Ultrastructure of hyphae and ascospores in the genus Eremascus eidam

Hyphae and ascospores of Eremascus fertilis and E. albus were studied in ultrathin sections. The lateral wall of the hyphae had a thick electron-light inner layer and a thin dark outer layer. The septa had a simple central pore with or without a plug, and there were Woronin bodies in the vicinity. The wall of the ascospores of E. fertilis showed a thick light inner layer and a thin dark outer layer. In the wall of the spores of E. albus a dark fibrillar layer was present between the light inner layer and the dark outer layer. The spores of this species germinated with a tube the wall of which was continuous with a newly formed layer inside the spore wall.This investigation was supported by the Netherlands Organization for the Advancement of Pure Research (Z. W. O.)     

Asymmetries in the Machband phenomena

The Mach bands are directly related to the size and the shape of on-center off-surround neural units in human vision. The effects of various stimulus parameters were studied on both bright and dark bands of equal plateau intensities. At low overall intensities, the dark band increases markedly in width, while the bright band does not. However, the bandwidth is more affected by the brightness slope, than by the plateau intensity per se. In this case, both bands vary approximately linearly and inversely with the log of the slope. The bright bands are slightly wider (4′) than the dark bands, for matched intensities. Both bands almost double in width with only a ±30′ para-foveal fixation. Optical blur enlarges the bands as predicted from the spread function. A comparable enlarging effect found with pupil size increase is not so readily understood. The apparent centers of the bright bands are positioned significantly more asymmetrically between the two edges than are the dark band centers. Eccentric neural units are considered as possible explanations for some of these non-linearities. Supported, in part, by Research Grant No. EY00319-05 from the National Eye Institute, National Institutes of Health, Bethesda, Maryland, and by a fight for Sight Grant-in-Aid G-428 from the National Council to Combat Blindness, Inc., New York, New York.     

Comparative cytology and taxonomy of theUlvaphyceae II. Ulvalean characteristics of the stephanokont flagellar apparatus ofDerbesia tenuissima

Summary The stephanokont flagellar apparatus of the zoospores ofDerbesia tenuissima (De Not.) Crouan is examined and compared to the flagellar apparatuses of other green algae. The flagella ofDerbesia are attached to two of three bands which lie at the junction of the body and papilla. Serial longitudinal and cross sections reveal that the basal bodies are attached to the bands along their sides and at their proximal ends. The bands are not striated in any plane. The lack of striation in the bands and the partial covering of the proximal end of the basal bodies by one of the bands closely resemble the type of flagellar connection system described as the Bryopsis-type byMelkonian (1980). Zoospores of ulvalean green algae also possess these features, suggesting that green siphons are phylogenetically related to theUlvales. It is proposed that green siphons be tentatively classified in theUlvaphyceae rather than in theChlorophyceae orCharophyceae.This work supported by NSF Grant DEB 78-03554.     

Ultrastructure of the ascospores of some species of the Torulaspora group

Development and germination of the ascospores in species of the Torulaspora group of yeasts have been described. Most species had warty spores which, in sections, showed a dark outer layer consisting of the outer unit membrane of the prospore wall and a layer underneath formed at an early stage of development of the spores. In mature spores the light inner layer of the wall was delimited at the outside by a thin dark layer. The warts often contained dark material. The ascospores of two Pichia and three Debaryomyces species were studied for comparison; they differed in sections from the Torulaspora spores. The taxonomic implications of the ultrastructural observations have been discussed.     

Vacuolar protein in apical and flower-petal cells

Summary Vegetative apices, floral apices and flower petals of five Solanaceae (potato, tomato, tobacco, petunia and nightshade) and of corn and Nigella were examined with an electron microscope for the presence of protein bodies in the cell vacuoles. Electron-dense bodies were found in vacuoles of all plants investigated but not in every tissue examined. The bodies observed in the apices are similar to the protein bodies previously found in tomato leaves where they appear to be related to the presence of chymotrypsin inhibitor I protein (Shumway et al., 1970). The bodies appeared in very young cells in small vacuoles, disappearing as the cell matured. They are apparently related to the growth and development of the new cells. The results suggest that plants may regulate specific proteins within the apical region through selective synthesis and degradation of proteins accompanied by compartmentalization in the vacuole.Scientific Paper No. 3822, College of Agriculture, Washington State University, Pullman, Project 1791. This investigation was supported in part by the State of Washington Initiative Measure 171 funds, the Graduate School Research funds, by the U.S. Department of Agriculture, Cooperative State Research Service Grant 915-15-29, and U.S. Public Health Service Grant 2K3-GM-17059.Program in Genetics and Department of Botany.Program in Genetics.     

