大鼠脑皮质微血管内皮细胞培养和形态学观察

为获取较纯净脑微血管内皮细胞进行血脑屏障的病理生理研究，我们采用脑组织匀浆、过滤和酶消化技术分离大鼠脑微血管，对分离的脑微血管内皮细胞进行了体外培养和形态学观察。倒置显微镜下，细胞具有单层“卵石样”排列的典型特征、电镜观察可见细胞间连接，免疫酶技术显示，95％以上的细胞为第Ⅷ因子相关抗原反应阳性，进一步证实为血管内皮细胞。     

Distribution of the largest neuron in mouse caudate-putamen nucleus: Its position in large-cell — Medium-cell clusters

Summary The position of the largest striatal neuron within territories delimited by medium-sized clustered neurons was charted in Nissl-stained sections through the mouse caudate-putamen nucleus. Medium-sized neuron somata occur in close proximity to this large cell at some point in the anteroposterior, mediolateral or dorsoventral extent of its soma. The size of the network of medium-sized neurons associated with the large cell may vary from two to 15 neurons. Even when this network is extensive, the large neuron is never completely surrounded. Most often, this cell also borders a fascicle of internal capsule fibers, and the entire cellular island may be aligned either parallel to or perpendicular to the orientation of these fibers. These findings suggest the hypothesis that cellular territories in the caudate-putamen nucleus have a very specific orientation in three dimensional space.     

Towards the phenotyping of soft tissue tumours by cell surface molecules

Summary This study is aimed at the characterization of soft tissue tumours (STT) by means of cell surface molecules. To achieve this, normal mesenchymal tissues were extensively examined for expression of leucocyte differentiation (CD) antigens and HLA molecules. The panel of antigens finally examined in STT comprised CD10, CD13, CD24, CD34, CD36, CD56, CD57, HLA-A,B,C, ₂-microglobulin, HLA-DR, -DP, and -DQ and the HLA-D-associated invariant chain (Ii). STT were determined by conventional histomorphological and immunohistochemical criteria. The immunohistological analysis was based on serial frozen sections, one of which was used to demonstrate CD53 antigen. This very broadly distributed leuco/histiocyte-restricted antigen allowed for the distinction between the background of interstitial stromal cells and the neoplastic population. In some STT, the expression pattern of the cell surface molecules corresponded to that in their non-neoplastic counterparts. The majority of STT, however, showed considerable changes in the cell surface immunophenotype compared to their cells of origin. These alterations consisted mainly in an aberrant induction/neoexpression and, to a much lesser extent, in an aberrant down-regulation/loss of cell surface antigens. Nevertheless, some immunophenotype configurations are described which, for the time being, can be considered to be useful supplements in the differential diagnosis of this complex class of tumours. The data also indicate considerable changes in cell surface antigen expression occurring in the course of neoplastic transformation of mesenchymal cells. Detailed analysis of alterations in the functional repertoire of neoplastic mesenchymal cells might provide new insights into the biology of STT, possibly leading to new concepts for therapeutic intervention.This study was supported by the Tumorzentrum Heidelberg/ Mannheim and by the Dr. Mildred-Scheel-Stiftung für Krebshilfe (W50/89/Mö2)     

NIH 3T3细胞转化前后细胞骨架及细胞表面纤维粘连蛋白的免疫荧光观察

本实验用抗纤维粘连蛋白(FN)亲和层析纯抗体和抗管蛋白抗体及鬼笔环肽,以免疫荧光组织化学方法,对NIH3T3细胞转染人肺腺癌细胞系AGZY基因组DNA前后细胞表面FN及细胞内骨架系统进行染色观察。结果表明,细胞在发生转化后,微丝及微管均表现出明显受损,细胞骨架结构不清,呈现为弥散样荧光;细胞表面FN大量减少,仅及正常NIH3T3细胞的1/9,其分布也由细丝形成的网状,变成斑点或斑块状。这一结果进一步证实,细胞恶变是涉及到细胞骨架系统及膜表面糖蛋白变化的复杂过程,并预示这些变化可能就是导致细胞形态发生变化、细胞失去正常生长调控的原因之一。     

Protective effect of thymidine against cytostatic action of cyclic-AMP in mammalian cell cultures

Under the influence of 1 mM cyclic-adenosine-3,5-monophosphate (cyclic-AMP) the degree of survival and rate of reproduction of Chinese hamster cells in culture were reduced to 27 and 42% of the control level, respectively. Addition of 0.02 mM thymidine along with the cyclic-AMP almost completely abolished the cytostatic effect of the latter. Thymidine also prevented the cytostatic effect of noncyclic 5-AMP, but did not affect death of the cells due to the action of dibutyryl cyclic-AMP and theophylline. Thymidine did not prevent the inhibitory action of cyclic-AMP on a mutant line of mouse cells deficient in thymidine kinase. It is concluded that, in the concentrations used, the cytostatic action of exogenous cyclic-AMP on mammalian cells is the result of its splitting to 5-AMP in the culture medium, and that it acts by blocking one of the stages of TMP biosynthesis.Department of Genetics and Plant Breeding, Faculty of Biology, M. V. Lomonosov Moscow University. (Presented by Academician S. E. Severin.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 80, No. 7, pp. 93–95, July, 1975.     

