人重组白细胞介素-6对大鼠海马神经元Bcl-2表达的影响

观察重组人白细胞介素－6（rhIL－6）对体外培养大鼠海马神经元Bcl－2表达的影响。方法取培养3、7、14、21和28天（d）的两组（对照组和rhIL－6组）培养神经元,分别观察其生长发育和神经元活存数,并用抗Bcl－2抗血清进行免疫组化染色,观察Bcl-2免疫反应（Bcl-2－IR）阳性和阴性神经元数目,计算Bcl-2－IR阳性神经元所占百分率,并在图像分析仪上对Bcl-2－IR神经元作平均光密度的色谱分析。结果培养3、7、14、21和28d时,rhIL－6组神经元活存数、Bcl－2－IR阳性神经元数和Bcl-2-IR阳性神经元的平均光密度均明显高于对照组。结论rhIL－6能增强生长发育过程中海马神经元Bcl-2的表达,减少神经元的退化死亡,表明rhIL-6对体外培养的海马神经元具有神经营养作用。     

人重组白细胞介素—6对大鼠海马神经元Bcl—2表达？…

目的 观察重组人白细胞介素－６（ｒｈＩＬ－６）对体外培养大鼠海马神经元Ｂｃｌ－２表达的影响。方法 取培养３、７、１４、２１和２８天（ｄ）的两组（对照组和ｒｈＩＬ－６组）培养神经元,分别观察其生长发育和神经元活存数,并用抗Ｂｃｌ－２抗血清进行免疫组化染色,观察Ｂｃｌ－２免疫反应（Ｂｃｌ－２－ＩＲ）阳性和阴性神经元数目,计算Ｂｃｌ－２－ＩＲ阳性神经元所占百分率,并在图像分析仪上对Ｂｃｌ－２－ＩＲ神经元     

人重组白细胞介素-6可减少缺氧-复氧后大鼠海马培养神经元的DNA损伤

用原位末端标记（ＴＵＮＥＬ）法观察人重组白细胞介素－６（ｒｈＩＬ－６）对缺氧－复氧后大鼠海马培养神经元的ＤＮＡ损伤的影响。结果显示,海马神经元缺氧－复氧后ＤＮＡ损伤神经元百分率明显增高,经ｒｈＩＬ－６ 预处理的海马神经元缺氧－复氧后ＤＮＡ损伤神经元百分率明显低于对照组。本结果表明,缺氧－复氧能使体外培养海马神经元发生ＤＮＡ损伤,ｒｈＩＬ－６ 可减少缺氧－复氧后海马神经元的ＤＮＡ损伤。提示ｒｈＩＬ－６ 对海马神经元缺氧－复氧损伤可能具有一定保护作用。     

黄芪穴位注射对慢性精神分裂症血可溶性白细胞介素2受体的影响

目的:探讨足三里穴位注射黄芪液对慢性精神分裂症患者免疫功能的影响。方法:对30例患者足三里穴位注射黄芪液,分别于注射前、注射4周和8周用酶联免疫法检测血可溶性白细胞介素2受体(sIL-2R)变化,并与30例健康者比较,进行相关因素分析。结果:穴位注射前患者sIL-2R水平显著高于健康者,注射后显著降低;老年组的sIL-2R水平明显高于青年组及中年组;氯丙嗪组sIL-2R水平明显高于氯氮平组及联合用药组;不同临床亚型组间的sIL-2R水平无显著差异。结论:足三里穴位注射黄芪液对慢性精神分裂症患者异常偏高的sIL-2R有显著改善作用。     

侧脑室注射α-干扰素及白细胞介素-2对大鼠丘脑束旁核痛兴奋神经元放电的影响

目的 从神经元放电的角度进一步研究α-干扰素(IFN-α)及IL-2的中枢镇痛作用。方法 以串脉冲刺激右侧坐骨神经和有齿镊子夹尾作为伤害性刺激,诱发大鼠丘脑束旁核痛兴奋神经元放电,记录在不同大鼠侧脑室注射IFN-α和IL-2对大鼠丘脑束旁核痛兴奋神经元电活动的影响。结果 IFN-α使大鼠丘脑束旁核痛兴奋神经元的放电频率降低,在给药前:PEN诱发放电秒净增值16.33±4.03;注射后6min、12min、18min,分别为7.92±0.64(P<0.01)、5.59±0.47(P<0.01)、7.44±0.59(P<0.01)。注射后20min开始恢复,24min基本恢复至注射前水平。IL-2也可使PEN放电频率降低,给药前,PEN诱发放电频率净增值为(17.16±3.94)Hz,注射后4min、8min、12min、16min 分别为(5.86±0.91)Hz、(2.81±0.96)Hz、(2.67±0.45)Hz、(4.11±0.46)Hz。此外,脑室注射IFN-α和IL-2还能使诱发放电的潜伏期延长,注射前后相比差异显著。结论IFN-α和IL-2均能使PEN的放电频率降低,潜伏期延长。     