Ultrastructural observations on spermiogenesis in the peanut worm,Themiste pyroides (Chamberlin, 1920) (Sipuncula; Sipunculidea)

Summary

The process of spermiogenesis and the ultrastructure of the spermatozoa in the peanut worm, Themiste pyroides, from the Sea of Japan were observed with electron microscopy (SEM and TEM). The testes are composed of groups of spermatogonia and are covered by peritoneal cells. Clusters of spermatocytes are released from the testes into the coelomic fluid. Connected by intercellular bridges, the spermatocytes within a given cluster develop asynchronously. Proacrosomal vesicles and a flagellum appear in spermatocytes. Spermatids in the clusters retain the intercellular connections. During spermiogenesis, the acrosomal vesicle, formed by coalescence of small proacrosomal vesicles in the basal part of the spermatid, migrates to the apical part of the cell to form a conical-shaped acrosome. The basal concavity lying above the nucleus is filled with subacrosomal substance. The midpiece contains four mitochondria, two centrioles, and some residual cytoplasm with dark glycogen-like granules. A peculiar annulus structure develops around the base of the flagellum. The distal centriole has a pericentriolar complex consisting of radially oriented elements. Before the spawning process, the spermatozoa are filtered throughout the ciliary nephrostomal funnel into the excretory sac of paired nephridia where they are stored for a short time. The sperm are released into the sea water via nephridiopores. Spermatozoa remaining in the coelomic fluid after spawning are resorbed by amoebocytes. This species from Vostok Bay is characterized by a prolonged spawning period from June to early October. The reproductive strategy of T. pyroides is discussed in comparison with that of Thysanocardia nigra, the latter having a unique pattern of packaging of the spermatozoa, resulting in the formation of spermatozeugmata, as a reproductive adaptation to the very short spawning period.     

A comparison of banding patterns in salivary gland chromosomes of two species ofDrosophila

An attempt was made to compare the details of chromosome structure between two distantly related species ofDrosophila, D. melanogaster andD. hydei, which belong to different subgenera. Several short stretches of salivary gland chromosomes were selected on the basis of presumed homologous markers, either mutants associated with chromosome breaks, or experimental puffs. Banding patterns were compared in map diagrams, compiled from photographs. It proved possible to correlate with fair accuracy at least a short sequence of bands in each of the examples studied. The map regions were however very different otherwise. It was not possible to judge in how far this is a consequence of major or very small rearrangements, or other types of change. The chromosomal location of one permanent puff and of several others some of which are formed normally during pupation, and some of which appear after a period of oxygen deprivation, was found to be in complete agreement with genetical data which indicate homology of chromosomes 2 (hydei) with3R (melanogaster), and of 4 with3L. On the other hand, relative position and sequence of these puffs within the chromosome are different in the two species. Supported by the Netherlands Organization for the Advancement of Pure Research, Grant 913-28.     

Studies of pollen maturation in cotton: the storage reserve accumulation phase

Summary This study follows the maturation of the pollen grain of cotton (Gossypium hirsutum L.), particularly the development of the vegetative cytoplasm and the various storage products formed. CTEM, HVEM, stereoscopy, and cyto-histochemistry were used to examine the events occurring during the 9 days before anthesis. Starch began to accumulate in plastids at anthesis minus 9 days and reached a peak concentration shortly before anthesis; lipid deposition followed a similar pattern, but started at 6 days before anthesis. Lipid bodies were always seen closely oppressed to the endoplasmic reticulum (ER). Dictyosomes appear active during the entire 9 days; first producing vesicles involved in the formation of the intine and, later, producing vesicles stored in the pollen grain. The dictyosome vesicles appear to contain polysaccharides and concentrate in layers around the lipid bodies. Ribosomes increase in number from 6 days before anthesis and are particularly numerous in the mature pollen. From anthesis minus 6 days until anthesis, the ER cisternae become increasingly inflated and, in the hours immediately before pollen release, form pockets filled with lipid bodies and dictysosome vesicles. The mature pollen has a core region filled with ER pockets and a peripheral cytoplasm in which such pockets are generally lacking.This research was supported in part by NSF Grant BMS575-22-23 and Grant N.RR-00592 from the Division of Research Resources, National Institutes of Health     

Determination of age in the Japanese monkey from growth layers in the dental cementum

The teeth of 14 Japanese monkeys (Macaca fuscata) were examined to establish an exact method of determining age by histological observation of dental cementum. The cementum showed annual growth layers, which were especially remarkable in the incisor root and in the molar cementum deposited at the junction of the roots. The layer of cementum formed in winter appears as a dark layer in stained sections and as a translucent layer in unstained ground sections. In the incisor the first dark and light layers are formed at the age of three years, whereas in the molar they do not appear at a definite age. The layers are thick and clear in the upper medial incisor. As a result, the age of a Japanese monkey can be determined by adding two to the number of dark layers and an outer light layer. It is interesting that the formation of the cementum of the first molar begins a few years after its eruption. The relation between this fact and the pressure of occlusion is discussed.     

