Fine Structure of Sea Urchin Sperm Membranes

Author(s):  
S. Inoue ◽  
A. Buday ◽  
G.H. Cousineau

From electron microscope studies of thin sections it is known that the entire surface of a spermatozoon of sea urchin is covered by a plasma membrane, or sperm membrane, of an approximate thickness of 100Å (1). In these experiments the surface replica method was applied for the study of the fine structure of the sperm membrane.Spermatozoa from Strongylocentrotus purpuratus (sea urchins supplied by the Pacific Biomarine Supply Company, Venice, Calif.) were washed several times by centrifugation in Millipore-filtered sea water. After fixation in a 2.5% glutaraldehyde-paraformaldehyde (sea water mixture at 4°C) for an hour, spermatozoa were washed with sea water and then with distilled water for several times. A few drops of specimen were dried on a glass slide and the surface replica was prepared according to the method previously described (2) with the exception that the spermatozoa were decomposed in 18 N H2SO4 for about 20 hours at room temperature. The replica films were examined with a JEM-7A electron microscope.

1958 ◽  
Vol 4 (5) ◽  
pp. 575-578 ◽  
Author(s):  
J. J. Pasteels ◽  
P. Castiaux ◽  
G. Vandermeerssche

Fertilized, but still unsegmented, eggs of the sea urchin Paracentrotus lividus have been centrifugated at 30,000 gravity. A comparison has been made between the cytochemical reactions of the different layers (RNA, polysaccharides, acid phosphatase) and the fine structure as shown in thin sections studied with the electron microscope.


1965 ◽  
Vol 25 (2) ◽  
pp. 81-100 ◽  
Author(s):  
Luther E. Franklin

Sea urchin gametes predominate in molecular studies of fertilization, yet relatively little is known of the subcellular aspects of sperm entry in this group. Accordingly, it seemed desirable to make a detailed examination of sperm entry phenomena in sea urchins with the electron microscope. Gametes of the sea urchins Arbacia punctulata and Lytechinus variegatus were used in this study. Samples of eggs containing 2 to 8 per cent oocytes were selected and fixed with osmium tetroxide in sea water at various intervals after insemination. Fixed specimens were embedded in Epon 812, sectioned, and examined with an electron microscope. An apical vesicle was observed at the anterior end of the acrosome. The presence of this structure, together with other observations, suggested that initiation of the acrosome reaction in sea urchin sperm involves dehiscence of the acrosomal region with the subsequent release of the acrosomal granule. Contact and initial fusion of gamete membranes was observed in mature eggs and oocytes and invariably involved the extended acrosomal tubule of the spermatozoon. Only one spermatozoon normally enters the mature egg. The probability of locating such a sperm in ultrathin sections is exceedingly low. Several sperm do normally enter oocytes. Consequently, observations of sperm entry were primarily restricted to the latter. The manner of sperm entry into oocytes did not resemble phagocytosis. Organelles of the spermatozoon were progressively divested of their plasma membrane as they entered the ground cytoplasm of the oocyte fertilization cone. Initiation of the acrosome reaction, contact and initial fusion of gamete membranes, and sperm entry into oocytes of sea urchins conform to the Hydroides-Saccoglossus pattern of early fertilization events as described by Colwin and Colwin (13).


Author(s):  
Roberta M. Bruck

An unusual structure in the cochlea is the spiral limbus; this periosteal tissue consists of stellate fibroblasts and collagenous fibers embedded in a translucent ground substance. The collagenous fibers are arranged in vertical columns (the auditory teeth of Haschke). Between the auditory teeth are interdental furrows in which the interdental cells are situated. These epithelial cells supposedly secrete the tectorial membrane.The fine structure of interdental cells in the rat was reported by Iurato (1962). Since the mouse appears to be different, a description of the fine structure of mouse interdental cells' is presented. Young adult C57BL/6J mice were perfused intervascularly with 1% paraformaldehyde/ 1.25% glutaraldehyde in .1M phosphate buffer (pH7.2-7.4). Intact cochlea were decalcified in .1M EDTA by the method of Baird (1967), postosmicated, dehydrated, and embedded in Araldite. Thin sections stained with uranyl acetate and lead citrate were examined in a Phillips EM-200 electron microscope.


1955 ◽  
Vol 1 (1) ◽  
pp. 69-88 ◽  
Author(s):  
Sanford L. Palay ◽  
George E. Palade

1. Thin sections of representative neurons from intramural, sympathetic and dorsal root ganglia, medulla oblongata, and cerebellar cortex were studied with the aid of the electron microscope. 2. The Nissl substance of these neurons consists of masses of endoplasmic reticulum showing various degrees of orientation; upon and between the cisternae, tubules, and vesicles of the reticulum lie clusters of punctate granules, 10 to 30 mµ in diameter. 3. A second system of membranes can be distinguished from the endoplasmic reticulum of the Nissl bodies by shallower and more tightly packed cisternae and by absence of granules. Intermediate forms between the two membranous systems have been found. 4. The cytoplasm between Nissl bodies contains numerous mitochondria, rounded lipid inclusions, and fine filaments.


