Visual cells of zebrafish optic tectum: Mapping with small spots

The zebrafish optic tectum is anatomically similar to those of goldfish and other teleosts, both in its laminar structure and the morphology of intrinsic neurons as studied with Golgi stains. We have applied standard electrophysiological techniques to study the visual properties of tectal cells, utilizing a computer system for stimulus control and data recording. All tectal cells have very large receptive fields, averaging 25–39 degrees in linear dimensions. Retinal receptive fields are smaller, averaging 7–13 degrees. In many cases the receptive fields of tectal cells, but never of retinal cells, consist of two parts (main field and accessory field) separated by tens of degrees. The two parts are differentially adapted by background illumination, accessory fields becoming unresponsive under lit conditions while main fields do not. This may reflect separate retinal input channels.Four types of tectal cells are described, which differ in their spontaneous activity in the dark and response to stationary spots. Type I are not spontaneously active in the dark, but respond phasically at ON and OFF. Type T are tonically active and give more prolonged phasic responses to ON and OFF. They may also have pure-inhibitory receptive fields in which spot ON suppresses the spontaneous firing with no phasic excitation. Type S are also silent in the dark, but give sustained firing as long as a spot is ON in the receptive field. Cells of type B fire spontaneously in bursts; the burst rate may be raised or lowered by stationary spots, but there is no phasic response. Each of the four physiological types is found to occur among the cells of the periventricular layer, all of which share a stereotyped overall morphology. Tectal cells do not exhibit spatially separated ON and OFF areas or orientation specificity.     

血红素氧合酶-1对人胃癌SGC7901细胞5-氟尿嘧啶化疗敏感性的影响

目的:体外观察血红素氧合酶-1(heme oxygenase-1,HO-1)对人胃癌细胞SGC7901 5-氟尿嘧啶(5-FU)化疗敏感性的影响及其可能的机制?方法:应用不同浓度锌原卟啉(ZnPP)?氯化血红素(Hemin)单独或联合5-FU作用于人胃癌SGC7901细胞不同时间,四甲基偶氮唑盐(MTT)法观察药物对细胞生长抑制作用;Western blot分析细胞HO-1蛋白表达变化;流式细胞术检测细胞凋亡率;活性氧(ROS)检测试剂盒分析其凋亡机制?结果:ZnPP和5-FU均可明显抑制人胃癌SGC7901细胞生长,呈剂量依赖性;5-FU联用ZnPP(10 μmol/L)与单用药相比,明显提高了细胞生长抑制率和凋亡率(P < 0.05),联用Hemin(10 μmol/L)则明显降低细胞生长抑制率和凋亡率(P < 0.05);人胃癌SGC7901细胞中可见HO-1表达,单用5-FU或Hemin后均能使其表达增强,联用ZnPP可逆转此现象;单用5-FU与对照组比较,细胞内ROS活性明显增高,联用后ZnPP可进一步提高细胞内ROS活性(P < 0.05),而联用Hemin明显降低细胞内ROS活性水平(P < 0.05)?结论:ZnPP抑制HO-1表达,能够增强人胃癌SGC7901细胞对5-FU化疗敏感性,这一作用可能与增加细胞内ROS活性有关?     

蝙蝠葛酚性碱对胃癌细胞株 SGC -7901 Fas 基因表达的影响

目的：探讨蝙蝠葛酚性碱（PAMD）抗胃癌的作用机理。方法：通过实时定量PCR的方法观察PAMD对胃癌细胞株SGC-7901FasmRNA表达的影响,以此探讨PAMD抗肿瘤作用机理。结果：PAMD各剂量组均能上调FasmRNA的表达,较阳性对照组明显,但差异与空白组比较均不具有统计学意义（P〉0．05）。结论：PAMD抗胃癌的作用机理可能与其上调Fas基因表达有关。     

Protein expression and mRNA cellular distribution of the NKCC1 cotransporter in the dorsal root and trigeminal ganglia of the rat

Primary afferent neurons maintain depolarizing responses to GABA into adulthood. The molecular basis for this GABAergic response appears to be the Na+K+2Cl- cotransporter NKCC1 that contributes to the maintenance of a high intracellular chloride concentration. Recently, a role for NKCC1 has been proposed in nociceptive processing which makes it timely to gain a better understanding of the distribution of NKCC1 in sensory ganglia. Here, we describe that, in the rat, NKCC1 mRNA is predominately expressed by small and medium diameter dorsal root (DRG) and trigeminal (TG) ganglion neurons. The colocalization of NKCC1 mRNA with sensory neuron population markers was assessed. In the DRG, many NKCC1 mRNA-expressing neurons colocalized peripherin (57.0+/-2.5%), calcitonin-gene-related peptide (CGRP, 39.2+/-4.4%) or TRPV1 immunoreactivity (50.0+/-1.9%) whereas only 8.7+/-1.2% were co-labeled with a marker for large diameter afferents (N52). Similarly, in the TG, NKCC1 mRNA-expressing neurons frequently colocalized peripherin (50.0+/-3.0%), CGRP (35.4+/-2.6%) or TRPV1 immunoreactivity (44.7+/-1.2%) while 14.8+/-1.3% were co-labeled with the N52 antibody. NKCC1 mRNA was also detected in satellite glial (SGCs) in both the DRG and TG. Colocalization of NKCC1 protein with the SGC marker NG2 confirmed the phenotype of these NKCC1-expressing glial cells. In contrast to in situ hybridization experiments, we did not observe NKCC1 immunoreactivity in primary afferent somata. These findings suggest that NKCC1 is expressed in anatomically appropriate cells in order to modulate GABAergic responses in nociceptive neurons. Moreover, these results suggest the possibility of a functional role of NKCC1 in the glial cells closely apposed to primary sensory afferents.     

