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.     

The synaptology of parvalbumin-immunoreactive neurons in the primate prefrontal cortex.

Electron microscopy and immunocytochemistry with a monoclonal antibody against parvalbumin (PV) were combined to analyze the distribution and morphology of PV-immunoreactive (PV-IR) neurons and the synaptology of PV-IR processes in the principal sulcus of the macaque prefrontal cortex. Parvalbumin-IR neurons are present in layers II-VI of the macaque principal sulcus (Walker's area 46) and are concentrated in a band centered around layer IV. PV-IR cells are exclusively non-pyramidal in shape and are morphologically heterogeneous with soma sizes ranging from less than 10 microns to greater than 20 microns. Well-labeled neurons that could be classified on the basis of soma size and dendritic configuration resembled large basket and chandelier cells. A novel finding is that supragranular PV-IR neurons exhibit dendritic patterns with predominantly vertical orientations, whereas infragranular cells exhibit mostly horizontal or oblique dendritic orientations. PV-IR cells within layer IV exhibit a mixture of dendritic arrangements. Vertical rows of PV-IR puncta, 15-30 microns in length, resembling the "cartridges" of chandelier cell axons were most dense in layers II, superficial III, and the granular layer IV but were not observed in the infragranular layers. Cartridges were often present beneath unlabeled, presumed pyramidal cells. PV-IR puncta also formed pericellular nests around pyramidal cell somata and proximal dendrites, suggestive of basket cell innervation. PV-IR axons were occasionally observed in the white matter underlying the principal sulcus. Electron microscopic analysis revealed that PV-IR somata and dendrites are densely innervated by nonimmunoreactive terminals forming asymmetric (Gray type I) synapses as well as by fewer terminals forming symmetric (Gray type II) synapses. The majority of terminals forming symmetric synapses with PV-IR post-synaptic structures were not immunolabeled; however, some of these boutons did contain PV-immunoreactivity. PV-IR boutons exclusively form symmetric synapses and heavily innervate layer II/III pyramidal cells. PV-IR axon cartridges formed numerous axo-axonic synapses with the axon initial segments of pyramidal cells 15-20 microns beneath the axon hillock and also terminated on large axonal spines of the initial segment. Furthermore, we failed to observe a mixture of PV-immunoreactive and non-immunoreactive boutons composing a single axon cartridge. Pyramidal cell somata and proximal dendrites were also heavily innervated by PV-IR boutons forming symmetric synapses, again, consistent with basket cell innervation. In addition, PV-IR axon terminals frequently formed symmetric synapses with dendritic shafts and spines of unidentified neurons.(ABSTRACT TRUNCATED AT 400 WORDS)     

Recognition of an extensive range of IgE-reactive proteins in cod extract

Allergy to fish is one of the most common food allergies. Gad c 1 is the only fish allergen which has been purified and characterized. Other allergens have been detected by Western blot in cod extracts. We have now improved the Western-blot procedure in order to characterize fish IgE-reactive proteins from extracts prepared under different conditions: pre-rigor mortis and postrigor mortis. EDTA addition or not. and DEAE ion-exchange chromatography. Several IgE-reactive protein bands have been identified over a wide molecular-weight range. In particular, the 104- and 130-kDa IgEreactive protein bands were detected. These new bands may correspond to aggregates, as EDTA increased the relative amount of the 60-, 67-, 104-, and 130-kDa IgE-reactive protein bands in Western blot. All these bands were also detected by an antiparvalbumin monoclonal antibody, specific to the first calcium-binding site. The longer period of storage increased the relative amounts of the 41-, 80-, 104-. and 130-kDa IgE-reactive protein bands. The 18-kDa band was detected only in fish stored for several days. In conclusion, we have described IgE-reactive protein bands over a wide molecular-weight range (12–130 kDa) in Western blot of cod extract, and shown that EDTA and storage conditions may influence the relative distribution of IgE-reactive protein bands.     

