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Biomedical subjects

P P Giorgi

Publications and source records attributed to P P Giorgi.

At least 19 recordsLinked to original sources

Primary olfactory terminations in the forebrain of amphibia: a comparative study with soybean agglutinin.

Ambystoma and Xenopus were used to investigate olfactory terminations in the forebrain by exploiting the selective binding of soybean agglutinin to primary olfactory projections. Lectin binding and tridimensional reconstruction provided new information on the distribution of olfactory glomeruli and on regional and interspecific differences. Ambystoma showed an uniform labelling of all olfactory terminations, which were localized in the rostro-lateral forebrain without obvious segregation of different terminal fields, except at dorsal levels. On the contrary, the binding pattern of Xenopus confirmed the existence of three sets of primary olfactory neurons characterized by clear differences in lectin binding. Selective soybean agglutinin binding and a detailed tridimensional reconstruction provided evidence for the existence of a set of neuronal projections, described for the first time, which terminate in a discrete bilateral region of the forebrain localized below the main olfactory bulb. The presence of three different sets of primary olfactory neurons, the experimental potentials and the possible functional significance of SBA binding sites are discussed.

Ambystoma mexicanum↗

Retinal wholemounts: a simple method for precise mapping.

A simple and inexpensive method to analyse retinal wholemounts is described. It allows the definition of regularly spaced sites to map regional specializations of the retina under light microscopy. After attaching a piece of translucent millimetre graph paper behind the slide holding the preparation, the two grid patterns of the graph paper and of the eye-piece grid are used in combination for the definition of sites for analysis and the return to specific areas as small as 50-100 microns2. The present method is simpler and offers several advantages when compared to others based on X-Y movements of the microscope stage or on photographic montage.

Evaluation Studies as Topic↗

Uptake and axonal transport of horseradish peroxidase isoenzymes by different neuronal types.

The uptake and transport of basic and acidic horseradish peroxidase isoenzymes was compared in the neuromuscular, visual and olfactory systems of Xenopus larvae and postmetamorphic frogs. The concentration (w/v) of the two preparations was corrected to compensate for their difference in enzymatic activity (unit/w), which was seven-fold higher in basic horseradish peroxidase. Uptake and transport of horseradish peroxidase isoenzymes could be demonstrated with 7% basic horseradish peroxidase, but not with equal amounts of 49% acidic horseradish peroxidase in all systems investigated: retrograde transport from terminals of retinal ganglion cells, isthmotectal neurons and spinal motoneurons, as well as anterograde transganglionic transport along olfactory neurons. A very weak labelling of the same neuronal pathways by acidic horseradish peroxidase was obtained only after increasing the amount injected by approximately two-fold. Basic horseradish peroxidase isoenzymes were also preferentially taken up and transported retrogradely by broken axons of the optic nerve. When tested, similar results were obtained in both larvae and frogs suggesting that preferential uptake and transport of basic horseradish peroxidase is a general feature of all neurons and of all developmental stages. Electron microscopical analysis of the outer layers of the optic tectum revealed that, in the same experimental conditions producing no retrotrade labelling of optic axons, acidic horseradish peroxidase was rarely found to enter nerve terminals. It appears that interactions between horseradish peroxidase and neuronal membranes occur during uptake and transport and that molecular charge plays an important role, beyond non-specific fluid-phase endocytosis. We suggest that differences between horseradish peroxidase isoenzymes as neuronal tracers reflect a process of adsorptive endocytosis related to general characteristics of neuronal membranes (regardless of age) and not to specific receptor-mediated interactions characteristic of neuronal specificity.

Animals↗

Soybean agglutinin binding to the olfactory systems of the rat and mouse.

The binding of the lectin soybean agglutinin (SBA) to the olfactory system of both the rat and mouse was investigated histochemically. SBA bound to fibers in the accessory olfactory nerve and to glomeruli in the accessory olfactory bulb. In addition, SBA binding sites were present in some, but not all, glomeruli in the ventrolateral and ventromedial portions of the main olfactory bulb of only the rat. Under standard experimental conditions, SBA did not bind to neurons in other regions of the olfactory system nor to any other neurons in the brain. This selective binding of SBA to only some glomeruli in the olfactory bulb provides additional support for the presence of, at least, two subclasses of olfactory receptor cells in the nasal cavity. Whether these neuronal subclasses are the same as those previously characterized by monoclonal antibodies in rabbit remains to be determined.

Animals↗

Selective binding of soybean agglutinin to the olfactory system of Xenopus.

The binding patterns of four different lectins conjugated to horseradish peroxidase were investigated in the nervous system of juvenile Xenopus borealis. Only the lectin soybean agglutinin revealed a very selective binding pattern, which was restricted to the olfactory system. The olfactory and vomeronasal epithelia, the olfactory and accessory olfactory nerves and the olfactory and accessory olfactory bulbs were all labelled. The ventral portions of the olfactory nerve and bulb were however more intensely labelled than their dorsal portions. The rest of the brain and spinal cord did not bind this lectin except for a small discrete set of unmyelinated axons travelling in the medial forebrain bundle. Ultrastructural investigations revealed that soybean agglutinin was confined to the cell surface of olfactory neurons. The selective binding of this lectin of olfactory neurons suggests that specific cell surface glycoconjugates binding soybean agglutinin may have either a functional or developmental role in the olfactory system of Xenopus.

