Unusual Intranuclear Structures in Chick Sympathetic Neurons

Author(s):  
E. B. Masurovsky ◽  
H. H. Benitez ◽  
M. R. Murray

Recent light- and electron microscope studies concerned with the effects of D2O on the development of chick sympathetic ganglia in long-term, organized culture revealed the presence of rod-like fibrillar formations, and associated granulofibrillar bodies, in the nuclei of control and deuterated neurons. Similar fibrillar formations have been reported in the nuclei of certain mammalian CNS neurons; however, related granulofibrillar bodies have not been previously described. Both kinds of intranuclear structures are observed in cultures fixed either in veronal acetate-buffered 2%OsO4 (pH 7. 4), or in 3.5% glutaraldehyde followed by post-osmication. Thin sections from such Epon-embedded cultures were stained with ethanolic uranyl acetate and basic lead citrate for viewing in the electron microscope.

Author(s):  
Roberta M. Bruck

An unusual structure in the cochlea is the spiral limbus; this periosteal tissue consists of stellate fibroblasts and collagenous fibers embedded in a translucent ground substance. The collagenous fibers are arranged in vertical columns (the auditory teeth of Haschke). Between the auditory teeth are interdental furrows in which the interdental cells are situated. These epithelial cells supposedly secrete the tectorial membrane.The fine structure of interdental cells in the rat was reported by Iurato (1962). Since the mouse appears to be different, a description of the fine structure of mouse interdental cells' is presented. Young adult C57BL/6J mice were perfused intervascularly with 1% paraformaldehyde/ 1.25% glutaraldehyde in .1M phosphate buffer (pH7.2-7.4). Intact cochlea were decalcified in .1M EDTA by the method of Baird (1967), postosmicated, dehydrated, and embedded in Araldite. Thin sections stained with uranyl acetate and lead citrate were examined in a Phillips EM-200 electron microscope.


Author(s):  
M.K. Bhatnagar ◽  
S.M. Snelgrove-Hobson ◽  
P.V.V. Prasada Rao

Cytpplasmic microtubules, ubiquitous cell organelles, lie free in the cytosol without being compartmented off by membranes. In rare instances, however, intracisternal microtubules (ICM) have been reported in ganglionic neurons of aging dogs and in the proximal tubule cells (PTC) of rabbit kidneys. We report here the presence of ICM in the PTC of the kidneys of Peking ducks (Anas platyrhynhos).A total of 106 Peking ducks (24 males and 24 females of 9 months, 48 females of 15 months, 6 males of 30 months and 2 males and 2 females of 48 months of age) were anesthetized and exsanguinated. Pieces of kidney cortex were fixed in 4% glutaraldehyde in 0.5M phosphate buffer (pH 7.3) at 29°C for four hours. Samples were post fixed in 2% osmium tetroxide for two hours. One micron sections of Epon-embedded cortices were stained with toluidine blue and thin sections (70-70 nm) contrasted with uranyl acetate and lead citrate were examined with a JEOL 100-S electron microscope at 80 kV.


Author(s):  
Annette M. Andrews ◽  
Alex M. Cameron ◽  
James W. Townsend ◽  
Winslow G. Sheldon

Pituitaries of 6 C3H/HEJ MTV+ mice about 660 days of age were fixed by immersion in cacodylate-buffered 4% glutaraldehyde, post-fixed in 1% osmium tetroxide, stained en block with aqueous uranyl acetate, dehydrated in a graded series of ethanol solutions, cleared in acetone, and embedded in an Epon-Araldite resin mixture. Semi-thin (1 μm) sections were taken of the entire face of the block, and stained with toluidine blue. Subsequently, thin sections (100 nm) were prepared from mesas of the two sides of the pars distal is and one mesa of the pars intermedia. Thin sections were stained with ethanolic uranyl acetate followed by Sato's lead citrate (1), then examined on a Philips EM201 electron microscope.


Author(s):  
C. N. Sun ◽  
C. Araoz ◽  
H. J. White

The ultrastructure of a cerebral primitive neuroectodermal tumor has been reported previously. In the present case, we will present some unusual previously unreported membranous structures and alterations in the cytoplasm and nucleus of the tumor cells.Specimens were cut into small pieces about 1 mm3 and immediately fixed in 4% glutaraldehyde in phosphate buffer for two hours, then post-fixed in 1% buffered osmium tetroxide for one hour. After dehydration, tissues were embedded in Epon 812. Thin sections were stained with uranyl acetate and lead citrate.In the cytoplasm of the tumor cells, we found paired cisternae (Fig. 1) and annulate lamellae (Fig. 2) noting that the annulate lamellae were sometimes associated with the outer nuclear envelope (Fig. 3). These membranous structures have been reported in other tumor cells. In our case, mitochondrial to nuclear envelope fusions were often noted (Fig. 4). Although this phenomenon was reported in an oncocytoma, their frequency in the present study is quite striking.


