CYTOCHEMICAL LOCALIZATION OF ACID PHOSPHATASE ACTIVITY IN GRANULE FRACTIONS FROM RABBIT POLYMORPHONUCLEAR LEUKOCYTES

When rabbit peritoneal exudates (97% polymorphonuclear [PMN] leukocytes, 2% mononuclear cells) were fractionated by zonal sedimentation or isopycnic centrifugation, four fractions (A, B, C, and D) were obtained, as reported earlier. "A" consisted largely of PMN azurophil granules, "B" of PMN specific granules, and "D" of membranous elements. The source of the more heterogeneous "C" fraction (containing acid hydrolases) was uncertain. To gain further information on the nature of this fraction, cytochemical tests for acid phosphatase (AcPase) were carried out on the starting cells and on the fractions. In intact PMN, lead phosphate reaction product was found in Golgi complexes, perinuclear cisternae, and some azurophil granules (immature forms or disrupted mature forms) of a few cells. The specifics and the intact azurophils were not reactive. Reaction product was also found within Golgi cisternae, secondary lysosomes, and some of the azurophil granules of mononuclear cells. Observations on the A and B fractions confirmed those in situ regarding the localization of reaction product in disrupted PMN azurophils, its absence from specifics, and the latency of the enzyme activity in intact azurophils. In the C fraction, AcPase was found in three structures (a) Golgi cisternae, (b) dense bodies, and (c) small pleomorphic granules Comparison with the starting cells indicates that the Golgi complexes are probably derived from both PMN leukocytes and mononuclear cells, whereas the remaining elements resemble (in size, shape, and density) secondary lysosomes and azurophil granules of mononuclear cells. The results indicate that the bulk of the cytochemically detectable AcPase present in the C fraction is derived from mononuclear cells, rather than from PMN leukocytes     

大豆子叶内酸性磷酸酶活性的超微结构定位

开花后３５～５０ ｄ 期间和萌发早期（播种后４～８ ｄ）的大豆（Ｇｌｙｃｉｎｅｍ ａｘ Ｌ．）种子中,酸性磷酸酶主要分布在子叶细胞中的蛋白体内;在内质网内也检测到酸性磷酸酶活性。此外,在萌发早期的部分子叶细胞的质膜外侧及其细胞壁基质中可见密集的酸性磷酸酶活性;而且在近质膜的胞质中常见到一些富含磷酸铅沉淀的胞质小泡,似与质膜融合     

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.     

小麦受精过程中酸性磷酸酶的超微细胞化学定位

小麦（Ｔｒｉｔｉｃｕｍ ａｅｓｔｉｖｕｍ ）受精前成熟胚囊,除胚囊中央细胞的合点端细胞质中有酸性磷酸酶外,其余部位均未发现酸性磷酸酶。受精时期,以下部位存在酸性磷酸酶活性：卵细胞的细胞核内一部分染色质和细胞质中大部分线粒体;精、卵核融合时两核的核周腔内;退化助细胞合点端细胞质和一些液泡内;进入雌性细胞中的两个精核;胚囊各成员细胞的细胞壁及胚囊周围珠心细胞的细胞壁。二细胞原胚中未见有酸性磷酸酶。早期胚乳游离核染色质上有酸性磷酸酶。小麦受精过程酸性磷酸酶的分布特点可能与卵细胞生理状态的变化和细胞质中线粒体的改组、助细胞的退化、精核的生理状态以及精核与卵核的核膜融合等有关。     

LOCALIZATION OF DEHYDROGENASE AND ACID PHOSPHATASE IN THE ABSCISSION ZONE OF BEAN LEAVES

Leaf abscission in Phaseolus vulgaris L. cv. ‘Contender’ is associated with enzymatic changes during and prior to separation. Deblading resulted in a localized increase in dehydrogenase and acid phosphatase in the abscission zone. Increased enzyme activities were observed 24–48 hr after deblading. In debladed plants separation was complete in 6–8 days. At separation, dehydrogenase activity appeared to decrease and localization was specific to the protective layer, while the petiole side had no activity. In contrast, acid phosphatase activity was observed in some layers of cells on the petiole side after separation. Ethylene treatment promoted abscission and separation occurred in 24–48 hr in both debladed and intact plants. No protective layer was formed during ethylene-induced abscission. Enzymatic changes similar to those observed in debladed control plants were observed with ethylene treatment. Ethylene induced an additional abscission layer between the pulvinus and petiole, where an abscission layer normally does not form. In this ethylene-induced abscission layer, similar enzyme activities were detected.     

