM-10 nM), it was found that mid-range concentrations of estradiol (100 pM) and THC (10 pM) tended to increase field potentials in CA1 of rodents. Higher dose levels of each agent were found to depress neuronal activity. In the context of prior findings, these results suggest that the two compounds share a common mechanism of action in the hippocampus.     

Dephosphorylation of eNOS on Thr495 after transient forebrain ischemia in gerbil hippocampus

We investigated phosphorylation of endothelial nitric oxide synthase (eNOS) at two major sites, Ser1177 and Thr495, which has a critical role to control its activity, in the gerbil hippocampal microvasculature after transient forebrain ischemia. Ser1177 phosphorylation was unchanged by 24 h after reperfusion, despite post-ischemic up-regulation of eNOS protein. However, Thr495 phosphorylation significantly and persistently decreased by 48 h. We here defined the changes in eNOS phosphorylation in vivo following brain ischemia/reperfusion. (ischemia).     

Stress differentially regulates synaptophysin and synaptotagmin expression in hippocampus.

BACKGROUND: In view of the effects of stress on synaptic plasticity, the regulation of synaptophysin and synaptotagmin expression by immobilization was analyzed by in situ hybridization. METHODS: Rats were exposed to immobilization stress, which induced typical behavioral alterations, such as reduced locomotor activity after stress exposure. Determination of mRNA levels of the integral synaptic vesicle proteins was performed immediately after acute or chronic immobilization. RESULTS: The results demonstrate that stress exposure leads to reduced expression of synaptophysin but increased expression of synaptotagmin in the hippocampus. CONCLUSIONS: This rapid and differential regulation of synaptic vesicle proteins could be responsible for some of the morphological, biochemical, and behavioral changes observed after stress exposure. These changes may be relevant to such clinical disorders as psychoses, depression, and posttraumatic stress disorder that are sensitive to stress and involve changes in neural and synaptic plasticity.     

小剂量伽玛刀照射对致癎大鼠脑神经元nNOS表达的影响

目的 通过观察低剂量伽玛刀照射对致痫大鼠大脑皮质及海马神经元nNOS表达的影响，初步探讨伽玛刀治疗癫痫的作用机制。方法 将44只青霉素致痫大鼠模型等分为实验组和实验对照组，对实验组进行伽玛刀照射（周边剂量12Gy），应用免疫组化方法检测两组大脑皮质及海马神经元nNOS表达的变化。结果 nNOS在两组动物的皮质和海马表达均有显著性差别．实验组显著性低于实验对照组；实验对照组表达呈双高峰现象。结论 nNOS在伽玛刀治疗癫痫的机制中具有重要作用。     

K+ M-current regulates the transition to seizures in immature and adult hippocampus

PURPOSE: Loss-of-function mutations in Kv7.2 or Kv7.3 K(+) channel subunits underlies the neonatal epilepsy benign familial neonatal convulsions (BFNC). These two subunits interact to form a functional K(+) channel that underlies the M-current (I(M)), a voltage-dependent noninactivating K(+) current. In BFNC, seizures begin shortly after birth, and spontaneously remit in the first few months of life. The nature of this window of vulnerability is unclear. We address this issue using a hippocampal slice model, to study the effects of I(M) blockade or augmentation on epileptiform activity. METHODS: We used the Mg(+)(+)-free seizure model in adult and immature (P8-P15) acute rat hippocampal slices. We recorded from both CA1 and CA3 regions using extracellular and intracellular methods. RESULTS: When M-channels are blocked pharmacologically, the transition from interictal to ictal bursting becomes much more likely, especially in immature brain. We also show augmentation of I(M) is effective in stopping ictal events in immature brain, at the developmental age that approximates a human newborn in cortical development. I(M) appears to counter the sustained N-methyl-D-aspartate (NMDA) receptor-mediated depolarizations needed to trigger an ictal event. The increased likelihood of ictal bursting by I(M) blockade is not shared by other selective K(+) channel blockers that increase hippocampal excitability. CONCLUSIONS: Voltage-dependent M-channels are activated during interictal bursts and contribute to burst termination. When these channels are compromised, interictal burst duration becomes sufficient to trigger the sustained depolarizations that underlie ictal bursts. This transition to ictal bursts upon I(M) blockade is especially likely to occur in immature hippocampus. This selective function of M-channels likely contributes to the transient window of vulnerability to seizures that occurs with BFNC.     

