THE LOCALIZATION OF ENZYME ACTIVITIES IN THE RAT BRAIN

Studies with rat brain illustrate the usefulness of formol-calcium-fixed tissue for studying both enzymatic "chemoarchitectonics" and intracellular organelles. Unembedded frozen sections and polyvinyl alcohol-embedded sections may be used to demonstrate the activities of DPNH-tetrazolium reductase localized in mitochondria and ergastoplasm, TPNH-tetrazolium reductase localized in mitochondria, ATPase (and/or apyrase or ADPase) in cell membranes, and acid phosphatase in lysosomes.¹ Among the observations recorded are: (1) the presence of lysosomes in all cells of the brain; (2) the presence of numerous large lysosomes near the nuclei of capillary endothelial cells; (3) a polarized arrangement of large lysosomes in epithelial cells of the ependyma and choroid plexus; (4) the presence of ATPase activity in the cell membranes of some neurons; (5) the presence of either an apyrase or combination of ATPase and ADPase in the cell membranes of neuroglia and capillaries; (6) the presence of both DPNH- and TPNH-tetrazolium reductase activities in neuroglia; (7) the presence of DPNH- and TPNH-tetrazolium reductase activities in mitochondria and of DPNH-tetrazolium reductase activity in Nissl substance. The possible functional significance of these localizations is briefly discussed, as is their relation to "quantitative histochemistry" data available in the literature.     

POLYRIBOSOMES IN PROLIFERATING AND NONPROLIFERATING ROOT MERISTEM CELLS

Pea root meristem cells which are actively dividing contain higher proportions of polyribosomes than do cells whose progression through the mitotic cycle has been arrested by starvation. The initiation of cell division which follows the addition of sucrose to starved roots is accompanied by an increase in polyribosome content. This reformation of polyribosomes is insensitive to actinomycin D; it is suggested that preexisting mRNA molecules, synthesized prior to starvation, participate in polyribosome reformation, but that those mRNA species necessary for movement of cells through the mitotic cycle are gradually lost during the starvation period.     

THE CYTOCHEMICAL LOCALIZATION OF ASCORBIC ACID IN ROOT TIP CELLS

The intracellular distribution of ascorbic acid was studied in frozen-dried root tips of Allium cepa and Vicia faba by the silver nitrate procedure. The sites of the ascorbic acid as indicated by the deposited silver appear as spherical (0.2 to 0.6 µ in diameter) cytoplasmic particles. The site appears to have small amounts of lipides and to be rich in ribonucleic acid. These particles are concluded to be submicroscopic in size and associated, in the elongating cell, with the cell surface. In the meristematic cells they appear fewer in number and are distributed throughout the cytoplasm.     

扬子鳄消化道黏液细胞和5种酶的组织化学定位

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FINE STRUCTURE STUDIES OF PHLEUM ROOT MERISTEM CELLS. I. MITOCHONDRIA

Avers , Charlotte J. (Douglass Coll., Rutgers—The State U., New Brunswick, N.J.) Fine structure studies of Phleum root meristem cells. I. Mitochondria. Amer. Jour. Bot. 49(9): 996–1003. Illus. 1962.—Mitochondrial numbers were computed from thick sections embedded in paraffin and stained with iron hematoxylin, from methacrylate embedded, 0.5 μ-thick sections stained with osmium, and from electron micrographs of ultrathin sections fixed in KMnO₄. The mean mitochondrial counts per average cell (20 × 20 × 10μ) were: 121 ± 4 in paraffin sections, 600 ± 40 in methacrylate sections, and 991 ± 96 from electron micrographs. These numbers were higher than the Janus green B count of 91 ± 2 per cell found during an earlier study. In each case, arithmetical conversion factors were used to calculate total numbers of mitochondria per cell since section thicknesses were less than that of whole cell length or depth. Determinations of numbers of mitochondria probably varied depending on section thickness and specific staining procedures used, but the higher count from electron micrographs was assumed to be reasonable on a volumetric basis. The cytoplasmic volume of the average cell is about 2500 μ³ and the volume of a single average mitochondrion is about 0.1μ^3. On this basis, the variety of structures and their relatively sparse distribution seems possible despite the apparently high numbers found. In addition to mitochondria, the numbers of various cellular components per unit cytoplasmic area were determined. These data showed that mitochondria were about 6 times more numerous than proplastids, about 4.5 times more numerous than Golgi bodies, and even more frequent when compared with vesicles, dense bodies, or microbodies. No correlation was found between cytoplasmic area and numbers of organelles per unit area. Photographic evidence was presented for the occurrence in plant cells of the hepatic cell, polymorphic “lysosome” described by Ashford and Porter. The controversial nature of the lysosome is discussed.     

