Osmiophilic Core Research Articles

An Introduction to Loricifera, Cycliophora, and Micrognathozoa

Loriciferans, cycliophorans and micrognathozoans are amongst the latest groups of animals to be discovered. Other than all being microscopic, they have very different body plans and are not closely related. Loriciferans were originally assigned to the Aschelminthes. However, both new molecular and ultrastructural researches have shown that Aschelminthes consist of two unrelated groups, Cycloneuralia and Gnathifera. Cycloneuralia may be included in the Ecdysozoa, including all molting invertebrates, and Gnathifera are more closely related to Platyhelminthes. The phylum Loricifera shares many apomorphic characters (e.g., scalids on the introvert) with both Priapulida and Kinorhyncha, and can be included in the taxon Scalidophora, a subgroup of Cycloneuralia. Cycliophora was originally allied to the Entoprocta and Ectoprocta (Bryozoa) based on ultrastructual research. Subsequent molecular data show they may be related to Rotifera and Acanthocephala, within the taxon Gnathifera. The phylogenetic position of Cycliophora is therefore not settled, and more ultrastructural and molecular data are needed. Micrognathozoa is the most recent major group of animals to be described. They show strong affinities with both Rotifera and Gnathostomulida (within the taxon Gnathifera), especially in the fine structure of the pharyngeal apparatus, where the jaw elements have cuticular rods with osmiophilic cores. Furthermore the micrognathozoans have two rows of multiciliated cells that form a locomotory organ, similar to that seen in some gastrotrichs and interstitial annelids. This character is never seen in Rotifera or in the monociliated Gnathostomulida. Rotifera and Acanthocephala always have a syncytial epidermis (Syndermata). Micrognathozoa lack this characteristic feature. Therefore, they are postulated to be placed basally in the Gnathifera, either as a sister-group to Gnathostomulida or as a sister-group to Rotifera + Acanthocephala.

Micrognathozoa: A new class with complicated jaws like those of Rotifera and Gnathostomulida

A new microscopic aschelminth-like animal, Limnognathia maerski nov. gen. et sp., is described from a cold spring at Disko Island, West Greenland, and assigned to Micrognathozoa nov. class. It has a complex of jaws in its pharynx, and the ultrastructure of the main jaws is similar to that of the jaws of advanced scleroperalian gnathostomulids. However, other jaw elements appear also to have characteristics of the trophi of Rotifera. Jaw-like structures are found in other protostome taxa as well—for instance, in proboscises of kalyptorhynch platyhelminths, in dorvilleid polychaetes and aplacophoran mollusks—but studies of their ultrastructure show that none of these jaws is homologous with jaws found in Gnathostomulida, Rotifera, and Micrognathozoa. The latter three groups have recently been joined into the monophylum Gnathifera Ahlrichs, 1995, an interpretation supported by the presence of jaw elements with cuticular rods with osmiophilic cores in all three groups. Such tubular structures are found in the fulcrum of all Rotifera and in several cuticular sclerites of both Gnathostomulida and Micrognathozoa. The gross morphology of the pharyngeal apparatus is similar in the three groups. It consists of a ventral pharyngeal bulb and a dorsal pharyngeal lumen. The absence of pharyngeal ciliation cannot be used as an autapomorphy in the ground pattern of the Gnathifera because the Micrognathozoa has the plesiomorphic alternative with a ciliated pharyngeal epithelium. The body of Limnognathia maerski nov. gen. et sp. consists of a head, thorax, and abdomen. The dorsal and lateral epidermis have plates formed by an intracellular matrix, as in Rotifera and Acanthocephala; however, the epidermis is not syncytial. The ventral epidermis lacks internal plates, but has a cuticular oral plate without ciliary structures. Two ventral rows of multiciliated cells form a locomotory organ. These ciliated cells resemble the ciliophores present in some interstitial annelids. An adhesive ciliated pad is located ventrally close to a caudal plate. As in many marine interstitial animals—e.g., gnathostomulids, gastrotrichs, and polychaetes—a special form of tactile bristles or sensoria is found on the body. Two pairs of protonephridia with unicellular terminal cells are found in the trunk; this unicellular condition may be the plesiomorphic condition in Bilateria. Only specimens with the female reproductive system have been found, indicating that all adult animals are parthenogenetic females. We suggest that 1) jaws of Gnathostomulida, Rotifera, and the new taxon, Micrognathozoa, are homologous structures; 2) Rotifera (including Acanthocephala) and the new group might be sister groups, while Gnathostomulida could be the sister-group to this assemblage; and 3) the similarities to certain gastrotrichs and interstitial polychaetes are convergent. J. Morphol. 246:1–49, 2000 © 2000 Wiley-Liss, Inc.

