Anatomy and fine structure of neurons in the deutocerebral projection pathway of the crayfish olfactory system.

Golgi impregnation and neurobiotin injection were used to examine details of the neural pathways in the olfactory system of the freshwater crayfish, Procambarus clarkii. Deutocerebral projection neurons (globuli cells) were directly injected with neurobiotin. These neurons have dendritic arborizations in the ipsilateral olfactory and accessory lobes, and they project axons to the lateral protocerebrum, where they terminate in microglomeruli of the hemi-ellipsoid body. The axons of the deutocerebral projection neurons are readily impregnated by Golgi procedures, and they terminate as an expanded membranous knot about 5 microns in diameter. Electron microscopy on Golgi-stained terminals has revealed that each knot makes several hundred synapses with small spine-like or shaft-like processes of postsynaptic neurons. Injection of neurobiotin into local interneurons of the hemi-ellipsoid body and subsequent examination of stained preparations with the electron microscope reveals that these cells are a major postsynaptic target of the deutocerebral projection neurons. Furthermore, the local interneurons make extensive efferent synaptic connections with unidentified neurons in the terminal medulla.     

Distribution of radioactive leucine following uptake by olfactory sensory neurons in normal and heteromorphic crayfish antennules

Surgical excision of one of the compound eyes from juvenile crayfish leads to the regeneration of a heteromorphic antennule in 30% of the cases. Most of the heteromorphic antennules generated this way are bifurcate appendages possessing morphologically distinct medial and lateral branches. These structures are identical to the internal and external flagella of the normal antennules, and the homolog of the external flagellum bears aesthetascs supplied by olfactory sensory neurons. Autoradiographic analysis of the brain following exposure of heteromorphic antennules to tritiated leucine indicates that the supernumerary sensory axons transport the labeled amino acid into their central terminals at appropriate target locations within the ipsilateral olfactory lobe. The data suggest that olfactory input from heteromorphic antennules is incorporated into the organized central projection of olfactory afferents from the normal antennule.     

Low birth weight – is it associated with few and small glomeruli in normal subjects and NIDDM patients?

Summary These studies were undertaken to ascertain if there is any association between low birth weight, and low kidney weight, few and/or small glomeruli, in kidneys from a control group and a group of non-insulin-dependent diabetic (NIDDM) patients. The background for this study comes from findings suggesting a correlation between low birth weight and the development of NIDDM and high blood pressure. Furthermore, Brenner has postulated that humans born with a low number of glomeruli, thereby having a low glomerular filtration surface area, have a greater tendency to develop high blood pressure. We examined 79 autopsy kidneys, with known weight from normal and NIDDM patients, which had previously been used for studies of glomerular number and volume. In the archives of the Danish midwives we were able to find birth weight for 26 NIDDM patients and an age- and sex-matched sample of 19 control persons. The kidney weight (g) (Control: 137 ± 36; NIDDM: 150 ± 38; 2p = 0.26), glomerular number (10³) (Control: 670 ± 176; NIDDM: 673 ± 200; 2p = 0.95), glomerular volume (10⁶μm³) (Control: 6.25 ± 1.48; NIDDM: 5.71 ± 1.74; 2p = 0.28) or birth weight (g) (Control: 3577 ± 400; NIDDM: 3489 ± 429; 2p = 0.49) were not different between the groups. There was no significant correlation between birth weight and glomerular number (Control: 2p = 0.80; r = 0.06 and NIDDM: 2p = 0.10; r = –0.33), glomerular volume (Control: 2p = 0.43; r = 0.19 and NIDDM: 2p = 0.78; r = 0.06) or kidney weight (Control: 2p = 0.56; r = 0.14 and NIDDM: 2p = 0.81; r = 0.05). Our results on a limited number of subjects in Denmark do not support the hypothesis that there is any association between low birth weight and low kidney weight or low birth weight and few and/or small glomeruli in NIDDM patients. [Diabetologia (1996) 39: 1634–1637]     

