Distribution and size of lipid droplets in oocytes recovered from young lamb and adult ovine ovaries

2020 ◽  
Vol 32 (11) ◽  
pp. 1022
Author(s):  
Amirhossein Abazarikia ◽  
Federica Ariu ◽  
Mahsa Rasekhi ◽  
Mahdi Zhandi ◽  
Sergio Ledda
Keyword(s):  
Lipid Droplets ◽  
Nile Red ◽  
The Other ◽  
Pattern Distribution

This study evaluated the distribution and size of lipid droplets (LDs) in oocytes recovered from young and adult ovine ovaries. Collected oocytes were categorised on the basis of their major diameter (small (SO), 70–90 µm; medium (MO), >90–110 µm; large (LO), >110–130µm) and were stained with Nile red to detect LDs. In adult and young oocytes, a diffuse pattern distribution of LDs was dominant in all classes except adult LO and young SO and LO. Larger LDs (i.e. >3µm) were mostly present in young SO and LO, whereas smaller LDs (1–3µm) were detected in the other adult and young oocyte categories.

An ultrastructural study on the shark liver

1990 ◽  
Vol 48 (3) ◽  
pp. 512-513
Author(s):  
Masako Yamada ◽  
Yutaka Tanuma
Keyword(s):  
Portal Vein ◽  
Kupffer Cells ◽  
Ultrastructural Study ◽  
Lipid Droplets ◽  
Microscopic Level ◽  
The Other ◽  
Fish Stock ◽  
Structural Studies ◽  
Light Microscopic Level ◽  
Perfusion Fixation

Although many fine structural studies on the vertebrate liver have been reported on mammals, avians, reptiles, amphibians, teleosts and cyclostomes, there are no studies on elasmobranchii liver except one by T. Ito etal. (1962) who studied it on light microscopic level. The purpose of the present study was to as certain the ultrastructural details and cytochemical characteristics of normal elasmobranchii liver and was to compare with the other higher vertebrate ones.Seventeen Scyliorhinus torazame, one kind of elasmobranchii, were obtained from the fish stock of the Ueno Zoo aquarium, Ueno, Tokyo. The sharks weighing about 300-600g were anesthetized with MS-222 (Sigma), and the livers were fixed by perfusion fixation via the portal vein according to the procedure of Y. Saito et al. (1980) for 10 min. Then the liver tissues were immersed in the same fixative for 2 hours and postfixed with 1% OsO4-solution in 0.1 Mc acodylate buffer for one hour. In order to make sure a phagocytic activity of Kupffer cells, latex particles (0.8 μm in diameter, 0.05mg/100 g b.w.) were injected through the portal vein for one min before fixation. For preservation of lipid droplets in the cytoplasm, a series of these procedure were performed under ice cold temperature until the end of dehydration.

Neurosecretory Cells in the Optic Tentacles of Certain Pulmonates

1962 ◽  
Vol s3-103 (62) ◽  
pp. 211-226
Author(s):  
NANCY J. LANE
Keyword(s):  
Cell Body ◽  
Lipid Droplets ◽  
Cell Types ◽  
Neurosecretory Cells ◽  
The Other ◽  
Oval Cells

Cells considered to be neurosecretory have been observed in the optic tentacles of certain stylommatophoran pulmonates. Such cells are divisible into three distinct types, of which those called the ‘collar’ cells surround the central digitate ganglion and eye. The other two types, the ‘lateral oval’ and the ‘lateral processed’ cells, lie laterally in the tentacle, on the inner edge of the outer dermo-muscular sheath. All three cell-types have branching dendritic processes, containing granules. The dendrites of the collar and of the lateral cells apparently extend from the cell-body to the surface of the epithelium. The axonal processes of all three types are thick and contain granules. The ground cytoplasm of these cells is scarcely visible owing to the great number of homogeneous, spheroidal granules that are present. The granules are sudanophil, and the ones in the collar cells contain phospholipid (probably cerebroside as well). All three types of cells contain a much smaller number of lipid droplets, with sudanophil and osmiophil externum and sudanophobe and osmiophobe internum; these are dispersed through the cytoplasm. Special ‘perinuclear bodies’, also binary in structure, are present in the collar cells and lateral oval cells. Cells of the types described in this paper have not been found in other sub-classes of the Gastropoda, nor in the Basommatophora, but only in the pulmonate order, Stylommatophora. They appear to form an area of active neurosecretion in the retractile tentacles of these animals.

