西瓜柱头乳突细胞分泌活动期间ATP酶活性超微结构定位

研究了西瓜柱头乳突细胞ＡＴＰ酶活性的超微结构定位。分泌活动旺盛的细胞中,质膜、内质网、质体的内部片层、胞间连丝以及多数大液泡的膜上面都有大量ＡＴＰ酶活性反应产物,线粒体和小泡上只有少量酶活性反应产物。分泌活动停止后处于解体状态的细胞内,反应产物主要定位于液泡膜上。分泌旺盛的乳突细胞质膜具有高的ＡＴＰ酶活性表明分泌物运出需要大量能量,内质网ＡＴＰ酶活性强可能意味着该细胞参与分泌物合成。     

甘蔗叶不同部位ATP酶活性细胞化学定位

甘蔗叶片,叶鞘和肥厚带韧皮部 ATP 酶活性定位于筛管、伴胞的质膜、内质网和某些伴胞细胞基质、小囊泡和发育成熟的液泡上;叶片韧皮部薄壁细胞、厚壁细胞和厚壁通道细胞质膜及小囊泡中亦显示有 ATP 水解产物;维管束鞘细咆与厚壁细胞或厚壁通道细胞所构成的细胞间隙上也存在有 ATP 酶活性反应产物沉淀。甘蔗叶片大、中、小三种维管束,从小维管束到大维管束,面向细胞间隙的细胞表面上的 ATP 酶活性逐渐增强,而维管束鞘细胞质膜上的 ATP 酶活性则趋于减弱;同一维管束内则以韧皮部细胞的 ATP 酶活性最强。维管束鞘细胞与叶肉细胞之间存在很多的胞间连丝,并表现出高的 ATP 酶活性。讨论了 ATP 酶活性的分布状态与叶肉细胞的光合产物向韧皮部运输的关系。     

杂交鹅掌楸体胚发生过程中ATP酶活性的超微细胞化学定位

利用透射式电镜,通过胚性细胞的超微切片观察,对杂交鹅掌楸体细胞胚胎发生和发育过程中ATP酶活性进行了超微细胞化学定位.结果表明,非胚性细胞的质膜、液泡膜等膜系统当中存在ATP酶活性,质体、核膜、细胞壁以及细胞间隙上有少许沉积;早期胚性细胞ATP酶反应产物主要沉积于质膜、液泡膜上、淀粉粒、细胞壁加厚处;胚性细胞后期ATP酶活性从质膜逐渐转移入细胞内,细胞质、壁旁体、胞间连丝、细胞膜与细胞间隙、细胞核等处均有ATP酶活性反应.随着胚性细胞的发育及分裂,包裹细胞的厚壁、细胞核、核仁与染色质等处也出现ATP酶活性反应沉淀物.说明杂交鹅掌楸体细胞胚胎发生及发育过程中存在丰富的能量代谢.     

云杉针叶三磷酸腺苷酶活性的超微细胞化学定位

用磷酸铅沉淀技术研究了云杉幼龄针叶及成龄树针叶细胞的三磷酸腺苷酶(ATPase)活性定位。从形态结构上可以看到,种子萌发的幼龄针叶细胞的细胞壁较薄, ATP酶活性反应产物磷酸铅颗粒主要分布于细胞质和细胞质膜上。成龄针叶细胞壁较厚,内质网等细胞器发达,ATP酶主要在细胞壁有较高的活性反应。两者细胞的液泡及细胞核中也有少量定位。幼龄针叶ATP酶活性较成龄针叶的活性较弱, 这种活性变化与不同发育时期叶的生理功能有关。     

西瓜花粉壁超微结构与花粉ATP酶细胞化学定位

研究了西瓜花粉壁超微结构以及单核花粉液泡化时期ATP酶活性超微细胞化学定位。花粉壁的外壁分为外层和内层,外层包括覆盖层、基粒棒和基足层等三层,内层只包含一层。外层电子密度相对较小,内层电子密度相对较大;外层与内层之间有缝隙。ATP酶活性反应产物主要分布在细胞质基质、质体、内质网和花粉内壁中。     

