scholarly journals STUDIES ON MICROPEROXISOMES II. A CYTOCHEMICAL METHOD FOR LIGHT AND ELECTRON MICROSCOPY

1972 ◽  
Vol 20 (12) ◽  
pp. 1006-1023 ◽  
Author(s):  
ALEX B. NOVIKOFF ◽  
PHYLLIS M. NOVIKOFF ◽  
CLEVELAND DAVIS ◽  
NELSON QUINTANA

A modification of the Novikoff-Goldfischer alkaline 3,3'-diaminobenzidine medium for visualizing peroxisomes is described. It makes possible light microscopic as well as electron microscopic studies of a recently described class of peroxisomes, the microperoxisomes. Potassium cyanide (5 x 10–3 M) is included in the medium to inhibit mitochondrial staining, the pH is 9.7 and there is a high concentration of H2O2 (0.05%). Two cell types have been chosen to illustrate the advantages of the new procedure for demonstrating the microperoxisomes: the absorptive cells in the human jejunum and the distal tubule cells in the guinea pig kidney. Suggestive relations of microperoxisomes and lipid are described in the human jejunum. The microperoxisomes are strategically located between smooth endoplasmic reticulum that radiates toward the organelles and contains lipid droplets and "central domains" of highly specialized endoplasmic reticulum which do not show the lipid droplets. The microperoxisomes are also present at the periphery of large lipid-like drops. In the guinea pig kidney tubule there is a striking difference between the thick limb of Henle and distal tubule. The distal tubule has a population of cells with large numbers of microperoxisomes readily visible by light microscopy; these cells are not present in the thick limb of Henle. Other differences between the two are also described.

Author(s):  
F. G. Zaki

Addition of lithocholic acid (LCA), a naturally occurring bile acid in mammals, to a low protein diet fed to rats induced marked inflammatory reaction in the hepatic cells followed by hydropic degeneration and ductular cell proliferation. These changes were accompanied by dilatation and hyperplasia of the common bile duct and formation of “gallstones”. All these changes were reversible when LCA was withdrawn from the low protein diet except for the hardened gallstones which persisted.Electron microscopic studies revealed marked alterations in the hepatic cells. Early changes included disorganization, fragmentation of the rough endoplasmic reticulum and detachment of its ribosomes. Free ribosomes, either singly or arranged in small clusters were frequently seen in most of the hepatic cells. Vesiculation of the smooth endoplasmic reticulum was often encountered as early as one week after the administration of LCA (Fig. 1).


1975 ◽  
Vol 18 (1) ◽  
pp. 1-17
Author(s):  
A. Pleshkewych ◽  
L. Levine

A prominent cytoplasmic inclusion present in living mouse primary spermatocytes has been observed by both light and electron microscopy. It began to form at prometaphase and continued to increase in thickness and length as the cells developed. By metaphase it was a distinct sausage-shaped boundary that enclosed a portion of the cytoplasm between the spindle and the cell membrane. At the end of metaphase, the inclusion reached its maximum length. At telophase, it was divided between the daughter secondaries. The inclusion persisted as a circular contour in the interphase secondary spermatocyte. Electron microscopy of the same cultured cells that were previously observed with light microscopy revealed that the inclusion was a distinctive formation of membranes. It consisted of agranular cisternae and vesicles, and was therefore a membranous complex. Many of the smaller vesicles in the membranous complex resembled those found in the spindle. The cisternae in the membranous complex were identical to the cisternal endoplasmic reticulum of interphase primary spermatocytes. Nevertheless, the organization of vesicles and cisternae into the membranous complex was unique for the primaries in division stages, since such an organization was not present in their interphase stages.


1997 ◽  
Vol 3 (S2) ◽  
pp. 55-56
Author(s):  
H.J. Finol ◽  
D.D. Mondragón ◽  
Y.M. González ◽  
C. Paradisi ◽  
N. González ◽  
...  

Although liver function tests could be abnormal in humans taking cocaine the histopathological basis for this disorder has not been well established. Light microscopic studies have shown the existence of peripheral, centrilobular or diffuse necrosis. The only electron microscopic investigation we could find reports hepatocyte alterations including dilated rough endoplasmic reticulum, hypertrophy of smooth endoplasmic reticulum, and existence of phagolysosomes. In this work we report the liver ultrastructural pathology in chronic cocaine users.Liver biopsies were obtained in five male patients, 25-44 years old. These patients had consumed cocaine and other drugs (marihuana, alcohol, amphetamines, etc..) for 7-30 years. All of them had altered liver function tests. Tissue samples were processed with routine techniques for transmission electron microscopy and observed in a Hitachi H-500 electron microscope.Abnormalities observed included those previously reported as swollen rough and smooth endoplasmic reticulum, presence of autophagic vacuoles and lipid deposition.