155 CHARACTERIZATION OF LIPID DROPLETS IN BOS INDICUS AND BOS TAURUS EMBRYOS

2013 ◽  
Vol 25 (1) ◽  
pp. 226 ◽  
Author(s):  
E. P. López-Damián ◽  
T. Fiordelisio ◽  
M. A. Lammoglia ◽  
M. Alarcón ◽  
M. Asprón ◽  
...  
Keyword(s):  
Lipid Droplets ◽  
Embryo Quality ◽  
Developmental Stages ◽  
Bos Taurus ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Bos Indicus ◽  
Nile Red ◽  
Blastocyst Stage ◽  
Transfer Program

Accurate evaluation of bovine embryos for assessing developmental stage and quality is critical to the success of any embryo transfer program. However, this evaluation process has been reported to be highly subjective in Bos indicus (BI) and can vary as much as 23% compared with that of Bos taurus (BT). These differences in assessment may be related to the quantity of lipid droplets (LD) within the embryo, which has been shown to have a negative effect in cryopreserving embryos. The aim of the present study was to characterize the number and size of LD in different developmental stages of fresh embryos from BI and BT and to compare LD across the three different embryo quality grades (1 = excellent or good, 2 = fair, and 3 = poor). Nonsurgical embryo collection was performed 7 days post-insemination in 10 BI and 10 BT females. Forty-eight embryos were evaluated for stage and grade using stereoscopic microscopy, processed for transmission electron microscopy, and stained with Nile red. Digitalized images were analyzed with ImageJ (National Institutes of Health, Bethesda, MD, USA), contour of lipid droplets were designed, and values of perimeter, area, and fluorescence intensity were assessed. Nonparametric statistical analysis (Mann–Whitney) was utilized. There was no difference in LD number for BT or BI for morulae and blastocyst; however, BI morulae presented larger LD compared with blastocyst stage embryos (286 µm2 v. 223 µm2; P < 0.05). Likewise, BI TF cells had more LD compared with inner cell mass (ICM) cells (48 v. 36; P < 0.05). BT TF cells exhibited larger LD compared with ICM cells (149 µm2 v. 128 µm2; P < 0.05), while BI embryos exhibited a larger area of LD in the ICM compared with the TF (591 µm2 v. 472 µm2; P < 0.05). In all embryos, BI contained more lipid droplets than BT (78 v. 49; P < 0.05). Across all quality grades (good, fair, and poor) there was no difference in the number of LD in BT embryos; however, BI grade-3 embryos presented more LD than grade-1 (36 v. 25). BT embryos LD were larger than BI LD (907 µm2 v. 625 µm2; P < 0.05). Fluorescence images showed higher arbitrary units of fluorescence (auf) for LD in BI. Compared with BT embryos (386 auf v. 280 auf; P < 0.05). These results suggest that BI embryos contain more and smaller LD than BT embryos and the LD described for BI embryo quality grade 1 are larger than those of quality grades 2 and 3, and even though the number of LD in morulae and blastocyst stage embryos are not different LD size is reduced as development occurs. Research funding provided by UNAM-DGAPA-PAPIIT IN200810.

scholarly journals Assessing the Cadmium Effects on the Benthic Foraminifer Ammonia cf. parkinsoniana: An Acute Toxicity Test

Water ◽  
2020 ◽  
Vol 12 (4) ◽  
pp. 1018 ◽  
Author(s):  
María Teresa Losada Ros ◽  
Eqbal Al-Enezi ◽  
Erica Cesarini ◽  
Barbara Canonico ◽  
Carla Bucci ◽  
...  
Keyword(s):  
Lipid Droplets ◽  
Nile Red ◽  
Natural Seawater ◽  
Benthic Foraminifer ◽  
Complementary Information ◽  
Direct Evaluation ◽  
Quality Guidelines ◽  
Bioassay Experiment ◽  
Confocal Images ◽  
Hazardous Pollutants

Heavy metals are one of the most hazardous pollutants in marine environments because of their bioaccumulation and biomagnification capabilities. Among them, cadmium (Cd) has been considered as one of the most dangerous for marine organisms. Here we incubated Ammonia cf. parkinsoniana specimens, a benthic foraminiferal taxon used in previous experiments, for up to 48 h in natural seawater with different concentrations of Cd to unravel the physiological change. We document a reduced pseudopodial activity of the Cd-treated specimens at concentrations >10–100 ppb in comparison with the control specimens. Moreover, confocal images of Cd-treated specimens using Nile Red as a fluorescent probe reveal an enhanced intracellular neutral lipid accumulation in the form of lipid droplets at 6 h and 12 h. This bioassay experiment allows for the direct evaluation of Cd-dose to A. cf. parkinsoniana-response relationships under laboratory controlled conditions and provides complementary information to field observations as well as to water quality guidelines and thresholds.

scholarly journals Nile red: a selective fluorescent stain for intracellular lipid droplets.

