A mitochondrion as the structural basis of the formation of a cell-type-specific organelle inDictyostelium development

Summary The mitochondrion has been mainly given attention as a self-reproductive and respiratory organelle. We report here that the mitochondrion may participate in the formation of a cell-type-specific organelle, coupling with the Golgi complex. During the development ofDictyostelium discoideum, the two types of cells, i.e., the anterior prestalk cells and the posterior prespore cells form a polarized cell mass. Prespore differentiation is characterized by the presence of unique vacuoles named PSVs (prespore-specific vacuoles) in the cytoplasm. Thus the PSV is the most essential organelle to understand the structural basis of cell differention in this organism. In differentiating prespore cells, the mitochondrion exerts a remarkable transformation to form a sort of vacuole (M-vacuole). Using a PSV specific antibody, it was immunocytochemically shown that a PSV antigen (C-10) is localized in the M-vacuole as well as in the lining membrane of PSV. Interestingly, the C-10 antigen was also noticed in the Golgi cisternae that had fused with M-vacuole. Based on these findings, we propose here a promising model which suggests how both mitochondria and Golgi cisternae may be coordinately involved in the PSV formation. This model will provide a new aspect of mitochondrial functions in cell differentiation.     

Acid phosphatase activity during spore differentiation of the red algaeGigartina teedii andChondria tenuissima

The acid phosphatase activity during carposporogenesis inGigartina and tetrasporogenesis inChondria was studied using the Gomori technique. During the first steps of gonimoblast maturation ofGigartina, portions of cytoplasm are ensheathed by ER cisternae with acid phosphatase activity, giving rise to autolysosomal concentric membrane bodies. In a similar way large mucilage sacs are severed. They extrude their contents in a kind of exocytosis. Multivesicular bodies, concentrically arranged cisternae and extracytoplasmic compartments, each with acid phosphatase activity, remain in young carpospores for some time, probably as remnants of the autophagocytotic and exocytotic events. The Golgi apparatus is poorly developed in gonimoblast cells and young carpospores. It becomes a prominent cell component in maturing carpospores and then participates in cell wall formation. Only some of the dictyosomal cisternae contain acid phosphatase; these are irregularly distributed in the dictyosome. — In pre- and postmeiotic tetraspore mother cells ofChondria massive lead deposits are found in the dictyosomes and in adjacent Golgi vesicles. Finer lead precipitates occur in ER cisternae, especially in those which are sequestering starch-grain-containing portions of the cytoplasm to give rise to autolysosomes. During cell cleavage, the dictyosomes aggregate. They become devoid of acid phosphatase activity with the exception of vesicles at the trans face. Later, Golgi stacks associate and have common, Gomori positively reacting, narrow cisternae at the cis face. The Golgi apparatus derived cored vesicles do not contain lead precipitates whereas the Golgi cisternae in the final stage of tetrasporogenesis show acid phosphatase activity. Variations in acid phosphatase distribution are explained in the light of current models of membrane flow.Dedicated to Univ.-Prof. DrO. Härtel on the occasion of his 80th birthday.     

Cytochemical localization of terminal N-acetyl-D-galactosamine residues in cellular compartments of intestinal goblet cells: implications for the topology of O-glycosylation

The O-linked oligosaccharides of mucin-type glycoproteins contain N- acetyl-D-galactosamine (GalNAc) that is not found in N-linked glycoproteins. Because Helix pomatia lectin interacts with terminal GalNAc, we used this lectin, bound to particles of colloidal gold, to localize such sugar residues in subcellular compartments of intestinal goblet cells. When thin sections of low temperature Lowicryl K4M embedded duodenum or colon were incubated with Helix pomatia lectin- gold complexes, no labeling could be detected over the cisternal space of the nuclear envelope and the rough endoplasmic reticulum. A uniform labeling was observed over the first and several subsequent cis Golgi cisternae and over the last (duodenal goblet cells) or the two last (colonic goblet cells) trans Golgi cisternae as well as forming and mature mucin droplets. However, essentially no labeling was detected over several cisternae in the central (medial) region of the Golgi apparatus. The results strongly suggest that core O-glycosylation takes place in cis Golgi cisternae but not in the rough endoplasmic reticulum. The heterogenous labeling for GalNAc residues in the Golgi apparatus is taken as evidence that termination of certain O- oligosaccharide chains by GalNAc occurs in trans Golgi cisternae.     