Author(s):  
Julio Sepúlveda-Saavedra ◽  
Beatriz González-Corona ◽  
Víctor A. Tamez Rodríguez ◽  
Ma. Victoria Bermúdez de Rocha ◽  
Alfredo Piñeyro López

It has been shown in previous studies that the toxin T-514 isolated from K. humboldtiana induces severe damage to the lung in treated rodents. Histopathological findings include edema, and alveolar hemorrage. However, the ultraestructure of the lesion has not been investigated. In this study we used two species of rodents: Hamster and guinea pig, and a primate: Macaca fascicularis. Animals received different single dosis of the toxin via intraperitoneal. Control animals received only the vehicle (propylen glycol). Inmediately after spontaneous death, lung samples were fixed in Karnovsky-Ito fixative, post fixed in osmium tetroxide and embedded in epon. Thin sections were prepared with an Ultratome V LKB, stained with uranly acetate and lead citrate, and studied in an electron microscope Zeiss-EM109.


Author(s):  
Hsin-Kan Wu ◽  
Mei-Chu Chung

In one of our earlier papers (Wu et al. 1978), we suggested that glutelin is the major composition of the round storage protein bodies although they also contain relatively more prolamine than the angular one does. Immunochemical studies of Krishnan et al. (1986) later showed the presence of glutelin in the irregularly-shaped (angular) protein bodies while the prolamines were found in the round ones. Our recent experiment using protein A-gold technique found that prolamine is mainly deposited into the angular protein bodies.Small blocks (1 mm3) of 7 DAF (days after flowering) caryopsis of Orvza perennis were fixed with 3% paraformaldehyde and 3% glutaraldehyde in 0.1M sodium phosphate, pH7.4, dehydrated in a graded ethanol series and infiltrated with Spurr’s resin. Thin sections, after gold labeling, were stained with uranyl acetate and lead citrate. Rabbit antibodies were raised against purified prolamine. Protein A-gold sol complex was prepared based on the technique of Horisberger et al. (1977).


Author(s):  
W. P. Wergin ◽  
S. Roy ◽  
E. F. Erbe ◽  
C. A. Murphy ◽  
C. D. Pooley

Larvae of the nematode, Steinernema carpocapsae Weiser strain All, were cryofixed and freezesubstituted for 3 days in acetone containing 2% osmium tetroxide according to established procedures. Following chemical fixation, the nematodes were brought to room temperature, embedded in Spurr's medium and sectioned for observation with a Hitachi S-4100 field emission scanning electron microscope that was equipped with an Oxford CT 1500 Cryotrans System. Thin sections, about 80 nm thick, similar to those generally used in conventional transmission electron microscope (TEM) studies were mounted on copper grids and stained with uranyl acetate for 30 min and lead citrate for 5 min. Sections about 2 μm thick were also mounted and stained in a similar fashion. The grids were mounted on an Oxford grid holder, inserted into the microscope and onto a cryostage that was operated at ambient temperature. Thick and thin sections of the larvae were evaluated and photographed in the SEM at different accelerating voltages. Figs. 4 and 5 have undergone contrast conversion so that the images would resemble transmitted electron micrographs obtained with a TEM.


1974 ◽  
Vol 22 (10) ◽  
pp. 929-944 ◽  
Author(s):  
DAVID B. NICHOLS ◽  
HAZEL CHENG ◽  
C. P. LEBLOND