Differentiation of neuroblasts in the chick optic tectum up to eight days of incubation: A Golgi study

Tectal histogenesis up to the 8th day of incubation is described, according to findings in Golgi-impregnated material. Emphasis is placed on the mode of initial differentiation of the different types of neuroblasts and evidence has been obtained that there are essentially two kinds of neuroblasts within the optic tectum.Type I cells initiate their differentiation process, starting from an interphase epithelial morphology, by losing their ventricular attachment while they keep the subpial one. This subpial attachment subsequently transforms into a sprouting formation, whose growth produces the axon of the neuroblast. The cell body is transported by translocation of the nucleus within the peripheral cylinder of cytoplasm. Periventricular, multipolar and shepherd's crook neurons of the tectum are shown to differentiate out of type I neuroblasts.Type II cells lose their ventricular attachment shortly after mitosis, and they are thus free unattached cells. They migrate radially to the superficial tectal strata, forming the cortical plate of the tectum. Their leading processes transform into dendrites, and the axons grow finally out of the inner poles of the cell bodies. These neuroblasts differentiate into a set of known adult neuronal types (piriform radial, horizontal and stellate neurons of the stratum griseum et fibrosum superficiale).     

SGC7901细胞对化疗药物的敏感性研究

目的 了解胃腺癌细胞株SGC7901细胞对不同化疗药物的敏感性,为临床选择化疗药物提供信息. 方法 分别用5氟尿嘧啶、阿霉素、顺铂、表阿霉素、丝裂霉素、环磷酰胺的常规用药峰浓度孵育SGC7901,采用MTT法,测量孵育前后细胞的吸光度,计算各化疗药物对SGC7901细胞的抑制率. 结果 在常规峰浓度时对SGC7901细胞的抑制率分别为: 5氟尿嘧啶57.4%、阿霉素56.4%、顺铂39.8%、表阿霉素38.0%、丝裂霉素92.2%、环磷酰胺15.0%. 结论 SGC7901对化疗药物的敏感性依次为丝裂霉素、5氟尿嘧啶、阿霉素、顺铂、表阿霉素及环磷酰胺.达到50%以上抑制率的药物只有丝裂霉素、5氟尿嘧啶、阿霉素.     

A Study of Tumor Apoptosis Induced by Medicine Serum of Traditional Chinese Medicine

Objective Adopting serological methods of compound Chinese medicine that including ＇Yi-Fei-Yin＇ to treat lung cancer, ＇Yi-Chang-Tai＇ to treat lung cancer and ＇Wei-Ai-Ning＇ to treat gastric cancer to explore the induced effect on the tumor cell apoptosis. Methods （1） Culture conventionally A549 human lung cancer cell lines, CCL229 human colorectal carcinoma cell lines and gastric cancer cell line SGC7901, and observe under the inverted microscope. （2） Three Chinese medicine compound which were decoction, three consecutive days of intragastric administration to rats, last administration was two hours before the animals were killed, according to the serum drug Preparation prepared medicinal serum. （3） Three drugs were cultured in serum of A549 human lung cancer cell lines, CCL229 human colorectal carcinoma cell lines and cell line SGC7901 48 hours. （4） With rubber scraper scraped cells, cell suspension will be transferred to the centrifuge tube centrifuge 2000 rpm/min, 15min, Agar processing into pieces, washed with PBS, 1% osmium acid fixed, gradient ethanol and acetone dehydration, epoxy-embedded mesh, the conventional ultra thin section, electron stain. （5） To observe and take a picture under the JEM1200 TEM. Results （1） The parapodium of tumor cells decreased even disappeared, endocytoplasmic reticulum, mitochondria and other cell organelles for Poly IC, chromatin condensation, margination a ＇crescent＇. （2） It is apoptosis characteristic structure--the membrane parcels, containing some of the broken device and cell chromatin of apoptotic bodies. Conclusion Compound Chinese medicine serum has the function to induce tumor cells apoptosis, widely used for clinical medicine compound against lung cancer, gastric cancer and colorectal cancer provide reliable experimental evidence.     

