The ultrastructure of mitosis during sporangiogenesis inWoronina pythii (Plasmodiophoromycetes)

Summary Mitotic divisions during sporangiogenous plasmodial cleavage inWoronina pythii were studied with transmission electron microscopy. We conclude that these nuclear divisions (e.g., transitional nuclear division, and sporangial mitoses) share basic similarities with the cruciform nuclear divisions inW. pythii and other plasmo-diophoraceous taxa. The major distinction appeared to be the absence of nucleoli during sporangial mitosis and the presence of nucleoli during cruciform nuclear division. The similarities were especially evident with regard to nuclear envelope breakdown and reformation. The mitotic divisions during formation of sporangia were centric, and closed with polar fenestrae, and characterized by the formation of intranuclear membranous vesicles. During metaphase, anaphase, and telophase, these vesicles appeard to bleb from the inner membrane of the original nuclear envelope and appeared to coalesce on the surface of the separating chromatin masses. By late telophase, the formation of new daughter nuclear envelopes was complete, and original nuclear envelope was fragmented. New observation pertinent to the mechanisms of mitosis in thePlasmodiophoromycetes include a evidence for the incorporation of membrane fragments of the original nuclear envelope into new daughter nuclear envelopes, and b the change in orientation of paired centrioles during sporangial mitosis.     

The ultrastructure of mitosis inIsochrysis galbana parke (Prymnesiophyceae)

Summary Mitosis and cytokinesis have been studied in the flagellate algaIsochrysis galbana Parke (Prymnesiophyceae). Nuclear division is preceded by replication of the flagella and haptonema, the Golgi body and the chloroplast; fission in the chloroplast occurs in the region of the pyrenoid. During prophase, spindle microtubules radiating from two ill-defined poles are formed. The nuclear envelope breaks down and the chromatin condenses. At metaphase the spindle is fully developed, some pole-to-pole microtubules passing through the well-defined chromatin plate, others terminating at it. No kinetochores or individual chromosomes were observed. By late metaphase, many Golgi-derived vesicles may be seen against the two poleward faces of the metaphase plate. During anaphase, the two daughter masses of chromatin move towards the poles. In early telophase, the nuclear envelope of each daughter nucleus is complete only on the side towards the adjacent chloroplast, remaining open on the interzonal side. However, during telophase each nucleus becomes reorientated so that it lies lateral to the long axis of the spindle and with its open side towards the chloroplasts. By late telophase, each new nuclear envelope is complete and confluence with the adjacent chloroplast ER established.Cytokinesis and subsequent segregation of the daughter cells are effected by the dilation of Golgi- and ER-derived vesicles in the interzonal region. No microtubular structures are involved. Comparisons with the results from other studies of mitosis in members of thePrymnesiophyceae show that they all have a number of features in common, but that there are differences in detail between species.     

Fine structure of division in ciliate protozoa. I. Micronuclear mitosis in Blepharisma

The mitotic, micronuclear division of the heterotrichous genus Blepharisma has been studied by electron microscopy. Dividing ciliates were selected from clone-derived mass cultures and fixed for electron microscopy by exposure to the vapor of 2% osmium tetroxide; individual Blepharisma were encapsulated and sectioned. Distinctive features of the mitosis are the presence of an intact nuclear envelope during the entire process and the absence of centrioles at the polar ends of the micronuclear figures. Spindle microtubules (SMT) first appear in advance of chromosome alignment, become more numerous and precisely aligned by metaphase, lengthen greatly in anaphase, and persist through telophase. Distinct chromosomal and continuous SMT are present. At telophase, daughter nuclei are separated by a spindle elongation of more than 40 µ, and a new nuclear envelope is formed in close apposition to the chromatin mass of each daughter nucleus and excludes the great amount of spindle material formed during division. The original nuclear envelope which has remained structurally intact then becomes discontinuous and releases the newly formed nucleus into the cytoplasm. The micronuclear envelope seems to lack the conspicuous pores that are typical of nuclear envelopes. The morphology, size, formation, and function of SMT and the nature of micronuclear division are discussed.     

CRUCIFORM NUCLEAR DIVISION IN SOROSPHAERA VERONICAE

Sorosphaera veronicae Schroet. is an endobiotic, holocarpic, obligately parasitic fungus presently classified in the Plasmodiophoromycetes. The ultrastructure of nuclear envelope formation in somatic nuclear division in cystosoral plasmodia was studied. The inner membrane of the nuclear envelope during prophase appears to invaginate and blebb off intranuclear membranous vesicles. The intranuclear membranous vesicles become associated with the surface of the separating chromatin in anaphase and eventually are involved in the formation of daughter nuclear envelopes within the original nuclear envelope. The sequence of nuclear envelope breakdown and reformation in S. veronicae is noteworthy because it emphasizes alternate methods of nuclear envelope formation other than the generally considered “typical” formation described in Allium cepa L.     

