Fine Structural Changes in the Adenohypophyses of Rats Following Destruction of the Pituitary Stalk

Author(s):  
K. Kovacs ◽  
E. Horvath ◽  
J. M. Bilbao ◽  
F. A. Laszlo ◽  
I. Domokos

Electrolytic lesions of the pituitary stalk in rats interrupt adenohypophysial blood flow and result in massive infarction of the anterior lobe. In order to obtain a deeper insight into the morphogenesis of tissue injury and to reveal the sequence of events, a fine structural investigation was undertaken on adenohypophyses of rats at various intervals following destruction of the pituitary stalk.The pituitary stalk was destroyed electrolytically, with a Horsley-Clarke apparatus on 27 male rats of the R-Amsterdam strain, weighing 180-200 g. Thirty minutes, 1,2,4,6 and 24 hours after surgery the animals were perfused with a glutaraldehyde-formalin solution. The skulls were then opened and the pituitary glands removed. The anterior lobes were fixed in glutaraldehyde-formalin solution, postfixed in osmium tetroxide and embedded in Durcupan. Ultrathin sections were stained with uranyl acetate and lead citrate and investigated with a Philips 300 electron microscope.

Author(s):  
E. Horvath ◽  
K. Kovacs ◽  
I. E. Stratmann ◽  
C. Ezrin

Surgically removed human pituitary glands as well as pituitary tumors fixed in glutaraldehyde, postfixed in osmium tetroxide, embedded in epon resin, stained with uranyl acetate and lead citrate have been investigated by electron microscopy in order to correlate ultrastructure with functional activity. In the course of this study two distinct types of microfilaments have been identified in the cytoplasm of adenohypophysiocytes.Type I microfilaments (Fig. 1) were found in the cytoplasm of anterior lobe cells of five female subjects with disseminated mammary cancer and two patients with severe diabetes mellitus. The breast cancer patients were treated pre-operatively for various periods of time with different doses of oxysteroids. The microfilaments had an average diameter of JO A, formed parallel bundles, were scattered irregularly in the cytoplasm and were frequently located in the perikaryon. They were not membrane-bound and failed to show any periodicity.


Author(s):  
M. E. Bayer

Cultures of Escherichia coli CIA (Bertani) growing logarithmically in nutrient medium were infected with 5 to 200 bacteriophage §X 174 per cell. At various times after infection the cultures were fixed for 1 hour in nutrient medium containing 5% formaldehyde pH 7, then pelleted and resuspended in 1% OSO4 in L-medium pH 7 for 1 hour; the material was pelleted again and resuspended in a mixture of 1% OsO4 and 1% uranyl-acetate in water; in this mixture the cells were fixed for 10 hours at 20° C; after fixation they were dehydrated in acetone and embedded in Vestopal W. In ultrathin sections the first structural changes became visible 15 minutes after infection when some of the cells seemed to swell and round up. In the cells small aggregates of electron dense material were observed in the chromosomal areas, and sometimes virus particles were also seen within the area of the cells’ chromosome.


Author(s):  
G. E. Visscher ◽  
R. L. Robison ◽  
R. G. Engstrom

It was our goal to evaluate the reliability and reproducibility of semi-automated morphometric techniques in the analysis of structural changes observed during drug safety assessment. Studies are presented to correlate the ultrastructural, morphometric and biochemical effects that the two hypolipidemic agents, clofibrate and probucol, produce in rats.Charles River CD Sprague-Dawley derived male rats (200-225 g body wt.) were used for the three studies performed. In studies I and II, clofibrate was administered as a dietary admixture to approximate a dosage of 300 mg/kg/day for six days. In study III, clofibrate and probucol were given as dietary admixtures to approximate dosages of 300 and 250 mg/kg/day respectively for fourteen days. Processing of hepatic specimens for electron microscopy included fixation in 1.3% sym-collidine buffered osmium tetroxide, ethanol dehydration and Epon embedment. Thin sections (600Å) were stained with uranyl acetate and lead citrate. Survey and photography was performed in the manner according to Weibel. Final prints (14,400x) were analyzed with a Zeiss MOP for morphometric quantitation.


1972 ◽  
Vol 54 (2) ◽  
pp. 227-237 ◽  
Author(s):  
H. G. BURGER ◽  
G. FINK ◽  
V. W. K. LEE

SUMMARY The presence of luteinizing hormone releasing factor (LH-RF) activity was investigated in pituitary stalk and systemic blood collected from rats ovariectomized at least 3 weeks previously, and in stalk blood from male rats in which electrodes had been implanted in the medial preoptic area of the brain. Most of the assayable luteinizing hormone (LH) present in the blood samples was eliminated by acid-ethanol extraction followed by ultrafiltration. The ultrafiltrates were injected into ovariectomized rats treated with oestrogen and progesterone, and increments in the concentration of LH in the sera of these animals, estimated by radioimmunoassay, were taken as an indication that the filtrate was able to release LH from the anterior pituitary gland. The ultrafiltrates of both the stalk and systemic plasma from the ovariectomized rats exhibited LH-RF activity as did the ultrafiltrates of blood collected from the pituitary stalk of the male rats during electrical stimulation of the preoptic area; stalk blood collected from these animals before the current was applied appeared to be inactive. The LH-RF activity of the ultrafiltrates of systemic and pituitary stalk plasma taken from ovariectomized rats was similar, and, therefore, the possibility is raised that the response of the pituitary glands in ovariectomized rats treated with oestrogen and progesterone is of an all or none type. The presence of appreciable quantities of LH-RF in the systemic plasma of ovariectomized rats may explain the discrepancy between bioassay and immunoassay estimates of LH in the plasma of these animals. The rapid increase in the concentration of serum LH and in the LH-RF activity of pituitary stalk plasma which followed stimulation of the preoptic area suggests that this region of the brain may be important in the control of the secretion of LH in the male as well as in the female animal.