侧脑室注射α—干扰素及白细胞介素—2对大鼠丘脑束旁核痛兴奋神经元放电的影响

为了便于大家查询、利用Internet上的有关行为学和智力缺陷杂志文献资源,作者收集、整理和编写了一个“Internet行为学和智力缺陷杂志文献搜索引擎”,仅供参考。希望同行们在使用中发现问题,提出宝贵意见。1 收集范围 收集了32种国际著名的有关行为学的杂志和5种有关智力缺陷杂志。包括行为生物学、行为遗传学、行为药理学、行为医学、行为矫正等。     

重组白喉毒素-人白细胞介素13嵌合毒素诱导胶质瘤细胞凋亡作用的初步研究

目的探讨重组白喉毒素-人白细胞介素13嵌合毒素DT389-IL13对胶质瘤细胞的杀伤作用机制.方法在U251细胞中加入不同浓度的嵌合毒素DT389-IL13,作用72 h后分别进行H-33342染色,观察胶质瘤细胞核形态,电镜观察细胞超微结构,用流式细胞仪计数亚二倍体细胞百分数等.结果3种检测结果均表明,嵌合毒素DT389-IL13可诱导U251胶质瘤细胞发生凋亡,1×10-9mol/L嵌合毒素作用72 h后亚二倍体细胞百分比为38%,1×10-8mol/L作用后亚二倍体细胞百分比为99%,明显高于对照组(3.63%),与荧光染色观察的结果一致;电镜下还可见固缩的线粒体.结论嵌合毒素DT389-IL13可诱导胶质瘤细胞凋亡.     

白细胞介素-2及其受体在癫痫患者发作中的变化和对癫痫大鼠的致惊作用

目的探讨白细胞介素-2(interluekin-2, IL-2)及其可溶性受体在癫痫患者发作中的变化和对癫痫大鼠的致惊作用.方法采用ELISA方法检测58例癫痫患儿和23例健康儿血清IL-2及其可溶性受体的浓度,并进行分组比较;观察侧脑室注射IL-2对遗传性癫痫易感大鼠(P77PMC)惊厥发作的影响.结果癫痫组血清IL-2及其可溶性受体浓度[(25.86±5.21) ng/mL和(758.26±78.49) U/mL ]明显高于对照组[(13.78±3.24) ng/mL和(325.67±34.58) U/mL](P<0.01),且发作期[(30.31±6.77) ng/mL和(806.25±112.35) U/mL]高于间歇期[(20.27±4.66) ng/mL和(584.15±57.85) U/mL ](P<0.01),脑电图异常者[(27.18±4.35) ng/mL和(724.48±78.56) U/mL]高于正常者[(20.04±3.55) ng/mL和(621.85±77.45) U/mL],但与发作类型、病程长短及经治疗与否无关;侧脑室注射IL-2(5 000 U/L)5～10 μL后大鼠惊厥评分从26升高到36(P<0.05),惊厥持续时间从32 s延长到65 s(P<0.01),但对潜伏期无影响.结论 IL-2在癫痫患者处于高水平状态,直接侧脑室注射IL-2可加重大鼠惊厥程度,提示其参与癫痫发病过程.     