1983 ◽  
Vol 31 (8) ◽  
pp. 987-999 ◽  
Author(s):  
J Roth

A method is described for the electron microscopic detection of lectin-binding sites in different cellular compartments and extracellular structures that uses thin sections from resin-embedded tissues. Various lectins (Ricinus communis lectin I and II, peanut lectin, Lotus tetragonolobus lectin, Ulex europeus lectin I, Lens culinaris lectin, Helix pomatia lectin, and soybean lectin) were bound to particles of colloidal gold and used for direct staining of thin sections or glycoprotein--gold complexes were prepared and applied in an indirect technique (concanavalin A and horseradish peroxidase--gold complex; wheat germ lectin and ovomucoid--gold complex). The details for preparation of such complexes from 14 nm gold particles are reported. The conditions of tissue processing that gave satisfactory staining results and good fine structure preservation were mild aldehyde fixation without osmification and low temperature embedding with the hydrophilic resin Lowicryl K4M. None of the so-called etching procedures was necessary prior to labeling of Lowicryl K4M thin sections. Examples of the use of this approach for detection of glycoconjugates in the rough endoplasmic reticulum, Golgi apparatus, and mucin of intestinal goblet cells as well as plasma membrane and various intracellular structures of absorptive intestinal and renal tubular cells are shown. A comparison is made with preembedding staining results on Concanavalin A-binding site localization in rat liver which shows that problems of penetration common in such a technique are circumvented by the postembedding approach described here. Concanavalin A-binding sites were not only consistently found in nuclear envelope, rough and smooth endoplasmic reticulum, plasma membranes, and collagen fibers, but also in mitochondria, glycogen, ribosomes, and nucleus. These data and those of a previous investigation (Roth J, Cytochem 31:547, 1983) prove the applicability of this cytochemical technique for postembedding localization of glycoconjugates by light and electron microscopy.


2001 ◽  
Vol 170 (1) ◽  
pp. 99-111 ◽  
Author(s):  
K Toda ◽  
K Takeda ◽  
T Okada ◽  
S Akira ◽  
T Saibara ◽  
...  

Aromatase P450 (CYP19) is an enzyme catalysing the conversion of androgens into oestrogens. We generated mice lacking aromatase activity (ArKO) by targeted disruption of Cyp19 and report the characteristic features of the ArKO ovaries and uteri as revealed by histological and biochemical analyses. ArKO females were totally infertile but there were as many developing follicles in their ovaries at 8 weeks of age as in wild-type ovaries. Nevertheless, no typical corpus luteum was observed in the ArKO ovaries. Electron microscopy revealed the presence of well-developed smooth endoplasmic reticulum, few lipid droplets and mitochondria with less organized tubular structures in the ArKO luteinized interstitial cells. These ultrastructural features were different from those of the wild-type interstitial cells, where there are many lipid droplets and mitochondria with well-developed tubular structures, characteristic of steroid-producing cells. When ArKO mice were supplemented with 17beta-oestradiol (E(2); 15 microg/mouse) every fourth day from 4 weeks of age for 1 month, increased numbers of follicles were observed in the ovaries as compared with those of untreated ArKO mice, although no typical corpus luteum was detectable. Ultrastructural analysis revealed the disappearance of the accumulated smooth endoplasmic reticulum in the luteinized interstitial cells after E(2 )supplementation. Transcripts of pro-apoptotic genes such as p53 and Bax genes were markedly elevated in the ArKO ovaries as compared with those of wild-type mice. Although E(2) supplementation did not cause suppression of the elevated expression of p53 and Bax mRNAs, it caused marked enhancement of expression levels of lactoferrin and progesterone receptor mRNAs in the uteri as well as increases in uterine wet weight. At 8 months of age, ArKO mice developed haemorrhages in the ovaries, in which follicles were nearly depleted, while age-matched wild-type females still had many ovarian follicles. Furthermore, macrophage-like cells were occasionally observed in the ArKO ovarian follicles. These results suggested that targeted disruption of Cyp19 caused anovulation and precocious depletion of ovarian follicles. Additionally, analysis of mice supplemented with E(2) demonstrated that E(2) apparently supports development of ovarian follicles, although it did not restore the defect in ovulation.


1973 ◽  
Vol 51 (12) ◽  
pp. 2307-2314 ◽  
Author(s):  
Saeed R. Khan ◽  
Henry C. Aldrich

Termitaria snyderi Thaxter forms small discoid lesions on the exoskeleton of different species of termites. Its conidiogenesis has been studied by light and electron microscopy. The phialides are oriented parallel in a closely packed sporodochium. The conidia are produced endogenously in basipetal succession from a fixed conidiogenous locus and are liberated when the tip is broken off the phialide as a result of the force applied by the formation of new conidia. The area of the phialide beyond the locus forms a tubular collarette. The conidium initial buds out at the locus and after it has received its organelles and reached a certain size it is delimited by a centripetally growing transverse septum. The region of the growing septum has many vesicles which may be involved in cross wall synthesis. Conidia are cylindrical, uninucleate, and double-walled. They have mitochondria, endoplasmic reticulum (ER), conspicuous lipid droplets, and vacuoles. Each conidiophore has long mitochondria, elongate nuclei, and much endoplasmic reticulum. The plasmalemma of the conidiophore is highly convoluted.


