Cytoprotective actions of 2,4-dimethoxybenzylidene anabaseine in differentiated PC12 cells and septal cholinergic neurons

The potential cytoprotective actions of a novel nicotinic agent 2,4-dimethoxybenzilidene anabaseine (DMXB) were investigated in differentiated PC12 cells and transected rat septal cholinergic neurons in vivo. In NGF-differentiated PC12 cells, removal of both NGF and serum led to cell loss, a reduced % of cells expressing neurites, the release of lactate dehydrogenase, and a decrease in total cellular protein. Cell loss was apparent within 24 h, and remained constant between 4–8 days post-NGF removal. NGF alone (100 ng/ml), DMXB (10 μM), but not nicotine (10 μM), prevented these cell and neurite losses. DMXB-induced cytoprotection was blocked by 1 μM mecamylamine. DMXB (1 mg/kg, ip) injected twice but not once per day protected cholinesterase-staining septal neurons from retrograde degeneration following unilateral fimbrial transections. The twice per day DMXB injection-protocol also decreased cell roundness among cholinesterase-staining cells in the lesioned septal hemisphere compared to saline-injected animals. These studies suggest that DMXB may exert cytoprotective activity in NGF-sensitive neuronal populations. © 1994 Wiley-Liss, Inc.     

Morphological types of ganglion cells in the dog and wolf retina.

The morphological types of ganglion cells in the dog and wolf retina were studied by intracellular staining with Lucifer Yellow. These retinae contain a range of ganglion cell types that closely correspond to those found in cat retina: alpha cells with large somata and large, relatively densely branched dendritic trees; beta cells with medium-sized somata and small, densely branched dendritic trees; and a variety of other types with smaller somata and varying dendritic branching patterns and dendritic field sizes. The correspondence of canine and cat ganglion cell types strengthens the view that there is a common set of ganglion cell types in carnivores. Alpha and beta cell dendritic trees of dog and wolf are monostratified in either the inner or the outer part of the inner plexiform layer, suggesting an on/off dichotomy in the response to light. Dendritic field sizes of dog alpha and beta cells increase from the central area to peripheral retina: alpha cell fields from 160-200 microns to about 1,100 microns diameter, and beta cell fields from 25 microns to about 360 microns diameter. These sizes are quantitatively very similar to those found in cat retina. The close qualitative and quantitative morphological correspondence of cat and dog ganglion cells suggests that they are also functionally very similar. It is likely that dog alpha cells have brisk-transient (Y), and dog beta cells brisk-sustained (X) concentric receptive fields. From the smallest beta cell sizes it is concluded that the visual acuity of the dog may be as good as that of the cat.     

Rapid effect of stress concentration corticosterone on glutamate receptor and its subtype NMDA receptor activity in cultured hippocampal neurons

Objective: To study the rapid effect of glucocorticoids (GCs) on NMDA receptor activity in hippocampal neurons in stress and to elucidate its underlying probable membrane mechanisms. Methods: Whole-cell patch-clamp recording was used to assess the effect of stress concentration corticosterone (B) on the responses of cultured hippocampal neurons to glutamate and NMDA (N-methy-D-asparatic acid). To make clear the target of B, intracellular dialysis of B(10μmol/L)through patch pipette and extracellular application of bovine serum albumin-conjugated corticosterone (B-BSA, 10μmol/L)were carried out to observe their influence on peak amplitude of NMDA-evoked current. Results: B had a rapid, reversible and inhibitory effect on peak amplitude of GLU-or NMDA-evoked current in cultured hippoeampal neurons. Furthermore, B-BSA had the inhibitory effect on INMDA as that of B, but intraeeUularly dialyzed B had no significant effect on INMDA. Conclusion: These results suggest that under the condition of stress, GCs may rapidly, negatively regulate excitatory synaptic receptors-glutamate receptors (GluRs), especially NMDA receptor (NMDAR) in central nervous system, which is mediated by rapid membrane mechanisms, but not by classical, genomic mechanisms.     