Transport of rosettes from the golgi apparatus to the plasma membrane in isolated mesophyll cells ofZinnia elegans during differentiation to tracheary elements in suspension culture

Summary Rosettes of six particles have been visualized by freeze-fracture in the protoplasmic fracture (PF) faces of: a) the plasma membrane, b) Golgi cisternae, and c) Golgi-derived vesicles in mesophyll cells ofZinnia elegans that had been induced to differentiate synchronously into tracheary elements in suspension culture. These rosettes have been observed previously in the PF face of the plasma membranes of a variety of cellulose-synthesizing cells and are thought to be important in cellulose synthesis. InZinnia tracheary elements, the rosettes are localized in the membrane over regions of secondary wall thickening and are absent between thickenings. The observation of rosettes in the Golgi cisternae and vesicles suggests that the Golgi apparatus is responsible for the selective transport and exocytosis of rosettes in higher plants, as has been previously indicated in the algaMicrasterias (Giddings et al. 1980). The data presented indicate that the Golgi apparatus has a critical role in the control of cell wall deposition because it is involved not only in the synthesis and export of matrix components but also in the export of an important component of the cellulose synthesizing apparatus. The rosettes are present in the plasma membrane and Golgi vesicles throughout the enlargement of the secondary thickening, suggesting that new rosettes must be continually inserted into the membrane to achieve complete cell wall thickening.Abbreviations EF Golgi vesicles, exoplasmic fracture; the plasma membrane, extracellular fracture - PF protoplasmic fracture     

Effects of induced pinocytotic activity and extreme temperatures on the morphology of golgi bodies in Amoeba proteus

Summary The morphology of the Golgi apparatus of Amoeba proteus can be influenced by substances inducing pinocytotic activity as well as by extreme temperatures. During the ingestion of a solution of 0.5% egg white the number of Golgi bodies decreases from 100% measured in control cells to 82% measured in cells showing induced pinocytosis. Simultaneously the ratio of the surface area of the cisternae at the proximal face to that of the vesicles at the distal face of single dictyosomes remains constant (1.74–1.72).The decrease and increase of the temperature of the culture medium to 4° C and 30° C respectively, causes the disappearance of most of the dictyosomes. After keeping the cells for 3–10 h at these temperatures the number of Golgi bodies was only 5–10% of the controls. A continued treatment with cold or warm culture medium leads to a partial reorganization of dictyosomes. After 15 h the number of Golgi bodies counted per cell returned to 57% at 4° C and 38% at 30° C. The ratio of the surface area of the Golgi cisternae to the surface area of the Golgi vesicles also alters under the influence of extreme temperatures. The values measured after treating the cells for 3 h, 4 h 10 h and 15 h at 4° C and 30° C amounted to 0.75, 0.85, 1.14 1.53 and 0.93, 0.38, 0.88, 1.60, respectively, compared to 1.72 of control amoebae.The different values of the ratio of the surface area of cisternae to that of vesicles indicate that there are strong morphological changes of single dictyosomes.     

The localization of thiamine pyrophosphatase activity in Meckel's cartilage cells during endochondral ossification

Summary The cytochemical distribution of thiamine pyrophosphatase (TPPase) activity in Meckel's cartilage cells of the mouse embryo has been studied during the endochondral ossification. All the cartilage cells contain reaction product within the Golgi apparatus. In immature chondrocytes, at the reserve cell zone, TPPase activity is restricted to several inner cisternae of independent Golgi apparatus. In mature cells at the proliferative cell zone, several Golgi complexes form a Golgi network connecting with each other by the TPPase positive tubular stalks. Golgi cisternae, condensing vacuoles and vesicles also contain reaction product. In the hypertrophic chondrocytes located in the calcifying zone, their disorganized Golgi apparatus still retain reaction product. Some chondrocytes, even those located within calcified or opened lacunae, exhibit intact structures and normal cytochemical enzyme distribution. These data indicate the possibility that some chondrocytes may survive and contribute the formation of mandible.     

