THE FLAGELLAR APPARATUS OF OXYRRHIS MARINA (PYRROPHYTA)1

The three-dimensional structure of the flagellar apparatus in the dinoflagellate Oxyrrhis marina has been reinvestigated and found to consist of several previously unknown components and component combinations that appear strikingly similar to those of some gymnodinoid taxa. The flagellar apparatus of this dinoflagellate is asymmetric and extremely complex consisting of a longitudinal and a transverse basal body that gives rise to eight structurally different components. The only posteriorly directed component is the large microtubular root that consists of 45–50 microtubules at its origin and is attached proximally to a perpendicularly oriented striated fibrous component. Arising from each basal body, two striated fibrous roots with different periodicities extend to the cell's left. A single stranded microtubular root with associated electron dense material emanates from the transverse basal body and also extends to the cell's left. A striated fibrous connective arises from the longitudinal basal body and extends toward the cell's right ventral surface and terminates near the sub-thecal microtubular system. A compound root consisting of microtubules and electron dense material also originates from the longitudinal basal body and extends ventrally into the anterior region of the tentacle. Structural similarities between the parallel striated fibrous roots of Oxyrrhis and Polykrikos are discussed as are flagellar apparatus similarities among other gymnodinoid dinoflagellates. A diagrammatic reconstruction of the Oxyrrhis flagellar apparatus is also presented.     

THE FLAGELLAR APPARATUS OF THE ZOOSPORE OF UROSPORA PENICILLIFORMIS(CHLOROPHYTA)1

The flagellar apparatus of Urospora penicilliformis (Roth) Aresch. is unique, or at least very unusual among green algae. The flagellar axonemes are rigid, and contain wing-like projections. There are no central microtubules in the most proximal part of the axoneme. The transition region contains a series of electron dense transverse lamellae rather than a single septum, and lacks a stellate pattern. There is no cartwheel pattern in the proximal part of the basal bodies. The latter are associated with four different types of fibrous elements: ascending striated fibers that attach to an electron dense plate in the papillar center, lateral striated fibers that parallel microtubular roots, fibrous elements that link adjacent basal bodies, and finally two massive striated fibers that descend into the cell, passing closely along the nucleus (system II fibers, or rhizoplasts). Each of the four microtubular flagellar roots is sandwiched between two system I striated structures. The roots are probably equal; they contain proximally four, and distally up to eight microtubules. Based on the zoospore flagellar apparatus, it is concluded that the multinucleate U. penicilliformis is related to the Ulvaphyceae. Finally, a possible explanation in functional terms is given for the peculiar external morphology and behavior of the zoospore.     

FEEDING APPARATUS OF THE COLORLESS FLAGELLATE KATABLEPHARIS (CRYPTOPHYCEAE)1

The feeding apparatus of Kalablepharis ovalis (isolated from a freshwater impoundment in Colorado) and Katablepharis clone G-2 (isolated from the littoral of the Black Sea near Yalta in the Crimea) consists of inner and outer oval-shaped arrays of microtubules that begin at the anterior end of the cell and pass into the posterior of the cell. Each array of microtubules contains groups of microtubules with two to eight microtubules per group depending on the position of the array in the cell. A specialized area of the plasma membrane, the mouth, occurs at the anterior end of the cell. The mouth is oval with the long axis oriented dorsoventrally and consists of a raised ridge surrounding a central depression. The anterior end of the microtubules of the inner and outer arrays supports the raised ridge of the mouth. In freeze-fracture replicas, the protoplasmic face of the plasma membrane contains intramembrane particles on the raised ridge of the mouth. Three small membrane-cisternae occur on the protoplasmic side of the plasma membrane in the area of the mouth. Katablepharis clone G-2 also has five or six additional large membrane-cisternae associated with the inner microtubular array in the anterior portion of the cell. These larger membrane-cisternae do not occur in K. ovalis. Vesicles with electron-dense contents occur in association with the microtubular arrays. Katablepharis ovalis has a second type of vesicle containing a single-membrane profile associated with the microtubule arrays. The structure of the microtubular arrays in Katablepharis is compared with similar structures in suctorian ciliates and dinoflagellates.     

