An electron microscopic study of vitellogenesis and egg membrane formation in Lytta nuttalli Say (ColeopterarMeloidae)

1970 ◽  
Vol 48 (4) ◽  
pp. 651-657 ◽  
Author(s):  
P. R. Sweeny ◽  
N. S. Church ◽  
J. G. Rempel ◽  
Wendy Frith
Keyword(s):  
Electron Microscopy ◽  
Plasma Membrane ◽  
Follicle Cells ◽  
Vitelline Membrane ◽  
Electron Microscopic ◽  
Yolk Formation ◽  
Membrane Formation ◽  
Electron Dense Material ◽  
Egg Membrane ◽  
Pass Through

Vitellogenesis and egg membrane formation in the terminal ovarian follicles of Lytta nuttatii were investigated by electron microscopy. Three kinds of yolk globules are produced. They apparently are composed predominantly of carbohydrates, lipids, and proteins, respectively. The "carbohydrate" and "lipid" yolk are assembled in the ooplasm, the former by rough endoplasmic reticulum and the latter by Golgi complexes. Their production begins early in oogenesis. "Proteid" yolk formation begins somewhat later. The "proteid" yolk globules evidently are formed from exfraovarian materials that pass through large spaces that develop between the follicular epithelial cells, then through the oocyte plasma membrane by pinocytosis. Fairly late in development, glycogen granules appear in the inner ooplasm. In the nearly fully grown follicle, the "membranous system" of the vitelline membrane is elaborated. It probably is formed largely from an electron-dense material of undetermined origin that accumulates outside the bases of the oocyte plasma membrane microvilli. Immediately after completion of the vitelline membrane, the chorion is laid down, presumably from dense globules of material produced by Golgi complexes in the follicle cells.

The relationship between oöcyte and follicle in the hen's ovary as shown by electron microscopy

Development ◽  
1965 ◽  
Vol 13 (2) ◽  
pp. 215-233
Author(s):  
Ruth Bellairs
Keyword(s):  
Electron Microscopy ◽  
Cell Membrane ◽  
Raw Materials ◽  
Follicle Cell ◽  
Follicle Cells ◽  
Vitelline Membrane ◽  
Zona Radiata ◽  
Golgi Bodies ◽  
Pass Through ◽  
The Relationship

In the adult hen each oöcyte is surrounded by a capsule of follicle cells and all the raw materials that enter the oöcyte must pass through this capsule. It is not surprising, therefore, that the morphological relationships between the follicle and the oöcyte are of a highly specialized nature. Several workers have studied them, mainly by light microscopy, but their findings have not been unanimous, largely because of difficulties in resolving fine details. For instance, although it has frequently been suggested that certain structures pass from the follicle cell into the oöcyte, these structures have been interpreted by different authors as Golgi bodies, as mitochondria or as fat drops. Similarly, there have been several different theories about the relationship between the cell membrane of the oöcyte, the zona radiata and the vitelline membrane.

Immunocytochemical localization of p65 with one-nanometer gold particles following silver development

1989 ◽  
Vol 47 ◽  
pp. 1042-1043
Author(s):  
Richard W. Burry ◽  
Diane M. Hayes
Keyword(s):  
Plasma Membrane ◽  
Bovine Serum Albumin ◽  
Calf Serum ◽  
Specific Protein ◽  
Immunocytochemical Localization ◽  
Electron Microscopic ◽  
Gold Particles ◽  
Pass Through ◽  
Label Distribution ◽  
Phosphate Buffered Saline

Electron microscopic (EM) immunocytochemistry localization of the neuron specific protein p65 could show which organelles contain this antigen. Antibodies (Ab) labeled with horseradish peroxidase (HRP) followed by chromogen development show a broad diffuse label distribution within cells and restricting identification of organelles. Particulate label (e.g. 10 nm colloidal gold) is highly desirable but not practical because penetration into cells requires destroying the plasma membrane. We report pre-embedding immunocytochemistry with a particulate marker, 1 nm gold, that will pass through membranes treated with saponin, a mild detergent.Cell cultures of the rat cerebellum were fixed in buffered 4% paraformaldehyde and 0.1% glutaraldehyde (Glut.). The buffer for all incubations and rinses was phosphate buffered saline with: 1% calf serum, 0.2% saponin, 0.1% gelatin, 50 mM glycine 1 mg/ml bovine serum albumin, and (not in the HRP labeled cultures) 0.02% sodium azide. The monoclonal #48 to p65 was used with three label systems: HRP, 1 nm avidin gold with IntenSE M development, and 1 nm avidin gold with Danscher development.