The localization of thiamine pyrophosphatase activity in Meckel's cartilage cells during endochondral ossification

The cytochemical distribution of thiamine pyrophosphatase (TPPase) activity in Meckel's cartilage cells of the mouse embryo has been studied during the endochondral ossification. All the cartilage cells contain reaction product within the Golgi apparatus. In immature chondrocytes, at the reserve cell zone, TPPase activity is restricted to several inner cisternae of independent Golgi apparatus. In mature cells at the proliferative cell zone, several Golgi complexes form a Golgi network connecting with each other by the TPPase positive tubular stalks. Golgi cisternae, condensing vacuoles and vesicles also contain reaction product. In the hypertrophic chondrocytes located in the calcifying zone, their disorganized Golgi apparatus still retain reaction product. Some chondrocytes, even those located within calcified or opened lacunae, exhibit intact structures and normal cytochemical enzyme distribution. These data indicate the possibility that some chondrocytes may survive and contribute the formation of mandible.     

The Golgi apparatus is a primary site of intracellular damage after photosensitization with Rose Bengal acetate

The aim of the present investigation was to elucidate whether the Golgi apparatus undergoes photodamage following administration of the fluorogenic substrates Rose Bengal acetate (RBAc) and irradiation at the appropriate wavelength. Human HeLa cells were treated in culture and the changes in the organization of the Golgi apparatus were studied using fluorescence confocal microscopy and electron microscopy, after immunocytochemical labeling. To see whether the cytoskeletal components primarily involved in vesicle traffic (i.e., microtubules) might also be affected, experiments of tubulin immunolabeling were performed. After treatment with RBAc and irradiation, cells were allowed to grow in drug-free medium for different times. 24 hr after irradiation, the cisternae of the Golgi apparatus became packed, and after 48-72 hr they appeared more fragmented and scattered throughout the cytoplasm; these changes in the organization of the Golgi cisternae were confirmed at electron microscopy. Interestingly enough, apoptosis was found to occur especially 48-72 h after irradiation, and apoptotic cells exhibited a dramatic fragmentation of the Golgi membranes. The immunolabeling with anti-tubulin antibody showed that microtubules were also affected by irradiation in RBAc-treated cells.     

Structure,differentiation, and multiplication of Golgi apparatus in fungal hyphae

Summary Golgi apparatus in subapical regions of hyphae consist of paranuclear dictyosomes with 4–5 cisternae each. Transverse and tangential sections provide ultrastructural evidence for a three-dimensional architectural model of the Golgi apparatus and a stepwise mechanism for dictyosome multiplication. The dictyosomes are polarized, with progressive morphological and developmental differentiation of cisternae from the cis to the trans pole. Small membrane blebs and transition vesicles provide developmental continuity between the nuclear envelope and the adjacent dictyosome cisterna at the cis face. Cisternae are formed as fenestrated plates with extended tubular peripheries. The morphology of each cisterna depends on its position in the stack, consistent with a developmental gradient of progressive maturation and turnover of cisternae. Mature cisternae at the trans face are dissociated to produce spheroid and tubular vesicles. Evidence in support of a schematic sequence for increasing the numbers of dictyosomes comes from images of distinctive and unusual forms of Golgi apparatus in hyphal regions where nuclei and dictyosomes multiply, as follows: (a) The area of the nuclear envelope exhibiting forming-face activity next to a dictyosome expands, which in turn increases the size of cisternae subsequently assembled at the cis face of the dictyosome. (b) As subsequent large cisternae are formed and mature as they pass through the dictyosome, an entire dictyosome about twice normal size is built up. The number of cisternae per stack remains the same because of continuing turnover and loss of cisternae at the trans face, (c) This enlarged dictyosome becomes separated into two by a small region of the nuclear envelope next to the cis face that acquires polyribosomes and no longer generates transition vesicles, (d) As a consequence, assembly of new dictyosomes is physically separated into two adjacent regions, (e) As.the enlarged cisternae are lost to vesiculation at the trans pole, they are replaced by two separate stacks of cisternae with typical normal diameters, (f) The net result is two adjacent dictyosomes where one existed previously. Dictyosome multiplication is thus accomplished as part of the normal developmental turnover of cisternae, without interrupting the functioning of the Golgi apparatus as it continues to produce new secretory vesicles from mature cisternae at the trans face. Coordination of Golgi apparatus multiplication with nuclear division ensures that each daughter nucleus receives a complement of paranuclear dictyosomes.     

