scholarly journals LIGHT AND ELECTRON MICROSCOPE LOCALIZATION OF BINDING SITES OF ANTIBODIES AGAINST OVINE LUTEINIZING HORMONE AND ITS TWO SUBUNITS IN RAT ADENOHYPOPHYSIS USING PEROXIDASE-LABELED ANTIBODY TECHNIQUE

1973 ◽  
Vol 58 (3) ◽  
pp. 503-521 ◽  
Author(s):  
C. Tougard ◽  
B. Kerdelhue ◽  
A. Tixier-Vidal ◽  
M. Jutisz

The binding sites of antisera generated in the guinea pig against ovine luteinizing hormone (oLH) and its two subunits (oLHα and oLHß) have been localized in rat anterior pituitaries taken from normal or castrated males and from ovariectomized females with the peroxidase-labeled antibody method, using light and electron microscopy. With the light microscope, the cells positive with antiserum to ovine luteinizing hormone (A-oLH) were violet after the Alcian blue-periodic acid-Schiff (AB-PAS) staining; they were also positive for A-oLHα and for A-oLHß and, from castrated males, they displayed an increased affinity for A-oLHß. Another cell type which was blue after the AB-PAS method reacted with the A-oLHα only; these cells, presumably thyrotropic cells, were retracted after castration and, besides their affinity for A-oLHα, acquired an affinity for A-oLHß. As seen through the electron microscope, two cell types were positive for A-oLH, A-oLHß, and A-oLHα and may be identified as luteinizing hormone-secreting cells. Type A cells were characterized by two classes of rounded, secretory granules. Type B cells were smaller and contained only small secretory granules. 1 mo after the rats were castrated the type A cells were hypertrophied and vacuolized. In both cases the secretory granules were the main sites of the antigenicity with the three antisera. A positive reaction was also found in the cytoplasm, particularly in hypertrophied cells from ovariectomized females and with A-oLHß. The cisternae of the rough endoplasmic reticulum were usually negative, except in highly degranulated cells from ovariectomized females and with A-oLHß.

1980 ◽  
Vol 28 (2) ◽  
pp. 101-114 ◽  
Author(s):  
C Tougard ◽  
R Picart ◽  
A Tixier-Vidal

The binding sites of antisera (anti) to the beta (beta) subunits of rat follicle-stimulating hormone (rFSH), rat luteinizing hormone (rLH), and rat thyroid-stimulating hormone (rTSH) have been localized in rat anterior pituitaries by immunocytochemistry using light and electron microscopy. With the light microscope, LHbeta and FSHbeta were found in the same cells, which were violet after the alcian blue-periodic acid Schiff (AB-PAS) staining. TSHbeta was found in polygonal or stellate cells that were blue after AB-PAS. With the electron microscope, the thyrotropic cells contained very small secretory granules. LHbeta and FSHbeta were found in various types of cells (types A and B and their intermediate forms), which had previously been identified as gonadotropic cells. On serial ultrathin sections using the postembedding method the same cells and even some granules inside these cells were stained by both anti-rLHbeta and anti-rFSHbeta. A comparison of binding sites of anti-rLHbeta was performed using the preembeeding and the postembeeding methods. Antigenicity was observed on secretory granules whatever the method used. However, binding sites of anti-rLHbeta were detected inside the cisternae of the rough endoplasmic reticulum only with the preembedding method.