Studies on the Ovarian Follicle Cells of Drosophila

1963 ◽  
Vol s3-104 (67) ◽  
pp. 297-320
Author(s):  
R. C. KING ◽  
ELIZABETH A. KOCH
Keyword(s):  
Ovarian Follicle ◽  
Protein A ◽  
Follicle Cell ◽  
Follicle Cells ◽  
Vitelline Membrane ◽  
Membrane Material ◽  
Adjacent Layer ◽  
Membrane Formation ◽  
Precursor Material ◽  
Egg Shell

Studies are described of the ultrastructure of the follicle cells which invest the oocyte of Drosophila melanogaster at the time of vitelline membrane formation. Of particular interest are organelles made up of endoplasmic reticulum organized into a husk of concentric lamellae which surround lipidal droplets. These epithelial bodies are seen only at the time the vitelline membrane is being formed, and it is assumed therefore that the lipidal material of the epithelial body may be utilized somehow in the fabrication of the vitelline membrane. Cytochemical studies have shown this membrane to contain at least 5 classes of compounds; a protein, two lipids (which may be distinguished by differences in their resistance to extraction by various solvents), and 2 polysaccharides (1 neutral and 1 acidic). Studies were made of vitelline membrane formation in the ovaries of flies homozygous for either of 2 recessive, female-sterile genes (tiny and female sterile). In the case of the ty mutation vitelline membrane material is sometimes secreted between follicle and nurse cells, while in the mutant fes vitelline membrane is observed in rare instances to be secreted between follicle cells and an adjacent layer of tumour cells. In the latter case the vitelline membrane shows altered cytochemical properties. The fact that vitelline membrane can be secreted by follicle cells not adjacent to an oocyte demonstrates that it is the follicle cell rather than the oocyte that plays the major role in the secretion of the precursor material of the vitelline membrane. Subsequently the follicle cells secrete the egg-shell, or chorion, which is subdivided into a dense, compartmented, inner endochorion, and a pale, outer exochorion. A description is given of the ultrastructure of the follicle cells during the secretion of the endochorion and the exochorion. The endochorion contains a protein, a polysaccharide, and a lipid, all of which may be distinguished cytochemically from the vitelline membrane compounds. The exochorion contains large amounts of acidic mucopolysaccharides. Specialized follicle cells form the micropylar apparatus and the chorionic appendages. The formation of the chorion and chorionic appendages is discussed in the light of information gained from abnormalities of the chorions and chorionic appendages seen in ty and fs 2.1 oocytes. Subsequent to the time the egg leaves the ovariole a layer of waterproofing wax is secreted between the vitelline membrane and the chorion.

scholarly journals Microvillus-specific Mr 75,000 plasma membrane protein of human choriocarcinoma cells.

1987 ◽  
Vol 35 (8) ◽  
pp. 809-816 ◽  
Author(s):  
R Pakkanen ◽  
K Hedman ◽  
O Turunen ◽  
T Wahlström ◽  
A Vaheri
Keyword(s):  
Electron Microscopy ◽  
Plasma Membrane ◽  
Membrane Protein ◽  
Cell Surface ◽  
Polyclonal Antibodies ◽  
Apical Cell ◽  
Antigenic Site ◽  
Electron Microscopic ◽  
Permeabilized Cells ◽  
Choriocarcinoma Cells