西瓜花粉壁超微结构与花粉ATP酶细胞化学定位

研究了西瓜花粉壁超微结构以及单核花粉液泡化时期ATP酶活性超微细胞化学定位。花粉壁的外壁分为外层和内层 ,外层包括覆盖层、基粒棒和基足层等三层 ,内层只包含一层。外层电子密度相对较小 ,内层电子密度相对较大 ;外层与内层之间有缝隙。ATP酶活性反应产物主要分布在细胞质基质、质体、内质网和花粉内壁中     

枸杞胚性细胞分化的超微结构和ATP酶的细胞化学定位研究

枸杞的胚性细胞多由愈伤组织表层的薄壁细胞分化而来,与愈伤组织中未分化的细胞相比,胚性细胞呈卵圆形,细胞核大,核仁明显,细胞质浓厚并含有丰富的细胞器,细胞壁较薄,细胞间有胞间连丝相通;胚性细胞发育到晚期细胞壁加厚,胞间连丝逐渐消失,细胞核向一端偏移,有大液泡形成;胚性细胞的第一次分裂多为均等分裂,形成二细胞原胚,继续分裂形成多细胞原胚;组成多细胞原胚胚体的细胞核大,核形状不规则,细胞质浓厚,细胞器丰富,在质体中出现淀粉的积累。在胚性细胞发育的早期,ATP酶活性主要位于质膜上,随后在液泡内和细胞核中都出现ATP酶活性的分布;随着胚性细胞壁的加厚,细胞壁加厚处和细胞间隙中也出现ATP酶活性反应;当多细胞原胚形成后,ATP酶活性反应主要定位于液泡膜上。由此分析了结构特征、ATP酶活性定位变化与胚性细胞分化的关系。     

黄瓜雌花发育过程中柱头的腺特征

利用透射电镜技术研究了黄瓜(Cucumis sativus L.)雌花柱头发育过程中传递组织、分泌组织和乳突细胞的超微结构.在整个发育过程中,乳突细胞和分泌组织细胞的细胞质内密布很多管状及槽库膨大的内质网,产生很多分泌囊泡;在成熟柱头的传递组织和分泌组织细胞间观察到大量的胞间连丝;乳突细胞和分泌细胞高度液泡化,质膜内折;在柱头发育过程中分泌组织细胞的核周腔扩大形成裂瓣状核,到柱头成熟阶段裂瓣状核更加明显.进一步的研究显示,在成熟柱头的不同组织细胞中,ATPase的活性呈现在质膜和液泡膜上,随着柱头的发育,PM-H -ATPase的比活性明显增强.结果表明,黄瓜雌花柱头的腺特征随发育进程而趋于显著.     

黄瓜雌花发育过程中柱头的腺特征(英文)

利用透射电镜技术研究了黄瓜（Ｃｕｃｕｍｉｓ　ｓａｔｉｖｕｓ　Ｌ．）雌花柱头发育过程中传递组织、分泌组织和乳突细胞的超微结构。在整个发育过程中,乳突细胞和分泌组织细胞的细胞质内密布很多管状及槽库膨大的内质网,产生很多分泌囊泡;在成熟柱头的传递组织和分泌组织细胞间观察到大量的胞间连丝;乳突细胞和分泌细胞高度液泡化,质膜内折;在柱头发育过程中分泌组织细胞的核周腔扩大形成裂瓣状核,到柱头成熟阶段裂瓣状核更加明显。进一步的研究显示,在成熟柱头的不同组织细胞中,　ＡＴＰａｓｅ的活性呈现在质膜和液泡膜上,随着柱头的发育,ＰＭ－Ｈ＋－ＡＴＰａｓｅ的比活性明显增强。结果表明,黄瓜雌花柱头的腺特征随发育进程而趋于显著。     