The enteroendocrine cells of the mouse duodenum were examined in the light and electron microscopes by comparing Epon sections stained by Gomori's silver methenamine with adjacent sections stained by Masson's ammoniacal silver nitrate, iron hematoxylin or uranyl acetate and lead citrate. Furthermore, the granules present in the enteroendocrine cells stained with silver methenamine were investigated in the electron microscope: their shape was defined as either spherical or nonspherical and, in either case, their density was measured by cytophotometry. The comparison of adjacent sections of mouse duodenum by different techniques indicates that the granules of all enteroendocrine cells are stained by iron hematoxylin, whereas those of about 94% of the cells are stained by silver methenamine. Since silver methenamine stains exactly the same cells as ammoniacal silver nitrate, it may be used as a test of argentaffinity. It is concluded that about 94% of the enteroendocrine cells are argentaffin and about 6% are nonargentaffin. The argentaffin cells display a wide spectrum of granule shape. Nearly all of the granules of a cell may be spherical or nearly all nonspherical or, more commonly, there is a fair number of both kinds. In the few cells which are not argentaffin, over 70% of the granules are spherical. After silver methenamine staining, the mean density of spherical and nonspherical granules is statistically equal within any given cell. Yet, when cells are compared to one another, the mean density of the granules, and therefore the argentaffinity, increases with the proportion of nonspherical granules. It is concluded that, in the mouse duodenun, argentaffinity is most pronounced in the cells with irregular nonspherical granules, but it is by no means confined to such cells. Moreover, the argentaffinity as well as the shape of the granules varies within wide limits.


1959 ◽  
Vol 6 (1) ◽  
pp. 77-84 ◽  
Author(s):  
Stuart W. Smith

Sympathetic ganglia of the horned lizard, Phrynosoma cornutum, were fixed in OsO4 and imbedded in methacrylate. Thin sections were cut for electron microscopy. Some adjacent thick sections were cut for light microscopy and were stained in acidified, dilute thionine both before and after digestion by RNase. In the light microscope two types of Nissl bodies are found, both removable by RNase: (1) a deep, diffuse, indistinctly bounded, metachromatic variety, and (2) a superficial, dense, sharply delimited, orthochromatic sort. Electron microscopically, the former ("reticular" Nissl bodies) corresponds to the granulated endoplasmic reticular structure of Nissl material previously described by others, whereas the latter ("areticular" Nissl bodies) comprises compact masses of particles of varying internal density and devoid of elements of endoplasmic reticulum. The constituent particles of the areticular Nissl material are 4 to 8 x the diameter of single ribonucleoprotein granules of the reticular Nissl substance and seem, near zones of junction with the reticular type, to arise by clustering of such granules with subsequent partial dispersion of the substance of the granules into an added, less dense material. It is suggested that the observed orthochromasia of the areticular Nissl substance is due to accumulation of a large amount of protein bound to RNA and, further, that these Nissl bodies may represent storage depots of RNA and protein.


2001 ◽  
Vol 281 (2) ◽  
pp. R635-R644 ◽  
Author(s):  
Azeez A. Aileru ◽  
Aline De Albuquerque ◽  
John M. Hamlyn ◽  
Paolo Manunta ◽  
Jui R. Shah ◽  
...  

Altered sympathetic nervous system activity has been implicated often in hypertension. We examined short-term potentiation [posttetanic potentiation (PTP)] and long-term potentiation (LTP) in the isolated superior cervical ganglia (SCG) from Sprague-Dawley (SD) rats given vehicle, digoxin, or ouabain by subcutaneous implants as well as in animals with ouabain-induced hypertension (OHR), and inbred Baltimore ouabain-resistant (BOR) and Baltimore ouabain-sensitive (BOS) strains of rats. Postganglionic compound action potentials (CAP) were used to determine PTP and LTP following a tetanic stimulus (20 Hz, 20 s). Baseline CAP magnitude was greater in ganglia from OHR than in vehicle-treated SD rats before tetanus, but the decay time constant of PTP was significantly decreased in OHR and in rats infused with digoxin that were normotensive. In hypertensive BOS and OHR, the time constants for the decay of both PTP and LTP ( t L) were increased and correlated with blood pressure (slope = 0.15 min/mmHg, r = 0.52, P < 0.047 and 6.7 min/mmHg, r = 0.906, P < 0.0001, respectively). In BOS and OHR, t L (minutes) was 492 ± 40 ( n = 7) and 539 ± 41 ( n = 5), respectively, and differed ( P < 0.05) from BOR (257 ± 48, n = 4), SD vehicle rats (240 ± 18, n = 4), and captopril-treated OHR (370 ± 52, n = 5). After the tetanus, the CAP at 90 min in BOS and OHR SCG declined less rapidly vs. SD vehicle rats or BOR. Captopril normalized blood pressure and t L in OHR. We conclude that the duration of ganglionic LTP and blood pressure are tightly linked in ouabain-dependent hypertension. Our results favor the possibility that enhanced duration of LTP in sympathetic neurons contributes to the increase in sympathetic nerve activity in ouabain-dependent hypertension and suggest that a captopril-sensitive step mediates the link of ouabain with LTP.


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