MICROMORPHOLOGY OF A SMALL DINOFLAGELLATE PROROCENTRUM MARIAE-LEBOURIAE (PARKE & BALLATINE) COMB. NOV.1,2,3

The surface structures of the bivalvate dinoflagellate Prorocentrum mariae-lebouriae are described in detail. It has an almost spheroidal shape in face-view, a compressed saucer-shape in side view, with a distinct striated band at the edge of the cell. Its surface is covered with small spines in a regular pattern, with 450 nm distance between pairs. The spines are 100–120 nm wide and 200–300 nm long. There are 600–700 spines on each valve. At the anterior cell end, one of the values has a V-shaped depression which contains a specialized structure accommodating the 2 flagellar pores. The flagellar pores are-enclosed by 8 small, thick plates held together and to the values by sutures. The flagellar pore area consists of 2 distinct structures: an apical collar possessing a curved forked plate and a larger structure composed of an unbranched, plate. There are 2 flagellar canals located between the flagellar pore plates. Beneath each flagellar canal lies a row of 11 microtubules. A row of microtubules forming a microtubular cylinder is situated adjacent to the oblong flagellar canal near a simple pusule. The microtubular cylinder encircles electron dense bodies. The bases of the longitudinal and transverse flagella appear to lie at an angle to each other. The above features are illustrated with transmission and scanning electron micrographs.     

ELECTROMAGNETIC ACTIVITY OF YEAST CELLS IN THE M PHASE

Electromagnetic activity around yeast mitotic cells (Saccharomyces cerevisiae) was measured in the frequency range 8–9 MHz and special care was taken to extract reliable information from the raw signals. The characteristic of cold-sensitive tubulin mutants tub2-401 and tub2-406, which come to arrest before mitosis at a restrictive temperature (14°C) and which re-enter mitosis upon a shift back to a permissive temperature (28°C), was used to prepare synchronized mitotic cells. Immunofluorescence microscopy using an antitubulin antibody was used to analyze microtubular structures. The arrested tub2-401 mutant that had back-shifted to permissive temperature displayed no microtubules and no electromagnetic activity around the cells. In contrast, the arrested cells of the mutant tub2-406 displayed developed, but aberrant, nonfunctional microtubules and a high electromagnetic activity around the cells. The electromagnetic activity around the arrested mutant tub2-401 back-shifted to permissive temperature peaks at four time points which may coincide with (i) formation of the mitotic spindle, (ii) binding of chromatids to kinetochore microtubules, (iii) elongation of the spindle in anaphase A, and (iv) elongation of the spindle in anaphase B. The profile of the electromagnetic activity around the synchronized mutant tub2-406 at permissive temperature seems to be delayed by the time required for aberrant nonfunctional microtubules to be depolymerized. Experimental results presented in this paper support Pohl's idea of existence of the electromagnetic field around yeast cells.     

Study of Dientamoeba fragilis Jepps & Dobell I. Electronmicroscopic Observations of the Binucleate Stages II. Taxonomic Position and Revision of the Genus*