1963 ◽  
Vol 18 (2) ◽  
pp. 429-440 ◽  
Author(s):  
Ryan W. Drum

The cytoplasmic fine structure of the motile, pennate diatom, Nitzschia palea was studied in thin sections viewed in the electron microscope. The cells were fixed in OsO4, embedded in methacrylate, and immersed in 10 per cent hydrofluoric acid (HF) for 36 to 40 hours to remove the siliceous cell wall prior to sectioning. The HF treatment did not cause any obvious cytoplasmic damage. The dictyosome complex is perinuclear, and located only in the central cytoplasm. Mitochondria are sparse in the central cytoplasm, but abundant in the peripheral cytoplasm, and fill many of the transvacuolar cytoplasmic strands. Characteristic, amorphous oil bodies fill certain cytoplasmic strands and probably are not leucosin. The pyrenoid appears to be membrane limited, and oil droplets are found adjacent to the pyrenoid. The pyrenoid of another diatom, Cymbella affinis, is also membrane-limited. The membrane limiting the pyrenoid may be a composite of the terminal portions of chloroplast discs, facilitating rapid movement of photosynthate into the pyrenoid matrix, where the characteristic oil droplets may be formed. Carinal fibrils are found singly in each carinal pore, and may be involved in the locomotion of Nitzschia palea.


Author(s):  
S. Inoue ◽  
E. C. Preddie ◽  
P. Guerrier

From electron microscope studies of thin sections the sea urchin egg is known to be surrounded by the peripheral membrane system which is made up of the outer coat (vitelline membrane), which elevates from an egg surface after fertilization and becomes a part of the fertilization membrane, and the plasma membrane. In these experiments an effort has been made to isolate plasma membranes of sea urchin eggs and these isolated membranes were observed in the electron microscope.The vitelline membrane of the eggs from the sea urchin Strongylocentrotus purpuratus was at first digested away by the treatment with 0.02% trypsin in 0.01 M Tris-HCl buffer (pH 8.0) for 5 minutes at 28°C. The plasma membranes were then isolated according to the method of Song et al. which was used for the isolation of rat liver plasma membranes. The vitelline membrane-free eggs were gently homogenized in 10-3 M NaHC03 (pH 7.5) and freed membranes were collected by centrifugation over a discontinuous sucrose gradient preparation.


1970 ◽  
Vol 6 (3) ◽  
pp. 655-667
Author(s):  
L. F. LA COUR ◽  
B. WELLS

The 1-4 chromocentres seen in nuclei of Fritillaria lanceolata, which derive from fusion of heterochromatic segments situated proximal to the centromere in all but two of the 24 chromosomes, were studied with the electron microscope in thin sections of pollen mother cells at zygotene and pachytene, in respect of the synaptinemal complex. Prophase stages of meiosis in two plants were also surveyed briefly with the light microscope. The latter observations revealed that the timing of the separation of heterochromatic segments from chromocentres is genetically controlled. In one plant the segments were still contained in chromocentres at pachytene, whereas in the other they were free at zygotene. At this time they could be identified by a near-surface position in the nucleus and an even condensation concomitant with an absence of chromomeres. In thin section, the fine structure of the chromocentres in zygotene nuclei was distinctive in that the chromatin fibrils were less condensed and more widely dispersed than those in euchromatic regions. The fibrillar network was also interspersed with ‘clear areas’ or channels. After further chromosome condensation, the condensation of fibrils in the chromocentres became equivalent at pachytene to those in euchromatic regions. Synaptinemal complexes were seen at zygotene and pachytene both in euchromatic regions and chromocentres. Their presence in the chromocentres signifies that homologous chromosomes must have been closely paired in regions extending from the centromeres to the distal ends of the heterochromatic segments already at telophase of the last pre-meiotic mitosis. Configurations involving entangled pairs of axial cores, peculiar to zygotene and chromocentres and parts of euchromatic regions proximal to them, are interpreted as resulting from restricted movement.


1966 ◽  
Vol 12 (6) ◽  
pp. 1125-1136 ◽  
Author(s):  
Alice Reyn ◽  
A. Birch-Andersen ◽  
S. P. Lapage

The line structure of Haemophilus vaginalis (Gardner and Dukes 1955) was compared with that of four, possibly related species (Butyribacterium rettgeri, Corynebacterium diphtheriae var. mitis, Lactobacillus acidophilus, Haemophilus influenzae) and an unrelated species, Neisseria haemolysans, which had shown a similar degree of Gram-variability as that of H. vaginalis. Although H. vaginalis was first described as a Gram-negative rod, its fine structure, particularly that of cell wall and septa, was more like that of Gram-positive organisms. Also N. haemolysans had a fine structure close to that of Gram-positive organisms, and its typical Gram-positive cell wall varied in. thickness from one cell to another.The study did not solve the problem of the classification of the so-called H. vaginalis, but the appearance of the few strains studied in the electron microscope suggests that it: should be included in Corynebacterium or Butyribacterium rather than in Lactobacillus.


1972 ◽  
Vol 18 (7) ◽  
pp. 997-1002 ◽  
Author(s):  
I. L. Stevenson ◽  
S. A. W. E. Becker

Methods have been developed for the rapid, reproducible induction of high-density populations of F. oxysporum chlamydospores. On transferring washed pregerminated conidia to a simple two-salts medium, chlamydospore morphogenesis was evident by 12 h and masses of mature spores could be harvested at the end of 4 days. Electron-microscope studies of thin sections of mature chlamydospores reveal a thick triple-layered cell wall. The cytoplasm contains, in addition to large lipid deposits, a nucleus, mitochondria, and endoplasmic reticulum all typical of fungal cells. Chlamydospores of F. oxysporum exhibit two distinct types of cell surface in thin section. The outer wall layer of two of the isolates studied was smooth-surfaced while the outer layer of the two other isolates was distinctly fibrillose. Some evidence is presented suggesting that the fibrillose material arises through the partial breakdown of the original hyphal wall.


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