重组人促红细胞生成素对慢性脑缺血大鼠学习记忆能力及凋亡相关蛋白P53和Bcl-2表达的影响

目的 研究重组人促红细胞生成素(rhEPO)对慢性脑缺血大鼠学习记忆能力及凋亡相关蛋白P53和Bcl-2表达的影响. 方法 16只健康雄性SD大鼠结扎双侧颈总动脉(2VO)建立永久性慢性脑缺血模型,将其按照随机数字表法分为对照组和实验组,每组各8只.模型建立3d后,实验组每7天经鼻腔给予150 U/125 μL rhEPO,至建模后8周末;对照组于同一时间鼻腔给予等量的生理盐水.8周后应用Morris水迷宫观测2组大鼠的运动及空间学习记忆能力.采用HE染色观察大脑皮层及海马CA1区神经元形态学的变化,并采用图像分析软件对大脑皮层厚度与神经元数目进行比较.免疫组化法检测P53及Bcl-2蛋白的表达水平.原位末端标记(TUNEL)法检测凋亡细胞数. 结果 (1)行为学测试结果显示:实验组平均逃避潜伏期及穿越平台次数均较对照组有所改善,差异有统计学意义(P＜0.05).(2)HE染色结果显示:与实验组相比,对照组大鼠皮层及海马CA1区锥体神经元胞体萎缩、核固缩且皮层厚度变薄,数目减少,差异有统计学意义(P＜0.05).(3)免疫组化结果显示:与对照组相比,实验组Bcl-2的表达明显增强,P53平均灰度值明显增加,差异有统计学意义(P＜0.05).(4)TUNEL染色结果显示:对照组凋亡细胞数较实验组明显增加,差异有统计学意义(P＜0.05). 结论 rhEPO能改善慢性脑缺血大鼠运动、记忆及空间定向能力,这种神经功能保护作用的机制可能与减少神经元凋亡相关.     

Retinal ganglion cell death in a DBA/2J mouse model of glaucoma Microglial activation and intraocular pressure

BACKGROUND: Retinal microglia has been shown to reactivate in a murine model of pigmentary glaucoma. However, the relationship between microglial activation and intraocular pressure (IOP) elevation and retinal ganglion cell (RGC) death is still unclear. OBJECTIVE: To verify that microglial activation and tumor necrosis factor alpha (TNF-α) expression is involved in RGC death with elevated IOP and prolonged time of glaucomatous optic nerve lesion in a DBA/2J mouse model of glaucoma. DESIGN, TIME AND SETTING: This randomized, controlled, animal experiment was performed at the Peking University Third Hospital, Peking University Eye Center, China between December 2006 and May 2008. MATERIALS: DBA/2J mice and C57BL/6J mice (Jackson Laboratory, USA), rat anti-mouse CD11b monoclonal antibody (Serotec, UK), and goat anti-TNF-α polyclonal antibody (Sigma, USA) were used in this study. METHODS: A total of 100 female, DBA/2J mice at 3, 6, 9,12, and 14 months of age (20 mice per age group) were used for the glaucoma model, and 18 C57BLV6J mice at 3, 9,14 months of age (6 mice per age group) were used as normal controls. The anterior segment of the eye was observed using a slit-lamp biomicroscope. IOP was measured using a microneedle system. Morphology and number of retinal microglia were observed using immunohistochemistry. RGCs were quantified using Nissl staining. Co-localization of TNF-a and microglia was observed using double-labeling immunofluorescence. Excavation of the optic nerve head was observed utilizing hematoxylin-eosin staining. MAIN OUTCOME MEASURES: The following parameters were measured: IOP levels, numbers of RGCs and activated microglia, and TNF-α expression. RESULTS: In 6-month-old DBA/2J mice, dispersed pigment was observed, and some mice developed increased IOP. At 9 months of age, IOP levels reached a peak. In 3-month-old DBA/2J mice, microglia were activated. In 6-month-old DBA/2J mice, the number of activated microglia was significantly increased and migrated to the outer retinal layer. In 9-month-old mice, TNF-α expression was co-localized with microglia. Significant RGC loss occurred in mice aged 9 to 14 months, with the presence of optic nerve fiber loss and optical nerve head excavation. IOP returned to normal levels at 12 months of age, but microglia remained activated, which was consistent with RGC loss. CONCLUSION: Retinal microglial activation was partially attributed to increased IOP. Activated microglia might be mainly responsible for RGC loss. TNF-α expression was evident in the inner retinal layer. However, the relationship between TNF-α and RGC loss remains poorly understood.     

Influence of the visual cortex on responses of retinal ganglion cells in the rat

The objective of the present investigation was to answer the following question: Does the visual cortex affect the neuronal firing of retinal ganglion cells in the rat? To test this hypothesis, the visual cortex was inactivated by a reversible cryoblockade. Action potentials of a ganglion cell were recorded from its axon at the optic tract level prior to, during, and following cortical blockade. The results indicated that indeed the visual cortex influenced the retinal output since its inactivation led to a modification of the firing pattern evoked in response to a flash of light. In most cases the modification was an increase of the bursting pattern of the evoked discharges. By contrast cooling nonvisual areas failed to modify ganglion cells' discharge. A comparison between cortico-geniculate and cortico-retinal feedback loops seems to suggest that the first path is involved mostly with the spatial organization of center-surround receptive fields, whereas the second path is associated with temporal aspects of the retinal responses in the rat.     