We have previously purified from cultured JEG-3 choriocarcinoma cells an Mr 75,000 protein, originally detected using antibodies to a retrovirus-related synthetic peptide. Using polyclonal antibodies, we have now localized this protein immunocytochemically in JEG-3 cells at both light and electron microscopic levels. In immunofluorescence microscopy of saponin-permeabilized cells, the antigen appeared as dots and short strands at the apical cell surface. In pre-embedding immunoperoxidase electron microscopy, the Mr 75,000 protein was specifically localized to microvilli on the apical cell surface. Immunoferritin electron microscopy was used to assess more quantitatively the antigen distribution in the plane of the plasma membrane, and to define the position of the antigenic site(s) with respect to the membrane. The immunoferritin results confirmed the microvillus specificity of the Mr 75,000 protein and showed that the antigenic portion of the protein is within a few nanometers from, and on the cytoplasmic side of, the lipid bilayer. In detergent extraction experiments, the Mr 75,000 antigen was highly enriched in the soluble fractions. These results demonstrate that the Mr 75,000 protein is a membrane protein highly specific for microvilli.

The eggshell of Drosophila melanogaster. II. New staging characteristics and fine structural analysis of choriogenesis

1986 ◽  
Vol 64 (10) ◽  
pp. 2152-2175 ◽  
Author(s):  
Lukas H. Margaritis
Keyword(s):  
Electron Microscopy ◽  
Light And Electron Microscopy ◽  
Dark Field ◽  
Posterior Pole ◽  
Follicle Cells ◽  
Vitelline Membrane ◽  
Simultaneous Formation ◽  
Last Stage ◽  
Flocculent Material ◽  
Factor C

The characteristics of the stages of choriogenesis have been identified using light and electron microscopy. Nine stages have been discerned (11A, 11B, 12A, 12B, 12C, 13A, 13B, 14A, 14B), replacing the four stages used so far (11, 12, 13, 14). Characteristics used to determine the stage of the choriogenesis include (a) the size of oocyte as compared with the whole follicle, (b) the length of the chorionic appendages, and (c) the fine structure of the chorionic layers at the main shell and at the specialized regions. Factors a and b were detected by dark-field light microscopy on living follicles, whereas factor c was studied with electron microscopy. At stage 11A the vitelline membrane has just been completed. At stage 11B the follicle cells secrete the wax layer and the respiratory appendages start to form. Stage 12A follicles secrete endochorion at the anterior pole and the appendages elongate, whereas at stage 12B the main shell follicle cells start to secrete endochorion complex. Stage 12C shows initiation of pillar formation at the main shell and 150 μm long appendages. Stage 13A is characterized by 200 μm long appendages and formation of endochorionic cavities at the main shell, through the participation of a "flocculent" material. At stage 13B the endochorionic "roof is formed, which is completed at stage 14A by the simultaneous formation of the "roof network." The last stage, 14B, exhibits 300 μm long appendages and the secretion of exochorion over the entire follicle. The above stages are accompanied by region-specific formation of specialized structures which include the respiratory appendages, the operculum, the posterior pole, the micropyle, and the collar.

scholarly journals Immunogold Electron Microscopic Demonstration of Distinct Submembranous Localization of the Activated γPKC Depending on the Stimulation

2007 ◽  
Vol 56 (3) ◽  
pp. 253-265 ◽  
Author(s):  
Miho Oyasu ◽  
Mineko Fujimiya ◽  
Kaori Kashiwagi ◽  
Shiho Ohmori ◽  
Hirotsugu Imaeda ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Plasma Membrane ◽  
Laser Scanning ◽  
Fluorescent Protein ◽  
Fluorescent Microscopy ◽  
Confocal Laser ◽  
Immunogold Electron Microscopy ◽  
Subcellular Targeting ◽  
Electron Microscopic ◽  
Proper Distance