1985 ◽  
Vol 100 (3) ◽  
pp. 965-973 ◽  
Author(s):  
P Greenspan ◽  
E P Mayer ◽  
S D Fowler
Keyword(s):  
Lipid Droplets ◽  
Low Density Lipoprotein ◽  
Nile Red ◽  
Density Lipoprotein ◽  
Peritoneal Macrophages ◽  
Intracellular Lipid ◽  
Fluorescence Excitation ◽  
Flow Cytofluorometry ◽  
Aortic Smooth Muscle ◽  
Red Fluorescence

We report that the dye nile red, 9-diethylamino-5H-benzo[alpha]phenoxazine-5-one, is an excellent vital stain for the detection of intracellular lipid droplets by fluorescence microscopy and flow cytofluorometry. The specificity of the dye for lipid droplets was assessed on cultured aortic smooth muscle cells and on cultured peritoneal macrophages that were incubated with acetylated low density lipoprotein to induce cytoplasmic lipid overloading. Better selectivity for cytoplasmic lipid droplets was obtained when the cells were viewed for yellow-gold fluorescence (excitation, 450-500 nm; emission, greater than 528 nm) rather than red fluorescence (excitation, 515-560 nm; emission, greater than 590 nm). Nile red-stained, lipid droplet-filled macrophages exhibited greater fluorescence intensity than did nile red-stained control macrophages, and the two cell populations could be differentiated and analyzed by flow cytofluorometry. Such analyses could be performed with either yellow-gold or red fluorescence, but when few lipid droplets per cell were present, the yellow-gold fluorescence was more discriminating. Nile red exhibits properties of a near-ideal lysochrome. It is strongly fluorescent, but only in the presence of a hydrophobic environment. The dye is very soluble in the lipids it is intended to show, and it does not interact with any tissue constituent except by solution. Nile red can be applied to cells in an aqueous medium, and it does not dissolve the lipids it is supposed to reveal.

The role of intermediate filaments in adrenal steroidogenesis

1990 ◽  
Vol 97 (4) ◽  
pp. 679-687
Author(s):  
G. Almahbobi ◽  
P.F. Hall
Keyword(s):  
Electron Microscopy ◽  
Intermediate Filaments ◽  
Cholesterol Ester ◽  
Lipid Droplets ◽  
Specific Binding ◽  
Protein A ◽  
Nile Red ◽  
Hormone Synthesis ◽  
Adrenal Cells ◽  
Adrenal Steroidogenesis

Cholesterol is stored in adrenal cells as ester in lipid droplets, which are transported to mitochondria to provide a substrate for steroid hormone synthesis. Using mouse adrenal tumour cells (Y-1), we show here that approximately 33% of the adrenal cell cholesterol ester is bound tightly to intermediate filaments while the rest is either loosely attached or free in the cytosol. Specific binding of droplets to intermediate filaments was demonstrated by immunofluorescence and electron microscopy. Immunofluorescence was based upon Nile Red to stain lipid and antibodies to vimentin, actin and tubulin. Electron microscopy, including immunoelectron microscopy with protein A conjugated to gold particles (5 nm), was used to examine whole mounts of cytoskeletons and intermediate filaments. Immunofluorescence reveals that bound droplets are surrounded by a capsule containing vimentin and can be removed from the filaments by extraction with ethanol or 6 M urea. Negative staining of the urea extracts revealed isolated droplets. To the extent that cholesterol ester is the storage form of steroidogenic cholesterol, the knowledge that lipid droplets containing such esters are attached to intermediate filaments may prove important in unravelling the complex process of the transport of cholesterol to mitochondria.

scholarly journals X-ray microanalysis of aldehyde-fixed glycogen contrast-stained by OsVI . FeII and OsVI . RuIV complexes.

1980 ◽  
Vol 28 (11) ◽  
pp. 1242-1250 ◽  
Author(s):  
W C de Bruijn ◽  
J M van Buitenen
Keyword(s):  
Electron Microscope ◽  
Complex Formation ◽  
Rat Liver ◽  
Lipid Droplets ◽  
Liver Glycogen ◽  
The Other ◽  
Glutaraldehyde Fixation ◽  
Fixed Tissue ◽  
X Ray ◽  
The Mean