THE ULTRASTRUCTURE OF CRUCIFORM NUCLEAR DIVISION IN SOROSPHAERA VERONICAE (PLASMODIOPHOROMYCETE)

Vegetative nuclear divisions in cystosoral Plasmodia from the shoot system of Sorosphaera veronicae Schroeter were studied with standard transmission electron microscopy. Each metaphase nucleus forms a cruciform configuration as the persistent nucleolus elongates perpendicularly to chromatin aligned on the equatorial plate. The nuclear envelope remains intact during metaphase and anaphase. Each spindle pole consists of a fenestrated nuclear envelope with an exteriorly situated centriole and closely associated endoplasmic reticulum. Intranuclear membranous vesicles occur within metaphase and anaphase nuclei and are closely associated with chromatin and nuclear envelope. Microtubules pass from centrioles into the nucleus and are also attached to chromatin at kinetochores.     

Light and electron microscopic studies of meiosis in the basidia ofPholiota terrestris

Summary The two division of meiosis that occur in the distal portion of the basidia ofPholiota terrestris were studied with light and electron microscopy. A diglobular spindle pole body (SPB), consisting of two globular elements and a connecting, electron-dense middle piece, is closely attached to the nuclear envelope of the fusion nucleus. During prometaphase I the globular elements separate and pass to the opposite poles as the chiastic spindle is formed. Evidently, the middle piece also separates with each resulting half persisting as an eccentric, electron-dense portion of the monoglobular SPB of meta-, ana-, and telophase nuclei. Also during prometaphase I, the nuclear envelope becomes discontinuous, especially in the lower region of the spindle. Light microscopic evidence of nucleolar extrusion at prometaphase I and II was observed. At metaphase I the SPB's move away from the condensed chromatic mass as the chromatids move asynchronously along the expanding spindle, evidently, due both to the elongation of the continuous fibers and the shortening of the chromosomal fibers. Two images resembling typical kinetochroes are illustrated in anaphase I nuclei, and others were seen during the study. At early telophase I and II the nuclear envelope is present laterally, is then formed in the interpolar region, and eventually appears between the chromatin and monoglobular SPB. A perforated ER cap, which is penetrated by microtubules, delimits the SPB. The nucleus enlarges, the chromatin becomes diffused except adjacent to the SPB, and the perinuclear ER becomes uniformly oriented around the nuclear envelope. At interphase I a diglobular SPB was not clearly documented. During interphase I the ER cap disappears but the perinuclear ER persists. Division II, with the exception of prophase, is essentially identical to division I. The postmeiotic, haploid nuclei migrate to the median or proximal region of the basidium. The diglobular SPB reappears. The meiotic apparatus inP. terrestris is considered to have the same fundamental features as those of plants and animals and in detail conforms to the pattern described in several light and electron microscopic studies of other Homobasidiomycetes.     

MITOSIS IN THE AQUATIC FUNGUS RHIZOPHYDIUM SPHEROTHECA (CHYTRIDIALES)

The structure of centric, intranuclear mitosis and of organelles associated with nuclei are described in developing zoosporangia of the chytrid Rhizophydium spherotheca. Frequently dictyosomes partially encompass the sides of diplosomes (paired centrioles). A single, incomplete layer of endoplasmic reticulum with tubular connections to the nuclear envelope is found around dividing nuclei. The nuclear envelope remains intact during mitosis except for polar fenestrae which appear during spindle incursion. During prophase, when diplosomes first define the nuclear poles, secondary centrioles occur adjacent and at right angles to the sides of primary centrioles. By late metaphase the centrioles in a diplosome are positioned at a 40° angle to each other and are joined by an electron-dense band; by telophase the centrioles lie almost parallel to each other. Astral microtubules radiate into the cytoplasm from centrioles during interphase, but by metaphase few cytoplasmic microtubules are found. Cytoplasmic microtubules increase during late anaphase and telophase as spindle microtubules gradually disappear. The mitotic spindle, which contains chromosomal and interzonal microtubules, converges at the base of the primary centriole. Throughout mitosis the semipersistent nucleolus is adjacent to the nuclear envelope and remains in the interzonal region of the nucleus as chromosomes separate and the nucleus elongates. During telophase the nuclear envelope constricts around the chromosomal mass, and the daughter nuclei separate from each end of the interzonal region of the nucleus. The envelope of the interzonal region is relatively intact and encircles the nucleolus, but later the membranes of the interzonal region scatter and the nucleolus disperses. The structure of the mitotic apparatus is similar to that of the chytrid Phlyctochytrium irregulare.     