Light and electron microscopy of virus inclusions inAmaranthus lividus cells

Summary Amaranthus plants infected with a virus of rod-shaped particles showed under the light microscope intracytoplasmic amorphous and crystalline inclusions.The submicroscopic organization of mesophyll cells from infectedAmaranthus leaves by electron microscopy is described. Besides big crystalline inclusions, long dark inclusions correspondent to needle-like inclusions observed by light microscopy are definable in the cytoplasm. The amorphous inclusion bodies were formed by an overgrown protrusion of vacuolate cytoplasm containing virus particles, long very dark stained inclusions forming dense bands and rings, normal elements of the cytoplasm such as mitochondria, endoplasmic reticulum and ribosomes, and some spherosomes. Inclusions and virus particles were not found in chloroplasts, mitochondria or nuclei of infected cells.     

Disintegration of H3-labelled spermatocytes in Melanoplus differentialis

Summary Whole cysts in the testicular tubes of Melanoplus differentialis disintegrate giving rise to large Feulgen positive bodies. The disintegration affects all the nuclei of a cyst.The Feulgen positive bodies are strongly labelled with tritiated thymidine, a confirmation of their DNA content.Spectrophotometric measurements reveal that the bodies contain 3.5 times more DNA per unit area than the autosomes of normal spermatocyte nuclei and an equal amount of DNA per unit area as the sex chromosome of the same nuclei.This regular disintegration of spermatocytes is not considered as a pathological condition but as an adaptation by which large amounts of DNA are easily released at a convenient time of development.Supported by a research grant from the Swedish Natural Science Research Council.     

Dark bands on pyritic internal moulds of the Early Jurassic ammonites Oxynoticeras and Cheltonia from Gloucestershire,England: interpretation and significance to ammonite growth analysis

Abstract: Thin, radiating, darker bands occur on pyritic internal moulds of the Early Jurassic ammonites Oxynoticeras and Cheltonia from Bishop’s Cleeve, Gloucestershire. They closely resemble true colour patterns preserved in Early Jurassic Calliphylloceras from Kutch, India, and false colour patterns reported in Carboniferous and Triassic ammonoids. Up to five dark bands occur within the body chamber, suggesting that they do not represent serially repeated anatomical structures, but the same feature repeatedly formed during growth. Dark bands are interpreted as traces of black bands deposited on the inside of the shell at the aperture during pauses in growth. The angles between dark bands and between septa correlate strongly in Cheltonia, suggesting that pauses in growth coincided with septal secretion during the chamber formation cycle. There are, however, no other indications that growth was episodic in either genus.     

Comparison of developmentally controlled glucoamylase from normal and mutant strains of the basidiomycete Schizophyllum commune

During the formation of fruit bodies, the basidiomycete Schizophyllum commune produces glucoamylase activity. DEAE-cellulose chromatography resolves the enzymic activity into a major peak found early in development and a minor peak which appears when the fruit bodies are mature. A mutant homokaryon has been isolated which constitutively produces glucoamylase activity without any fruiting. When purified, the mutant enzyme and the major fruit body enzyme appear to be identical by several physical and kinetic parameters including molecular weight, temperature sensitivity, and K_m.Supported in part by NIH Research Grant AI 09779-03 from the National Institute of Allergy and Infectious Diseases.     

Distribution of constitutive heterochromatin in mammalian chromosomes

Using a special staining technique, a survey of the chromosomes of many mammalian species showed that constitutive heterochromatin is present in all cases and that the heterochromatin pattern appears to be specific and consistent or each chromosome and each taxon. Usually heavy heterochromatin is found in the centromeric areas, but terminal heterochromatin is not uncommon. Occasionally interstitial heterochromatin bands occur. In some species, such as the Syrian hamster and Peromyscus, many chromosome arms are completely heterochromatic.Supported in part by Research Grant GB-13661 from the National Science Foundation.     