Are the most numerous sensilla of terrestrial isopods hygroreceptors? Ultrastructure of the dorsal tricorn sensilla of Porcellio scaber

The ultrastructure of the tricorn sensilla of the woodlouse Porcellio scaber was investigated in cryofixed and freeze-substituted, or chemically fixed specimens. The tricorn sensilla have a foramenized triangular-shaped outer hair and bear a poreless rod-like inner hair. The conical base of the inner hair is connected to the base of the outer hair by a complex cuticular structure. Each sensillum contains three sensory cells. The tip of one of the three dendrites contains a tubular body and is clamped between two bulges of the dendritic sheath. The two other dendrites protrude to the tip of the inner hair, flush against the cuticular wall. The microtubules in the ciliary segments are arranged in nine double tubuli that have neither osmiophilic cores nor arms. The ciliary rootlets are small. The inner segment of the largest dendrite wraps around the two smaller dendrites and one of seven enveloping cells in a mesaxon-like manner. Although this ultrastructure deviates considerably from most crustacean mechanosensitive sensilla, it nevertheless suggests a mechanosensitive function, at least for one of the sensory cells. In many aspects, the tricorn sensilla resemble the thermo-hygrosensilla of insects. However, our results suggest that the structural criteria for thermo-hygro-sensitivity used in insects cannot simply be applied to crustaceans.

Endosperm Development in Solanum nigrum L. Formation and Distribution of Lipid Bodies

The formation and distribution pattern of lipid bodies was investigated in spring and summer-grown seeds of Solanum nigrum. L. Lipid bodies were first formed in endosperm cells abutting the young globular embryo. The initial distribution pattern resembled an inverted cone with the 'tail' of the cone pointed away from the embryo. As the seed developed, lipid body formation commenced in the endosperm cells immediately adjacent to the cone and then in the remaining endosperm cells. RER cisternae were frequently found appressed to one side of the lipid body and sometimes continuous with the electron-lucent zone which separated the electron dense osmiophilic core from the surrounding half-unit membrane. The electron-lucent zone was of variable width and more noticeable in spring-grown seeds. In summer-grown seeds only, the osmiophilic cores of adjacent lipid bodies were joined by bridges of similar osmiophilic material. When the cotyledons of the torpedo-stage embryo had curved (approx. 14 d after anthesis), the osmiophilic stainability of the lipid bodies began to disappear as they were converted into oleosomes. Glyoxysome-like organelles were found only in the endosperm's 'Zone of Separation and Secretion' and arose earlier in the spring-grown seeds than in the summer-grown seeds. The early formation of lipid bodies, the spatial and temporal deposition pattern and the occurrence of glyoxysome-like organelles only in the Zone of Separation and Secretion suggests that the lipid bodies act as a reserve to be utilized during periods of uneven or low transfer of photosynthetic products.

Compartmentalization of monoaminergic synaptic vesicles in the storage and release of neurotransmitter.