Fine structure of the macronucleus during the cell division cycle of Euplotes eurystomus,a Ciliate Protozoon

This paper reports new observations obtained from a study of macronuclear fine structure throughout various stages of the cell division cycle of Euplotes. Study of the ultrastructural organization of the macronuclear chromatin indicates that much of the chromatin is organized into continuous masses, portions of which appear to be attached to the nuclear envelope. The macronuclear envelope appears unchanged in the region of a replication band, and apparent attachments of the chromatin to the inner membrane of the nuclear envelope are maintained in the reticular and diffuse zones. Intranuclear helices were never observed in the diffuse zone. During macronuclear division, linear elements (fibrils or microtubules) were observed in close association with both chromatin bodies and nucleoli. The ultrastructural data suggest that the intranuclear linear fibrils have two functions: elongation of the dividing nucleus, and attachment of chromatin bodies and nucleoli to the envelope. The significance of these observations for macronuclear division and chromatin segregation is considered.     

The ultrastructure of mitosis in myxamoebae and plasmodia of Physarum flavicomum

Electron microscopy of glutaraldehyde-osmium-fixed samples of haploid myxamoebae and diploid plasmodia of the myxomycete Physarum flavicomum Berk. reveal dissimilar spindle apparatus during mitosis in the two cell types. Myxamoebae exhibit an astral type of mitosis with centrioles at the poles and nuclear envelope breakdown during prophase. Plasmodial nuclei lack centrioles at mitosis and have an intranuclear spindle, with nuclear envelope persisting during the entire division. Coated vesicles are noted during prophase and telophase in myxamoebae and their role in spindle formation and dispersion is suggested.     

Substructural morphogenesis of the spindle apparatus in meiotic basidia ofCoprinus micaceus (Agaricales)

The spindle apparatus ofCoprinus micaceus begins to develop from the diglobular polar body outside the nucleus. During both meiotic divisions it operates inside the nuclear envelope and consists of two amorphous poles, a central bundle of interpolar microtubules, and chromosomal microtubules. A metaphase plate cannot exist because the interpolar strand of fibers is persistent throughout the division process. Within the spindle axis more than 100 microtubules can be estimated. They are encircled by a ring of chromatic structures. During the telophase the former spindle pole is evaginated from the nuclear envelope and contacts the plasmalemma near the cell wall.     

Ultrastructure of the nuclei during different phases of the cell cycle in the antheridial filaments ofChara vulgaris L.

Summary Synchronously dividing nuclei of the antheridial filaments ofChara vulgaris at the 32-celled stage have different structure depending on the period of interphase.During S phase which begins as early as at the start of telophase (coincidently with the nuclear envelope formation and chromosome decondensation) one can observe a gradual reduction in the content of condensed chromatin, having the appearance of an indistinct network. During the middle S period the area of condensed chromatin decreases to the lowest level of about 29% of nuclear profile and the nuclear envelope becomes folded. At the end of S phase the condensed chromatin forms a more distinct and thicker reticulum which covers an area of about 52%.During the early G₂ phase, the area occupied by the condensed chromatin was about 41% and it was found to assume the shape of large and iregular clusters localized mainly near the nucleoli. The reticulate form of chromatin, characteristic of the S period, disappears almost completely. During the next period of interphase the condensed chromatin disperses considerably and covers now 24% of the area. At the end of the G₂ phase the condensed chromatin reappears and transforms into chromosomes. Then the condensed chromatin removes from the nuclear envelope.This work was supported by the Polish Academy of Sciences within the project 09.7.3.1.4.     

Ultrastructural changes in major organelles during spermatial differentiation inBangia (Rhodophyta)

Summary The major organelles within the cells of maleBangia atropurpurea (Roth) C. Ag. filaments undergo a series of ultrastructural transformations during the production of spermatia. Initially, thylakoids within the large axial chloroplast develop a reticulate pattern commencing at the central pyrenoid region. Subsequent changes involve loss of lobes and diminution of volume through division; chloroplasts in final stages contain a few dilated, distorted thylakoids and many plastoglobuli. During differentiation the large nucleolus disappears from the nucleus and four masses of chromatin aggregate near the nuclear envelope. Furrows originating from the nuclear envelope form double membranes around each of the chromatin masses and most of the nucleoplasm is eliminated. Several types of fibrillar vesicles are formed during the process and large floridean starch reserves are utilized. Multilamellar bodies and microbody-like structures occur within the cells during certain phases of spermatiogenesis.     

