scholarly journals Ultrastructural and molecular characteristics of crayfish photoreceptor membranes.

1976 ◽  
Vol 69 (3) ◽  
pp. 721-732 ◽  
Author(s):  
H R Fernandez ◽  
E E Nickel
Keyword(s):  
Freeze Fracture ◽  
Photoreceptor Cells ◽  
Molecular Characteristics ◽  
Thin Sections ◽  
Freeze Fracturing ◽  
Mean Diameter ◽  
Hexagonal Arrangement ◽  
Photoreceptor Membranes

The ultrastructure of photoreceptor cells of the crayfish (P. clarkii) has been examined by means of thin sections and freeze-fracturing. The study reveals that in the photoreceptor membranes there are particles associated primarily with the A faces of freeze-fracture preparations which have a mean diameter of 80-84 A and a density of 6,600 per per micrometer2. Treatment of the retina with digitonin (a substance capable of extracting visual photopigments) in Ringer's causes marked disruption of the hexagonal arrangement of the microvilli, breakdown of the microvilli into smaller segments, and gradual removal of the particles. The estimated photopigment concentration in the microvillus is 4,000 per micrometer. It is suggested that the observed particles represent the photopigment in situ.

scholarly journals Membrane asymmetry and enhanced ultrastructural detail of sarcoplasmic reticulum revealed with use of tannic acid.

1978 ◽  
Vol 79 (3) ◽  
pp. 601-616 ◽  
Author(s):  
A Saito ◽  
C T Wang ◽  
S Fleischer
Keyword(s):  
Sarcoplasmic Reticulum ◽  
Tannic Acid ◽  
Freeze Fracture ◽  
Membrane Vesicles ◽  
Thin Sections ◽  
Membrane Asymmetry ◽  
The Asymmetry ◽  
First Time ◽  
Negative Staining Electron Microscopy

Fixation of purified sarcoplasmic reticulum (SR) membrane vesicles, using glutaraldehyde supplemented with 1% tannic acid, reveals newly visualized ultrastructure in thin sections. The trilaminar appearance of the membrane is highly asymmetric; the outer electron-opaque layer is appreciably wider (70 A) than the inner layer (20 A). The asymmetry is not referable to lack of penetration of the tannic acid since: (a) SR vesicles made permeable with 1 mM EDTA, pH 8.5, show similar asymmetry; (b) treatment of SR with trypsin results in progressive loss in protein content and decrease in the thickness of the outer layer, until in the limit the trilayer has a symmetric appearance; (c) within the same muscle section, the SR membrane appears highly asymmetric whereas the sarcolemma has a more symmetric appearance; (d) reconstituted SR vesicles have a symmetric appearance with equally broad inner and outer layers (approximately 70 A); the symmetric structure is confirmed by freeze-fracture and negative staining electron microscopy. Heavy and light SR vesicles obtained by isopycnic density sedimentation of purified SR have the same asymmetric appearance of the membrane and seem to differ mainly in that the heavy vesicles contain internal contents consisting largely of Ca++-binding protein. The asymmetry of the SR membrane is referable mainly to the unidirectional alignment of the Ca++ pump protein, the major component (90% of the protein) of the membrane. The asymmetry of the SR membrane can be visualized now for the first time in situ in thin sections of muscle.

A freeze-fracture study of the cuticle of adult Nippostrongylus brasiliensis (Nematoda)

Parasitology ◽  
1993 ◽  
Vol 107 (5) ◽  
pp. 545-552 ◽  
Author(s):  
D. L. Lee ◽  
K. A. Wright ◽  
R. R. Shivers
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Freeze Fracture ◽  
Surface Coat ◽  
Nippostrongylus Brasiliensis ◽  
Freeze Fracturing ◽  
Transmission Electron ◽  
Fracture Study ◽  
20 Nm