Annexin A2的结构和功能及其与疾病的关系

Annexins是一类钙依赖性膜磷脂结合蛋白。Annexin A2是Annexins家族中的一个重要成员,存在于细胞膜、胞质和胞外。Annexin A2蛋白在细胞膜构架的形成、膜聚合、内吞和胞吐中均扮演重要角色。Annexin A2还参与某些病理过程,与心血管疾病、血液病、肿瘤等多种疾病相关。文章简要综述了Annexin A2分子的结构、生化特征、生物学功能及其与疾病的关系。     

宫颈病变组织中E-cad和β-cat的表达及其意义

目的:研究上皮性钙黏附蛋白 (epithelial cad,E cad)和β 连接素(β catenin,β cat)在宫颈上皮内瘤样病变 (cervicalintraepithelialneoplasia,CIN)和 宫颈鳞癌中的表达及其意义。方法:应用 SP法检测E cad和β cat在正常宫颈上 皮、CINⅠ级、CINⅡ～Ⅲ级及宫颈鳞癌组 织的表达。结果:正常宫颈中E cad和β cat为膜表达。E cad表达在正常宫颈 CINⅠ级、CINⅡ～Ⅲ级和宫颈鳞癌中分别 为86%(12/14)、82%(9/11)、54%(7 13)和21%(8/38);β cat膜表达率分别为 79%(11/14)、64%(7/11)、23%(3/13)和 11%(4/38);E cad表达在正常宫颈与鳞 癌之间、CINⅠ级与鳞癌之间的差异有统 计学意义,P<0.05;在鳞癌的不同病理学 分级和肌层浸润深度之间的差异有统计 学意义,P<0.05。β cat表达在正常宫颈 与CINⅡ～Ⅲ级、宫颈鳞癌以及CINⅠ级 与鳞癌之间差异有统计学意义,P<0.05 在宫颈鳞癌的不同病理分级之间差异有 统计学意义,P<0.05。宫颈鳞癌中,E cad与β cat的表达形式具相关性,P< 0.001。结论:E cad和β cat异常表达是 宫颈上皮性肿瘤进展的早期事件;在宫颈 浸润性鳞癌中,它们的异常表达可能与肿 瘤的侵袭潜能有关。     

Repeated Baclofen treatment ameliorates motor dysfunction,suppresses reflex activity and decreases the expression of signaling proteins in reticular nuclei and lumbar motoneurons after spinal trauma in rats

The interruption of supraspinal input to the spinal cord leads to motor dysfunction and the development of spasticity. Clinical studies have shown that Baclofen (a GABA_B agonist), while effective in modulating spasticity is associated with side-effects and the development of tolerance. The aim of the present study was to assess if discontinued Baclofen treatment and its repeated application leads antispasticity effects, and whether such changes affect neuronal nitric oxide synthase (nNOS) in the brainstem, nNOS and parvalbumin (PV) in lumbar α-motoneurons and glial fibrillary acidic protein in the ventral horn of the spinal cord. Adult male Wistar rats were exposed to Th9 spinal cord transection. Baclofen (30 mg/b.w.) diluted in drinking water, was administered for 6 days, starting at week 1 after injury and then repeated till week 4 after injury. The behavior of the animals was tested (tail-flick test, BBB locomotor score) from 1 to 8 weeks. Our results clearly indicate the role of nitric oxide, produced by nNOS in the initiation and the maintenance of spasticity states 1, 6 and 8 weeks after spinal trauma. A considerable decrease of nNOS staining after Baclofen treatment correlates with improvement of motor dysfunction. The findings also show that parvalbumin and astrocytes participate in the regulation of ion concentrations in the sub-acute phase after the injury.     

慢性心房颤动右心房肌细胞内钙蛋白酶含量及细胞凋亡水平的变化

[目的]探讨风湿性二尖瓣病变伴心房颤动患者右心房心肌细胞中钙蛋白酶(Calpain)的含量变化及细胞凋亡水平的变化及相关关系.[方法]选择风湿性二尖瓣病变行二尖瓣置换手术的心脏病患者共30例,其中窦性心律(对照组)15例,慢性房颤(观察组)15例.在手术中取右心房组织,应用蛋白免疫印迹法测定各病人心肌细胞内Calpain I、Calpain Ⅱ的含量;应用TUNEL法检测各病人心房肌细胞凋亡指数(AI),并分析比较它们之间的关系.[结果]与对照组比较,观察组Calpain I含量明显上升(1.585±0.260 vs 1.191±0.099,P＜0.01);Calpain Ⅱ含量无明显变化(O.845±0.071 vs 0.823士0.078,P=0.22);凋亡水平上升(34.00%±10.87%vs 15.53%±6.52%,P＜0.01).慢性房颤组的Calpain I含量AI呈明显正相关(r=0.894,P＜0.01).Calpain Ⅱ含量与AI无明显相关(r=0.24,P=0.39).[结论]Calpain I和凋亡可能在风湿性房颤的发展及维持中起重要作用,且二者关系密切.     