丁苯酞对血管性痴呆大鼠认知功能、海马神经元结构及nNOS的影响

目的研究丁苯酞对血管性痴呆（VD）大鼠认知功能、海马神经元结构、及神经元型一氧化氮合酶（nNOS）的影响。方法采用结扎双侧颈总动脉方法制备慢性前脑缺血动物模型,100只老龄大鼠随机分5组,应用水迷宫、透射电镜及免疫组化方法对各组大鼠学习记忆、神经元结构、nNOS表达进行观察。结果与假手术组比较,大鼠学习记忆能力在造模后差异有显著性意义（P〈0.05）;大鼠海马区神经元在造模后变性水肿明显,与模型组比较,丁苯酞不同剂量治疗1月后,大鼠学习记忆能力明显改善（P〈0.05）,神经元变性水肿减轻,nNOS阳性神经元表达减少（P〈0.05）。结论丁苯酞能显著改善VD大鼠学习记忆能力;减轻神经元变性水肿;抑制nNOS阳性神经元表达。     

低剂量伽玛刀照射对致(癎)大鼠皮质及海马神经元c-fos和nNOS表达的影响

目的 通过观察低剂量伽玛刀照射对致(癎)大鼠大脑皮质及海马神经元c-fos和脑型一氧化氮合酶(nNOS)表达的影响,探讨伽玛刀治疗癫(癎)的作用机制.方法 将44只青霉素致(癎)大鼠模型等分为实验组和实验对照组大鼠各22只,另取4只正常大鼠作为正常对照组.实验组行伽玛刀照射(周边剂量12 Gy)后,应用免疫组化方法,观察大脑皮质及海马神经元c-fos和nNOS表达的变化.结果 无论是皮质还是海马,c-fos和nNOS在实验组与实验对照组动物之间,表达均有明显的差别,实验组表达明显少于实验对照组,而后者呈现双高峰现象.结论 c-fos和nNOS在伽玛刀治疗癫(癎)的机制中发挥了重要作用.     

低剂量伽玛刀照射对致大鼠皮质及海马神经元c-fos和nNOS表达的影响

目的 通过观察低剂量伽玛刀照射对致痫大鼠大脑皮质及海马神经元c—fos和脑型一氧化氮合酶（nNOS）表达的影响，探讨伽玛刀治疗癫痫的作用机制。方法将44只青霉素致痫大鼠模型等分为实验组和实验对照组大鼠各22只，另取4只正常大鼠作为正常对照组。实验组行伽玛刀照射（周边剂量12Gy）后，应用免疫组化方法，观察大脑皮质及海马神经元c-fos和nNOS表达的变化。结果无论是皮质还是海马，c—fos和nNOS在实验组与实验对照组动物之间，表达均有明显的差别，实验组表达明显少于实验对照组，而后者呈现双高峰现象。结论c—fos和nNOS在伽玛刀治疗癫痫的机制中发挥了重要作用。     

Mnemonic correlates of unit activity in the hippocampus

The role of the hippocampus in memory processing was examined by recording single unit activity while rats performed two different types of memory tasks. The same apparatus was used for all tasks; it consisted of two goal boxes, side by side, on the end of a runway. One goal box was white, the other was black. Experiment I used a working memory, delayed match-to-sample (DMTS) task. A trial began with a sample phase in which the rat was forced to a goal box containing a reward. The rat was then placed at the beginning of the runway again for the choice phase and allowed to enter either of the two goal boxes. Entering the goal box with the same color as that entered during the sample phase was rewarded. Experiment II used a within-subjects, within-units, design to test rats in two reference memory tasks, a cue task and a spatial task. During the cue task, the rat was rewarded for choosing the same colored goal box on each trial regardless of its spatial location. During the spatial task, the rat was rewarded for choosing the goal box in a specific location on each trial regardless of its color. During all tasks, the location of the goal boxes was changed between trials in a pseudorandom, counterbalanced fashion so that each colored goal box was on the right for half of the trials and on the left for half of the trials. During performance of the DMTS task, activity of most units was correlated with a combination of factors such as color and location, or color and phase. For example, most units showing differential activity in one of the colored goal boxes fired more when that box was in a certain spatial location, or during either the sample or choice phase. During performance of the reference memory tasks, the activity of most units was not correlated with behavior. However, the rate for some units changed between the cue and spatial tasks. When unit activity was correlated with behavior, it was dependent on a combination of dimensions such as color and spatial location. These results demonstrate that units in the hippocampus respond to combinations of stimulus dimensions such as color and spatial location, and to the temporal context necessary to solve a working memory task.