Monoaminergic nerves are characterized by the presence of a population of small synaptic vesicles (40-60 nm in diameter) containing a few large vesicles (80-90 nm in diameter). Thus, although both types of vesicles contain monoamines, the small vesicles must be considered as the organoid responsible for the storage and release of the neurotransmitter, whereas the large ones possibly are involved in the modulation of the process. The small vesicles are electron-lucent or have an osmiophilic electron-dense core that is always linked to the vesicle membrane. Considering morphological and histochemical evidence under different experimental conditions, we proposed the existence of two compartments in the small vesicles: the core and the matrix, corresponding respectively to the electron-dense core and the electron-lucent space between the core and the vesicle membrane in osmium tetroxide fixations. The sizes of both compartments are inversely related, i.e., the smaller the core, the larger the matrix and vice versa. The core even disappears, giving way to a small electron-lucent vesicle made exclusively by the matrix. Thus, the matrix is a constant component of the vesicle, whereas the core is a transient one. Each compartment has a different pool of amine: a loosely bound, easily releasable pool in the matrix and a tightly bound, more resistant pool in the core. These two pools subserve, respectively, a tonic or phasic release of the neurotransmitter, correlated with a tonic or phasic stimulation of the receptor. The core may be considered as a storage or reserve pool. Experimental evidence from our laboratory supports the concept that different mechanisms are operative in both compartments in the release of the neurotransmitter. For instance, a Ca2(+)-independent release would be primarily concerned with the neurotransmitter contained in the matrix, and a Ca2(+)-dependent efflux would be primarily related with the neurotransmitter stored in the core. However, it still must be established that a simple relationship exists between each kind of stimulus and each vesicle compartment, rather than both compartments being integrated in a dynamic functional unit.

Heat shock induced ultrastructural alterations in Stylonychia mytilus cells.

Fine structural observations on heat shocked cells of S. mytilus reveal that cell organelles undergo structural alterations. Mitochondria show distorted shapes with disorganized cristae and vacuolar spaces. Pulse heat shock results in dilated rough endoplasmic reticulum, abundant polysomes as well as smooth endoplasmic reticulum. Heat shocked cells show membrane bound bodies containing osmiophilic cores. In macronuclei, dense chromatin breaks up into discrete bodies accompanied by the appearance of bundles of fine filaments and clustering of nuclear pores. The most prominent changes are noticed in nucleoli. Within 15 min of heat shock, nucleoli show hypertrophy and fine fibrillar zone which gradually replaces the granular zone by 120 min giving the nucleoli ring shaped configuration. In S phase cells, macronuclei show the arrested replication band in which the diffused zone (the site of DNA replication) is absent.

Ultrastructural and immunohistochemical study of endocrine cells in the midgut of the cockroach Blaberus craniifer (Insecta, Dictyoptera)

The midgut of Blaberus craniifer is principally made up of columnar epithelial cells which are derived from small regenerative cells found grouped in nidi. Between them, small sparsely granulated cells with clear cytoplasm can be observed lying on the basal lamina. Mainly based on the size, shape and texture of their secretory granules, at least ten types of such endocrine cells have been identified. Five cell types contain a uniform population of dense granules: (1) medium-sized, round to oval granules; (2) small elongated granules; (3) large irregular granules; (4) oval granules with a highly osmiophilic core; (5) oval, haloed granules. Five others are characterized by a heterogeneous population of granules: (6) small, round to oval, variably electron-dense granules; (7) oval medium-sized granules of variable electron density; (8) large irregular granules of variable electron density; (9) small dense granules and large vesicles with filamentous material; (10) small dense granules and very large pale vesicles.

Are the funnel-canal organs the 'campaniform sensilla' of the shore crab, Carcinus maenas (Decapoda, Crustacea)? II. Ultrastructure.

The funnel-canal organs on the dactyls of the shore crab, Carcinus maenas, are innervated by 3-24 sensory cells with unbranched dendrites, which attain a length of 500-1400 micron. The outer dendritic segments are enclosed in a dendritic sheath and pass through the cuticle within a canal. Two dendrite types can be distinguished according to ultrastructural criteria: Type I has a long ciliary segment, A-tubules with an osmiophilic core and arms, and a thick ciliary rootlet. Type II possesses only a short ciliary segment and a thin ciliary rootlet. Each funnel-canal organ contains two type-I dendrites. Their ciliary bases appear a few micron distal to those of the type-II dendrites (1 to 22 in number). Two inner and two to eight outer enveloping cells belong to a sensillum. The innermost enveloping cell contains a large scolopale. In the second enveloping cell single scolopale rods are present. Thus, the funnel-canal organs are characterized by structural features typical for mechanosensitive scolopidia, on the one hand, and for chemoreceptors, on the other. Therefore, the funnel-canal organs are very likely bimodal sensilla (contact chemoreceptors). A comparison with other arthropod sensilla shows that cuticular mechanoreceptors of aquatic crustaceans generally exhibit a 'scolopidial' organization.