CIC-3氯离子通道在谷氨酸诱导视网膜神经节细胞RGC-5凋亡中的作用：视神经保护的新靶点？

摘要 氯离子通道在神经细胞凋亡中所起的作用受到越来越多学者的关注,但目前国内外尚无其在视网膜神经节细胞中的作用研究及探讨。以往人们多用氯离子通道阻滞剂研究其功能,但是由于阻滞剂的非特异性使实验结果值得商榷,本文采用cDNA转染及RNA干扰的方法特异性的针对视网膜神经节细胞RGC-5中ClC-3氯通道进行研究,并探讨了TGF-β/Smads信号转导通路在此过程中所起的作用。结果显示：RGC-5内存在ClC-2、 ClC-3、ClC-5的表达;ClC-3 cDNA转染谷氨酸诱导的RGC-5细胞凋亡后,细胞存活率明显升高、凋亡率明显减少;而应用RNAi技术阻断ClC-3氯通道后,细胞存活率下降、凋亡率明显增加,并且TGF-β2、Smads2、Smads3、Smads4、Smads7含量均不同程度的升高。由此认为,ClC-3氯离子通道在谷氨酸诱导的视网膜神经节细胞RGC-5凋亡中起保护作用,TGF-β/Smads信号转导通路在RGC-5凋亡和保护过程中起到了一定的作用。     

Alpha B-crystallin improved survival of retinal ganglion cells in a rat model of acute ocular hypertension

Increased endogenous αB-crystallin protein levels have been shown to reduce cell apoptosis,although the effects of exogenous αB-crystallin protein remain poorly understood.The present study established an acute ocular hypertension model in the right eye of Sprague-Dawley rats.Fluorogold retrograde tracing and immunofluorescence methods showed that the number of retinal ganglion cells decreased in the right eyes and caspase-3 expression increased following acute ocular hypertension.Intravitreal injection of αB-crystallin in the right eye increased the number of retinal ganglion cells and reduced caspase-3 expression.Results demonstrated that exogenous αB-crystallin protein inhibited caspase-3 expression and improved retinal ganglion cell survival following acute ocular hypertension.     

Topographic specializations in the retinal ganglion cell layer of Australian passerines

Thornbills, honeyeaters, and silvereyes represent an abundant group of Australian passerines, whose diversity in niche differentiation suggests a pivotal role for vision. Using stereological methods and retinal wholemounts, we studied the topographic distribution of neurons in the ganglion cell layer of insectivorous, nectarivorous, and frugivorous species occupying terrestrial and arboreal microhabitats. All species studied have a central convexiclivate fovea (peak densities from 130,000 to 160,000 cells/mm²), which is shallow in the terrestrial/insectivorous yellow‐rumped thornbill and deep in the arboreal/nectarivorous honeyeaters and frugivorous silvereye. Surrounding the fovea, neuronal densities in the ganglion cell layer form a broadly ovoid and asymmetric plateau in the yellow‐rumped thornbill and a more restricted, circular and symmetric plateau in the other species. These differences in the plateau organization may reflect specific needs to locate food on the ground or among dense vegetation. We also found a temporal area (peak densities from 43,000 to 54,000 cells/mm²) across species, which increases spatial resolution in the frontal visual field and assists with foraging. Using microtubule‐associated protein 2 (MAP2) immunohistochemistry, we detected a higher concentration of giant ganglion cells forming an area gigantocellularis in the temporal retina of all species. Giant ganglion cell densities also form a horizontal streak in all species, except in the yellow‐rumped thornbill, which has an unusual increase toward the retinal periphery. In the yellow‐rumped thornbill and silvereye, giant ganglion cells also peak in the nasal retina. We suggest that these topographic variations afford differential sampling of motion signals for the detection of predators. J. Comp. Neurol. 522:3609–3628, 2014. © 2014 Wiley Periodicals, Inc.     