Sheathed nuclear division in primary spermatocytes of Orgyia antiqua (Lepidoptera, Insecta)

The restructuring of spermatocytes during the first meiotic division is examined in the moth Orgyia antiqua (Lymantriidae, Lepidoptera) using transmission electron microscopy. Particular emphasis was placed on the behaviour of the perispindle membrane system. These membranes develop from layers of smooth endoplasmic reticulum wrapped around the prophase I nucleus and are retained until early telophase I. The original nuclear envelope is dissolved in metaphase I. Polar fenestrae in the perispindle membrane stacks are filled with numerous irregular membrane elements. The formation of new nuclear envelopes around the daughter nuclei takes place inside the perispindle membrane system. Finally, the membrane stacks rupture concomitantly with spindle elongation in late telophase I. Thus, division of primary spermatocytes in Orgyia antiqua has a surprising degree of similarity with the so-called closed mitosis. This mode of division is typical for many protozoa, algae and fungi. In the pertinent cells, the original nuclear envelope persists around the spindle area during nuclear division. In order to distinguish the closed mitosis from the situation in Orgyia antiqua spermatocytes, the term 'sheathed nuclear division' is suggested for the latter.     

MITOSIS IN THE OCTAFLAGELLATE PRASINOPHYTE,PYRAMIMONAS AMYLIFERA (CHLOROPHYTA)1

Cell division is described in the octaflagellate prasinophyte Pyramimonas amylifera Conrad and is compared in related genera. Basal bodies replicate at preprophase and move toward the poles. Cells remain motile throughout division. The nuclear envelope disperses and chromosomes begin to condense at prophase. Pairs of multilayered kinetochores are evident on the chromosomes of the metaphase plate. Spindle microtubules extending from the region of the basal bodies and rhizoplasts attach to the kinetochores or extend from pole to pole. Numerous vesicles and ribosomes have entered the nuclear region and the incipient cleavage furrow invaginates. The chromosomes move toward the poles at anaphase leaving a broad interzonal spindle between the two chromosomal plates. The nuclear envelope reforms first around the chromatin on the side adjacent to the spindle poles and later on the interzonal side. The cleavage furrow progresses into the interzonal spindle at telophase. By late telophase the nucleoli have reformed and the chromosomes have decondensed. The interzonal spindle has not been observed late in telophase. As the cleavage furrow nears completion the cells begin to twist and contort, ultimately separating the two cells.     

Nuclear envelope localization of an adenovirus tumor antigen maintains the integrity of cellular DNA.

The adenovirus early-region 1B 19,000-molecular-weight tumor antigen is required for oncogenic transformation of cells by adenovirus. We have demonstrated that this tumor antigen is located in the nuclear envelope of infected and transformed cells and that a fraction of the protein within the nuclear envelope is associated with the nuclear lamina. During cell division in the transformed cells, the nuclear envelope containing the tumor antigen dissociates at metaphase and then reforms around the separated daughter chromosomes at telophase. Adenovirus mutants carrying lesions in the gene encoding this tumor antigen cause degradation of host cell chromosomal DNA, and in these mutants, the intracellular localization of the 19,000-dalton protein is altered. These results demonstrate that components of the nuclear envelope function in the organization of chromatin in infected and transformed cells and that a virus-encoded protein plays a critical role in this process.     

ELECTRON MICROSCOPIC STUDIES OF MITOSIS IN AMEBAE : I. Amoeba proteus

Individual organisms of Amoeba proteus have been fixed in buffered osmium tetroxide in either 0.9 per cent NaCl or 0.01 per cent CaCl₂, sectioned, and studied in the electron microscope in interphase and in several stages of mitosis. The helices typical of interphase nuclei do not coexist with condensed chromatin and thus either represent a DNA configuration unique to interphase or are not DNA at all. The membranes of the complex nuclear envelope are present in all stages observed but are discontinuous in metaphase. The inner, thick, honeycomb layer of the nuclear envelope disappears during prophase, reappearing after telophase when nuclear reconstruction is in progress. Nucleoli decrease in size and number during prophase and re-form during telophase in association with the chromatin network. In the early reconstruction nucleus, the nucleolar material forms into thin, sheet-like configurations which are closely associated with small amounts of chromatin and are closely applied to the inner, partially formed layer of the nuclear envelope. It is proposed that nucleolar material is implicated in the formation of the inner layer of the envelope and that there is a configuration of nucleolar material peculiar to this time. The plasmalemma is partially denuded of its fringe-like material during division.     