Ultrastructure of the pyloric endocrine cells in the cartilaginous fishScyliorhinus stellaris

Abstract In the pyloric mucosa of Scyliorhinus stellaris at least eight cell types can be identified, showing well-differentiated ultrastructural features, especially in the secretory granules. With semithin/ultrathin techniques only some of these cells are easily demonstrable. Type I and Type II cells can be considered as enterochromaffin (EC) cells since both are positive to argentaffin reaction, but the ultrastructural features of the granules are quite dissimilar. Type III cells could not be found in the semithin sections but the aspect of the granules is like that in the intestinal EC cells. Type IV cells contain round vesicular granules with an eccentric, highly osmiophilic core in which the number and size of the granules can vary from cell to cell. This cell type is positive to the Grimelius silver reaction and may be equivalent to the gastrin storing or gastrin-cholecystokinin storing cell. Type V cells contain round granules with a clear halo showing a weakly positive Grimelius silver reaction. Histochemically this cell type resembles the pancreatic A cell but the granule size is slightly different. Type VI cells are characterized by argyrophilic and non-argentaffin granules, round or rod-shaped. Type VII and Type VIII cells show similarities with two pancreatic cell types. The Type VII may be a D cell. All pyloric endocrine cells except cell Type III are of Fujita's “open” type.

Ultrastructural evidence for monoamine uptake by vesicles of pineal sympathetic nerves immediately after their stimulation.

Bilateral electrical stimulation of the preganglionic fibers to the superior cervical ganglia of the rat markedly reduces the number of osmiophilic dense cores present in the nerve vesicles of the sympathetic fibers in the pineal gland. These cores owe their density to the presence of noradrenaline and serotonin in the vesicles. When sympathetic nerves of the pineal organ are exposed immediately after stimulation for a brief period to the false neurotransmitter 5-hydroxydopamine, either in vitro or in vivo, dense precipitates reappear in the electron-lucent vesicles. On the basis of these observations, it is concluded that the vesicles remaining in the nerves after releasing their neurotransmitter content have the capacity to take up and store monoamines. This provides a morphological correlate for the recent biochemical evidence suggesting that the vesicles in sympathetic nerves are reused after neurotransmitter release.

Intercellular granules and vesicles in prolonged cerebral vasospasm.

Prolonged vasospasm was produced in the dog basilar arteries by introduction of fresh arterial blood or norepinephrine into the chiasmatic cistern. Myonecrosis was limited to a small number of smooth-muscle cells, and a large number of muscle cells appeared intact. The most characteristic change was the appearance of aggregated granules and vesicles in the widened extracellular space between smooth-muscle cells, particularly near the adventitia and the elastic lamina. The granules were spherical, 50 to 100 nm in diameter, often surrounded by a single membrane, and contained a dense osmiophilic core, about 40 to 90 nm in diameter. In addition, the elastic lamina often became loose and corrugated or broken.

Morphological study of experimental cerebral vasospasm.