百合花粉母细胞间细胞融合期间腺苷三磷酸酶活性的细胞化学定位及其与染色质胞间转移的关系

用标准的磷酸铅沉淀的细胞化学方法,对百合花粉母细胞间染色质穿壁运动期间及其前后三个时期中的腺苷三磷酸酶(ATP 酶)活性进行了超微结构的定位。结果表明:(1)在穿壁前,ATP 酶活性主要定位于质膜、胞间连丝及细胞间隙;在内质网、高尔基体、质体和某些局部的基质(groundplasm)中,也表现有 ATP 酶活性反应的产物;但在染色质和核仁中,一般都没有这种反应。(2)在穿壁时,染色质从一个细胞穿壁转移到另一个相邻细胞,同时看到染色质和核仁内出现密集的 ATP 酶活性反应产物;在内质网和高尔基体的腔内以及质体的片层上也产生明显的 ATP 酶活性反应;而在质膜、胞间连丝及细胞间隙内 ATP 酶活性明显降低,甚至看不到明显的活性反应。(3)在穿壁后,质膜及细胞间隙中又产生明显的 ATP 酶活性反应产物,但核内染色质上的 ATP 酶活性则显著降低,而核仁内则仍有较高的活性。同前二个时期一样,内质网、高尔基体和质体上的 ATP 酶仍表现明显的活性反应。最后讨论了三个不同发育时期 ATP 酶活性及其分布部位的改变与染色质胞间转移的关系。     

ATPase in mature and differentiating phloem and xylem

A cytochemical study using a lead precipitation technique has been made of the distribution of adenosine triphosphatase (ATPase) in mature and differentiating phloem and xylem cells of Nicotiana tabacum and Pisum sativum. The sites of ATPase localization in tobacco phloem were the plasma membrane, endoplasmic reticulum, mitochondria, dictyosomes, plasmodesmata, and the dispersed P proteins of mature sieve elements. In pea phloem sieve elements ATPase was localized in the endoplasmic reticulum, but was not associated with the P proteins or plasma membranes at any stage of their differentiation. In pea transfer cells ATPase activity was associated with the endoplasmic reticulum at all stages of their differentiation and with the plasma membrane of transfer cells that had formed wall ingrowths. In xylem cells of both tobacco and pea the patterns of ATPase activity was similar. At early stages of differentiation ATPase activity was associated with the plasma membrane and the endoplasmic reticulum. At intermediate stages of differentiation ATPase activity continued to be associated with the endoplasmic reticulum, but was no longer associated with the plasma membrane. At later stages of xylem element differentiation ATPase activity was associated with disintegrating organelles and with the hydrolyzing cell walls.     

Ultrastructural localization of glucose-6-phosphatase activity in proximal convoluted tubule cells of rat kidney

Summary The ultrastructural localization of glucose 6-phosphatase activity was investigated in the proximal convoluted tubule cells of the rat kidney. The reaction product for the enzyme activity was present in the endoplasmic reticulum and nuclear envelope, as reported for the hepatic enzyme and others, but was absent from the brush border, plasma membrane and other organelles. The metabolic significance of the association of this enzyme with the endoplasmic reticulum and the role of the enzyme in the active reabsorption and transport of glucose in the renal tubules are discussed.     