薏苡种子萌发过程中盾片上皮细胞超微结构的变化及酸性磷酸酶的动态

利用酶电镜技术,观察了薏苡种子萌发过程中其盾片上皮细胞超微结构的变化及酸性磷酸酶的动态分布。发现蛋白质体的降解发生在萌发旱期,接着发生了超微结构的明显变化包括:(1)脂肪体的大量降解;(2)各种细胞器如内质网、核糖体、线粒体、质体等的形成、发育及增殖。萌发早期,酸性磷酸酶主要是在种子形成时业已合成的,被贮存在蛋白质体中、细胞核内及内壁上。后期,酸性磷酸酶主要是由内质网新合成的。所有这些存在于细胞内的酸性磷酸酶可能是通过质膜或胞间连丝进入胞间进而释放入胚乳细胞内。     

甜菊愈伤组织中的质膜内陷:超微结构和酸性磷酸酶细胞化学定位

对在分化条件下的甜菊 (Stevia rebaudiana)愈伤组织分生区域细胞的质膜内陷进行了超微结构和酸性磷酸酶细胞化学研究。结果表明 ,在不同液泡化状态的细胞中均有质膜内陷存在。在原生质浓密的细胞中 ,质膜呈起伏的波纹状 ,某些部位发生明显内陷 ,大小不等 ,多呈圆球状。在部分液泡化细胞中 ,质膜内陷体积增大 ,内含物增多且结构复杂。在液泡化细胞中 ,质膜内陷嵌入中央液泡 ,但彼此间以一膜间隙隔开。质膜内陷中的内含物以小泡和卷绕的膜结构形式存在。酸性磷酸酶活性定位结果显示 ,质膜及其内陷含高的酶活性。推测质膜内陷在功能上与液泡相似 ,构成了这些细胞水解空间的一部分。     

LOCALIZATION OF ACID PHOSPHATASE IN LIPOFUSCIN GRANULES AND POSSIBLE AUTOPHAGIC VACUOLES IN INTERSTITIAL CELLS OF THE GUINEA PIG TESTIS

The intracellular localization of acid phosphatase in guinea pig testicular interstitial cells was investigated by incubating nonfrozen thick sections of glutaraldehyde-perfused testis in a modified Gomori medium and preparing the tissue for electron microscopy. Lipofuscin pigment granules in these cells contain dense pigment, granular matrix, and often a lipid droplet. Reaction product is seen in the matrix of the pigment granules, and they may therefore be called residual bodies. At least some of the dense pigment appears to be derived from myelin figures and membrane whorls, since suitable intermediates can be seen. Lipid droplets found free in the cytoplasm are another possible source of pigment. In both cases the chemical mechanism is presumed to be autoxidation of unsaturated lipid. Acid phosphatase is present in the inner cisterna of Golgi elements. Enzyme activity also appears in possible autophagic vacuoles bounded by double membranes; the reaction product lies between the membranes. Consideration of the enzyme as a tracer suggests that the autophagic vacuoles are derived from the Golgi complex. Possible stages in the formation of these vacuoles by the inner Golgi cisternae are observed.     

ELECTRON MICROSCOPIC LOCALIZATION OF ACID PHOSPHATASE AND THIAMINE PYROPHOSPHATASE ACTIVITY IN HYPOTHALAMIC NEUROSECRETORY CELLS OF THE RAT

Supraoptic nuclei in the hypothalamus of rats were fixed for the electron microscope by vascular perfusion with solutions of glutaraldehyde followed by post fixation with osmium tetroxide. Cytochemical methods for detection of acid phosphatase and thiamine pyrophosphatase activity have been applied to glutaraldehyde-fixed frozen sections containing the neurosecretory cells. The enzyme activities have been localized to certain Golgi cisternae. Acid phosphatase activity is present in the large (0.4 µ to 1.0 µ) granules or dense bodies which are surrounded by a single limiting membrane; both features characterize these structures as lysosomes. Smaller (0.1 µ) granules also present in the perikarya are generally unreactive towards enzyme activity and resemble in form the neurosecretory granules in the neurohypophysis.     

萌发前后燕麦种子内酸性磷酸酶的细胞化学研究

组织化学定位显示,在燕麦盾片和糊粉层细胞内,酸性磷酸酶是普遍地存在着的。种子萌发0,1,2天时,酶活性基本局限在细胞质内;细胞核和壁内则无酶活性。但当种子萌发至3—4天时,酶活性在胞质内增加,显示细胞内开始有新的酶产生。这些新产生出来的酶,还被分泌出细胞,进入胞壁。用组织等电点聚焦,基本上反映了与盾片和糊粉层的组织定位一致的酶变化情形。但从这两种组织所得到的同工酶带谱则并不一样。因为从糊粉层可以得到10条清晰的同工酶带(其中6条在糊粉层不清晰)。其次,胚乳的同工酶带谱,与盾片和糊粉层的同工酶带谱也是不完全一样的。     

RADIOAUTOGRAPHIC LOCALIZATION OF SODIUM PUMP SITES IN RABBIT INTESTINE

Direct demonstration of the cellular location of sodium pumping constitutes a key problem in the solution of intestinal sodium absorption. Utilizing silicone-impregnated epoxy sections of freeze-dried, osmium-fixed tissue, ouabain-³H and inulin-³H light microscope radioautographs have been produced which show that: lateral but not brush border membranes of rabbit small intestine bind ouabain-³H (high specific activity) with an affinity so great that a subsequent washing in ouabain-free medium has little effect on binding; lateral membrane binding is not apparent with low specific activity ouabain-³H, and inulin-³H and ouabain-³H (low specific activity) in the cores of the villi do not equilibrate with the intercellular spaces. Preliminary tracer measurements of ouabain-³H and inulin-¹⁴C spaces also agree with these findings As ouabain is a specific inhibitor of active sodium transport, these observations provide direct support for the view that lateral membrane pumping of sodium into the intercellular spaces causes, through osmotic forces on water, a flow of fluid out of these spaces into the interstitium.     