Author(s):  
S. K. Aggarwal ◽  
P. McAllister ◽  
R. W. Wagner ◽  
B. Rosenberg

Uranyl acetate has been used as an electron stain for en bloc staining as well as for staining ultrathin sections in conjunction with various lead stains (Fig. 1). Present studies reveal that various platinum compounds also show promise as electron stains. Certain platinum compounds have been shown to be effective anti-tumor agents. Of particular interest are the compounds with either uracil or thymine as one of the ligands (cis-Pt(II)-uracil; cis-Pt(II)-thymine). These compounds are amorphous, highly soluble in water and often exhibit an intense blue coloration. These compounds show enough electron density to be used as stains for electron microscopy. Most of the studies are based on various cell lines (human AV, cells, human lymphoma cells, KB cells, Sarcoma-180 ascites cells, chick fibroblasts and HeLa cells) while studies on tissue blocks are in progress.


Author(s):  
Kosuke Ueda ◽  
Hiroto Washida ◽  
Nakazo Watari

IntroductionHemoglobin crystals in the red blood cells were electronmicroscopically reported by Fawcett in the cat myocardium. In the human, Lessin revealed crystal-containing cells in the periphral blood of hemoglobin C disease patients. We found the hemoglobin crystals and its agglutination in the erythrocytes in the renal cortex of the human renal lithiasis, and these patients had no hematological abnormalities or other diseases out of the renal lithiasis. Hemoglobin crystals in the human erythrocytes were confirmed to be the first case in the kidney.Material and MethodsTen cases of the human renal biopsies were performed on the operations of the seven pyelolithotomies and three ureterolithotomies. The each specimens were primarily fixed in cacodylate buffered 3. 0% glutaraldehyde and post fixed in osmic acid, dehydrated in graded concentrations of ethanol, and then embedded in Epon 812. Ultrathin sections, cut on LKB microtome, were doubly stained with uranyl acetate and lead citrate.


Author(s):  
S.L. Asa ◽  
K. Kovacs ◽  
J. M. Bilbao ◽  
R. G. Josse ◽  
K. Kreines

Seven cases of lymphocytic hypophysitis in women have been reported previously in association with various degrees of hypopituitarism. We report two pregnant patients who presented with mass lesions of the sella turcica, clinically mimicking pituitary adenoma. However, pathologic examination revealed extensive infiltration of the anterior pituitary by lymphocytes and plasma cells with destruction of the gland. To our knowledge, the ultrastructural features of lymphocytic hypophysitis have not been studied so far.For transmission electron microscopy, tissue from surgical specimens was fixed in glutaraldehyde, postfixed in OsO4, dehydrated and embedded in epoxy-resin. Ultrathin sections were stained with uranyl acetate and lead citrate and examined with a Philips 300 electron microscope.Electron microscopy revealed adenohypophysial cells of all types exhibiting varying degrees of injury. In the areas of most dense inflammatory cell infiltration pituitary cells contained large lysosomal bodies fusing with secretory granules (Fig. 1), as well as increased numbers of swollen mitochondria, indicating oncocytic transformation (Fig. 2).


Author(s):  
Taber A. Ba-Omar ◽  
Philip F. Prentis

We have recently carried out a study of spermiogenic differentiation in two geographically isolated populations of Aphanius dispar (freshwater teleost), with a view to ascertaining variation at the ultrastructural level. The sampling areas were the Jebel Al Akhdar in the north (Group A) and the Dhofar region (Group B) in the south. Specimens from each group were collected, the testes removed, fixed in Karnovsky solution, post fixed in OsO, en bloc stained with uranyl acetate and then routinely processed to Agar 100 resin, semi and ultrathin sections were prepared for study.


Author(s):  
Arya K. Bal

In the course of studies in the root meristem tissue of Rubus chamaemorus L. some important changes in the ultrastructural morphology were observed during the initiation of senescence at the end of the growing season.Root meristems were collected from naturally growing healthy populations of Cloudberry plants, and fixed in Karnovsky's mixture or in 2.5% glutaraldehyde in phosphate buffer. The samples were osmicated, dehydrated following usual methods and embedded in Epon. Ultrathin sections were stained in uranyl acetate and lead citrate.Figure 1 shows part of a dense cell in the meristem. The electron density of these cells is due to large amounts of a particulate material in the cytoplasmic matrix. The smallest particle seen in electron micrographs is about 40 A, although larger aggregates are also found, which remain randomly distributed in association with various cell organelles. Dense substance has been found associated with golgi membranes, proplastids, vacuoles and microtubules (Fig. 2).


Author(s):  
Daniel C. Pease

It is reasonable to think that phospholipid micelles should be visible and identifiable in electron micrographs of ultrathin sections if only they can be preserved throughout the embedding process. The development of highly polar, water-containing, aminoplastic embedments has made this a likely possibility. With this in mind, an investigation of the lecithin-secreting, Type II pneumocytes of the lung is underway.Initially it has been easiest to recognize phospholipid micelles in lung tissue fixed first with glutaraldehyde, and then secondarily exposed to osmium tetroxide. However, the latter is not a necessary concomitant for micellar preservation. Conventional uranyl acetate and lead citrate staining is finally applied. Importantly, though, the micelles have been most easily seen in tissue embedded in 507. glutaraldehyde polymerized with urea, as described in detail by D.C. Pease and R.G. Peterson (J. Ultra- struct. Res., 41, 133, 1972). When oriented appropriately, the micellar units are seen as tiny, bilayer plates.


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