Canine basilar artery was studied in vivo and in vitro. Cell membranes of vascular smooth muscle were differentiated into caveolae intracellulares zones and caveolae-free zones, both of which were generally arranged in an alternate manner and parallel to the long axis of the smooth muscle cell. The caveolae intracellulares, about 60 nm in diameter, were neatly oriented in one to several rows with a center-to-center distance of about 80 nm. Adherens, gap, and tight junctions were occasionally found in the smooth muscle. In vitro study of canine basilar artery suggested the necessity of extracellular Ca supply for prolonged contraction, and strontium was selectively deposited in sarcoplasmic reticulum and mitochondria. Prolonged vasospasm was produced in the canine basilar artery by the injection of fresh blood or norepinephrine into the chiasmatic cistern. The caveolae intracellulares of vascular smooth muscle were found to be increased in number in vasospasm. In thin sections, myonecrosis was found in a small number of muscle cells, while a large number of muscle cells appeared intact. The most characteristic change was the appearance of aggregated granules and vesicles in the widened extracellular space between smooth muscle cells, particularly in the periphery of muscular layer and beneath the elastic lamina. The granules were spherical, 50-100 nm in diameter and were often surrounded by a single membrane and contained a dense osmiophilic core of about 40-90 nm diameter. In addition, the elastic lamina often became loose and corrugated or broken.

Light and Electron Microscopical Characterization of Heterophilic Granulocytes in the Intestinal Wall and Islet Parenchyma of the Hagfish,Myxine glutinosa(Cyclostomata)

Heterophilic granulocytes were studied in the blood, intestinal wall, and islet parenchyma of the Atlantic hagfish (Myxine glutinosa) by light and electron microscopical methods. The granulocytes are pseudoeosinophils and show a PAS-positive cytoplasmic reaction. Ultrastructurally, the cells contain evenly distributed pleomorphic cytoplasmic granules with the granule membrane close to the osmiophilic core. Emigrated blood granulocytes are found extra-vascularly in the submucous connective tissue, and obviously they can pass the basal lamina and migrate into the epithelium of the intestine, bile duct, and islet parenchyma. Though the staining characteristics of hagfish granulocytes are different from those of endocrine cells in the intestinal mucosa and islet parenchyma, intraepithelial granulocytes in some locations may sometimes be difficult to distinguish ultrastructurally from insulin-containing B-cells, since heterophil granules have both a size and a shape close to those of secretion granules in B-cells. However, in contrast to B-cells the granulocytes show the following ultrastructural features: a lobated nucleus with peripherally arranged electron-dense chromatin; cytoplasmic processes and often rod-like granules with no clear space between the granule membrane and core; prominent cytoplasmic vacuoles and microtubules; and sparse mitochondria and endoplasmic reticulum. Furthermore, immigrated granulocytes lack desmosomes and annulate lamellae. Some of the intraepithelial granulocytes in the mucosa show signs of disintegration and cell death. Degenerative cell processes are also described in the islet parenchyma.

Ultrastructure of early development of mouse parthenogenones.

AbstractMouse eggs were parthenogenetically stimulated with hyaluronidase and osmotic shock and the resulting two‐, four‐ and eight‐cell stage embryos examined with the electron microscope. The ultrastructure of parthenogenones during cleavage showed numerous differences when compared with normal development. Cortical reaction was absent and there were no microvilli on the cell surface of developing blastomeres. The cytoplasm contained numerous bundles of fibrous material, vacuoles and lipid droplets, multivesicular bodies, and agglomerates of vesicles sometimes connected with cortical granules. Cortical granules and vacuolated mitochondria were numerous and usually situated near the cell membrane. The number of free ribosomes was very small and agglomerates of ribosomes or rough endoplasmic reticulum were not observed. Nucleoli in almost all cells were so‐called primary nucleoli consisting of a dense, osmiophilic core and narrow ring of granular material. In only a few cells was reticulation of nucleoli and formation of fibrillar and granular parts noticed. Nuclear fragments and nuclei with broken membranes were present in some cells. Primary nucleoli persisted during mitosis and were in close contact with chromosomes. Cytokinesis was irregular and small and large fragments of cytoplasm surrounded by membrane were often observed between dividing cells. Our results suggested that failure of parthenogenones to develop normally is probably caused by numerous disturbances during early cleavage. Incomplete cortical reaction, persistence of primary nucleoli, a probable disturbance in RNA and protein synthesis, and irregularities during nuclear and cellular division were observed in all embryos.

