Search PubMedSearch

Biomedical subjects

D Tolivia

Publications and source records attributed to D Tolivia.

33 records · Page 2Linked to original sources

Ultrastructural study of the cellular types in the pineal organ of Gambusia affinis (teleost).

The pineal organ of Gambusia affinis was studied via light and electron microscopy. The cell types studied included photoreceptor cells, supporting cells, and a third cell type. The photoreceptor cells, which appear to form clusters, are divided into four regions: outer segment, inner segment, cell soma, and synaptic pedicle. Synaptic ribbons are commonly observed in the synaptic pedicle. The supporting cells separate the photoreceptor cells from the thick basal lamina that surrounds the entire pineal organ. The supporting cells show highly organized membrane formations, some lipid-like inclusions, and a diplosome. One of the centrioles gives rise to an invaginated cilium. The third cell type is observed infrequently and appears to be located mainly in the vicinity of the outer segments. The morphological characteristics of this cell type are similar to those of phagocytic cells. The ultrastructural features of the pineal organ of G. affinis are compared with those of other teleosts.

Animals

Ultrastructure of the blood vessels in the Harderian gland of the hamster (Mesocricetus auratus): existence of sinusoids.

The Harderian gland blood supply of female and male hamsters was studied using light and electron microscopy. A profuse vascularization surrounding secretory acini was observed. Among the blood vessels, the existence of large and irregular sinusoidal capillaries was apparent. These sinusoids appeared in close association to the basal aspect of the secretory cells. Typical, small, fenestrated capillaries were also observed within the connective tissue. The existence of this particular vascularization together with other morphological features of the secretory cell basal pole suggest a possible endocrine function of these orbital glands.

Animals

Differential thionin block staining of nerve cells and fibers for paraffin-embedded material in mammalian central nervous system.

A differential staining method of myelinated fibers and nerve cell bodies applicable to whole blocks of mammalian central nervous tissue is described. Experimental material fixed by perfusion or necropsy material fixed in block can be used. Blocks of 2 mm in thickness were obtained with a vibratome, immersed in 50% ethanol for 7 h and stained for 1 week in the following solution: 0.3% thionine, 5% formaldehyde and 5% acetic acid. After the staining period the blocks were washed in distilled water, dehydrated through graded alcohols, cleared in butyl acetate and infiltrated in paraffin. Sections of 10 microns in thickness were obtained, attached to slides, dewaxed in xylene and coverslipped with mounting media in the usual manner.

Animals

Membrane formations in the pineal cells of the teleost Gambusia affinis.

Highly organized membrane formations in the cytoplasm of photoreceptor and supporting cells of the pineal organ of Gambusia affinis are described. These membranous structures show different characteristics depending on the cell type where they are located. Some of the morphological characteristics of these membranous formations are similar to those of myeloid bodies, but their presence in the photoreceptor cells raises some doubt about their being myeloid bodies. The three-dimensional interpretation of these structures is discussed.

Animals

New technique for differential staining of myelinated fibers and nerve cells on paraffin sections.

A simple, rapid method for the differential staining of myelinated nerve fibers and nerve cell bodies, applicable to sections of central nervous system pieces embedded in paraffin, is described. Experimental material fixed by perfusion with mixed aldehydes or necropsy material fixed in formaldehyde can be used. Constant and homogeneous results are obtained with this technique, and the most important characteristic is the absence of differentiation in either of the steps: staining of myelinated fibers and staining of nerve cell bodies. Sections 15 microns thick were attached to slides, dewaxed, and hydrated. After hydration, sections are mordanted (30 min) in 2.5% iron alum (SO4)2FeNH4, and rinsed (1 min) in distilled water. Staining is for 180 min in the following solution: 5 ml freshly made 20% alcoholic hematoxylin diluted with 25 ml of distilled water and 25 ml of absolute ethanol to which 10 ml of 1% Li2CO3 is added. The sections are washed in distilled water (5 min) and stained during 5 min in the following solution: 0.2% pyronine, 20% formaldehyde in distilled water. The sections are dehydrated through 96% and absolute ethanol, cleared in eucalyptol, and mounted in Eukitt. Myelinated fibers appear dark blue, whereas nerve cell bodies are stained red and the cell nucleoli dark blue. This procedure provides an adequate contrast for observation and photography.

Animals

Differential technique to stain nerve cells and fibers in methacrylate sections.