Light- and electronmicroscopic observations on Dientamoeba fragilis strain A (Bi) 1 dealing primarily with the binucleate (arrested telophase) stages, predominant in all populations, revealed the microtubular nature of the extranuclear spindle which extends between the 2 polar complexes each adjacent to one of the nuclei. The spindle microtubules originate in paired, nonperiodic structures apparently homologous to the “atractophores” described from hypermastigotes. To the external surface of the atractophores are applied periodic elements, which extend laterally as the parabasal filaments. Extensive Golgi complexes overlie the filaments, these structures corresponding to the components of the parabasal apparatus known from trichomonads and hypermastigotes. The 2-layered structures, consisting of the atractophores and periodic layers, together with the proximal parts of the Golgi complex and the spindle microtubules constitute the polar complex. No kinetosome- or centriole-like organelles have been found in the polar complexes or elsewhere in the organism. The extranuclear spindle is composed of 2 microtubule bundles, each with ～30-40 microtubules. One of the bundles always appears at some distance from the nucleus; the other is juxtanuclear and is seen often to course within a groove of the nuclear envelope. A 3rd bundle of ～35-45 microtubules is seen on occasion to arise from the atractophores and to pass toward the nucleus at a wide angle to the other parts of the spindle. In some instances these microtubules traverse the nucleus within channels delimited by the nuclear envelope. The double-layered nuclear envelope contains numerous pores. Two morphologic types of rounded inclusions, one microbody-like, and the other with a more electron-translucent matrix, as well as digestive vacuoles containing rice starch, bacteria, and/or myelin configurations are distributed in the cytoplasm, which abounds also in glycogen granules. The fine structure of Dientamoeba is compared with those of trichomonads and of Entamoeba spp. The taxonomic position of Dientamoeba is discussed and emended; in view of its affinities, this genus is placed among trichomonads in the family Dientamoebidae Grassé, emend.     

A Comparative electron microscopic study of cytoplasmic microtubules and axial unit tubules in a spermatozoon and a protozoan

Cytoplasmic microtubules and axial unit tubules were studied in both sectioned and negatively-stained material. Walls of microtubules of frog lung-fluke (Haematoloechus medioplexus) spermatozoa have a helical substructure, while those of the flagellate, Trypanosoma lewisi, are composed of ten longitudinally-oriented filaments. Cross-bridges occur between some filaments of trypanosome microtubules. Doublet tubules of axial units in both cell types are structurally similar to the trypanosome microtubules, which may indicate similarity of function. Microtubules of fluke spermatozoa appear to be somewhat rigid, are resistant to sonication, and are considered to be mainly supportive. Circular profiles of wall subunits are seen in transverse sections of microtubules of both cell types and in doublet tubules of the trypanosome. Comparisons are made between sectioned and negatively-stained material; while negative-staining better reveals the fundamental substructure of microtubular elements, some distortion appears to occur. In connection with this research, a brief preliminary article demonstrated the presence of subunits in the walls of cytoplasmic microtubules of fluke spermatozoa (Burton, '66). Also, it was shown that the wall of these tubular elements possesses a helical structure, and a diagrammatic representation of the wall structure was set forth.     

Fine Structure of the Contractile Vacuole Pore in Paramecium*

The pore through which a Paramecium contractile vacuole communicates with the external environment is a 1.2 μm long and 1 μm diameter cylindrical orifice in the pellicle. During diastole, the vacuole:pore junction is closed by a substantial diaphragm which parts to the side at systole. The diaphragm is composed of inner and outer membranes continuous with the vacuole and pore membranes, respectively, and an intervening cytoplasmic layer containing filaments and irregular membranous tubules and vesicles. Microtubules, organized into 2 sets, are an important component of the pore apparatus. One set of ～ 16 microtubules forms an annulus around the pore. These microtubules are organized into a right-handed helix with a pitch of 0.5-0.6 μm, and thus complete slightly more than 2 turns in their course from the level of the diaphragm to the pore outer lip. They appear to be embedded in a layer of dense material immediately adjacent to the pore membrane. The other set consists of 5 or more bands of 10–20 microtubules which radiate in a slight left-handed helix from an insertion at the pore out over the vacuole surface to the ampullae.     

Ultrastructure of the flagellar apparatus inPleurochrysis (classPrymnesiophyceae)