Storage of ATP and 5-hydroxytryptamine in blood platelets of guinea-pigs.

1. By density gradient centrifugation of homogenized blood platelets of guinea-pigs, subcellular organelles have been isolated which exhibit much higher concentrations (mumole/mug protein) of ATP than the whole platelets.2. The organelles also store endogenous 5-HT whose concentration is, however, considerably inferior to that of ATP.3. Investigated by electron microscopy most of the isolated organelles look empty and only very few show dense osmiophilic cores indicating the presence of 5-HT. Accordingly the occurrence of dense osmiophilic organelles in the whole platelets is also very rare.4. Exogenous 5-HT taken up by the platelets markedly accumulates in the subcellular fraction containing the organelles storing ATP. Concomitantly, the platelets show a rise in the number of osmiophilic organelles and the frequency of osmiophilic cores in the isolated organelles is increased.5. From these and previous in vitro experiments it is concluded that the so called 5-HT storage organelles of blood platelets primarily store ATP and only subsequently accumulate 5-HT.

The innervation of the frog's heart. 3. Electronmicroscopy of the autonomic nerve fibres and their vesicles.

The electronmicroscopie appearance of the autonomic nerve fibres in the frog’s heart has been examined. The parasympathetic postganglionic fibres are typically cholinergic and have only a few cored vesicles amongst numerous small agranular ones. The sympathetic postganglionic fibres contain adrenaline and it is present in two types of vesicle, one is large (90 nm) and has an osmiophilic core, whilst the other is smaller (40 nm) and has no core unless the adrenaline is stained by the sodium dichromate/acrylic aldehyde method. There are also two types of noradrenaline-containing vesicles in mammalian nerves. These are compared with the vesicles in the frog nerves: there appear to be no differences other than the catecholamine stored.

Electron microscopic observations of visna virus-infected cell cultures

Electron microscopic observations of three cell lines infected with visna virus revealed two types of extracellular particles. The smaller of these was 65–110 mμ in diameter and contained a 20–30 mμ electron-dense core. Ordered arrays of the latter type of particle occurred rarely in the cytoplasm. After cesium chloride density gradient centrifugation of the virus, the band that contained maximal infectivity was composed of numerous particles with osmiophilic cores similar to those found in infected cell cultures. This finding suggests that such particles represent the infective agent. The second type of extracellular particle was larger (100–140 mμ in diameter), lacked an electron-dense core, and contained material similar in appearance to cellular cytoplasm. This form appeared to develop by budding from the cell surface.

Auerbach's plexus of mammals and man: electron microscopic identification of three different types of neuronal processes in myenteric ganglia of the large intestine from rhesus monkeys, guinea-pigs and man.