ONTOGENETIC REORGANIZATION OF THE ROOT MERISTEM IN THE COMPOSITAE

The organization of the root meristem in selected Compositae was investigated to determine whether changes in the pattern of cell arrangement occurred during root growth in species other than Helianthus annuus. Embryonic, short, and long primary roots of one species of each of twelve genera were prepared for microscopic examination. Additional intermediate growth stages were prepared for Echinacea pallida. The meristem of embryonic roots showed layers of initials typical for dicotyledons. The meristem in many of the short roots of eight species was reorganized by the development of a secondary columella. The long roots showed patterns similar to the embryonic roots. In three species which maintained closed meristems, two layers of cortical initials were common in the embryonic root, and as a general trend, a single layer of cortical initials became more common during root elongation. The cellular changes that resulted in the initiation of a secondary columella are characterized by the conversion of cortical initials to secondary columella initials by a shift in their plane of cell division. It is proposed that the size and shape of the quiescent center changes as the conversion takes place. No intermediate stages were observed which could account for the reduction of two layers of cortical initials to one layer.     

蚕豆细胞核仁和核质中剪接因子SC35的定位研究

本文以蚕豆(Vicia faba L.)根端分生组织细胞为材料,以抗SC35抗体为探针,在电镜下对SC35在高等植物细胞中的存在与否和分布特点进行了研究,发现经抗SC35抗体标记后,标明SC35位置的胶体金颗粒主要分布于核仁的致密纤维组分(DFC)、核质的染色质间颗粒(IGs)和染色质周边纤维处(PFs),而核仁的纤维中心(FC)、核仁液泡和集缩染色质团块中央部位的金颗粒很少。DFC, IGs和PFs处的金颗粒平均密度分别为65.89个/μm~2和36.28个/μm~2,远远高于集缩染色质团块中央部位以及FC和核仁液泡处的金颗粒平均密度(分别为5.90个/μm~2和6.26个/μm~2)。说明蚕豆细胞核仁的DFC,核质的IGs和PFs处富含剪接因子SC35。本文研究结果表明,SC35或SC35类蛋白在蚕豆细胞核质中的分布与其在哺乳动物细胞核质中的分布规律相似。同时本文首次报道了SC35或SC35类蛋白存在于核仁中。     

HISTOCHEMICAL LOCALIZATION OF ALCOHOL DEHYDROGENASE IN DEVELOPING BEAN SEEDS

Alcohol dehydrogenase activity can be readily localized within bean seeds using the tetrazolium method. In the seed coat, prior to the cotyledon stage of embryo development, staining was seen throughout, especially in the branched parenchyma cells and the integumentary tapetum. As the embryo entered the maturation stage, a gradual decrease in the staining intensity was observed throughout the seed coat. Embryos stained well at all stages of development. In the course of this investigation, a high activity of “nothing dehydrogenase” was observed. Since its activity could be inhibited by methylpyrazole, an ADH inhibitor, it is concluded that the nothing dehydrogenase activity is caused by alcohol dehydrogenase.     