SUMMARYThe surface of the cuticle of adult Nippostrongylus brasiliensis has been studied by means of the freeze-fracture technique and by transmission electron microscopy. Some of the surface coat appears to have been shed from the surface of the cuticle of adults fixed in situ in the intestine of its host and from the surface of individuals removed from the intestine and freeze-fractured. Freeze-fracturing the cuticle of individuals removed from the host has shown that this surface coat varies in thickness from 30 to 90 nm. The epicuticle is about 20 nm thick and cleaves readily to expose E- and P-faces. The P-face of the epicuticle possesses a small number of particles, similar to intra-membranous particles, whilst the E-face possesses a few, widely scattered depressions. Despite the presence of these particles the epicuticle is not considered to be a true membrane. Freeze-fracturing the remainder of the cuticle has confirmed its structure as described by conventional transmission electron microscopy. Clusters of particles on the P-face of the outer epidermal (hypodermal) membrane and corresponding depressions on the E-face of the membrane are thought to be associated with points of attachment of the cuticle to the epidermis (hypodermis). No differences in appearance of the cuticle and its surface layers were observed in individuals taken from 7-, 10-, 13- and 15-day infections.

scholarly journals THE SPLITTING OF HEPATOCYTE GAP JUNCTIONS AND ZONULAE OCCLUDENTES WITH HYPERTONIC DISACCHARIDES

1974 ◽  
Vol 61 (3) ◽  
pp. 575-590 ◽  
Author(s):  
Daniel A. Goodenough ◽  
Norton B. Gilula
Keyword(s):  
Portal Vein ◽  
Gap Junctions ◽  
Gap Junction ◽  
Freeze Fracture ◽  
Chloride Ion ◽  
Thin Sections ◽  
Zonulae Occludentes ◽  
The Times

Mouse livers were perfused in situ through the portal vein with the disaccharides sucrose, lactose, maltose, and cellobiose in hypertonic concentrations (0.5 M). This treatment resulted in plasmolysis of the hepatocytes and splitting of the gap junctions and zonulae occludentes. The junctions split symmetrically, leaving a half-junction on each of the two separated cells. The process of junction splitting is followed using the freeze-fracture technique, since the junctional membranes are indistinguishable from the nonjunctional membranes in thin sections once the splitting occurs. The split junctions are also studied using the freeze-etch technique, allowing a view of the gap junction extracellular surface normally sequestered within the 2-nm "gap." The monosaccharides sorbitol and mannitol did not split the junctions during the times studied (2 min), but substitution of the chloride ion with propionate in the perfusion mixture did result in junction splitting. An envelope of morphologically distinct particles surrounding freeze-fractured gap junctions is also described.

Photoreceptor disk membrane substructures by means of the complementary freeze-fracture-replica methods

1978 ◽  
Vol 36 (2) ◽  
pp. 156-157
Author(s):  
A. Tonosaki ◽  
M. Yamasaki ◽  
H. Washioka ◽  
J. Mizoguchi
Keyword(s):  
Cell Membranes ◽  
Freeze Fracture ◽  
Purple Membrane ◽  
Remarkable Case ◽  
Single Face ◽  
Disk Membrane ◽  
Associated Proteins ◽  
Photoreceptor Membranes

A vertebrate disk membrane is composed of 40 % lipids and 60 % proteins. Its fracture faces have been classed into the plasmic (PF) and exoplasmic faces (EF), complementary with each other, like those of most other types of cell membranes. The hypothesis assuming the PF particles as representing membrane-associated proteins has been challenged by serious questions if they in fact emerge from the crystalline formation or decoration effects during freezing and shadowing processes. This problem seems to be yet unanswered, despite the remarkable case of the purple membrane of Halobacterium, partly because most observations have been made on the replicas from a single face of specimen, and partly because, in the case of photoreceptor membranes, the conformation of a rhodopsin and its relatives remains yet uncertain. The former defect seems to be partially fulfilled with complementary replica methods.

A TEM study of the microstructure of avian eggshells

1992 ◽  
Vol 50 (2) ◽  
pp. 1102-1103
Author(s):  
S. Q. Xiao ◽  
S. Baden ◽  
A. H. Heuer
Keyword(s):  
Electron Microscope ◽  
Calcium Carbonate ◽  
Nucleation And Growth ◽  
Growth Mechanisms ◽  
Shell Surface ◽  
Thin Sections ◽  
Polycrystalline Metals ◽  
Transmission Electron ◽  
Nucleation And Growth Mechanisms