The Golgi apparatus ofPhytophthora cinnamomi makes three types of secretory or storage vesicles concurrently

Summary The formation of three types of vesicles in the oomycetePhytophthora cinnamomi was investigated using ultrastructural and immunocytochemical techniques. All three vesicles are synthesised at the same time; one type serves a storage role; the others undergo regulated secretion. A monoclonal antibody Lpv-1 that is specific for glycoproteins contained in the storage vesicles labelled the endoplasmic reticulum (ER), elements in the transition region between ER and Golgi stack, and cis, medial and trans Golgi cisternae. Cpa2, a monoclonal antibody specific for glycoproteins contained within secretory dorsal vesicles labelled the transition region, cis cisternae and a trans-Golgi network. Vesicles possessing a structure characteristic of mature secretory ventral vesicles were observed in close association with the trans face of Golgi stacks. The results suggest that all three vesicles are formed by the Golgi apparatus. Double immunogold labelling with Lpv-1 and Cpa-2 showed that these two sets of glycoproteins occurred within the same Golgi cisternae, indicating that both products pass through and are sorted concurrently within a single Golgi stack.     

Effects of brefeldin A on the Golgi apparatus, the nuclear envelope, and the endoplasmic reticulum in a green alga,Scenedesmus acutus

Summary The effects of brefeldin A (BFA) on the structure of the Golgi apparatus, the nuclear envelope, and the endoplasmic reticulum (ER), and on the thiamine pyrophosphatase (TPPase) activity in these organelles were examined in a green alga,Scenedesmus acutus, to obtain evidence for the existence of a retrograde transport from the Golgi apparatus to the ER via the nuclear envelope. InScenedesmus, Golgi bodies are situated close to the nuclear envelope throughout the cell cycle and receive the transition vesicles not directly from the ER, but from the nuclear envelope. BFA induced the disassembly of Golgi bodies and an increase in the ER cisternae at the trans-side of decomposed Golgi bodies in interphase cells and multinuclear cells before septum formation. The accumulated ER cisternae connected to the nuclear envelope at one part. TPPase activity was detected in all cisternae of Golgi bodies, but not in the nuclear envelope or the ER in nontreated cells. On the contrary, in BFA-treated cells, TPPase activity was detected in the nuclear envelope and the ER in addition to the decomposed Golgi bodies. When septum-forming cells were treated with BFA, the disassembly of Golgi bodies was less than that in interphase cells, and TPPase activity was detected in the Golgi cisternae but not in the nuclear envelope or the ER. These results suggest mat BFA blocks the anterograde transport from the nuclear envelope to the Golgi bodies but does not block the retrograde transport from the Golgi bodies to the nuclear envelope in interphase and multinuclear cells.Abbreviations BFA brefeldin A - ER endoplasmic reticulum - TPPase thiamine pyrophosphatase     

Distribution of golgi apparatus-associated polyribosomes across the polarity axis of dictyosomes of wild carrot (Daucus carota L.)

Summary Endoplasmic reticulum-polyribosome-Golgi apparatus associations were a general feature of cells of suspension cultures of wild carrot (Daucus carota L.). Free polyribosomes occurred within the Golgi apparatus zone for all dictyosomes and with equal frequency at all levels within the stack including the most mature or trans face. When evaluated and quantified from electron micrographs, approximately 60% of the dictyosome profiles were characterized by a system of transition elements consisting of part smooth-part rough endoplasmic reticulum. These were encountered most frequently in the immediate vicinity of the immature, forming or cis face, usually toward the periphery of the stacked cisternae. Analysis of serial sections showed that those dictyosome profiles not exhibiting this characteristic did so primarily because of an unfavorable plane of sectioning. All dictyosomes examined in 5 or more serial sections revealed some type of close association with endoplasmic reticulum. Some of the associations were so close that direct connections between Golgi apparatus and endoplasmic reticulum tubules could not be excluded. Also present, especially at the forming or cis face, were small 600 nm transition vesicles with nap-like surface coats on nearly 90% of the dictyosomes examined. More than 50% exhibited spiny (clathrin-)coated vesicles at the mature or trans face.     

Origin of the Golgi system in amoebae

Summary An electron microscopic study of the Golgi apparatus in the giant amoeba, Pelomyxa illinoisensis, has been presented. Studies of normally feeding, dividing, starving, and refeeding amoebae were made. The major finding is that plasmalemma vesicles, formed via pinocytosis and phagocytosis, either flatten or invaginate and form the cisternae of the Golgi apparatus. Plasmalemma vesicles are also a source of new cisternae during the lifetime of a given Golgi apparatus. The cisternae migrate through the Golgi system, but before being released they either inflate, or segment into smaller vesicles. It is postulated that they later empty into the contractile vacuole and into certain other vacuoles. No evidence was found for the fusion of smooth Golgi vesicles or fringed vesicles of any kind with the plasmalemma.Dedicated to Professor Friedrich Wassermann with admiration and affection on the occasion of his eightieth birthday.Work supported by U. S. Atomic Energy Commission. A part of the work was reported at the XVI International Congress of Zoology, Washington, D. C., in 1963.     