Summary The ultrastructure of the flagellar apparatus of aPleurochrysis, a coccolithophorid was studied in detail. Three major fibrous connecting bands and several accessory fibrous bands link the basal bodies, haptonema and microtubular flagellar roots. The asymmetrical flagellar root system is composed of three different microtubular roots (referred to here as roots 1,2, and 3) and a fibrous root. Root 1, associated with one of the basal bodies, is of the compound type, constructed of two sets of microtubules,viz. a broad sheet consisting of up to twenty closely aligned microtubules, and a secondary bundle made up of 100–200 microtubules which arises at right angles to the former. A thin electron-dense plate occurs on the surface of the microtubular sheet opposite the secondary bundle. The fibrous root arises from the same basal body and passes along the plasmalemma together with the microtubular sheet of root 1. Root 2 is also of the compound type and arises from one of the major connecting bands (called a distal band) as a four-stranded microtubular root and extends in the opposite direction to the haptonema. From this stranded root a secondary bundle of microtubules arises at approximately right angle. Root 3 is a more simple type, composed of at least six microtubules which are associated with the basal body. The flagellar transition region was found to be unusual for the classPrymnesiophyceae. The phylogenetic significance of the flagellar apparatus in thePrymnesiophyceae is discussed.     

THE FLAGELLAR APPARATUS OF CHRYSOLEPIDOMONAS DENDROLEPIDOTA (CHRYSOPHYCEAE), A SINGLE-CELLED MONAD COVED WITH ORGANIC SCALES1

The flagellar apparatus of Chrysolepidomonas dedrolepidota Peters et Andersen is similar to that of other members of the Ochromonadales, Chrysophyceae. there are four microtubular roots (R_1-4) and a system II fiber (= rhizoplast). the R₁ root consists of three microtubules that nucleate many cytoplasmic microtubules. One compressed band of 10 or more cytoplasmic microtubules is directed black along the R1 root in an anti-parallel direction. The R₂ root consists of one to two microtubules, and it extends toward the distal end of the R₁ root. The R₃ root consists of six (?seven) microtubules near its proximal end. The “a” and “f” microtubules of the R₃ root are under the short flagellum, and the “f” microtubule loops back and under the basal body, extending down to the nucleus. The R₄ root consists of one to two microtubules extending along the left side of the shot flagellum and curving under the short flagellum where it terminates near the “a” microtubule of R₃ Both flagella have a transitional plate and a transitional helix with five gyres. There is a thin, second plate in the basal body at the level of the distal end of the “c” tubules of the basal body triplets. The tripartite flagellar hairs have long lateral filaments but lack short lateral filaments. We compare the flagellar apparatus with that of other members of the Ochromonadales and members of the Hydrurales and Hibberdiales.     

Microtubular cytoskeletons of the trophic cells of five eumycetozoans

Summary The trophic cells of the protostelids,Ceratiomyxella tahitiensis, Cavostelium apophysatum, Planoprotostelium aurantium, andClastostelium recurvatum, and the reduced myxomycete,Echinostelium bisporum, were examined with indirect immunofluorescence to determine the overall structure of the microtubular cytoskeletons of each type of cell. All five species have a distinct flagellar apparatus in the amoeboflagellate state, but they vary with respect to the presence of body microtubules, those microtubules which do not focus on the flagellar apparatus. The obligate amoebae ofC. tahitiensis, C. apophysatum andC. recurvatum all have extensive microtubular cytoskeletons, but each is unique to its respective species. The obligate amoeba ofC. tahitiensis has scattered microtubules which often describe curved paths. The microtubules never appear to focus on MTOCs. The microtubular cytoskeleton ofC. apophysatum consists of relatively straight, rigid appearing microtubules. Small subpopulations of these microtubules radiate from MTOCs near the nucleus. The obligate amoeba ofC. recurvatum has a cytoskeleton consisting of numerous microtubules which radiate from a perinuclear MTOC and fill the body of the cell. These results correlate well with earlier ultrastructural observations which suggest that the amoeboflagellate state is homologous in these mycetozoans and that the obligate amoebae, when present, are unique to the various lineages in which they appear.     

THE CORTICAL MICROTUBULAR CYTOSKELETON OF OXYRRHIS MARINA (DINOPHYCEAE) OBSERVED WITH IMMUNOFLUORESCENCE AND ELECTRON MICROSCOPY1

The cortical microtubular cytoskeleton of Oxyrrhis marina Dujardin was investigated using indirect immunofluorescence and transmission electron microscopy. The cortical microtubular cytoskeleton is unlike that of other previously examined dinoflagellates because all cortical microtubules are oriented longitudinally and do not attach or abut tranverse microtubular arrays. This difference is considered along with other morphological and cytological variables as indicative of Oxyrrhis's phylogenetic position relative to the Dinophyceae.     