The avian eggshell is one of the most rapidly mineralizing biological systems known. In situ, 5g of calcium carbonate are crystallized in less than 20 hrs to fabricate the shell. Although there have been much work about the formation of eggshells, controversy about the nucleation and growth mechanisms of the calcite crystals, and their texture in the eggshell, still remain unclear. In this report the microstructure and microchemistry of avian eggshells have been analyzed using transmission electron microscope (TEM) and energy dispersive spectroscopy (EDS).Fresh white and dry brown eggshells were broken and fixed in Karnosky's fixative (kaltitanden) for 2 hrs, then rinsed in distilled H2O. Small speckles of the eggshells were embedded in Spurr medium and thin sections were made ultramicrotome.The crystalline part of eggshells are composed of many small plate-like calcite grains, whose plate normals are approximately parallel to the shell surface. The sizes of the grains are about 0.3×0.3×1 μm3 (Fig.l). These grains are not as closely packed as man-made polycrystalline metals and ceramics, and small gaps between adjacent grains are visible indicating the absence of conventional grain boundaries.

Gap junctions in the prothoracic glands of the tobacco hornworm, Manduca sexta

1992 ◽  
Vol 50 (1) ◽  
pp. 706-707
Author(s):  
Ji-da Dai ◽  
M. Joseph Costello ◽  
Lawrence I. Gilbert
Keyword(s):  
Gap Junctions ◽  
Small Molecules ◽  
Manduca Sexta ◽  
Freeze Fracture ◽  
Cell Communication ◽  
Cellular Level ◽  
Thin Sections ◽  
Prothoracic Glands ◽  
Polyhydroxylated Steroids ◽  
Stimulation Of

Insect molting and metamorphosis are elicited by a class of polyhydroxylated steroids, ecdysteroids, that originate in the prothoracic glands (PGs). Prothoracicotropic hormone stimulation of steroidogenesis by the PGs at the cellular level involves both calcium and cAMP. Cell-to-cell communication mediated by gap junctions may play a key role in regulating signal transduction by controlling the transmission of small molecules and ions between adjacent cells. This is the first report of gap junctions in the PGs, the evidence obtained by means of SEM, thin sections and freeze-fracture replicas.

Differentiation Processes of the Ocellar Retina of the Honeybee (Apis Mellifera L.)

1990 ◽  
Vol 48 (3) ◽  
pp. 430-431
Author(s):  
Maria Anna Pabst
Keyword(s):  
Apis Mellifera ◽  
Distal Part ◽  
Cell Layer ◽  
Single Layer ◽  
Photoreceptor Cells ◽  
Distal Segment ◽  
Compound Eyes ◽  
Thin Sections ◽  
Ultrastructural Studies ◽  
Adult Worker

In addition to the compound eyes, honeybees have three dorsal ocelli on the vertex of the head. Each ocellus has about 800 elongated photoreceptor cells. They are paired and the distal segment of each pair bears densely packed microvilli forming together a platelike fused rhabdom. Beneath a common cuticular lens a single layer of corneagenous cells is present.Ultrastructural studies were made of the retina of praepupae, different pupal stages and adult worker bees by thin sections and freeze-etch preparations. In praepupae the ocellar anlage consists of a conical group of epidermal cells that differentiate to photoreceptor cells, glial cells and corneagenous cells. Some photoreceptor cells are already paired and show disarrayed microvilli with circularly ordered filaments inside. In ocelli of 2-day-old pupae, when a retinogenous and a lentinogenous cell layer can be clearly distinguished, cell membranes of the distal part of two photoreceptor cells begin to interdigitate with each other and so start to form the definitive microvilli. At the beginning the microvilli often occupy the whole width of the developing rhabdom (Fig. 1).

Freeze-fracture observations on changes in the distribution of Filipin-sterol complexes during epididymal maturation and capacitation of boar spermatozoa

1990 ◽  
Vol 48 (3) ◽  
pp. 90-91
Author(s):  
N. Seki ◽  
Y. Toyama ◽  
T. Nagano
Keyword(s):  
Plasma Membrane ◽  
Essential Role ◽  
Freeze Fracture ◽  
Final Concentration ◽  
Freeze Fracturing ◽  
Physiological Conditions ◽  
Epididymal Maturation ◽  
Cacodylate Buffer ◽  
Sperm Membrane ◽  
Boar Spermatozoa

It is believed that i ntramembra.nous sterols play an essential role in membrane stability and permeability. To investigate the distribution changes of sterols in sperm membrane during epididymal maturation and capacitation, filipin has been used as a cytochemical probe for the detection for membrane sterols. Using this technique in combination with freeze fracturing, we examined the boar spermatozoa under various physiological conditions.The spermatozoa were collected from: 1) caput, corpus and cauda epididymides, 2) sperm rich fraction of ejaculates, and 3)the uterus 2hr after natural coition. They were fixed with 2.5% glutaraldehyde in 0.05M cacodylate buffer (pH 7.4), and treated with the filipin solution (final concentration : 0.02.0.05%) for 24hr at 4°C with constant agitation. After the filipin treatment, replicas were made by conventional freeze-fracture technique. The density of filipin-sterol complexes (FSCs) was determined in the E face of the plasma membrane of head regions.