Ultrastructural Studies of a Vesicle System Associated with Endoplasmic Reticulum in Exo-Erythrocytic Forms of Plasmodium berghei1

ABSTRACT. Fine structural studies of a specialized vesicle system associated with the endoplasmic reticulum (ER) of exo-erythrocytic Plasmodium berghei suggest that this system may be the equivalent of a Golgi apparatus. Patches of ER, randomly distributed in the cytoplasm of developing parasites, are formed of smooth and ribosome-studded cisternae intermingled with each other. The vesicle systems are located between as well as at the edges of ER aggregates and appear to be in different stages of budding from the cisternae. Prolonged osmication reveals distinct staining of the nuclear envelope and ER of the parasites as well as part of the Golgi apparatus of the hepatocytes. However, the small vesicles associated with the parasite's ER are unstained, as are the coated vesicles in the Golgi region of the liver cell. These sites in the parasite cytoplasm seem comparable to the concave surface of the Golgi apparatus in liver cells. The pinched-off vesicles fuse with others to form the prominent peripheral vacuolization characteristic of the nearly mature exo-erythrocytic form. The formation of these peripheral vacuoles and their subsequent fusion with the parasite membrane may be an exocytosis mechanism supplying the rapidly expanding parasite with new plasma membrane material.     

Nachweis von saurer Phosphatase in gezüchteten Hühnerherzmyoblasten

Zusammenfassung Ein verbessertes Verfahren zur Durchführung der Gomori-Reaktion gewährleistet die lagegetreue Darstellung der sauren Phosphatase in gezüchteten Hühnerherzmyoblasten. Enzymaktiv sind die nach Verfütterung von eiweißhaltiger Nahrung regelmäßig auftretenden Proteinspeicher, hier Cytosomen genannt, sowie die Cisternen und Vesikel der Golgi-Apparate. Zu Beginn der Interphase weisen die Myoblasten in der Regel bedeutend weniger enzymaktive Cytosomen und Golgi-Apparate auf als in älteren Interphasestadien. Die Golgi-Vesikel trifft man nur in der Golgi-Zone an. Zwischen den Cytosomen und den Golgi-Apparaten besteht eine enge Lagebeziehung. Alle Cytosomen und auch die gelegentlich auftretenden Cytolysomen enthalten vesikuläre Elemente in der Größenordnung der Golgi-Vesikel. Aus diesen Befunden wird geschlossen, daß die Golgi-Cisternen saure Phosphatase produzieren oder aktivieren und durch Abschnürung von Vesikeln an die Cytosomen übermitteln, die nunmehr Lysosomen-Charakter annehmen.

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Summary An improved method of the Gomori-technique enables the exact localization of acid phosphatase activity within cultured chicken heart myoblasts. Enzym activity is found in the protein storing granules, the so-called cytosomen, which usually appear within the myoblasts, when albumin has been added to the culture medium, as well as in the cisternae and vesicles of the Golgi apparatus. In early stages of interphase, the cells generally contain much fewer enzym-active cytosomes and Golgi apparatus than in later stages.The Golgi vesicles are only found in the Golgi zone. There is a close, topographical relation between the cytosomes and the Golgi apparatus. All cytosomes as well as the cytolysomes which are occasionally found in the myoblasts, enclose vesicular elements of the size of the Golgi vesicles. From these observations it may be concluded that the Golgi cisternae produce or activate acid phosphatase which is transfered to the cytosomes by the vesicles originating from the Golgi cisternae. Thus the cytosomes assume the character of lysosomes.

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Die Arbeit wurde durch eine Sachbeihilfe der Deutschen Forschungsgemeinschaft ermöglicht. Herrn Prof. Dr. R. Danneel danke ich für beratende Hilfe, Frl. I. Rosocha, Frl. I. Spieckermann und Frl. S. Kliewer für technische Assistenz.     