We examined the precise intracellular translocation of γ subtype of protein kinase C (γPKC) after various extracellular stimuli using confocal laser-scanning fluorescent microscopy (CLSM) and immunogold electron microscopy. By CLSM, treatment with 12- O-tetradecanoylphorbol-13-acetate (TPA) resulted in a slow and irreversible accumulation of green fluorescent protein (GFP)-tagged γPKC (γPKC–GFP) on the plasma membrane. In contrast, treatment with Ca2+ ionophore and activation of purinergic or NMDA receptors induced a rapid and transient membrane translocation of γPKC–GFP. Although each stimulus resulted in PKC localization at the plasma membrane, electron microscopy revealed that γPKC showed a subtle but significantly different localization depending on stimulation. Whereas TPA and UTP induced a sustained localization of γPKC–GFP on the plasma membrane, Ca2+ ionophore and NMDA rapidly translocated γPKC–GFP to the plasma membrane and then restricted γPKC–GFP in submembranous area (<500 nm from the plasma membrane). These results suggest that Ca2+ influx alone induced the association of γPKC with the plasma membrane for only a moment and then located this enzyme at a proper distance in a touch-and-go manner, whereas diacylglycerol or TPA tightly anchored this enzyme on the plasma membrane. The distinct subcellular targeting of γPKC in response to various stimuli suggests a novel mechanism for PKC activation.

scholarly journals ELECTRON MICROSCOPIC STUDY OF THE ATPASE ACTIVITY OF THE BAI STRAIN A (MYELOBLASTOSIS) AVIAN TUMOR VIRUS

1962 ◽  
Vol 15 (3) ◽  
pp. 451-462 ◽  
Author(s):  
Alex B. Novikoff ◽  
Guy de Thé ◽  
D. Beard ◽  
J. W. Beard
Keyword(s):  
Electron Microscopy ◽  
Plasma Membrane ◽  
Atpase Activity ◽  
Light Microscopy ◽  
Microscopic Study ◽  
Electron Microscopic Study ◽  
Frozen Sections ◽  
Electron Microscopic ◽  
Tumor Virus

Thymus glands of chicks with leukemia induced by BAI strain A (myeloblastosis) virus were fixed in cold 4 per cent formaldehyde-sucrose. Frozen sections were incubated in the ATPase medium of Wachstein and Meisel and studied by light microscopy and electron microscopy. The ATPase activity of the virus is localized to the outermost membrane of the virus. The membrane of the blast-like cells of the thymus cortex from which the virus emerges, by budding, also possesses such activity. It appears likely that the outermost membrane of the virus is derived from the plasma membrane of these cells.

Aspects of the development of yolk spheres in the hen's oöcyte, studied by electron microscopy

Development ◽  
1967 ◽  
Vol 17 (2) ◽  
pp. 267-281
Author(s):  
Ruth Bellairs
Keyword(s):  
Electron Microscopy ◽  
Cell Nucleus ◽  
Raw Materials ◽  
Morphological Changes ◽  
Laying Hen ◽  
Follicle Cells ◽  
Balbiani Body ◽  
Hen’S Egg ◽  
Pass Through ◽  
Yolk Nucleus

The yolk of the hen's egg is composed mainly of proteins, lipids and water (see reviews by Bellairs, 1964; Williams, 1966). It consists essentially of yolk spheres floating in an aqueous protein medium (Grodziński, 1939; Bellairs, 1961). The raw materials from which the yolk is formed are synthesized in the liver of the laying hen and pass from there in the blood to the ovary (see reviews by Romanoff, 1960; Bellairs, 1964). Each oöcyte is enclosed in a capsule of follicle cells, and all the raw materials pass through this capsule before they enter the oöcyte. The morphological changes that take place within the oöcyte as the yolk spheres form have been described previously by light microscopists who have produced a variety of theories to explain their observations. Formerly, it was supposed that yolk arose in the so-called ‘yolk nucleus’, or Balbiani body, which lies alongside the cell nucleus, but few would subscribe to this theory now.

scholarly journals Canine Cutaneous Histiocytoma A Light and Electron Microscopic Study

1970 ◽  
Vol 7 (1) ◽  
pp. 12-27 ◽  
Author(s):  
D. F. Kelly
Keyword(s):  
Electron Microscopy ◽  
Endoplasmic Reticulum ◽  
Plasma Membrane ◽  
Acid Phosphatase ◽  
Microscopic Study ◽  
Mononuclear Cell ◽  
Electron Microscopic Study ◽  
Light And Electron Microscopy ◽  
Perinuclear Region ◽  
Electron Microscopic