Taxonomische Studien in der Gattung Xerula R. Mre. (IV)

The taxonomically important characteristics of Xerula melanotricha Dörfelt are constant in fruit bodies, also in such which have been formed in culture. Studies on the development of the fruit bodies in culture showed that the macrosetae are formed already in the primordium. At first they are hyaline and colourless. A dark fluid (guttation) is secreted by the macrosetae. This is a specific characteristic of Xerula melanotricha. The fluid has an influence on the colours of the fruit bodies. The American specimens of “Collybia longipes” are revised by Mitchel & Smith (1978). These autors consider all the American collections to be a special variety, “Oudemansiella longipes (St. Amans) Moser var. americana Mitchel et Smith”. Investigations of the holotypus and of other collections suggested that this variety is a well characterized, seperate species. This taxon is an endemit of the Southern Rocky Mountains. In this paper it is described as Xerula americana Dörfelt sp. nov.     

Histamine content of peritoneal and tissue mast cells of growing rats

Summary p-Phenylenediamine/pyrocatechol mixture (PPD-PC) was evaluated as a reagent for the ultracytochemical demonstration of retrograde axonal transport of horseradish peroxidase (HRP). HRP crystals were applied to the proximal stumps of the severed infraorbital nerves in rats. After 48 h the rats were sacrificed by perfusion, and the trigeminal ganglia ipsilateral to the severed nerves were processed for HRP cytochemistry and then prepared for electron microscopy. PPD-PC was rapidly oxidized in HRP-labeled neurons to form a dark brown-black osmiophilic reaction product which was more readily visible than the DAB product in the sections. This facilitated selection by light microscopy of areas in the epoxy wafers for ultrathin sectioning. In thin sections viewed under the electron microscope, the osmicated electron opaque PPD-PC reaction product was present in membrane-bound structures including smooth endoplasmic reticulum and granules of various sizes. The PPD-PC reaction product formed after 10-min incubation appeared to be more electron opaque than the DAB reaction product formed after 20 min. PPD-PC was found to be much less readily oxidized than DAB by endogenous hemoproteins. This methodology facilitated the ultracytochemical localization of HRP in neurons following retrograde axonal transport.Supported by NIH Grants DE 04730, DE 02668 and DE 00288 from the National Institute for Dental Research, NIH Grant RR 0533 from the Division of Research Facilities and Resources, and a grant to the Neurobiology Program from the Alfred P. Sloan Foundation     

Electron microscope study on meiosis

Summary The cycle of the nucleolus and sex chromosome was studied with the electron microscope during the following stages of spermatogenesis and spermiogenesis of Gryllus argentinus: (1) spermatogonia; (2) prophase cyte I, leptotene, part of pachytene, and end of diplotene till breakdown of the nuclear envelope; (3) division I, metaphase and anaphase; (4) cyte II, prometaphase; (5) division II, metaphase, anaphase and telophase; (6) early and late spermatids. Some observations were also carried out in the primary oocyte until beginning of the growth period.It was found that nucleolus and sex chromosome are associated, at first without mixture of their components (leptotene) and afterwards interchanging components (pachytene). The interchange takes place by the passage from one element to another of filamentous units ot low electron density, similar in appearance to those existing in the medial plane of tripartite groups (synaptinemal complexes).At pachytene the primary results of interchange are: (1) the nucleolus contains filaments of chromosomal nature; (2) the nucleolus emits a long rod-like prolongation containing a cylindrical bundle of filaments, and an axial unit of the same nature; two equidistant clear spaces separate the axial unit from the cylindrical bundle and the latter from the dark wall of nucleolar material. At the end of diplotene these components are found organized in two bodies and a prolongation. One of the bodies is formed by a number of alternatively dark and light bands, the other by a pack of tubular units each showing the structure of the former nucleolar prolongation. The prolongation is either formed as in the preceding stage or it is composed of five ribbons, two dark ones outside and three light ones between them. It is supposed that both bodies are united by the prolongation but no definite proof was obtained. It is assumed that the complex thus constituted represents the sex chromosome.The sex chromosome was found at any phase of both divisions as well as at the intermediate stages between them; at the division phase the chromosome is separated from the autosomes and moves independently of them.The element could not be traced at telophase II but it reappears within the reorganized nuclei of spermatids. Amorphous nucleolar-like material and chromosome-like material are found associated at this stage with banded complexes like those seen at the end of prophase I. All these components undergo involution during spermatid maturation. At the final step of maturation no traces of them are found.A similar association of nucleolus and chromosome was found at prophase of primary oocytes of the same species. The associated body is of the same structure as that described for primary spermatocytes. The structures existing in the primary oocytes disorganize at the beginning of growth. At this time the nucleolus has developed into a large body containing masses of chromatin-like material.This investigation was supported in part by U.S. Public Health Service, Research Grant No. GM-08337 from the National Institute of General Medical Sciences.     