Ganglia from Auerbach's plexus of the large intestine (caecum, appendix vermiformis, colon transversum and rectum) in man, rhesus monkey and guinea-pig are composed of nerve cells and their processes, typical Schwann cells and a vast neuropil. The neuropil consists of dendrites and axons of intrinsic nerve cell perikarya and axons of extrinsic neurons. Axonal profiles in large nerve fibre bundles are of uniform size and appearance, embedded in infoldings of Schwann cell cytoplasm and contain occasional large granular vesicles, mitochondria and neurotubules. Preterminal axons widen into vesicle filled varicosities, some of which establish synaptic contact with intrinsic nerve cell bodies. At least three different types of neuronal processes can be distinguished in the myenteric neuropil according to the size, appearance and commutual proportion of vesicles present in axonal varicosities, and their ability to accumulate exogenous 5- and 6-hydroxydopamine and 5-hydroxydopa: 1. Axonal enlargements containing a major population of small electron lucent “synaptic” vesicles (350–600 A in diameter) together with a small number of membrane-bound, opaque granules (800–1,100 A). These profiles have been identified as “cholinergic” axons. The boutons establish synaptic contacts with dendritic processes of intrinsic nerve cell bodies; membrane specializations are found at the preand postsynaptic sites. 2. Axonal beads of sometimes very large diameter, containing an approximately equal amount of large granular vesicles (850–1,600 A) and small, electron lucent or faintly opaque vesicles (400–600 A). The granular core of the large vesicles is of medium electron density and may either fill the entire vesicle or is separated from the limiting membrane by a more or less clear interspace. The fibres probably belong to intrinsic neurons, and because of the similarity of the large, membrane-bound vesicles with neurosecretory elementary granules, they have been designated “p-type fibres” (polypeptide fibres). The granular core of the vesicles in these fibres becomes more electron dense after treatment with 5-OH-dopa. The accumulation of an amine precursor analogue in combination with a possible storage of a polypeptide substance (or an ATP-like substance) resembles the situation in several diffusely distributed endocrine cell systems. 3. Varicosities of axons equipped with small (400–600 A) empty or sometimes granular vesicles, medium sized (500–900 A) vesicles with highly electron dense cores and occasional large (900–1,300 A) granular vesicles. Pretreatment with 5-OH-dopamine increases the electron density in almost all medium-sized granular vesicles and some of the large granular vesicles; an osmiophilic core develops in some small vesicles. 6-hydroxydopamine results in degenerative changes in the varicosities of this type of neurons. Concomitantly, both catecholamine analogues markedly reduce neuronal noradrenaline in the large intestine, as demonstrated by fluorescence histochemistry and in fluorimetric determinations. The ultrastructural features of these varicosities and their reaction to 5- and 6-OH-dopamine indicate that they belong to adrenergic, sympathetic nerves. No membrane specializations could be detected at sites of close contact of the adrenergic boutons with dendrites and cell bodies of intrinsic nerve cells.

Observations on the ultrastructure of nerve cells in the brain of the earthworm, Eisenia foetida, with special reference to neurosecretion

The submicroscopic structure of nerve cells in the brain of the earthworm Eisenia was studied. Six types of neurons containing morphologically different inclusions are identified. Types 1, 2 and 3 contain vesicles filled with homogeneous materials of high electron density. These are essentially similar to “elementary granules” in neurosecretory systems of vertebrates and invertebrates. Type 4 shows dense-cored vesicles which resemble in size catechol-containing granules as described, for example, in chromaffin cells of the adrenal gland. Type 5 has clear vesicles with a mean diameter of 400 A. Some of these vesicles have a dense osmium deposit. Type 6 contains electron lucent vesicles with diameters of 500–800 A. Occasionally these have osmiophilic cores. Clear vesicles of types 5 and 6 are similar to classical synaptic vesicles, while “granulated vesicles” resemble in size and appearance those described in adrenergic nerve endings. All six vesicle types have the same mode of origin from Golgi membranes. All of these vesicles are considered to be discharged from the perikarya into the axons entering the neuropil.

HISTOCHEMICAL AND ULTRASTRUCTURAL STUDIES ON HELA CELL CULTURES EXPOSED TO SPINDLE INHIBITORS WITH SPECIAL REFERENCE TO THE INTERPHASE CELL.

Mammalian cells of the HeLa (S3) strain, when exposed to spindle inhibitors, have been found to undergo several morphological transformations during interphase as well as during mitosis. Some of these have been studied both histochemically and ultrastructurally. The lysosomes, represented by the multivesicular bodies in HeLa cells, form clusters and become circumferentially disposed instead of occupying the polarized juxtanuclear position characteristic of these organelles. In the electron microscope it is seen that they have acquired a dense osmiophilic core that is separated from the bounding unit membrane by an electron lucent halo. The Golgi apparatus fragments under the influence of spindle inhibitors and also takes up a circumferential distribution in a pattern similar to that of the lysosomes. On the ultrastructural level, no significant modifications in this organelle are seen. Also noted in the interphase cell are numerous 60-80 Å fibrils coursing through the cytoplasm as well as a paucity of spindle our microtubules. A striking similarity has been pointed out between the behavior of the lysosomes in the drugtreated interphase cell and the untreated, normal mitotic cell. A possible explanation of some of the changes noted has been given in terms of an interruption of protoplasmic flow resulting from the disappearance of microtubules.