兔肾微血管内皮细胞的体外培养及鉴定方法

目的 探索肾微血管内皮细胞的体外培养方法，为骨组织工程再血管化提供理想的种子细胞；方法 采用三步梯度筛网法，先获得较高纯度的肾血管球，再应用肾血管球外植法体外培养肾微血管内皮细胞；并对肾微血管内皮细胞进行Ⅶ因子相关抗原免疫组化鉴定和透射电镜观察。结果 体外培养出纯度很高的血管内皮细胞并培养了10代；Ⅷ因子相关抗原免疫组化阳性，透射电镜检测出W-P小体。结论 该法可成功培养出肾微血管内皮细胞。     

Two types of GABA-containing axon terminals in cerebellar glomeruli of cat: An immunogold-EM study

Summary Immunogold demonstration of GABA was used in ultrathin sections of cerebellar cortex of cat to identify GABA(+) profiles in cerebellar glomeruli. In addition to small, GABA-containing axonal varicosities found at the periphery of all glomeruli, a few complex glomeruli were found to contain a second type of GABA(+) terminal, with a mossy ending appearance. GABA(+) type I axon terminals, which were identified as the axonal arborization of local Golgi cells, contained ovoid, small synaptic vesicles. GABA(+) type II terminals, however, exhibited large, spheroid synaptic vesicles. Experimental evidence is provided that type II GABA(+) mossy like terminals are the endings of nucleocortical fibers.     

异丙酚预先给药对内毒素诱导大鼠肾小球血管内皮细胞通透性升高的影响

目的 探讨异丙酚预先给药对脂多糖(LPS)诱导大鼠肾小球血管内皮细胞通透性升高的影响.方法 分离、培养SD大鼠肾小球血管内皮细胞,以1×106/ml的密度接种于24孔培养板(200 μl/孔)和transwell小室(100 μl/室),采用随机数字表法,将其随机分为6组(n=10),正常对照组(C组)不作任何处理;脂肪乳对照组(I 组)加入10%脂肪乳4 μg/ml;异丙酚组(P组)加入异丙酚4μg/ml;LPS组(L组):加入LPS 10μg/ml;LPS+脂肪乳组(L+I组)加入10%脂肪乳4 μg/ml及LPS 10μg/ml;LPS+异丙酚组(L+P组)加入异丙酚4μg/ml 及LPS 10 μg/ml.于加入LPS前30 min加入脂肪乳或异丙酚,药物的浓度均为终浓度.加入LPS后6 h,收集细胞,采用逆转录-聚合酶链反应测定血管内皮细胞生长因子(VEGF)mRNA表达水平;收集上清液,采用酶联免疫吸附法测定VEGF浓度;测定血管内皮细胞通透性.结果 与C组比较,L组、L+I组和L+P组VEGF mRNA表达上调,上清液中VEGF浓度和血管内皮细胞通透性增加(P＜0.05),而I组和P组上述指标差异无统计学意义(P＞O.05).与L组比较,L+P组VEGF mRNA表达下凋,上清液中VEGF浓度和血管内皮细胞通透性降低(P＜0.05),L+I组上述指标差异无统计学意义(P＞0.05).上清液中VEGF浓度与血管内皮细胞通透性呈正相关(r=0.833,P＜0.05).结论 异丙酚预先给药可抑制LPS诱导大鼠肾小球血管内皮细胞通透性升高,其机制与下调VEGF表达有关.