ULTRACYTOCHEMICAL LOCALIZATION OF PLASMA MEMBRANE-ASSOCIATED PHOSPHATASE ACTIVITY IN DEVELOPING TOBACCO SEEDS

Plasma membrane-associated phosphatase activity was found in integumentary cells of developing tobacco ovules from the megaspore tetrad stage to seed maturity. Enzyme activity is greatest in the innermost layers of the integument from the mature megagametophyte stage on. The egg, zygote, and synergids almost totally lack plasma membrane-associated reaction product, while the antipodals show some activity at their chalazal ends. The endosperm has much plasma membrane-associated phosphatase activity in most of its cells during development, but it is primarily the outermost plasma membranes of the surface cells of the embryo that have associated reaction product. It is concluded that the plasma membrane-associated phosphatase activity is related to active transport of assimilates and that the integument is the most important site of active transport in the young ovule. After fertilization, in addition to the innermost layers of the integument, the endosperm and the outermost cells of the embryo become involved in active transport, which continues to seed maturity.     

ULTRASTRUCTURE AND ACID PHOSPHATASE IN PEDICEL ABSCISSION OF HIBISCUS

Pedicel abscission in Hibiscus rosa-sinensis was investigated by light and electron microscopy. During the pre-abscission period endoplasmic reticulum declined somewhat, dictyosomes increased in number and apparent activity, and mitochondria maintained their numbers. The observations suggested that dictyosomal vesicles were migrating to and fusing with the plasma membrane. The enzyme acid phosphatase was associated with dictyosomes and dictyosomal saccules, with small vacuoles and invaginations of the plasma membrane, and in the paramural region between the plasma membrane and the cell wall. Our interpretation is that acid phosphatase, (and probably also the enzymes involved in cell wall dissolution) are transported via an endoplasmic reticulum-dictyosome-vesicle carrier system to the paramural regions of the cell. In more general terms, our observations support the view that the enzymes involved in the cell wall hydrolysis of abscission are synthesized within a compartmentalized, lysosomal system prior to their release and action.     

CYTOCHEMICAL LOCALIZATION OF ACID PHOSPHATASES IN EUGLENA GRACILIS

The localization of induced and constitutive acid phosphatase activity in Euglena was studied by light and electron microscopy, using two different cytochemical methods. Cells grown in high phosphate medium have constitutive acid phosphatase activity in three regions: in the Golgi complex, around the paramylum bodies, and in the peri-reservoir vesicles. Cells that have formed an induced acid phosphatase by exposure to a phosphate-deficient medium have, in addition to the constitutive activity localized exactly as in the uninduced cell, a strong activity in the pellicle. The induced activity is not uniformly distributed over the pellicle, but is localized at the notch of each pellicle complex, near a group of about four fibrils and near a characteristic vesicle of the endoplasmic reticulum. In the cytostome, where fission begins during division, there is an alternation of large and small pellicle complexes, both of which have induced phosphatase activity. A similar alternation is seen over the entire pellicle of dividing cells.     

兔前囟的变异及脑定位的校正

在113只1.8—3.0kg的浙江本地白兔,观察到前囱不够规则的占一半左右;而且即使前囱基本规则的兔,由于头型矢径的个体差异亦可明显影响脑立体定位时APO平面的准确性。本文提出测量两侧眶后上切迹连线与矢状缝交点至人字缝尖的距离(全距),以估计头型矢径;然后按全距与缝距(前囟中心至人字缝尖距离)的回归关系,可确定理论上标准的前囟中心位置约在全距中点向前1.6mm处。经此法校正或确定前囟中心组APO平面的定位准确率(82.9％)比未校正组(46.7％)显著提高(P<0.001)。     

不同代龄及生长状态下2BS细胞的酸性磷酸酶活性及氯酯醒对此的影响

实验以人胚肺二倍体成纤维细胞(2BS)为材料,分别测定了处于不同代龄及不同生长时期的2BS细胞酸性磷酸酶(ACPase)活性。结果发现随着代龄增高,细胞ACPase活性上升。处于同一代龄的细胞,则接触抑制期细胞的ACPase活性显著高于生长期细胞。接触抑制引起的酶活性增高甚至超过代龄增加而引起的ACPase活性上升。30μg/ml的氯酯醒有抑制细胞ACPase活性的作用。     

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.