Animals↗

Endurance exercise does not modify nerve fibre morphology in the rat soleus nerve.

The effects of the myelination and growth of axons in the nerve to the soleus muscle (NSM) was investigated in young male Wistar rats. Experimental animals were run on a treadmill for 13 weeks (70 min/day, 6 days/week), while sedentary weight-matched animals of the same age and sex served as controls. The activity of the enzymes, phosphofructokinase (PFK) and succinate dehydrogenase (SDH) were measured in the soleus muscle to assess the effects of training on the anaerobic glycolytic and oxidative capacities. Axon and myelin sheath cross-sectional areas were measured from electron micrograph montages of the whole NSM with the aid of a digitizing tablet. The exercise programme produced large adaptive increases in the capacity of the soleus muscle for both oxidative and anaerobic glycolytic metabolism. The specific activity of PFK and SDH increased in the soleus muscle of exercised animals by 42.8% and 68.2%, respectively. In the NSM, however, there were no differences between control and exercised animals concerning the total number of myelinated nerve fibers, the size of axons and myelin sheaths of nerve fibres and the degree of myelination of axons (myelin area divided by axon area). Comparison with other studies suggests that the intensity of exercise may be the critical parameter responsible for discrepancies in the literature. Reports suggesting that exercise increases the size of nerve fibres should not be generalized to all exercise programmes.

Animals↗

Topography of the retinal ganglion cell layer of Xenopus.

Cell distribution in the retinal ganglion cell layer of Xenopus laevis was studied by electron microscopy and the retinal wholemount method. Unlike the familiar stratification of the retina, the ganglion cell perikarya and their axons were localised in the same layer, but strictly segregated in alternate rows converging in a radial pattern on the optic disc. Rows of perikarya (retinal cords) were two to ten cells wide and clearly defined for a little more than half the distance between the optic disc and the ora serrata. Detailed analysis of 25% of the retinal surface showed higher cell density (about 3500 cells/mm2) in the centrodorsal region than in the periphery (about 2000 cells/mm2). The higher density regions represent a weak visual streak having no nasotemporal asymmetry. After staining with cresyl violet, the majority of cells appeared to be small (10 micron diameter) with very little Nissl substance. The cytoplasm was scant; this was confirmed by electron microscopy. The difference between the total cell count (68470 +/- 7360) and the total axon count in the optic nerve (Wilson, 1971) suggests a population of 15% non-ganglionic cells (glia and amacrine cells) in the retinal ganglion cell layer. Retinal ganglion cell topography in Xenopus may have relevance to studies on retinal growth and retinotopy. The functional significance of the weak visual streak in Xenopus is consistent with a limited need for visual acuity.

Animals↗

Cellular DNA content in different species of Xenopus.

1. Biochemical methods were applied to determine the DNA content per cell in ten species and subspecies of Xenopus. 2. Cell density and DNA concentration were measured in a suspension of blood cells. 3. DNA was determined by the diphenylamine method and values were compared with those obtained by the ultraviolet absorption method and by cytofluorometry. 4. A good degree of reproducibility was demonstrated by a simple biochemical method which yielded values compatible with cytofluorometry. 5. Results suggested that the DNA of Xenopus muelleri, and perhaps that of other species with a large genome size, may be specially protected from chemical degradation.

Animals↗

Labelling by axonal transport of myelin-associated proteins in the rabbit visual pathway.

After intraocular injections of [3H]leucine, six regions of the visual pathway of adult rabbit were used to study the spatio-temporal pattern of the slow anterograde axonal transport of radioactive proteins associated with the particulate fraction, the water-soluble fraction and the myelin fraction. Unlike other fractions, myelin-associated labelled proteins represented a time-constant (for a given region) percentage of total tissue radioactivity. This percentage increased from the first half to the second half of the optic nerve and remained high in the chiasma and tract. The peak specific radioactivity of myelin decreased in the same direction. Myelin proteins were separated by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis and the labelling patterns obtained in different regions and at different survival times were compared. At the peak of myelin radioactivity of a given region the label was typically associated with four protein bands, L1, L2, L3 and L4, of 40000, 44000, 62000, and 68000 mol.wts. respectively. The basic protein, the proteolipid protein and the W1 component (mol.wt. 51000-53000) of the Wolfgram proteins were not significantly labelled. The radioactivity associated with the W2 component (mol.wt 60000) of the Wolfgram proteins could be derived from the closely migrating L3 component. At shorter survival times no clear labelling pattern could be detected. At longer survival times radioactivity was almost totally localized around band L3. The results presented underline the importance of choosing appropriate experimental conditions to obtain a consistent labelling pattern of myelin-associated proteins and to investigate the possible mechanism responsible for this phenomenon.

Animals↗

Science and ethics.

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International Cooperation↗