1986 ◽  
Vol 64 (4) ◽  
pp. 875-884 ◽  
Author(s):  
Patricia Schulz ◽  
William A. Jensen

Ovules of Capsella bursa-pastoris at the dyad and tetrad stages of meiosis and at the megaspore and two-nucleate stages of the gametophyte were studied with the electron microscope. The cells of the dyad and tetrad are separated by aniline blue fluorescent cross walls and receive all types of organelles and autophagic vacuoles that were present in the meiocyte. Autophagic vacuoles enclose ribosomes and organelles and show reaction product for acid phosphatase. Autophagic vacuoles and some plastids are absorbed into the enlarging vacuoles of the growing megaspore. Other plastids appear to survive meiosis and there is no evidence for their de novo origin. Some mitochondria appear to degenerate in the enlarging megaspore but others look healthy and there is no evidence for the de novo origin of mitochondria. The nucleolus of the developing megaspore becomes very large and the cytoplasm is extremely dense with ribosomes. The cell wall is thickened by an electron-translucent, periodic acid – Schiff negative, aniline blue fluorescent material and contains plasmodesmata that link the megaspore with the nucellus. The plasmalemma of the growing megaspore produces microvilluslike extensions into this wall that disappear with the formation of the two-nucleate gametophyte. Plasmodesmata disappear from the cell wall at the four-nucleate stage.


2021 ◽  
Vol 71 (1) ◽  
pp. 11-17
Author(s):  
Muhammad Ilyas Iqbal ◽  
Muchtaruddin Mansyur ◽  
Pudji Sari ◽  
Dwi Anita Suryandari ◽  
Pramudianto

Intoduction: Acute and chronic exposure to toluene at high doses is known to affect all organs of the body including the spermatogenesis process. In the industrial sector, the use of toluene as a solvent is still widely used, up to 10 million tons per year. The control over health problems that may occur is carried out by applying work exposure threshold values. This research aims to explore the effect of toluene exposure at the threshold value range on spermatogenesis.Method: This research used laboratory experiment on 30 male Wistar rats which were divided into five groups of different exposure levels, namely 12.5 parts per million (ppm], 25 ppm, 50 ppm, 100 ppm, and no exposure (control). Exposure was given for 4 hours daily over 14 days through a hood with measured release in the glass cage. The toluene exposure markers observed were Malondialdehyde (MDA) in the blood tissue and testicles using the Thiobarbituric Acid Reactive Substances (TBARS) method. The effect on the spermatogenicity process was assessed by counting the spermatogonia A cells of male Wistar rats with Periodic Acid Schiff (PAS) staining and is calculated by the Abercrombie formula. Analysis of the correlation between the level of exposure and its effect on the increase in malondialdehyde, and spermatogenesis was carried out using the Spearman correlation analysis.Result: There was a moderately positive correlation between levels of toluene exposure and plasma MDA levels (r = 0.42; p = 0.025). Meanwhile, on [the issue of] the quantity of spermatogonia cells, a high level of negative correlation with exposure levels was obtained (r = -0.68; p = 0.001).Conclusion: Toluene exposure in male Wistar rats within the range of threshold values influenced the increase in plasma MDA levels and decreased the Spermatogenia A cells. However, toluene exposure did not affect the testicular MDA levels of male Wistar rats.


Neurosurgery ◽  
1990 ◽  
Vol 26 (3) ◽  
pp. 397-403 ◽  
Author(s):  
Kathryn K. Berg ◽  
Bernd W. Scheithauer ◽  
Ignacio Felix ◽  
Kalman Kovacs ◽  
Eva Horvath ◽  
...  

Abstract Eight surgical and one autopsy specimen of pituitary adenomas (six cases of Cushing's disease, two of Nelson's syndrome. and one of hypopituitarism) were studied by histochemical, immunohistocytological, and ultrastructural methods. Eight tumors showed the characteristic histochemical profile of corticotroph adenoma—amphophilic to basophilic, and periodic acid-Schiff-positive to some extent. In all tumors, immunohistochemical studies revealed adrenocorticotropic hormone (ACTH) and à-subunit in the cytoplasm of some adenoma cells. By electron microscopy, seven tumors were found to be monomorphous; six were typical corticotroph adenomas and one was a subtype II silent corticotroph adenoma. One unique lesion was bimorphous—i.e., composed of corticotrophs as well as cells resembling glycoprotein cells. Immunoelectron microscopy by the double-labeling immunogold technique, performed on one corticotroph adenoma, demonstrated the presence of ACTH and à-subunit not only within the same adenoma cells but also within the same secretory granules. The cytogenesis of ACTH à-subunit tumors, a rare form of plurihormonal adenoma. remains to be elucidated. The duration of disease associated with these tumors exceeded the duration in patients with ordinary corticotroph adenomas. Given the low frequency with which increases in serum à-subunit are detectable in patients with such tumors—13% in this series—hormone production is not recognized at preoperative evaluation.