Patterns of cell death in the ganglion cell layer of the human fetal retina

The distribution of dying cells in the ganglion cell layer (GCL) of retinae from human fetuses has been analysed. Both whole-mounted and sectioned retinae have been studied. Results suggest that cells are lost from the GCL between weeks 14 and 30 of the gestation period, approximately. This period corresponds to the period during which axons are lost from the developing optic nerve. Cell loss is greatest between weeks 16 and 21 of the gestation period. The pattern of cell loss is nonuniform, and between weeks 16 and 24, the relative frequency of pyknotic cells (pyknotic cells:viable cells) in peripheral retina is considerably higher than in central retina. This pattern of cell loss predominates during the period in which a distinct centroperipheral gradient of cell densities emerges in the GCL of the human fetal retina (between 18 and 23 weeks gestation). It is suggested that the regional loss of ganglion cells may contribute to the formation of the cell density gradient.     

Regularity changes of the retinal nerve fiber layer and macular ganglion cell complex in patients with the amnestic mild cognitive impairment

Purpose/aim: We investigated the regularity changes of the retinal nerve fiber layer (RNFL) and macular ganglion cell complex (mGCC) of the amnestic mild cognitive impairment (aMCI) patients in this prospective cohort study.

Materials and methods: Twenty-four aMCI patients and 30 health controls, who are more than 60 years old, were recruited into the study. The RNFL and the mGCC average thickness were measured with Fourier-domain optical coherence tomography (FD-OCT).

Results: Compared with that in the controls, the intraocular pressure (IOP) was significantly lower in the aMCI patients. A significant decrease in RNFL thickness in superior temporal, temporal upper (TU), and temporal upper and lower (TL) (TU+TL) quadrants was found in the aMCI patients than in the controls. The average thickness of the mGCC was also significantly thinner in the aMCI patients than in the controls.

Conclusions: Retinal degeneration in the aMCI patients detected by OCT together with lower IOP may indicate disease pathological progression.     

Characterization of multiple bistratified retinal ganglion cells in a purkinje cell protein 2‐Cre transgenic mouse line

Retinal ganglion cells are categorized into multiple classes, including multiple types of bistratified ganglion cells (BGCs). The recent use of transgenic mouse lines with specific type(s) of ganglion cells that are labeled by fluorescent markers has facilitated the morphological and physiological studies of BGCs, particularly the directional‐selective BGCs. The most important benefit from using transgenic animals is the capability to perform in vivo gene manipulation. In particular, the Cre/LoxP recombination system has become a powerful tool, allowing gene deletion, overexpression, and ectopic expression in a cell type‐specific and temporally controlled fashion. The key to this tool is the availability of Cre mouse lines with cell or tissue type‐specific expression of Cre recombinase. In this study we characterized the Cre‐positive retinal ganglion cells in a PCP2 (Purkinje cell protein 2)‐cre mouse line. We found that all of the Cre‐positive retinal ganglion cells were BGCs. Based on morphological criteria, we determined that they can be grouped into five types. The On‐ and Off‐dendrites of three of these types stratified outside of the cholinergic bands and differed from directional selective ganglion cells (DSGCs) morphologically. These cells were negative for Brn‐3b and positive for both calretinin and CART retina markers. The remaining two types were identified as putative On‐Off and On‐DSGCs. This Cre mouse line could be useful for further studies of the molecular and functional properties of BGCs in mice. J. Comp. Neurol. 521:2165–2180, 2013. © 2012 Wiley Periodicals, Inc.     

Age-dependent changes in the regulation mechanisms for intracellular calcium ions in ganglion cells of the mouse retina

The purpose of this study was to investigate the role of intracellular calcium buffering in retinal ganglion cells. We performed a quantitative analysis of calcium homeostasis in ganglion cells of early postnatal and adult mice by simultaneous patch-clamp recordings in sliced tissue and microfluorometric calcium measurements with Fura-2. Endogenous calcium homeostasis was quantified by using the 'added buffer' approach which uses amplitudes and decay time constants of calcium transients to give a standard for intracellular calcium buffering. The recovery phase of depolarization-induced calcium transients was well approximated by a mono-exponential function with a decay time constant that showed a linear dependence on dye concentration. Endogenous calcium binding ratios were found to be 575 (n = 18 cells) in early postnatal and 121 (n = 18 cells) in adult retinal ganglion cells. With respect to ganglion cell degeneration at early postnatal stages, our measurements suggest that neuroprotection of a majority of developing ganglion cells partially results from a specialized calcium homeostasis based on high buffering capacities. Furthermore, the dramatic decrease of the intracellular calcium buffering capacity during ganglion cell development may enhance their vulnerability to neurodegeneration.