Basal Cell Carcinoma of Prostate With MSMB–NCOA4 Fusion and a Probable Basal Cell Carcinoma In Situ: Case Report

2021 ◽  
pp. 106689692110173
Author(s):  
Vilde Pedersen ◽  
Katrine S. Petersen ◽  
Klaus Brasso ◽  
Olga Østrup ◽  
Anand C. Loya
Keyword(s):  
Prostate Cancer ◽  
Basal Cell Carcinoma ◽  
Cell Carcinoma ◽  
Basal Cell ◽  
Carcinoma In Situ ◽  
Pathological Examination ◽  
Molecular Characteristics ◽  
Histological Lesion ◽  
Basal Cell Carcinomas

Basal cell carcinomas of prostate (BCCP) are very rare. Most arise in the transition zone and thus are associated with lower urinary tract symptoms and rarely associated with elevated prostate-specific antigen (PSA). These features make diagnosis/early diagnosis difficult because of the routine protocols followed. Basal cell carcinomas have distinctive histopathological, immunohistochemical, and to some extent also different molecular characteristics. Basal cell carcinoma in situ (BCCIS) is a nonexistent histological lesion as per the current literature, but here is an attempt to describe it through this case. A 74-year-old man presented with hematuria and previous diagnosis of prostatic hyperplasia. Based on this history, he underwent a prostatectomy ad modum Freyer. Pathological examination surprisingly revealed a diffusely infiltrative tumor with nonacinar adenocarcinoma morphology and many glandular structures probably representing BCCIS. Tumor was diagnosed as BCCP. Patient presented with metastasis to the abdominal wall 8 months postprostatectomy. BCCP is an aggressive type of prostate cancer, which might be challenging to diagnose based on routine protocols. This results in delayed diagnosis and treatment and thus poor prognosis. Furthermore, patients with this subtype of prostate cancer need appropriately designed, and maybe a totally different follow-up regimen as PSA is of no use for BCCP patients. Finally, diagnosis of BCCIS, if agreed upon its existence needs to be studied in larger cohorts as a precursor lesion.

scholarly journals THE ULTRASTRUCTURE OF LIPID-DEPLETED ROD PHOTORECEPTOR MEMBRANES

1974 ◽  
Vol 63 (2) ◽  
pp. 587-598 ◽  
Author(s):  
Izhak Nir ◽  
Michael O. Hall
Keyword(s):  
Lipid Extraction ◽  
Fatty Acid Analysis ◽  
Major Protein ◽  
Rod Photoreceptor ◽  
Thin Sections ◽  
Retinal Rod ◽  
Layer Chromatography ◽  
Chloroform Methanol ◽  
Photoreceptor Membranes

The structure of lipid-depleted retinal rod photoreceptor membranes was studied by means of electron microscopy. Aldehyde-fixed retinas were exhaustively extracted with acetone, chloroform-methanol, and acidified chloroform-methanol. The effect of prefixation on the extractability of lipids was evaluated by means of thin-layer chromatography and fatty acid analysis. Prefixation with glutaraldehyde rendered 38% of the phospholipids unextractable, while only 7% were unextractable after formaldehyde fixation. Embedding the retina in a lipid-retaining, polymerizable glutaraldehyde-urea mixture allows a comparison of the interaction of OsO4 with lipid-depleted membranes and rod disk membranes which contain all their lipids. A decrease in electron density and a deterioration of membrane fine structure in lipid-depleted tissue are correlated with the extent of lipid extraction. These observations are indicative of the role of the lipid bilayer in the ultrastructural visualization of membrane structure with OsO4. Negatively stained thin sections of extracted tissue reveal substructures in the lipid-depleted rod membranes. These substructures are probably the opsin molecules which are the major protein component of retinal rod photoreceptor membranes.

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