Cutaneous histiocytomas from 4 dogs were examined by light and electron microscopy. A large (up to 10 μ in diameter) mononuclear cell with prominent filiform processes of the plasma membrane predominated. Its cytoplasm contained relatively small amounts of endoplasmic reticulum and mitochondria, only occasional lysosomes, fibrils, most obvious in the perinuclear region, and small amounts of cytoplasmic debris. Acid phosphatase was not detected. Fibroblasts and collagen formed a small part of the lesion, except at the junction with surrounding dermis, where fibers were plentiful. The morphologic features of the lesion are compatible with the suggestion that the predominant cell is of histiocytic type.

scholarly journals Memoirs: Yolk-Formation in Periplaneta Orientalis

1931 ◽  
Vol s2-74 (294) ◽  
pp. 257-274
Author(s):  
R.A. R. GRESSON
Keyword(s):  
Epithelial Cells ◽  
Nuclear Membrane ◽  
Neutral Red ◽  
Follicle Cells ◽  
Osmic Acid ◽  
Staining Reaction ◽  
Yolk Formation ◽  
Cover Slip ◽  
Pass Through ◽  
The Masses

1. The Golgi vacuoles and fatty yolk-formation in Peri-planeta orientalis were studied by means of Mann-Kopsch, Kolatschev, 2 per cent, osmic acid and neutral red preparations. 2. The Golgi vacuoles of the young oocytes are situated in the vicinity of the nucleus; later they pass to the periphery of the cell. In the older oocytes, towards the posterior end of the ovarioles, they become evenly distributed in the ooplasm, store up fat, increase greatly in size, and give rise to the fatty yolkspheres. In the older oocytes they darken much more rapidly in 2 per cent, osmic acid. 3. In neutral red preparations clear non-stained vacuoles are seen to occupy similar positions to those of the dark bodies of the osmic preparations; on introducing a few drops of 2 per cent, osmic acid under the cover slip the vacuoles develop an osmophilic rim. These Golgi vacuoles are not stained by neutral red. 4. In 2 per cent, osmic acid preparations the Golgi vacuoles are seen to consist of an osmophilic rim and a central clear substance. 5. The Golgi vacuoles of the follicle-cells are similar to those of the egg, except that they do not increase greatly in size and are not so rapidly darkened in 2 per cent, osmic acid. 6. The nucleoli of the early oocytes are spherical in shape and are amphiphil or slightly basophil in staining reaction; they may contain small vacuoles. In slightly older oocytes the nucleoli are non-vacuolated; they become strongly basophil, irregular in outline, and, at the same time, give rise to emissions which pass through the nuclear membrane to the ooplasm, where they ultimately disappear. In a certain few oocytes the nucleolus was seen to have broken up into several masses, some of the latter, in all probability, fragmenting to form nucleolar extrusions. In a certain oocyte one of the masses was observed to be vacuolated before the first type of extrusion had ceased. 7. In the more highly developed oocytes the first type of nucleolar emission ceases, and the nucleolus becomes more spherical in outline. Numerous vacuoles appear which give origin to nucleolar extrusions. The latter become vacuolated, either before extrusion through the nuclear membrane, or later in the ooplasm. 8. The second type of nucleolar extrusions pass to the periphery of the egg. Later they become evenly distributed in the ooplasm, where they fragment to forin homogeneous granules. The latter form clear spheres (Kolatschev material) which rapidly increase in size to form the albuminous yolk-globules. 9. Chromatin was not observed in the oocyte nuclei, nucleoli, or nucleolar extrusions (Feulgen's technique). The chromatin of the follicle-cells is in the form of granules connected by threads (which give the chromatin reaction). The chromatin of the follicular epithelial-cells was observed as granules scattered through the nuclei. 10. Bacteroid forms were observed in the ooplasm at the periphery of the older oocytes. 11. The method of yolk-formation is similar to that of Peri-planeta americanaas described by Nath and Piare Mohan. 12. The writer's conclusions regarding the shape and character of the Golgi vacuoles agree with tne findings of Nath and his co-workers and with the former conclusions of the present writer for oocyte Golgi vacuoles.

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