日本血吸虫胆碱酯酶的组织化学定位

血吸虫的胆碱酯酶(ChE)与血吸虫神经介质的传递、肌肉活动以及与其它物质代谢均有密切关系,也是抗血吸虫药物作用的靶子之一。1959年沈美玲等进行了药物对日本血吸虫乙酰胆碱酯酶(AChE)活力的研究。最近,姚民一等(1981)采用等电聚焦电泳分离出日本血吸虫胆碱酯酶的同功酶。然而有关血吸虫胆碱酯酶的组织化学定位研究主要限于曼氏血吸虫(Pepler,1958;Lewert等,1965;Fripp,1967;Bueding等,1967;Bruckner等,1974;Diconza等,1975)除Bueding简单述及日本血吸虫成虫的AChE外,迄今国内外尚无日本血吸虫CbE的组化资料,而且日本血吸虫的神经系统也未被专门观察。由于ChE的组化定位能细致地显示出血吸虫的神经系统构造,并能进一步提     

大鳞副泥鳅肠道黏液细胞及消化酶活性分布特征研究

采用阿利新兰-碘酸雪夫氏反应(AB-PAS)染色法及酶学方法研究了大鳞副泥鳅成熟个体肠道各段黏液细胞分布及消化酶活性。结果表明, 大鳞副泥鳅肠道黏液细胞分为Ⅰ、Ⅱ、Ⅲ和Ⅳ 4种类型。前肠至后肠, 黏液细胞数量逐渐减少。前肠主要分布Ⅲ和Ⅳ混合型黏液细胞, 后肠则以Ⅱ和Ⅳ型酸性黏液细胞为主。肠道胰蛋白酶活性显著高于淀粉酶和脂肪酶。且后肠消化酶活性显著低于前肠和中肠。根据黏液细胞及消化酶活性分布特点, 表明大鳞副泥鳅属于杂食性鱼类, 前肠为其主要的消化吸收场所, 后肠中性黏液细胞的数量较少以及消化酶活性较低, 表明其对食物的消化吸收功能较弱, 与其为辅助呼吸功能的特点相关。     

HISTOCHEMICAL STUDY OF ENZYME ACTIVITY IN THE SHOOT APICAL MERISTEM OF BRASSICA CAMPESTRIS DURING TRANSITION TO FLOWERING. IV. GLUCOSE-6-PHOSPHATASE

Glucose-6-phosphatase (G6P) activity was determined in fresh-frozen, cryostat sections in the shoot apical meristem of Brassica campestris L. Enzymatic activity was differentially distributed in a zonate pattern in the vegetative meristem, but not in the transition and floral meristem. Vegetative apices showed a heterogenous localization with the highest activity in the central zone and the pith-rib meristem zone. At the early transition stage of development, G6P activity in the peripheral zone increased slightly. At the late transitional (prefloral) stage, G6P activity was not localized within the peripheral zone in island-like areas of activity. This is the first demonstration of G6P in shoot apical meristem at the vegetative, transition, and floral stage. The results indicate that G6P activity 1) is an accompanying event of evocation, but 2) does not mark incipient floral primordia. G6P may play an important role in the maintenance of glucose-6-phosphate homeostasis in an evoked shoot apical meristem.     

HISTOCHEMICAL LOCALIZATION OF ENZYMES IN TAPINANTHUS BANGWENSIS: ACID PHOSPHATASE

The distribution of acid phosphatase in the tissues of Tapinanthus bangwensis, a semiparasitic member of the Loranthaceae, and some of its hosts was studied. It has beer possible to work out convenient routine methods of pretreating tissues for histochemical enzyme localization, to modify, where necessary, conventional histochemical techniques for the localization of acid phosphatase, and to evaluate the azo-dye and metal-salt techniques at the optical level. Histochemical localization showed generally widespread activity and similarity in distribution for this enzyme in the parasite and host tissues. Although there is no direct correlation between these localizations and host-parasite relationships, the bearing that the localizations may have on such relationships is discussed in the light of the distributional evidence and the role usually ascribed to this enzyme.     

吸虫类的酚及酚酶的组织化学定位

本文对代表不同分类位置和不同寄居部位的8种吸虫体内的酚类物质及酚酶进行了组织化学定位的研究。结果表明:仅在上述虫体的成熟卵黄细胞颗粒球和子宫中的卵壳含有丰富的酚及酚酶物质,这些物质在卵壳形成中具有一定的生理作用。本文列表比较了吸虫纲中酚酶的组织化学,并对吸虫类卵壳蛋白的化学性质进行了讨论。