The composition of the contrast-donating complex of rat liver glycogen, nucleoplasm, erythrocytes, and mitochondria was established by X-ray microanalysis. In these compartments the presence of osmium and iron was shown qualitatively in tissue after glutaraldehyde fixation, treated with OsVIIIO4 plus K4FeII(CN)6 and in similar tissue treated with a combination of K2OsVIO4 plus K4FeII(CN)6. Osmium and ruthenium were detected in these compartments, in aldehyde-fixed tissue treated with mixtures containing K2RuIVL(CN)6 rather than K4FeII(CN)6. The iron detected in the glycogen, nucleoplasm, erythrocytes, and mitochondria of tissue treated with K2RuIV(CN)6 mixtures proved to derive from sources inside the electron microscope, and had to be considered an artifact. Quantitatively, the mean atomic ratios of osmium-to-iron and osmium-to-ruthenium were determined from spectra obtained by point analyses of the same compartments (glycogen, nucleoplasm, mitochondria, lipid droplets, and erythrocytes). After correction of the spectra for the instrumental iron contribution, the osmium-to-iron and osmium-to-ruthenium ratios in the glycogen were about 1:3 for tissue treated with those combinations including K2OsVIO4. In the other compartments, the osmium-to-iron and osmium-to-ruthenium ratios were virtually 1:0. For Os-VIIIO4 in combination with potassium ferrouscyanide however the osmium-to-iron ratio was 1:7 in the glycogen and 1:5 in all other compartments. OsVIIIO4 was combined with potassium ruthenium-cyanide, the osmium-to-ruthenium ratio was 1:2 in the glycogen and 2:1 in the other compartments. These results support our view that the selective glycogen contrast is obtained by complex formation.

A Clinico-Pathologic Study of Clear-Cell Hepatocellular Carcinoma

1987 ◽  
Vol 73 (4) ◽  
pp. 389-395 ◽  
Author(s):  
Riccardo A. Audisio ◽  
Luigia Bombelli ◽  
Luciano Lombardi ◽  
Salvatore Andreola
Keyword(s):  
Hepatocellular Carcinoma ◽  
Tumor Cells ◽  
Clinical Features ◽  
Ultrastructural Study ◽  
Lipid Droplets ◽  
Clear Cell ◽  
Clinical Course ◽  
The Other ◽  
Hepatocellular Carcinomas

We reviewed the pathologic and clinical features of 7 cases of clear-cell hepatocellular carcinomas. Tumor cells had a typical clear, empty cytoplasm due to prominent accumulation of glycogen and lipid droplets, as demonstrated by ultrastructural study in one of our cases. Follow-up of the 7 patients did not confirm the better prognosis of this tumor reported by other investigators. The clinical course of our cases was more unfavorable than that of the other 130 cases of usual hepatocellular carcinomas treated at this Institute.

scholarly journals Evidence of foam cell and cholesterol crystal formation in macrophages incubated with oxidized LDL by fluorescence and electron microscopy.

1995 ◽  
Vol 43 (10) ◽  
pp. 1071-1078 ◽  
Author(s):  
A M Klinkner ◽  
C R Waites ◽  
W D Kerns ◽  
P J Bugelski
Keyword(s):  
Electron Microscopy ◽  
Lipid Droplets ◽  
Foam Cell ◽  
Oxidized Ldl ◽  
Nile Red ◽  
Foam Cells ◽  
Cholesterol Crystal ◽  
Atherosclerotic Lesions ◽  
Membrane Bound ◽  
Nile Red Staining

Macrophage-derived foam cells are a prominent component of developing atherosclerotic lesions. We describe an in vitro model of foam cell formation which mimics some aspects of the evolution of foam cells in mature atherosclerotic lesions. Thioglycollate-elicited mouse peritoneal macrophages were incubated with copper-oxidized LDL (ox-LDL) for periods up to 168 hr. Identifiable foam cells were present after incubation with ox-LDL at 24, 72, and 168 hr. Control cells incubated without ox-LDL did not form foam cells. Fluorescence microscopy after staining with Nile red exhibited progressive accumulation of lipids, and transmission electron microscopy (TEM) showed distinct ultrastructural changes over time. Macrophages at 24 hr had a few non-membrane-bound lipid droplets but were otherwise identical to control cells. These lipid droplets fluoresced yellow-gold after Nile red staining. After 72 hr of incubation with ox-LDL, in addition to increased numbers of non-membrane-bound lipid inclusions, macrophages contained membrane-bound multilamellar lipoid structures. These multilamellar structures corresponded to areas of reddish-orange fluorescence after Nile red staining. In macrophages incubated with ox-LDL for 168 hr, the amount of cellular lipid was further increased and cholesterol crystal profiles were apparent within some multilamellar lipoid structures. Biochemical analysis showed that the total cholesterol content steadily increased over 168 hr. The increase in total cholesterol was accompanied by a dramatic increase in free cholesterol between 72 and 168 hr. These results demonstrate that long-term incubation of macrophages with ox-LDL increased lipid deposition in cultured cells and that, under the conditions studied, cholesterol crystals formed in macrophage foam cells. Moreover, this system allows investigation of the evolution of foam cells showing some characteristics of those found in atherosclerotic lesions.

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