Abstract：

Objective To investigate the influence of propofol pretreatment on the increased glomerular endothelial cell permeability induced by lipopolysaccharide (LPS) in rats.Methods Glomerular endothelial cells isolated from SD rats were cultured in 24-well plates(200 μl/well) and transwell filters (100 μl/filter) at 1×106/ml and assigned into 6 groups (n=10 each):control group (group C) , introlipid group (group I), propofol group (group P) , LPS group (group L), LPS+introlipid group (group L+I) and LPS+propofol group (group L +P). In group I, 10% introlipid 4 μg/ml was added. In group P, 4 μg/ml propofol was added. In group L, 10 μg/ml LPS was added. In group L+I, 10% introlipid 4 fig/ml combined with 10 μg/ml LPS was added. In group L+ P, 4 μg/ml propofol combined with LPS 10 μg/ml was added. Introlipid or propofol was added 30 min before the administration of LPS and the corresponding concentrations mentioned above were all final concentrations.After 6 h incubation with LPS, the cells were collected for measurement of vascular endothelial growth factor (VEGF) mRNA expression using RT-PCR. The supernatant was collected for determination of the VEGF concentration by ELJSA. The endothelial cell permeability was determined. Results Compared with group C, the expression of VEGF mRNA was up-regulated and the VEGF concentration and endothelial cell permeability were significantly increased in L, L+I and L + P groups (P＜0.05 ) ,but no significant change was found in the parameters mentioned above in I and P groups (P＞0.05). Compared with group L, the expression of VEGF mRNA was downregulated and the VEGF concentration and endothelial cell permeability were significantly decreased in L+P group (P＜0.05), but no significant change was found in the parameters mentioned above in group L+I(P＞0.05). A positive correlation existed between the concentration of VEGF and the permeability of endothelial cells(r= 0.833,P＜0.05).Conclusion Propofol pretreatment can decrease the increased glomerular endothelial cell permeability induced by LPS probably through down-regulation of VEGF expression.     

他克莫司对人肾小球系膜细胞增殖的影响

目的探讨他克莫司对体外培养的人肾小球系膜细胞增殖的影响及可能机制。方法采用四甲基偶氮唑盐法(MTT)检测不同浓度的他克莫司处理后系膜细胞的增殖情况;用流式细胞仪检测不同浓度的他克莫司作用48、72h后系膜细胞细胞周期的改变;用流式细胞仪检测不同浓度的他克莫司作用72h后系膜细胞发生凋亡的情况。结果①浓度为1μmol/L的他克莫司对系膜细胞的增殖无明显抑制作用;作用24、48和72h后,浓度为5～20μmol/L的他克莫司能显著抑制系膜细胞的增殖;随着他克莫司浓度的增大,抑制作用更显著,呈一定的剂量—效应关系;随着他克莫司作用时间的延长,抑制作用更显著,呈一定的时间—效应关系。②5μmol/L的他克莫司作用48、72h,G0/G1期的系膜细胞百分比增高,S期的系膜细胞百分比降低,细胞周期阻滞于G0/G1期。③5μmol/L的他克莫司作用72h,对系膜细胞的凋亡无明显影响。结论他克莫司显著抑制体外培养的人肾小球系膜细胞增殖,且呈剂量和时间依赖性;其可能机制是使系膜细胞的细胞周期阻滞于G0/G1期。     

Distribution, morphology, and synaptic targets of corticothalamic terminals in the cat lateral posterior-pulvinar complex that originate from the posteromedial lateral suprasylvian cortex