In vivo shedding of apical plasma membrane in the thyroid follicle cells of the mouse

Summary Clusters of luminal dense bodies, limited by a triple-layered membrane, were found in all follicle lumina in thyroid glands of mice. After thyroxine treatment the number of luminal dense bodies increased, especially in the periphery of the lumen, where the intraluminal bodies often displayed a striking resemblance to microvilli. In hyperplastic goiters, obtained by feeding mice with propylthiouracil, luminal dense bodies were replaced by intraluminal vesicles. During goiter involution the vesicles were gradually replaced by luminal dense bodies; the presence of intermediate forms suggests that vesicles and dense bodies are basically the same formations. Luminal dense bodies were observed in colloid droplets indicating their removal by endocytosis. As demonstrated by electron-microscopic cytochemistry, luminal dense bodies contain a membranebound peroxidase, and electron-microscopic autoradiography after administration of ¹²⁵I indicate that they possess an iodinating capacity.Our observations on mouse thyroid glands suggest that the luminal dense bodies, which appear as vesicles in hyperplastic glands, are formed by shedding of the apical plasma membrane of the follicle cell. The shedding process might be of importance for the turnover of plasma-membrane material.This study was supported by Grant No. 12X-537 from the Swedish Medical Research Council.     

Variation in Drosophila acetylcholinesterase

We examined the Canton-S strain of Drosophila melanogaster for electrophoretic variation of the enzyme acetylcholinesterase. The pattern of bands obtained (stained with acetylthiocholine) depended on age, sex, and tissue (i.e., head vs. body part and hemolymph). However, through mixing experiments, it was concluded that most of these apparent differences were due to modification of the enzyme by unknown substances located in the fly's body. The electrophoretic pattern of head acetylcholinesterase was altered so that it became characteristic of the body which was present during extraction. For example, when heads of D. melanogaster were homogenized in an extract from D. lebanonensis bodies, the characteristic AChse bands of melanogaster were absent and instead the bands of lebanonensis were found. it was found that extraction of adult heads in 0.1 M potassium phosphate buffer alone or with a 2-min exposure to 1 mg/ml trypsin at 20 C gave the most reproducible results, independent of age and sex. Using these conditions, 25 strains of D. melanogaster and 30 strains of D. pseudoobscura were examined without finding any reproducible electrophoretic variant of acetylcholinesterase. In addition, 53 strains from 39 other Drosophila species produced a total of only six electrophoretic forms of the head enzyme. Additional electromorphs were found when whole flies were used, but these were not studied in detail because of the possibility that they could be due to postextraction modification.This study was supported by the National Research Council of Canada, Grant No. A0629 and A0235.     

Some observations on spermatogenesis in Gelastocoris oculatus (Hemiptera) with the aid of the electron microscope

The nuclear cap in the spermatogonial and early spermatocyte cells of Gelastocoris is an aggregate of closely packed mitochondria with their long axes perpendicular to the nuclear membrane. Eventually in the early growth period, the mitochondria move from the cap and appear to become more or less equally distributed in the cytoplasm where they remain until their fusion in the spermatid to form the nebenkern. The Golgi complex consists of clusters of lamellae and vesicles, the Golgi bodies. Granules form within the vesicles, increase in size, move from their place of origin and become distributed at random in the cytoplasm. They are the pro-acrosomal granules and at the end of the growth period fuse to form the proacrosome, about which Golgi bodies collect. The Golgi bodies, however, never fuse into an acroblast. At one end of the oval-shaped pro-acrosome is a small dark body and a less dense vesicle the future of which is uncertain. The dark body eventually occupies a position at the tip of the acrosome. The pro-acrosome, after moving to the side of the nucleus opposite the nebenkern, differentiates into the acrosome which elongates into a tail-like structure. The nuclear membrane of some spermatocytes may appear wave-like in cross section, with the crest and trough different in appearance. Near the membrane and in the troughs of the waves large clusters of granules are frequently present. Similar clusters may be found elsewhere in the cytoplasm. Presumably they had their origin near the membrane but this is not conclusive. Bodies of indeterminate origin and structure may be present in the cytoplasm. They could be lysosomes but evidence is lacking. In late spermatocytes and in spermatids, a group of ten or twelve granules is present. They are smaller than the pro-acrosomal granules, are always closely associated and pass as a group into the tail. Their significance is unknown. The endoplasmic reticulum is typical of cells in general. There are no granule accumulations within the vesicles as in some secretory cells. Vesicles of various shapes and sizes are present within the centrosphere of the first meiotic division. While their location is similar to that of the centriole, the identity of the vesicles is uncertain.