Alkaline phosphatase biosynthesis in the endoplasmic reticulum and its transport through the Golgi apparatus to the plasma membrane: cytochemical evidence

Enzyme induction of HeLa cell placental alkaline phosphatase with various agents such as prednisolone, sodium butyrate, hyperosmolality (NaCl), or combination of these inducers resulted in the appearance of enzyme activity in the rough endoplasmic reticulum, nuclear envelope, Golgi apparatus, and plasma membrane. In the Golgi apparatus, intense reaction product deposits tended to be concentrated on its trans side, with small vesicles and granules also being positively stained. Inhibition of protein synthesis with cycloheximide was followed by the disappearance of enzyme activity from these cytoplasmic organelles but not from the plasma membrane. Treatment with monensin, a secretory protein transport inhibitor, uniformly increased activity in the rough endoplasmic reticulum while causing marked dilatation of the intensely positive Golgi cisternae. These results suggest that intracellular alkaline phosphatase is newly synthesized in the endoplasmic reticulum and then passes en route through the Golgi apparatus to the plasma membrane. Accordingly, the present system could represent the biosynthesis, transport, and incorporation of the model cell surface enzyme protein to add to the vesicular stomatitus virus glyco-1 (VSV-G) protein and acetylcholine receptor model systems for studying the dynamics of cell surface protein genesis, transport, and membrane integration.     

Cytochemical Localization of Adenosine Triphosphataes Activity During Cytomixis in Pollen Mother Cells of David Lily and Its Relation to the Intercellular Migrating Chromatin Substance

Standard lead precipitation procedures have been used to examine the localization of ATPase activity during cytomixis in pollen mother cells of Lilium davidii var. willmottiae (Wilson) Roffill. Before cytomixis, cells at this stage of development show ATPase activity on plasma membrane, in the endoplasmic reticulum, dictyosomes, plastids, plasmodesmata, and in part of the groundplasm; however, there is no ATPase activity on the chromatin and nucleolus. During cytomixis, the chromatin substance begin to transfer from one cell to an adjacent cell, reaction product indicating ATPase activity is observed associated with the chromatin and nucleolus. ATPase activity is also found with the cistenae of both endoplasmic reticulum and dictyosomes, and some plastids. There is no deposition of ATPase reaction product associated with the plasm membrane and intercellular spaces. After cytomixis, the chromatin is little or no deposition of enzyme reaction product. ATPase activity, however, is consistenlly found within the intercellular space and on the plasm membrane, and also occur in the endoplasmic reticulum, dictyosome and plastid. The presence or absence of ATPase activity in the cell structure of pollen mother cells before, during or after eytomixis is discussed in relation to the active uptake or export of water for short-distance transport. It is also suggested that the intensive ATPase activity in the nucleus during cytomixis of pollen mother cells is evidence for a transport system involved in the active movement of the intercellular migrating ebromatin substance.     

Studies on vinblastine-induced autophagocytosis in mouse liver. IV. Origin of membranes

The origin of the limiting membranes of autophagic vacuoles (AV) in mouse hepatocytes was studied by cytochemical techniques. Autophagocytosis was induced by an intraperitoneal injection of vinblastine (50 mg/kg). The marker enzymes used were adenosine triphosphatase for the plasma membrane, glucose-6-phosphatase for the endoplasmic reticulum and thiamine pyrophosphatase for the Golgi apparatus and the endoplasmic reticulum. All the three enzymes showed a characteristic localization in both control and vinblastine-treated hepatocytes. The space between the limiting membranes of a few apparently newly formed AV's showed weak glucose-6-phosphatase activity. Neither adenosine triphosphatase nor thiamine pyrophosphatase activities were observed on or between the AV membranes. It was suggested that endoplasmic reticulum membranes may be used as a source of AV membranes in hepatocytes. The lack of glucose-6-phosphatase activity in the limiting membranes even of most of the newly formed AV's suggests a transformation process of the membranes destined to form AV, during which the enzyme activity characteristic for endoplasmic reticulum may disappear from them.     

