Gap junction associated filaments in testis interstitial cells of the rat

Author(s):  
J.F. David-Ferreira ◽  
K.L. David-Ferreira ◽  
M.H. Miranda ◽  
J.C. Lemos

The testis interstitial cells of the rat are closely associated forming small aggregates in the vicinity of the vessels. The finality of the present study is to analise the relationship between the interstitial cells and to describe some particularities of their junctions.The observations were made in thin sections from testis of 2 to 3 months old rats fixed by immersion or perfusion with 2 to 3% glutaral- dehyde in s-collidine or cacodylate buffer followed by 2% osmium tetro-xyde in the same buffers. Some specimens have been prepared following the technique of Shea. The thin sections obtained after embedding in Epon were double stained in uranyl acetate and lead citrate. For the cryofracture study the tissues were fixed in 3% glutaraldehyde in cacodylate buffer, impregnated in 28% glycerol, frozen in Freon 22 and stored in liquid nitrogen. Freeze fracture and platinum-carbon shadowing were done in a Balzers 300.

Author(s):  
A. Julio Martinez ◽  
Richard J. Duma ◽  
Doris G. Fultz ◽  
Ruth B. Finley

Ultrastructural components of trophozoites and cysts of Acanthamoeba-Hartmannella (A-H) and Naegleria amebas have been investigated. The present report describes a technique to study the morphological features of such protozoa.The A-H strains selected were cultured for 7 days in axenic amebic media, and the Naegleria strains were cultured for 7 days in axenic media with blood. In situ fixation was accomplished by adding an equal volume of 4% Osmic Acid in Cacodylate buffer at 4¶C for one hour. Following frequent agitation, the tubes were then spun at 2500 RPM for 10 minutes to obtain a pellet. The pellets were dehydrated in graded series of alcohols (from 50% to absolute) and propylene oxide and then embedded in EPON. One micron sections were stained with Paragon (PS 1301) for thirty seconds. Thin sections were cut with a diamond knife, double stained with 5% alcoholic uranyl acetate solution and lead citrate and observed in an Hitachi-HS-8F2 electron microscope.


Author(s):  
T. Shirahama ◽  
A. S. Cohen ◽  
O. G. Rodgers

Forty-micron nonfrozen sections of myocardium from several healthy young adult New Zealand white rabbits, without pretreatment or 10-15 minutes after an intravenous ferritin injection (100 mg ferritin/100 gm. body weight), were double fixed with 2% formaldehyde - 2.5% glutaraldehyde in 0.1M cacodylate buffer and 2% OSO4 in 0.1M cacodylate buffer in the presence of 1000, 100, 50 or 20 ppm ruthenium red (RR), and embedded in Epon. Thin sections of small blood vessels including capillaries, unstained or stained with uranyl acetate and/or lead citrate, were photographed at initial magnifications of 40,000 or 80,000. The results were analysed to delineate the ultrastructural relation of mucopolysaccharides (MPS) in the architecture of pinocytic vesicles and the MPS role in pinocytosis.


Author(s):  
S. S. Ren ◽  
J. C. Chen ◽  
Y. R. Chen

The plants produced from rice anther culture varied in ploidy level. Genetic analysis of anther-derived plants indicated genome multiplication occurred in microspore development in vitro. Cytological studies on early roicrospore division suggested endoreduplication and nuclear fusion may be related to the production of nonhaploid plants. In the present study, fine structures of callus ontogeny in anther culture were emphasized.Anthers of rice(Oryza sativa L. c. v. Hsinchu 4, 2n=24) containing mid-uninucleate microspores were excised and cultured in Ng liquid medium, supplimented with 60 g/l sucrose, 1 mg/l kinetin and 2 mg/l naphthalenacetic acid. Cultured anthers were collected at 2 days intervals for 20 days and fixed in 2.5% glutaraldehyde in 0.1M cacodylate buffer(pH=7.0), postfixed in osmium tetraoxide, and then embedded in Spurr's resin. Thin sections were stained with uranyl acetate and lead citrate, observed under Hitachi H-600 at 75 KV.


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):  
M. Kessel ◽  
R. MacColl

The major protein of the blue-green algae is the biliprotein, C-phycocyanin (Amax = 620 nm), which is presumed to exist in the cell in the form of distinct aggregates called phycobilisomes. The self-assembly of C-phycocyanin from monomer to hexamer has been extensively studied, but the proposed next step in the assembly of a phycobilisome, the formation of 19s subunits, is completely unknown. We have used electron microscopy and analytical ultracentrifugation in combination with a method for rapid and gentle extraction of phycocyanin to study its subunit structure and assembly.To establish the existence of phycobilisomes, cells of P. boryanum in the log phase of growth, growing at a light intensity of 200 foot candles, were fixed in 2% glutaraldehyde in 0.1M cacodylate buffer, pH 7.0, for 3 hours at 4°C. The cells were post-fixed in 1% OsO4 in the same buffer overnight. Material was stained for 1 hour in uranyl acetate (1%), dehydrated and embedded in araldite and examined in thin sections.


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):  
Patricia L. Jansma

The presence of the membrane bound vesicles or blebs on the intestinal epithelial cells has been demonstrated in a variety of vertebrates such as chicks, piglets, hamsters, and humans. The only invertebrates shown to have these microvillar blebs are two species of f1ies. While investigating the digestive processes of the freshwater microcrustacean, Daphnia magna, the presence of these microvillar blebs was noticed.Daphnia magna fed in a suspension of axenically grown green alga, Chlamydomonas reinhardii for one hour were narcotized with CO2 saturated water. The intestinal tracts were excised in 2% glutaraldehyde in 0.2 M cacodyl ate buffer and then placed in fresh 2% glutaraldehyde for one hour. After rinsing in 0.1 M cacodylate buffer, the sample was postfixed in 2% OsO4, dehydrated with a graded ethanol series, infiltrated and embedded with Epon-Araldite. Thin sections were stained with uranyl acetate and Reynolds lead citrate before viewing with the Philips EM 200.


Author(s):  
M. Ashraf ◽  
F. Thompson ◽  
S. Miki ◽  
P. Srivastava

Iron is believed to play an important role in the pathogenesis of ischemic injury. However, the sources of intracellular iron in myocytes are not yet defined. In this study we have attempted to localize iron at various cellular sites of the cardiac tissue with the ferrocyanide technique.Rat hearts were excised under ether anesthesia. They were fixed with coronary perfusion with 3% buffered glutaraldehyde made in 0.1 M cacodylate buffer pH 7.3. Sections, 60 μm in thickness, were cut on a vibratome and were incubated in the medium containing 500 mg of potassium ferrocyanide in 49.5 ml H2O and 0.5 ml concentrated HC1 for 30 minutes at room temperature. Following rinses in the buffer, tissues were dehydrated in ethanol and embedded in Spurr medium.The examination of thin sections revealed intense staining or reaction product in peroxisomes (Fig. 1).


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).


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