FINE STRUCTURE OF THE ZOOSPORE OF OEDOCLADIUM CAROLINIANUM (CHLOROPHYTA) WITH SPECIAL REFERENCE TO THE FLAGELLAR APPARATUS1, 2

Ultrastructure of the motile zoospore has been investigated in Oedocladium catolinianum & Hoffman. An unwalled zoospore is usually produced from the contents of a terminal vegetative cell and consists of two principal regions: a small anterior dome and a larger body region; a ring of flagella marks the juncture of these two areas. Chloroplast inclusions consist of thylakoids, mature and incipient pyrenoids, starch and striated microtubules; no eyespot has been observed. Zoospores appear to possess permanent contractile vacuoles with numerous accessory vacuoles, coated vesicles and occasionally coated tubules. The cytoplasm of the dome contains numerous mitochondria ER and golgi bodies, as well as two distinct types of vesicles. The first contains an electron-dense; granular core and is surrounded by a loose, sinuate membrane. The second vesicle is electron-opaque and is found at the apex of the dome: it contains mucopolysaccharides employed during zoospore adhesion. A complex flagellar apparatus encircles the lower region of the dome. It consists of ca. 30–65 flagella, a ring-shaped fibrous band, flagella roots and additional supporting material. The flagella and roots alternate with one another beneath the fibrous band. The compound flagellar roots consist of two superimposed components: an outer ribbon-like unit composed of three microtubular elements and a single striated inner component. A band of support material lies beneath the proximal end of the basal bodies. It is a continuous fibrous band, although it often appears as three distinct, repetitive units.     

The structure of the flagellar apparatus of the swarm cells ofPhysarum polycephalum

Summary Flagellation ofPhysarum polycephalum amoebae (Myxomycete) involves the formation around the two kinetosomes of a flagellar apparatus leading to a modification in the shape of the amoeba and its nucleus. A tridimensional ultrastructural model of the flagellar apparatus is proposed, based upon observation of the isolated nucleo-flagellar apparatus complex. The flagellar apparatus is composed of a non-microtubular structure (the posterior para-kinetosomal structure), five microtubular arrays and two flagella: a long anterior flagellum and a short flagellum directed backwards. The asymmetry of the flagellar apparatus is due mainly to the presence of the posterior para-kinetosomal structure on the right side of the posterior kinetosome and of the two asymmetrical microtubular arrays 3 and 4. Thus, the flagellar apparatus is right-handed. This asymmetry implies also some spatial constraints on two other microtubular arrays (2 and 5). Except in the case of the microtubular array 1 which links the proximal end of the anterior kinetosome to the nuclear membrane, the number of microtubules of each microtubular array seems to be well defined: 39, 5–6, 7–9, and 2+2 for the microtubular arrays 2, 3, 4, and 5 respectively. All the elements of the nucleo-flagellar apparatus complex are linked either directly or indirectly through bridges. Furthermore, the microtubules which composed the microtubular array 3 are linked through bridges while the microtubules of the microtubular arrays 2, 3, and 4 seem to be linked through a reticulate material. All these spatial relationships lead to a great cohesion of the nucleo-flagellar apparatus complex which appears to be a well defined structure. This suggests thatPhysarum amoebal flagellation can be a promising system to study the morphogenesis of an eucaryotic cell.Abbreviations PIPES Piperazine-N,N-bis [2-ethane-sulfonic acid] - EGTA [Ethylenebis(oxyethylenenitrile)]tetraacetic acid - DMSO Dimethyl sulfoxide     