Ultrastructure of meristematic epidermal cells of maize root under water deficit conditions

Summary Structural components of meristematicZea mays primary root epidermal cells were observed after osmotic stress of the nutrient medium (12.5 atm.) and after rehydration. After non-lethal osmotic stress (24 hours), aggregation of nuclear chromatin, parallel ER arrangement, and reduction of mitochondrial cristae were found. Increased number of Golgi cisternae indicated vesicle production inhibition, and microtubules were absent in treated cells, although plastid structure remained unchanged. After lethal osmotic stress (48 hours), fragmentation of cytoplasmic membranes and a more severe structure damage of all cellular components occurred. Structure of the nucleus, mitochondria, Golgi apparatus and microtubules reappeared in the cells after rehydration. The only new feature of these cells was occurrence of smooth ER, which may indicate that the ER system has acquired a different function in regenerated cells.     

Immunolocalization of the oligosaccharide trimming enzyme glucosidase II

We used immunoelectron microscopy to localize glucosidase II in pig hepatocytes. The enzyme trims the two inner alpha 1,3-linked glucoses from N-linked oligosaccharide precursor chains of glycoproteins. Immunoreactive enzyme was concentrated in rough (RER) and smooth (SER) endoplasmic reticulum but not detectable in Golgi apparatus cisternae. Transitional elements of RER and smooth membraned structures close to Golgi apparatus cisternae contained labeling for glucosidase II. Specific labeling was also found in autophagosomes. These results indicate strongly that glucosidase II acts on glycoproteins before their transport to, and processing in Golgi apparatus cisternae, and suggest that an important transitional region for glucosidase II exists between RER and Golgi apparatus cisternae. Degradation in autophagolysosomes could form a normal catabolic pathway for glucosidase II.     

Subfractionation of rat liver Golgi apparatus by free-flow electrophoresis

Using the technique of preparative free-flow electrophoresis, cisternae of unstacked rat liver Golgi apparatus were separated into a series of fractions of increasing content of sialic acid, thiamine pyrophosphatase and 5'-nucleotidase, markers regarded as being concentrated toward the mature Golgi apparatus face. These same fractions showed a decreasing content of nucleoside diphosphatase, an endoplasmic reticulum marker. Fractions enriched in sialic acid also were enriched in cisternae from the mature or trans face of the Golgi apparatus as deduced from cytochemical criteria. Those fractions least enriched in sialic acid contained cisternae that accumulated deposits of reduced osmium under standard conditions, a test used to mark the opposite, forming or cis-face. Thus subfractionation along the functional polarity axis of the Golgi apparatus with separation of cis and trans face cisternae has been achieved.     

Membrane-glycogen complexes in rabbit extraocular muscle

Analysis of 432 electron micrographs of membrane-glycogen complexes revealed that: (1) Golgi apparatus is closely associated with 4.2% of the complexes, such associations occurring irrespective of the degree of glycogen loading in the complex. (2) Apparent ribosomes are seen in association with about 30% of the complexes, either attached to membranes or enclosed between cisternae. (3) In longitudinal sections of the muscle fibers, complexes may form columns which extend for as much as 40 microns along the fiber. (4) Various cytoplasmic organelles may become enclosed within a complex. (5) Some cisternae of a complex may assume the form of randomly oriented tubules, in contrast to the typical systematic array of flattened cisternae. (6) Some cisternae of a complex may become distended in a wide and uneven manner, in contrast to the typical narrow and even distension.     

Ultrastructural alteration of cytolytic T lymphocytes following their interaction with target cells. I. Hypertrophy and change of orientation of the Golgi apparatus.

The ultrastructure of cytolytic T lymphocytes adhered to the surface of target cells was investigated at different periods after start of interaction. Fifteen-minute incubation led to increase of number of Golgi apparatus cisternae and vacuoles. After 30 min incubation Golgi apparatus become oriented to the contact area. If several lymphocytes adhered to one target cell the Golgi apparatus of each of them was oriented toward the contact area. If one lymphocyte adhered simultaneously to two target cells its Golgi apparatus was oriented toward both target cells. Giant Golgi apparatus vacuoles were formed 30 to 60 min later and then moved to plasma membrane of lymphocyte and then the content of those vacuoles moved to the intercellular space between a cytolytic T lymphocyte and a target cell. The period required for the hypertrophy and change of orientation of Golgi apparatus is supposed to represent the “mobilization” step of a medium-sized and small killer lymphocyte.