A simple method for the simultaneous staining of nerve cells and fibers, applicable to sections of pieces embedded in methacrylate, is described. Sections of 12 micron in thickness were attached to slides and stained for 10-18 hours in the following solution: 0.03% thionin, 0.5% formaldehyde, 0.5% acetic acid in distilled water. They were then rinsed in acetic water (0.5% acetic acid) for 30 seconds, washed in distilled water, dehydrated through 96% and absolute ethanol, cleared in eucalyptol, and mounted in Eukitt.

Animals

A new rapid silver impregnation for neuronal bodies.

Frozen sections of avian and rat brains routinely fixed in 6% glutaraldehyde/2% paraformaldehyde and left in phosphate buffer for 1 month are cut at 15-20 microns and collected in distilled water. Sections are placed in ammoniacal silver solution for 30 s, rinsed in 100% acetone and developed in a reducing solution at 75 degrees C. Sections are toned in 1% gold chloride solution and fixed in 50% sodium thiosulfate. After washing, the sections are dehydrated, cleared and mounted in the usual way for light microscopy.

Animals

The hypothalamic magnocellular system in the domestic fowl. Study on semithin sections.

Neuronal characteristics and location of the neurosecretory, magnocellular, fuchsin-paraldehyde-positive (FA+) system of the fowl are described at the light-microscopic level on serial semithin sections. Three nuclei make up this system, the nucleus supraopticus, n. magnocellularis interstitialis and n. paraventricularis. These nuclei display magnocellular neurons, not showing a parvocellular component. The neurons of the three nuclei showed a scattered pattern of distribution and a dense surrounding neuropil. Groups formed by magnocellular neurons were found in the three nuclei and groups formed by one magnocellular and a parvocellular neurons were only found in the n. magnocellularis interstitialis and in the n. paraventricularis. The presence of neurons in apposition to blood vessels was frequent in the magnocellular FA+ system of the domestic fowl.

Animals

A threedimensional reconstruction program for personal computers.

A reconstruction program is described for personal computers, permitting threedimensional image reconstructions from serial sections. This program also makes it possible to calculate the volume of structures from their twodimensional images in histological sections. In this paper we show a program written in BASIC and debugged on a Sharp MZ-700 personal computer, equipped with the Sharp MZ-1P01 plotter. The program is explained in detail and is easily adapted to run on any personal computer.

Computers

A new and permanent staining method for starch granules using fluorescence microscopy.

A fluorescence technique has been developed for observing starch granules in plant tissues. Sections are stained with a mixture of dyes which we have named F.A.S.G.A. from the initials of the Spanish names of its components (fucsina, alcian blue, safranina, glicerina, agua), and viewed by epifluorescence microscopy. The starch granules fluoresce greenish yellow, allowing the degradative state to be observed. Cell structures which do not fluoresce are also differentiated. The stain permits identification of other structures when examined by visible light microscopy and is relatively resistant to fading over time.

Fabaceae

A new technique for differential and simultaneous staining of nerve cells and fibers.

A new and simple method for the simultaneous staining of nerve cells and fibers is described. The use of formol-thionin under controlled conditions of concentration and pH provides a highly reliable method for the staining of nerve cells in blue and fiber tracts in red, making any posterior differentiation redundant. This technique provides constant and reproducible results allowing simultaneous staining of many sections.

Animals

Castration increases cell damage induced by porphyrins in the Harderian gland of male Syrian hamster. Necrosis and not apoptosis mediates the subsequent cell death.

It is known that the Harderian gland of male Syrian hamster synthesizes a much smaller amount of porphyrins than the gland of the female and that castration greatly increases this synthesis. We have studied in this experimental model the behavior of the different classes of secretory cells and their role in the synthesis of porphyrins, attempting to clarify the participation of these compounds in the cell damage leading to the formation of clear cells previously described in the gland of females. We have also investigated the mechanism underlying the death of these secretory cells after porphyrin accumulation (necrosis vs apoptosis). To achieve this, we have utilized the following techniques: (a) morphometrical; (b) ultrastructural; (c) biochemical (fluorescence spectrophotometry); and (d) molecular (DNA nick-end labeling in methacrylate sections and dot blot analysis). The glands from male hamsters (serving as control) present a very low rate of damaged cells that progressively rises after castration. This rise runs parallel to that of porphyrin synthesis, porphyrin deposits, and the decrease of Type II secretory cells. The damage and subsequent death of the secretory cells in the gland is produced by the deposit of porphyrins in the mitochondrial membrane. This porphyrin accumulation leads to a complete mitochondrial destruction that finally results in cell death and its secretion into the lumen. We finally conclude that this event is not a physiological cell death (apoptosis) but the consequence of the toxic accumulation of porphyrins (necrosis).

5-Aminolevulinate Synthetase