蕲蛇酶减轻大鼠局灶性脑缺血再灌注损伤的实验研究(Ⅱ)

目的 从病理形态学观察蕲蛇酶对大鼠局灶性脑缺血再灌注损伤的保护作用并探讨其机制。方法 采用线栓法制备大鼠大脑中动脉栓塞后再灌注模型，光镜及电镜观察缺血3h恢复血流再灌24h后脑组织形态学变化。TTC染色观察脑梗死面积，免疫组织化学方法检测脑组织碱性成纤维细胞生长因子（bFGF）的表达。用药组分别在缺血即刻或再灌即刻静脉给予不同剂量的蕲蛇酶，观察比较模型组和蕲蛇酶所有给药组之间上述指标的变化。结果 缺血即刻给蕲蛇酶4U／kg组较再灌即刻给蕲蛇酶同剂量组显著减小梗死灶（P〈0．001），减轻脑组织病理改变，并使脑组织中bFGF蛋白表达明显增多。结论 蕲蛇酶对脑缺血再灌注损伤的神经保护作用可能与脑组织中bFGF表达增高有关；提示该药早期使用更为有效。     

Localization of proopiomelanocortin mRNA in functional subsets of neurons defined by their axonal projections

In situ cDNA:mRNA hybridization is a technique that has been developed for the visualization of cDNA:mRNA hybrids in individual cells. To use this technique to answer questions of regulation in heterogeneous populations of cells in the brain, it must be combined with other procedures allowing for the identification of functional subgroups of neurons. We report here a procedure by which in situ cDNA:mRNA hybridization may be combined with retrograde axonal tracing using the fluorescent tracer fast blue. Using this technique, it now becomes possible to measure mRNA regulation in functional subsets of cells defined by their axonal projections.     

Nucleus A10 dopaminergic neurons in inbred mouse strains: Firing rate and autoreceptor sensitivity are independent of the number of cells in the nucleus

Inbred mouse strains have different numbers of midbrain dopaminergic neurons; for example, BALB/cJ mice have 20–25% more neurons than CBA/J mice. As the number of cells decrease, for example in Parkinson's disease and in animals with midbrain dopaminergic cell lesions, the activity of their remaining cells increases. The purpose of the present experiment was to determine whether the functional properties of dopaminergic neurons in the ventral tegmental area (nucleus A10) differ in inbred mouse strains which possess different numbers of cells. The firing rate and autoreceptor sensitivity of A10 dopaminergic cells were examined in the in vitro slice preparation in BALB/cJ, C3H/HeJ, CBA/J, and DBA/2J mouse strains. It was observed that the autoreceptors on mouse dopaminergic neurons exhibit pharmacological properties of dopamine autoreceptors; activation of the autoreceptor produced a marked inhibition (50–70%) in cell firing rate by quinpirole (10⁻⁸ M), LY-141865 (10⁻⁷ M), (+)-3-(3-hydroxyphenyl)-N-n-propyl-piperidine (10⁻⁶ M), propyl-norapomorphine (10⁻⁵ M) and dopamine (10⁻⁴ M), and this inhibition was blocked or reversed by specific dopamine D2 receptor antagonists [(−) sulphide and spiroperidol, 10⁻⁶ M]. The baseline firing rates of the A10 cells did not differ among the four inbred strains [range 2.5 ± 0.2 (C3H/HeJ)−3.4 ± 0.3 (CBA/J) spikes/s ±SEM], and there was no significant difference in autoreceptor sensitivity among the mouse strains as assessed either by superfused dopamine (inhibitory dose 50% ≈150 μM), or by superfused quinpirole (inhibitory dose 50% ≈10 nM). These data indicate that differences in A10 cell numbers of 30% do not significantly influence the baseline firing rate or autoreceptor sensitivity of the cells.     

A neosynthesis of sodium channels is involved in the evolution of the sodium current in isolated adult DUM neurons

Short-term culture of isolated adult dorsal unpaired median (DUM) neurons of the cockroach Periplaneta americana has been used to study the evolution of the sodium current during the time in culture after axotomy and deafferentation treatment. An increase in the maximum peak amplitude of the sodium current recorded under voltage-clamp conditions with the patch-clamp technique in the whole-cell recording configuration, was only observed between 24h and 72h (75%) without any modification of the kinetics and the voltage-dependence of the current. A decrease in the level of foetal calf serum in the culture medium reduces the amplitude of the sodium current on all days but does not affect its time-course of development which was on the contrary completely abolished by both protein synthesis inhibitors, actinomycin D and cycloheximide. The results obtained in these neurons strongly suggest that a neosynthesis of sodium channel proteins is involved in the evolution of the sodium current induced by axotomy and deafferentation.     