1983 ◽  
Vol 61 (1) ◽  
pp. 182-194 ◽  
Author(s):  
E. E. Dudenhausen ◽  
P. Talbot

The spermatophore of the crayfish Pacifastacus leniusculus consists of two main parts: a sperm mass composed of sperm embedded in a dense fibrillar matrix and an acellular wall which surrounds the sperm mass and is formed from secretions produced in the vas deferens. Following extrusion from the male, the spermatophore wall, which is initially soft and sticky, undergoes a hardening process. In this study, the structure of the spermatophore walls of unextruded (soft) and hardened spermatophores were compared using light and electron microscopy. The wall of the unextruded spermatophore is composed of three concentric layers: a thin primary spermatophore layer which directly surrounds the sperm mass; a thick middle layer composed primarily of electron-dense, spherical granules; and a thick outer layer formed from a dense globular secretion. The primary spermatophore layer and outer globular layer are positive for carbohydrate with the periodic acid – Schiff method. Following extrusion and hardening, the walls of spermatophores showed several structural changes. These are (i) division of the middle granular layer into a compact inner region and a highly reticulated outer region; (ii) the loss of the outer globular layer; and (iii) the formation of a thickened ridge along one side of the spermatophore wall. The thickened ridge is fibrillar in structure and is believed to be derived from a structural modification of the outer globular layer. No structural modifications in the primary spermatophore layer were observed. We interpret these observations to indicate that the outer globular layer functions in attachment of the spermatophore to the female and the middle layer is involved in spermatophore hardening and sperm protection during storage.


1981 ◽  
Vol 59 (1) ◽  
pp. 61-69 ◽  
Author(s):  
B. J. Crawford ◽  
Alex Yan ◽  
M. MacDonald

The changes in surface morphology during the reexpression of differentiation of chick retinal pigmented epithelial cells (RPE) in clonal culture have been studied using the scanning electron microscope (SEM) and transmission electron microscope (TEM) and compared with those described in vivo. Three-week-old colonies demonstrated a gradual change in apical surface morphology along any colony radius. At the outer edge, the cell surfaces were either smooth with a few small filamentous protrusions or showed a varying number of large blebs. Toward the centre of the colony the surfaces demonstrated a gradual increase in filamentous protrusions. The apical surfaces of the most densely pigmented cells at the centre of the colony consisted mainly of small rounded protrusions. The changes in surface morphology of cells in the centre of younger colonies during redifferentiation were similar to those found along the radius of a 3-week-old colony. The results show that older colonies have all of the morphological stages of the redifferentiation process (and possibly the biochemical ones as well) arranged along any radius.The basal surfaces of all the colonies were covered by a thin acellular membrane that stained positively with periodic acid–Schiff (PAS) and which may contain fibronectins and appears to be involved in cellular attachment.