1979 ◽  
Vol 91 (1) ◽  
pp. 134-149 ◽  
Author(s):  
Peter M. Robinson ◽  
Elisabeth J. Rowe ◽  
E. Marelyn Wintour

ABSTRACT The cortex of sheep foetal adrenal glands from 25 days gestation until newborn (term equals 147 ± 3 days) were examined by light and electron microscopy. Three stages of development are of particular importance in relating structure to function: 1) from 35 to 60 days, 2) from 60 to 120 days and 3) from 120 days to term. Between 35 and 60 days one cortical cell type predominated. It contained mitochondria with lamellar and vesicular cristae, scattered long strands of granular endoplasmic reticulum and only small amounts of smooth endoplasmic reticulum. After about 60 days two zones were apparent in the cortex and chromaffin cells became concentrated in the medulla. After 80 days the outer zone contained cells which resembled mature zona glomerulosa cells and the cells in the inner zone remained like those seen between 35 and 60 days, except they contained even less smooth endoplasmic reticulum. However, after about 90 days a small number of deep inner zone cells contained mitochondria with vesicular cristae which thus resemble mitochondria in the mature zona fasciculata. From about 120 days there was an increase in the number of cells in the inner zone that contained mitochondria with vesicular cristae. These cells also contained substantial quantities of smooth endoplasmic reticulum. At term most inner zone cells have this mature appearance. Thus there is no "foetal cortex" in the sheep analogous to that found in human adrenal development, i. e. there is no prominent zone of cells containing large amounts of smooth endoplasmic reticulum which is present throughout most of the foetal period of development, and which regresses at birth. The structure of the cells present between 35 and 60 days was unexpected because it has been shown previously that sheep foetal adrenals of this age are capable of producing relatively large quantities of steroid hormones. However, the appearance of cells resembling mature zona glomerulosa cells at about 80 days correlates with the previously demonstrated ability of sheep adrenal glands of this age to produce relatively large quantities of aldosterone. The rapid development of numbers of mature cells in the last 3 weeks of gestation correlates with the previously described ability of near term sheep foetal adrenals to produce very large quantities of steroid hormones.


1968 ◽  
Vol 16 (5) ◽  
pp. 337-345 ◽  
Author(s):  
ZOLTAN POSALAKI ◽  
TIBOR BARKA

The alterations of hepatic microsomal fractions were studied in fasting rats given porphyrogenic doses of allylisopropylacetamide. A single dose of allylisopropylacetamide caused significant enlargement of the liver within 14 hr and in 24 hr the liver was about 49% heavier. Microsomal phospholipids of liver more than doubled in 48 hr after the administration of allylisopropylacetamide. This increase was accounted for principally by an increase in phospholipids of the smooth endoplasmic reticulum. Incorporation of 32P indi cated that the increase in microsomal phospholipids was caused mainly by a reduction of the rate of catabolism rather than by an augmentation of the rate of synthesis. Electron microscopic observations in agreement with the fractionation studies revealed a hypertrophy of the smooth membranes of the endoplasmic reticulum.


1979 ◽  
Vol 27 (6) ◽  
pp. 1017-1028 ◽  
Author(s):  
B S Weakley

Osmium-pyroantimonate solutions for the precipitation of cations are unsuitable for use with delicate mammalian oocytes. A variant of the pyroantimonate technique employing a mixture of pyroantimonate and glutaraldehyde has been found to give successful and repeatable results if a fixation time of 4 hr is used. Calcium-containing antimonate precipitates were localized principally in nuclei, smooth endoplasmic reticulum, Golgi apparatus, mitochondria, and cytoplasmic processes of both oocytes and follicle cells, and along the plasma membrane in small oocytes. Deposits were also concentrated around the periphery of lipid droplets in the follicle cells. The presence of calcium in the precipitates was confirmed by x-ray microprobe analysis.


Author(s):  
Yan Qiao ◽  
Goro Asano ◽  
Izumi Kashiwado ◽  
Yasuo Hattori

Vertigo may be induced by various factors such as vascular changes including arteriosclerosis and hypercholesterolemia in the inner ear. In orter to clarify the mechanism of vertigo, the inner ear of guinea pig in hypercholesterolemia condition was investigated by electron microscopic techniques.The guinea pigs have served as experimental materials. They were divided into two groups: one received non-administration of cholesterol and the other received the administration of cholesterol. The vestibular macula in inner ear were observed at 1, 2 and 3 months after the administration of cholesterol and also vascular function was investigated by using horseradish peroxidase as tracer.Ultrastructural changes such as the depletion of microvilli on the cell surface and increased microvesicles were detected in the cytoplasm of sensory cell after cholesterol administration. After two months, lipid droplets were demonstrated in the cytoplasm, associated with intercellular edema. The cytoplasmic projection with vacuolization was demonstrated in the surface of sensory cells in the vestibular macula after three months of cholesterol administration.


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