胫骨前肌疲劳时比目鱼肌诱发肌电图H波的变化及其机制探讨

为了解主动肌疲劳时拮抗肌脊髓运动神经元兴奋性变化的规律 ,本研究采用踝关节背屈运动形式 ,对胫骨前肌 (主动肌 )疲劳状态下的比目鱼肌 (拮抗肌 )诱发肌电图H波成分进行了观察。并以压迫阻断胫骨前肌Ⅰa类神经纤维传导的方法 ,对比目鱼肌H波变化机制进行了分析探讨。结果发现 :(1)胫骨前肌疲劳后 ,比目鱼肌H波明显受到抑制 ,与安静时比较呈非常显著性差异 ;(2 )胫骨前肌Ⅰa类神经纤维传导被阻断后 ,比目鱼肌H波的抑制现象没有解除。表明 ,胫骨前肌疲劳时比目鱼肌H波被抑制的原因 ,可能是由于主动肌内的代谢产物激活了Ⅲ·Ⅳ类神经纤维的感受器 ,Ⅲ·Ⅳ类神经纤维的传入冲动增加 ,使Ⅰa抑制性中间神经元被激活 ,导致拮抗肌脊髓运动神经元的兴奋性受到了抑制     

Cytochemical characteristics of neurons in the trigeminal mesencephalic nucleus of hatchling chicks

The goal of the present study was to identify cytochemical markers characteristic of muscle afferents in hatchling chicks. To this end, we stained neurons in the trigeminal mesencephalic nucleus with a variety of markers that label subsets of neurons in avian dorsal root ganglia. We found that trigeminal mesencephalic neurons are surprisingly heterogeneous in their cytochemical make-up, expressing, to varying degrees, substance P, cholecystokinin, carbonic anhydrase, calbindin D-28k, parvalbumin, and S-100β. Calbindin D28k and S-100β appeared to be expressed equally in medial and lateral divisions of the trigeminal mesencephalic nucleus. In contrast, substance P- and cholecystokinin-immunoreactive neurons were more abundant in the medial division, whereas carbonic anhydrase activity and parvalbumin immunoreactivity were stronger in the lateral division. We were unable to detect met-enkephalin, neuropeptide Y, calcitonin gene-related peptide, vasoactive intestinal peptide, somatostatin, γ-aminobutyric acid, or tyrosine hydroxylase in the trigeminal mesencephalic nucleus. Moreover, these neurons did not appear to bind the lectin Dolichos biflorus agglutinin. The heterogeneity of expression of markers among trigeminal mesencephalic nucleus neurons, especially between neurons in the medial and lateral divisions, suggests that these neurons are functionally diverse.     

电刺激黑质对脑干伤害性单位放电的影响

本实验采用单细胞外记录神经元单位放电的方法，在Ｗｉｓｔａｒ大鼠上观察到网状巨细胞核。一区伤害性神经元５７个，其中伤害兴奋性神经元（ＮＥＮ）４２个，伤害抑制性神经元（ＮＩＮ）１５个。电刺激黑质对ＮＥＮ（２９）和ＮＩＮ（８的放电呈抑制作用，其反应百分率分别由刺激黑质第１分钟时的１．０±２．１％和－２０．４±６．２％，降到第５分钟时的－８３．６±１２．３％和－６５．４±１０．７％（Ｐ＜０．０１）。刺激黑质对少数ＮＥＮ（１３）和ＮＩＮ（７）的放电有兴奋作用，其值分别由第］分钟时的２０．５±７．３％和１．０±３．４％，升高到第５分钟时的７４．５±１０．７％和２１，５±８．６％（Ｐ＜０．０５）。氟哌啶醇注入ＰＡＧ腹外侧部可阻断刺激黑质的效应，这提示从黑质到网状巨细胞核α－区存在着一条痛调制通路，而且这种通路的递质是多巴胺能的。