MTEC 1分泌的趋化因子引起特定亚群胸腺细胞的定向迁移

分析胸腺髓质上皮样细胞系ＭＴＥＣ１分泌的化学趋化因子对胸腺细胞亚群的趋化作用。方法以抗体加补体杀伤结合免疫磁珠及ｐａｎｎｉｎｇ法，将小鼠胸腺细胞分离纯化，获得ＣＤ４＋ＣＤ８＋（ＤＰ），ＣＤ４－ＣＤ８－（ＤＮ），ＣＤ４＋ＣＤ８－（ＣＤ４ＳＰ）及ＣＤ４－ＣＤ８＋（ＣＤ８ＳＰ）四亚群细胞，用Ｂｏｙｄｅｎ小室分析ＭＴＥＣ１┐ＳＮ对四群胸腺细胞的趋化作用。结果ＭＴＥＣ１┐ＳＮ对ＤＰ及ＣＤ４ＳＰ胸腺细胞有趋化活性（ＣＩ＝６．６±１．０及６．１±１．８）；对ＣＤ８ＳＰ细胞有中度趋化活性（ＣＩ＝３．２±１．０）；对ＤＮ趋化活性微弱（ＣＩ＝１．３±０．６）。化学趋化因子ＭＣＰ┐１纯品对ＣＤ４ＳＰ胸腺细胞显示强趋化活性（ＣＩ＝５．６），对ＤＮ胸腺细胞则无可测出趋化活性。结论ＭＴＥＣ１分泌的化学趋化因子对ＤＰ，ＣＤ４ＳＰ及ＣＤ８ＳＰ胸腺细胞有显著趋化作用，对ＤＮ胸腺细胞几乎无趋化作用。提示此类化学趋化因子有趋使胸腺发育中后期阶段的细胞向胸腺髓质区迁移和定位的作用。     

Cerebral ischemia induces transient intracellular redistribution and intranuclear translocation of the raf proto-oncogene product in hippocampal pyramidal cells

Summary In this report we describe changes in the intracellular redistribution of raf serine/threonine protein kinase (product of the raf proto-oncogene family) in hippocampal neurons following cerebral ischemia in Mongolian gerbils. For immunohistochemical localization studies polyclonal antisera specific for each of the A, B, and Raf-1 isotypes of raf, as well as a pan-raf antisera, were employed. Of these, only sera recognizing B-raf, as well as the general v-raf (raised against the conserved C-terminal region) were positive, indicating that B-raf is the major isotype in this neuronal region. Three different ischemie models were used (repeated 3 times for two min and single 5 or 15 min occlusions, of the common carotid arteries) to demonstrate that ischemie insult causes redistribution of raf protein kinase into the cell nucleus of hippocampal neurons. Increased amounts of raf protein in the nuclei of pyramidal cells following ischemia was confirmed by Western blot analysis of isolated nuclear fractionations. Moreover, an elevation in the level of nuclear raf protein also was detected in the contralateral (i.e. non-occluded hemisphere) neurons of CA1 and CA3 subfields 4 days after the ischemie insult indicating a possible transsynaptic increase in the amount of raf protein along with redistribution. The intranuclear translocation of the immunoreactive material started from the perinucleolar rim and with time extended throughout the nucleus. Enhanced levels and altered redistribution of the raf polypeptide in the nuclei of pyramidal cells of the CA3 subfleld appears to be reversible and returns to the normal level 12 days following the ischemic insult. In addition to triggering the above changes in the intracellular redistribution of raf, ischemie insult also caused an increase in the level of B-raf protein in reactive astrocytes.     

Administration of ciprofibrate to lactating mothers induces PPARα-signaling pathway in the liver and kidney of suckling rats

It is well known that the hypolipidemic drug ciprofibrate induces peroxisome proliferation in rodent liver, which in turn leads to the oxidative stress, and modifies some parameters related to cell proliferation and apoptosis. The administration of ciprofibrate to rats during the lactating period determined in their pups significant modifications in hepatic peroxisome enzyme activities, induction of the PPARalpha-target gene, Cyp4a10, and perturbation in cell proliferation and apoptosis, which affected the size of the liver. Moreover, this modification was associated to about two-fold induction of mRNA-PPARalpha. On the contrary, in the kidney, although a similar two-fold up-regulation of PPARalpha was detected, the induction of both peroxisomal enzyme activities and Cyp4a10 were weak, and no alterations were detected, neither in cell cycle nor in the size of the tissue. Our results indicate that the response to ciprofibrate is stronger in the liver than in the kidney of newborn rats.