肾癌中促红细胞生成素产生肝细胞受体A2的表达及其与E-钙黏蛋白、微血管密度的关系

目的 探讨促红细胞生成素产生肝细胞受体A2(Eph A2)在肾癌组织中的表达及其与肿瘤血管生成、E-钙黏蛋白的关系.方法 利用免疫组织化学法检测肾癌组织68例、正常肾组织24例中EphA2、E-钙黏蛋白的表达,并采用CD34抗体标记微血管内皮细胞,计算微血管密度(MVD).分析EphA2、MVD、E-钙黏蛋白与肾癌临床病理特征的关系及其的相关性.结果 肾癌组织中EphA2阳性表达及MVD明显高于正常肾组织,E-钙黏蛋白阳性表达明显低于正常肾组织(均P＜0.01).EphA2在肾癌组织的高分级组、高分期组、淋巴结转移组显著高于低分级组、低分期组和无淋巴结转移组(P＜0.01或＜0.05);MVD在肾癌组织的高分级组、高分期组、淋巴结转移组的表达亦显著高于低分级组、低分期组和无淋巴结转移组(均P＜0.01);MVD值还与肿瘤直径有关(均P＜0.01);E-钙黏蛋白在肾癌组织的高分级组、高分期组、淋巴结转移组显著低于低分级组、低分期组和无淋巴结转移组(P＜0.01或＜0.05);Spearman等级相关分析表明,EphA2蛋白表达与MVD呈显著正相关(r=0.555,P＜0.01),与E-钙黏蛋白蛋白的表达呈显著负相关(r=-0.483,P＜0.01).结论 EphA2蛋白的高表达预示肿瘤恶性程度的增加,其机制可能通过影响或协同E-钙黏蛋白和微血管生成来实现的.     

Activation of microglia and induction of pro-inflammatory cytokines in the hippocampus of type 2 diabetic rats

Abstract

Objectives:

The majority of immune cells in the brain are comprised of microglia, which undergo morphological changes when activated to remove damaged neurons and infectious agents from the brain tissue. In this study, we investigated the effects of type 2 diabetes on microglial activation and the subsequent secretion of pro-inflammatory cytokines, such as interferon-gamma (IFN-gamma) and interleukin-1beta (IL-1beta), in the hippocampus using Zucker diabetic fatty (ZDF) rats and Zucker lean control (ZLC) rats at various diabetic stages.

Methods:

Zucker lean control and Zucker diabetic fatty rats were sacrificed at 12 (early diabetic stage), 20, or 30 weeks of age (chronic diabetic stage), and the hippocampus was obtained via transcardiac perfusion or dissection for immunohistochemistry and western blot analysis, respectively.

Results:

Zucker diabetic fatty rats demonstrated significantly higher glucose levels at 12 and 30 weeks of age compared to ZLC rats. Microglia immunoreactive to ionized calcium-binding adapter molecule 1 (Iba-1) had hypertrophied cytoplasm with retracted processes at 30 weeks of age. In contrast, Iba-1-immunoreactive microglia displayed similar morphology in ZDF and ZLC rats at 12 and 20 weeks of age. Similarly, IFN-gamma and IL-1beta protein levels were significantly increased in ZDF rats compared to ZLC rats at 30 weeks of age, but not at 12 and 20 weeks of age. Interleukin-1beta immunoreactivity in the ZDF rats predominantly increased in the dentate gyrus and CA1 region of the hippocampus compared to that of ZLC rats at 30 weeks of age. In addition, IL-1beta immunoreactive structures in ZDF rats at 30 weeks of age were detected near the astrocytes and microglia.

Conclusion:

These results suggest that chronic diabetes activates microglia and significantly increases pro-inflammatory cytokine levels in the hippocampus.