TRACHELOMONAS HISPIDA VAR. CORONATA (EUGLENOPHYCEAE). I. ULTRASTRUCTURE OF CYTOSKELETAL AND FLAGELLAR SYSTEMS1,2

Trachelomonas hispida var. coronata Lemm. has a fibrous, mucilaginous, ovoid, mineralized envelope (lorica), the ornamentation and coloration of which are capricious in culture. Cells exhibit a radial distribution of most organelles: (i) A cortical endoplasmic reticulum, (ii) parietal chloroplasts, and (iii) a median vacuolar region surrounded by several Golgi bodies and diverse vesicles. Associated with the emergent flagellum is a “paraflagellar complex” that consists of dense globules, cross-striated ribbon-like structures, a paraflagellar body, and an array of parallel striated filaments. The stigma consists of a single layer of pigmented granules that partially surrounds the canal/reservoir transition zone where microtubular bands intersect. A microtubular cytoskeleton consists of pellicular microtubules, peri-canal microtubules, stigma-associated microtubules and para-reservoir microtubules. The thickenings on the posterior, concave margins of the pellicular strips suggest that this pellicle is of intermediate complexity between those of Euglena spirogyra (Ehrenb. and Trachelomonas volvocina (Ehrenb.).     

The Cytology of Sheep Rumen Ciliates. I. Ultrastructure of Epidinium caudatum Crawley1

Epidinium caudatum has an anterior vestibulum containing the adoral zone syncilia (AZS) on an extrusible peristome. The cytopharyngeal structures include a funnel-shaped arrangement of nematodesmata, longitudinal and transversely oriented microtubular ribbons all of which are located in the peristome, a structure which also contains filamentous phagoplasm. The origins of the microtubular ribbons indicate affinities to the rhabdos type of cytopharynx. The peristomal base is continuous with the tubular esophagus, the region connecting the two being ensheathed by a fibrous layer and low density cytoplasm. The esophagus has a microtubular/microfilamentous wall. A distinct cytoproct with associated myonemal structures occurs posteriorly. The skeletal plates consist of a large number of interconnected, variably shaped platelets and may have dual skeletal and storage functions. The endoplasm is more vesicular than the ectoplasm, the two separated by a fibrous boundary layer. The five-layered cortex has an external glycocalyx, a plasma membrane with two subtending membranes, homogeneous, microtubular, and microfilamentous layers. The syncilia of the AZS are mounted in a U-shaped band on the peristome with transversely oriented kinetics consisting of kinetosomes linked by a sub-kinetosomal rod. There is a bifurcated kinetodesma, dense support material forming a lateral spur with associated transverse microtubules, and postciliary, interkinetal, and occasional basal microtubules, nematodesmata, and a subciliary reticulum. A barren, possibly vestigial, somatic infraciliature consists of non-ciliated kinetosomes and a basal striated fiber with associated basal and perpendicular (cortical) microtubules.     

COMPARATIVE ULTRASTRUCTURE OF ZOOSPORES WITH PARALLEL BASAL BODIES FROM THE GREEN ALGAE DICTYOCHLORIS FRAGRANS AND BRACTEACOCCUS SP.

The chlorococcalean algae Dictyochloris fragrans and Bracteacoccus sp. produce naked zoospores with two unequal flagella and parallel basal bodies. Ultrastructural features of the flagellar apparatus of these zoospores are basically identical and include a banded distal fiber, two proximal fibers, and four cruciately arranged microtubular rootlets with only one microtubule in each dexter rootlet. In D. fragrans, each proximal fiber is composed of two subfibers, one striated and one nonstriated, and each sinister rootlet is composed of five microtubules (4/1), decreasing to four away from the basal bodies. In Bracteacoccus sp., each proximal fiber is a single unit, the sinister rootlets are four (3/1) or rarely five (4/1) microtubules, and each basal body is associated with an unusual curved structure. The basic features of the flagellar apparatus of the zoospores of these two algae resemble those of Heterochlamydomonas rather than most other chlorococcalean algae that have equal length flagella, basal bodies in the V-shape arrangement, and clockwise absolute orientation. It is proposed that these algae with unequal flagella and parallel basal bodies have a shared common ancestry within the green algae.     