COLONY DEVELOPMENT IN EUDORINA ELEGANS (CHLOROPHYTA,VOLVOCALES)1

A detailed ultrastructure study was made of cell division and colony development in Eudorina elegans Ehrenberg. At the onset of cell division and prior to nuclear division the nucleus moved from the cell center to the cell surface. During nuclear division the nuclear membrane remained intact, except for openings occurring at the nuclear poles. The spindle microtubules appeared to arise from a MTOC-like (microtubule organizing centers) structure, while centrioles were absent from the nuclear poles. Following telophase, daughter nuclei formed which were separated by several distinct bands of endoplasmic reticulum. Cytokinesis occurred with formation of a cleavage furrow, associated with a typical phycoplast band of microtubules. However, cytokinesis was incomplete, resulting in formation of cytoplasmic bridges between the plakeal cells. Upon completion of up to five successive cell divisions, the plakea underwent inversion, which appeared to involve the production of colonial envelope material and rearrangement of cytoplasmic bridges. A new hypothesis concerning inversion is postulated based on these observations.     

A UNIQUE MITOTIC VARIATION IN THE MARINE DINOFLAGELLATE OXYRRHIS MARINA (PYRROPHYTA)1

Mitosis is described in the flagellate Oxyrrhis marina Dujardin and is compared in related genera. Dense plaques develop in the nuclear envelope at prophase and give rise to an intranuclear spindle. Some of the microtubules associate with the chromosomes while others extend across the nucleus. The basal bodies migrate toward the poles early in division and retain a position lateral to the nuclear poles throughout mitosis. Microtubules are not present between the nucleus and the basal bodies. The nucleolus is persistent and elongates throughout anaphase and telophase. Chromosomal separation is accomplished by sliding of non-chromosomal microtubules and by elongation of the nuclear envelope rather than by shortening of the spindle microtubules. The nuclear envelope begins to constrict in the center early in anaphase. Continued constriction of the envelope and elongation of the nucleus leads to the formation of a dumbbell-shaped nucleus by late telophase. Mitosis culminates by the constriction of the nucleus into two daughter nuclei. The taxonomic position of Oxyrrhis marina is discussed in light of these findings.     

Electron Microscopic Studies on the Silver-stained Nucleolar Cycle of Physarum polycephalum

The dynamic changes of nucleolar ultrastructure in the cell cycle of Physarum polycephalum Schw. were studied by an en bloc silver-staining method. The results showed that the nucleolus was large in size and situated in the center of the nucleus in late G2-phase, and the fibrillar centers, dense fibrillar components and granular components could be observed in the nucleolus. During prophase, the nucleolus moved towards the periphery of the nucleus and in late prophase disintegrated near the nuclear envelope. In metaphase, the disintegrated nucleolar components were dispersed in masses and located at the periphery of the chromosomal region of the nucleus. No specifically silver-stained area and argentophilic protein sheath were observed on the chromosomes, but there were some big dispersed silver particles within the chromosomes. During telophase the nucleolar components moved towards the two poles along with the chromosomes and co-existed with the decondensing chromatin in daughter nuclei. The nucleolar components then gradually converged with one another and separated from the chromatin. A big nucleolus was formed in the nucleus about 120 min after the completion of mitosis.     

Distribution of ubiquitin protein in meristematic mesophyll ceils of barley leaves

The distribution of ubiquitin protein in meristematic mesophyll cells of barley (Hordeum vulgare L.) leaves was investigated by using immunofluorescence microscopy. Simultaneous observation of nuclei was achieved byDAPI (4 6-diamidino-2-phenylindol-dihydrochloride) staining. A strong correlation between the chromatin organisation and the ubiquitin distribution could be observed. Interphase nuclei revealed an intense content of ubiquitin and accumulation of ubiquitin at the nuclear envelope, whereas condensed chromosomes of dividing cells excluded any ubiquitin appearance. During cell division, the aggregation of ubiquitin protein was detected in the area of the mitotic spindle in anaphase as well as the area of the cell plate in the late telophase.