透明质酸、透明质酸酶和RGD抑制胃癌细胞SGC7901黏附侵袭的实验研究

目的　探讨单独及联合应用透明质酸 (HA)、透明质酸酶 (Hase)和RGD对SGC790 1细胞黏附和侵袭细胞外基质 (ECM)的抑制作用。方法　采用间接免疫荧光法测定SGC790 1细胞表面CD4 4与integrinβ1蛋白表达。单独或联合应用HA、Hase和RGD处理体外培养的胃癌细胞SGC790 1;MTT法和Boyden小室模型分别测定其黏附力和侵袭力 ,同时观察细胞形态。结果　人胃癌细胞系SGC790 1表达CD4 4与integrinβ1蛋白。与对照组比较 ,Hase及RGD肽均明显抑制SGC790 1的黏附力(P <0 .0 0 1)和侵袭力 (P <0 .0 5 ) ,效果优于HA(P <0 .0 5 ) ;Hase HA或三者联合的抑制效果优于任何单一抑制效果 (P <0 .0 0 1)。形态学观察发现 ,未处理组的部分细胞已经开始铺展 ,穿过人工基底膜后 ,表现出纤维母细胞样形态及形态各异的伪足 ;而处理组细胞形态较圆 ,伪足数目相对较少。结论　人胃癌细胞系SGC790 1表达功能性CD4 4和integrinβ1蛋白。单独及联合应用HA、Hase和RGD均不同程度抑制SGC790 1细胞对ECM的黏附和侵袭 ,以联合三者阻断效果为优。     

mdr1基因在多药耐药相关的几种人胃癌细胞系的表达

目的探讨mdr1在多药耐药相关的几种人胃癌细胞系中的转录、翻译、P-gp功能变动趋势.方法培养SGC7901/VCR(人胃癌多药耐药细胞亚系)、SGC7901和BGC823(人胃腺癌细胞系)于RPMI1640,用RT-PCR和原位杂交检测mdr1mRNA的表达,用免疫印迹和免疫组化检测P-gp的表达,用流式细胞仪检测阿霉素在细胞内的蓄积.结果半定量RT-PCR显示,SGC7901/VCR mdr1和β-actin吸收峰面积之比为5.63,SGC7901为0.61,BGC823为0.85.杂交信号呈紫蓝色颗粒,分布于胞质.免疫印迹显示SGC7901/VCRP-gp的表达最强,SGC7901最弱.P-gp阳性呈棕黄色颗粒,位于细胞膜上.SGC7901中,少数细胞P-gp表达极强.SGC7901/VCR平均光密度为(1.8310±0.8401),SGC7901为(0.3590±0.2512),BGC823为(0.6260±0.4996)(P＜0.05).SGC7901/VCR阿霉素特异荧光强度为(6.59±50.30),SGC7901为(35.88±14.55),BGC823为(27.44±7.06)(P＜0.001).结论在多药耐药相关的人胃癌细胞系SGC7901/VCR、SGC7901和BGC823中,mdr1基因均表达,P-gp具有药物外排功能,SGC7901/VCR的mdr1表达最强,SGC7901最弱,BGC823居中.     

Specific cerebral heat shock proteins and histamine receptor cross-talking mechanisms promote distinct lead-dependent neurotoxic responses in teleosts

Recent interests are beginning to be directed towards toxic neurobiological dysfunctions caused by lead (Pb) in aquatic vertebrates. In the present work, treatment with a maximum acceptable toxic concentration of this heavy metal was responsible for highly significant (p<0.01) abnormal motor behaviors such as hyperactive movements in the teleost Thalassoma pavo and the same treatment accounted for significantly (p<0.05) enhanced hyperventilating states. On the other hand, greater abnormal motor behaviors were detected in the presence of the histamine (HA) receptor subtype 2 (H(2)R) antagonist cimetidine (Cim), as shown by the very robust (p<0.001) increases of the two behavioral states. Interestingly, elevated expression levels of stress-related factors, i.e. heat shock protein70/90 (HSP90/70) orthologs were reported for the first time in hypothalamic and mesencephalic areas of Pb-treated teleosts. In particular, an up-regulation of HSP70 was readily detected when this heavy metal was given concomitantly with Cim, while the histamine subtype 3 antagonist (H(3)R) thioperamide (Thio), instead, blocked Pb-dependent up-regulatory trends of both chaperones in mostly hypothalamic areas. Moreover, intense neuronal damages of the above brain regions coincided with altered expressions of HSP70 and HSP90 when treated only with Cim. Overall these first results show that distinct H(n)R are able to exert a net neuroprotective role arising from their interaction with chaperones in fish exposed to Pb-dependent stressful conditions making this a potentially key interaction especially for T. pavo, aquatic species which plays an important ecological role towards the survival of other commercially vital fishes.