Studies on the fine structure of teleost blood cells

Summary Electron microscopic study of nucleated erythrocytes of the goldfish, Carassius auratus, reveals the microtubular elements comprising the marginal band which encircles the cell. Six to ten units are visible at each pole of the cell, immediately within the plasmalemma. Each tubular unit is composed of an electron dense membrane enclosing a less dense core. Cross-sectional units average 264 Å outer diameter, whereas tubules measured in longitudinal sections average 237 Å.The functions of the microtubules of the marginal bands are analyzed in view of Meves' original interpretation of maintenance of the discoidal form of the nucleated erythrocyte, and the more recent investigations in cell physiology of Trotter and Tilney. It is proposed that the microtubules possess a dual function: the support of the cell which is attributed to the hydroelastic properties of the turgid microtubules resulting from intratubular hydrostatic pressures; and the intracellular transport of materials via the intratubular fluid. The microtubules may, therefore, be considered as a skeletal system and part of an intracellular circulatory system.This project was supported by grants 2 G-895 and 2 G-505 from the United States Public Health Service.     

THE FINE STRUCTURE OF THE AXON AND GROWTH CONE OF THE DORSAL ROOT NEUROBLAST OF THE RABBIT EMBRYO

The centrally directed neurite of the dorsal root neuroblast has been described from the period of its initial entrance into the neural tube until a well-defined dorsal root is formed. Large numbers of microtubules, channels of agranular reticulum, and clusters of ribosomes are found throughout the length of the early axons. The filopodia of the growth cone appear as long thin processes or as broad flanges of cytoplasm having a finely filamentous matrix material and occasionally small ovoid or elongate vesicles. At first the varicosity is a small expansion of cytoplasm, usually containing channels of agranular reticulum and a few other organelles. The widely dilated cisternae of agranular reticulum frequently found within the growth cone probably correspond to the pinocytotic vacuoles seen in neurites in tissue culture. The varicosities enlarge to form bulbous masses of cytoplasm, which may measure up to 5 µ in width and 13 µ in length. They contain channels of agranular reticulum, microtubules, neurofilaments, mitochondria, heterogeneous dense bodies, and a few clusters of ribosomes. Large ovoid mitochondria having ribonucleoprotein particles in their matrix are common. Dense membrane specializations are found at the basal surface of the neuro-epithelial cell close to the area where the early neurites first enter the neural tube.     

The 3-dimensional fine structure of the mitotic spindle in Trypanosoma cruzi

The fine structure of nuclear division in the hemoflagellate Trypanosoma cruzi has been studied with serial sections and three-dimensional reconstructions of each divisional stage. After a preliminary stage in which the chromatin becomes dispersed, there is an equatorial stage defined by the appearance of an arranged set of ten dense plaques located about the equatorial region of the nucleus. At this stage a regular microtubular spindle is formed in the nucleus. Each plaque has a symmetrical structure formed by transverse bands and the bands are formed by tightly packed fibrillar material. The wide faces of the plaques are associated with tangential microtubules coming from the poles while the front and rear edges are free to associate with chromatin. Although structural continuity between chromatin fibers and the material of the plaques is possible, this continuity has not been proved. The equatorial spindle is formed by about 120 microtubules arranged in two sets of about 60 microtubules running from each pole to the dense plaques and divided into discrete bundles which reach a single plaque. The microtubules of each bundle may pass tangential to the wide faces of the plaque and end about 0.2 m beyond it, or they may end at the pole-facing edges of the plaque. No continuous, interpolar microtubules were observed at this stage. At the beginning of the elongational stage the dense plaques split into halves and each set of half-plaques migrates to one pole. During mid-elongational stage the pole-converging microtubules and the polar bulges disappear and microtubules become rearranged between the two sets of half-plaques. During late elongational stages, continuous microtubules run between the two sets of half-plaques and maximum nuclear elongation is attained. Chromatin remains dispersed throughout nuclear division. Two main movements have been observed in these mitotic nuclei: the migration of half-plaques to the poles and the elongation of the nucleus. Both these movements are accompanied by large changes in the architecture of the microtubular spindle and are probably dependent on microtubular function. It is concluded that the dense plaques play a kinetochore-like role and thus T. cruzi would have ten chromosomal units.