The lateral posterior (LP) nucleus is a higher order thalamic nucleus that is believed to play a key role in the transmission of visual information between cortical areas. Two types of cortical terminals have been identified in higher order nuclei, large (type II) and smaller (type I), which have been proposed to drive and modulate, respectively, the response properties of thalamic cells (Sherman and Guillery [1998] Proc. Natl. Acad. Sci. U. S. A. 95:7121-7126). The aim of this study was to assess and compare the relative contribution of driver and modulator inputs to the LP nucleus that originate from the posteromedial part of the lateral suprasylvian cortex (PMLS) and area 17. To achieve this goal, the anterograde tracers biotinylated dextran amine (BDA) or Phaseolus vulgaris leucoagglutinin (PHAL) were injected into area 17 or PMLS. Results indicate that area 17 injections preferentially labelled large terminals, whereas PMLS injections preferentially labelled small terminals. A detailed analysis of PMLS terminal morphology revealed at least four categories of terminals: small type I terminals (57%), medium-sized to large singletons (30%), large terminals in arrangements of intermediate complexity (8%), and large terminals that form arrangements resembling rosettes (5%). Ultrastructural analysis and postembedding immunocytochemical staining for gamma-aminobutyric acid (GABA) distinguished two types of labelled PMLS terminals: small profiles with round vesicles (RS profiles) that contacted mostly non-GABAergic dendrites outside of glomeruli and large profiles with round vesicles (RL profiles) that contacted non-GABAergic dendrites (55%) and GABAergic dendritic terminals (45%) in glomeruli. RL profiles likely include singleton, intermediate, and rosette terminals, although future studies are needed to establish definitively the relationship between light microscopic morphology and ultrastructural features. All terminals types appeared to be involved in reciprocal corticothalamocortical connections as a result of an intermingling of terminals labelled by anterograde transport and cells labelled by retrograde transport. In conclusion, our results indicate that the origin of the driver inputs reaching the LP nucleus is not restricted to the primary visual cortex and that extrastriate visual areas might also contribute to the basic organization of visual receptive fields of neurons in this higher order nucleus.     

Genetic kidney diseases disclose the pathogenesis of proteinuria

The sieving of plasma components occurs in the kidney through the glomerular capillary wall. This filter is composed of three layers: endothelium, glomerular basement membrane (GBM), and podocyte foot processes connected by slit diaphragms. Defects in this barrier lead to proteinuria and nephrotic syndrome. Previously, defective GBM was regarded to be responsible for proteinuria. However, recent work on genetic diseases has indicated that podocytes and the slit diaphragm are crucial in restricting protein leakage. Congenital nephrotic syndrome of the Finnish type (NPHS1) is caused by mutations in a novel NPHS1 gene, which encodes for a cell adhesion protein, nephrin. This protein is synthesized by podocytes, and seems to be a major component of the slit diaphragm. In severe NPHS1, lack of nephrin leads to missing slit diaphragm. The role of nephrin in acquired kidney diseases remains unknown. In addition to nephrin, other podocyte proteins (podocin, α-actinin-4, CD2AP, FAT) have recently been identified and associated with the development of proteinuria. It seems that the slit diaphragm and its interplay with the podocyte cytoskeleton is critical for the normal sieving process, and defects in one of these components easily lead to proteinuria.     

成人特发性局灶节段性肾小球硬化症的临床及病理分析

目的:了解特发性局灶节段性肾小球硬化症(focal segmental glomerulosclerosis,FSGS)在肾脏疾病中的构成变化及其临床病理诊断要点和意义.方法:对1990年1月至2001年12月在我院肾科住院行肾活检确诊为特发性FSGS的65例患者的临床资料及病理改变进行总结分析.结果:(1)特发性FSGS占肾活检患者的2.2%,占原发性肾小球疾病的3.2%,占肾病水平蛋白尿患者的5.8%,12年中发病无上升趋势.(2)其病变类型为单纯门部病变占12.3%,单纯周缘部病变占23.1%,两者的混合型占60.0%,尖端型占1.5%,塌陷性占3.1%.同时伴随的病变有:足细胞增生、肥大和空泡变性(58.5%),球囊粘连(90.8%),肾小管间质病变(92.3%)以及节段性内皮和系膜细胞增生(20.0%).由于病变分布呈局灶节段的特点,且病变多样、重叠出现,在病理诊断中应警惕误、漏诊的可能.(3)伴节段性内皮和系膜细胞增生的患者常表现为肾病水平蛋白尿(92.3%).结论:在肾脏病理工作中,提高警惕性和检查方法对于正确诊断特发性FSGS具有重要意义.特发性FSGS不是构成本组肾活检患者的常见病理类型.足细胞病变以及节段性内皮和系膜细胞增生可能与节段性硬化形成有关.