Studies on vinblastine-induced autophagocytosis in mouse liver

Summary The origin of the limiting membranes of autophagic vacuoles (AV) in mouse hepatocytes was studied by cytochemical techniques. Autophagocytosis was induced by an intraperitoneal injection of vinblastine (50 mg/kg). The marker enzymes used were adenosine triphosphatase for the plasma membrane, glucose-6-phosphatase for the endoplasmic reticulum and thiamine pyrophosphatase for the Golgi apparatus and the endoplasmic reticulum. All the three enzymes showed a characteristic localization in both control and vinblastine-treated hepatocytes. The space between the limiting membranes of a few apparently newly formed AV's showed weak glucose-6-phosphatase activity. Neither adenosine triphosphatase nor thiamine pyrophosphatase activities were observed on or between the AV membranes. It was suggested that endoplasmic reticulum membranes may be used as a source of AV membranes in hepatocytes. The lack of glucose-6-phosphatase activity in the limiting membranes even of most of the newly formed AV's suggests a transformation process of the membranes destined to form AV, during which the enzyme activity characteristic for endoplasmic reticulum may disappear from them.     

ULTRASTRUCTURAL LOCALIZATION OF ADENOSINE TRIPHOSPHATASE IN THE OVULES OF SAINTPAULIA IONANTHA (GESNERIACEAE) AND ITS RELATION TO SYNERGID FUNCTION AND EMBRYO SAC NUTRITION

Ovules of African violet were analyzed for adenosine triphosphatase activity. Ovules from unpollinated flowers of three different ages were fixed in buffered, 3% paraformaldehyde, incubated in the Wachstein-Meisel medium, and processed for electron microscopy. Results showed a heavy reaction product in the endothelium and inner micropylar cells of the integument with decreasing amounts elsewhere. Reaction product was localized primarily on the plasma membrane, and occasionally in the nuclear membrane, endoplasmic reticulum, small vacuoles, and mitochondria. The synergids, egg, central cell, and antipodals were essentailly devoid of reaction product except for rare occurrences in the smaller vacuoles and mitochondria of the synergids, and fragmentary deposits on the plasma membrane of the antipodals. No differences were found in any of the floral stages examined. These results suggest that the integumentary cells nearest the embryo sac are equipped with the necessary enzymes for active translocation of solutes into the embryo sac and that the cells of the megagametophyte apparently function more passively in this regard.     

Further characterization of HeLa S3 plasma membrane ghosts

A plasma membrane fraction of HeLa S3 cells, consisting of ghosts, is characterized more fully. A simple procedure is described which permits light and electron microscope study of the plasma membrane fraction through the entire depth of the final product pellet and through large areas parallel to the surface. Contamination by nuclei is 0.14%, too little for DNA detection by the diphenylamine reaction. Contamination by rough endoplasmic reticulum and ribosomes is small, a single ghost containing about 3% of the RNA in a single cell. Mitochondria were not encountered. Electron microscopy also shows (a) small vesicles associated with the outer surface of the ghosts, and (b) a filamentous web at the inner face of the ghost membrane. Sodium dodecyl sulfate (SDS)-polyacrylamide gel analysis shows that of the many Coomassie Blue-stained bands two were prominent. One, 43,000 daltons, co-migrated with purified rabbit muscle actin and constituted about 7.5% of the plasma membrane protein. The other major band, 34,000 daltons, was concentrated in the plasma membrane fraction. Two major glycoproteins detected by autoradiography of [14C]fucose-labeled glycoproteins on the gels, had apparent molecular weights of 35,000 daltons and 32,000 daltons. These major bands did not stain with Coomassie Blue. There were many other minor glycoprotein bands in the 200,000- to 80,000-dalton range. Ouabain-sensitive, Na+, K+-adenosine triphosphatase (ATPase) activity of the ghost fraction is purified 9.1 (+/- 2.2) times over the homogenate; recover of the activity is 12.0 (+/- 3.8%) of the homogenate. Enrichment and recovery of fucosylglycoprotein parallel those for ouabain-sensitive Na+, K+-ATPase activity. Fucosyl glycoprotein is recovered more than the enzyme activity in a smooth membrane vesicle fraction probably containing the bulk of plasma membrane not recovered as ghosts.