1966 ◽  
Vol 14 (12) ◽  
pp. 884-897 ◽  
Author(s):  
SERGEI P. SOROKIN

Lungs from marsupials, bats and rodents were studied by light and electron microscopy. In all three groups, the great alveolar cells exhibit similar morphologic and cytochemical characteristics. Cytoplasmic vacuoles seen in these cells by light microscopy correspond to cytosomes that are demonstrable in them by electron microscopy. Such cytosomes are osmiophilic, periodic acid-Schiff-positive and stainable with Sudan black after acetone extraction. After fixation in a mixture of aldehydes, followed by extraction in chloroform-methanol and postfixation in osmium tetroxide, cytosomes lose their osmiophilia. The cytoplasm of the great alveolar cell is notable for a loosely ordered granular endoplasmic reticulum, an extensive Golgi apparatus and numerous multivesicular bodies. Many forms transitional in appearance between multivesicular bodies and cytosomes are present. In these, osmiophilic matter occupies the intervesicular space. It is proposed that these bodies are the precursors of cytosomes. The cytosomes are interpreted to be products of the "lysosomal" system in this cell. Ultimately they are secreted onto the alveolar surface.


Author(s):  
S. Yamashiro ◽  
D. Wilson ◽  
J. St. George ◽  
D. Hyde ◽  
C. Plopper ◽  
...  

In the past, ozone inhalation studies have focused on the lower airways and lung parenchyma. The purpose of this study was to evaluate the effects of ozone on submucosal glands of upper airways. Six adult male bonnet monkeys were exposed to 0.64 ppm ozone continuously for 7 days, and three were exposed to chamber conditions without ozone. The animals were exsanguinated under barbiturate anesthesia. The trachea and lung were fixed by airway infusion of Karnovsky's fixative, which was adjusted to pH 7.4 and 440 milliOsmols. Sagittal sections of ventral trachea were embedded in glycol methacrylate and Araldite 502 for light and electron microscopy. One micrometer methacrylate sections were stained with Alcian blue-periodic acid Schiff (AB/PAS). Selected areas of Araldite-embedded tissue were sectioned for transmission electron microscopy, stained with uranyl acetate and lead citrate and examined with a Zeiss EM 10. Volume percentages of the lumen, granular and nongranular regions of fhe gland and the duct wall, respectively, were estimated by stereologic methods on AB/PAS stained sections.


Author(s):  
B. Giammara ◽  
T. Romaine ◽  
W. Ambrose ◽  
J. Hanker

Many variations of the periodic acid-Schiff(PAS) reaction have been utilized for electron microscopy based on the Gomori periodic acid-silver methenamine reaction (1) or the periodic acid-thiocarbohydrazide-osmium tetroxide(PATCO) reaction (2,3). These reactions are widely employed and have been very useful for the demonstration of one or more biomacromolecules or structures such as glycogen, basement membranes, reticular fibers or lipopolysaccharide. However, these reactions have various drawbacks such as complexity of methodology, ability to stain only a limited number of these components, or lack of adaptability for both light and electron microscopy. Our newly devised PATS reaction is relatively easy to perform. A full description of the details must await the outcome of a pending patent application. It consists essentially of a stepwise treatment of the sample with periodic acid, thiocarbohydrazide(TCH) and silver methenamine.


Author(s):  
Paul L. Krupa ◽  
Arya K. Bal ◽  
Gilles H. Cousineau

The fine structure of various gland cells and their secretory products was studied in the invasive stage (cercaria) of the platyhelminth parasite, Cryptocotyle lingua. Secretory granules or droplets occur in several different specialized cell types, but those that we call attention to here are found in the (1) surface cytoplasmic tegument or “cuticle”, (2) ducts of cephalic (penetration) glands, and (3) epithelial lining of the “excretory bladder”.The tegumental granules appear as numerous, membrane-bounded circular or oval profiles of uniform density (Fig. 1). They are scattered more or less randomly among mitochondria and other inclusion bodies of the tegument. Some of the longer granules, with a length to width ratio of about 7 to 1, have their long axes oriented perpendicularly to the surface plasma membrane of the parasite. In cercariae tested for acid hydrolases with sodium β-glycerophosphate in Barka and Anderson's modification of Gomori's medium, clumps of reaction product appear in the vicinity of the granules and elsewhere within the tegument, but not within the granules themselves. As granules that stain with periodic acid-Schiff, they are seen in certain subsurface gland cells as well as in the tegument under the light microscope.


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