Browsing by Author "FAUNDEZ, V"

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    AXONS SPROUT AND MICROTUBULES INCREASE AFTER LOCAL INHIBITION OF RNA-SYNTHESIS, AND MICROTUBULES DECREASE AFTER INHIBITION OF PROTEIN-SYNTHESIS - A MORPHOMETRIC STUDY OF RAT SURAL NERVES
    (1991) BUSTOS, J; VIAL, JD; FAUNDEZ, V; ALVAREZ, J
    Drugs that inhibit RNA or protein synthesis are known to affect some functional properties of axons. In this context, we studied the ultrastructural effects of actinomycin-D, an inhibitor of RNA synthesis, and cycloheximide and emetine, inhibitors of protein synthesis, in rat sural nerves. A silicone sleeve (4 mm long) loaded with drug was placed around the nerves and left for about a week. The ultrastructural alterations of axons and Schwann cells progressed over this period. After cycloheximide and emetine, the cytoplasm of Schwann cells was enlarged and the rough endoplasmic reticulum was prominent. After actinomycin-D, the Schwann cells reached the stage of lysis. Nonmedullated were more affected than myelinated axons. After cycloheximide and emetine, the axoplasmic matrix decreased substantially but reversibly. Microtubules of nonmedullated fibres decreased by about 50%. Actinomycin-D determined sprouting of axons and a rise of axonal microtubules; in nonmedullated axons, the normal inverse correlation between microtubular density and calibre gave way to a high and constant density for all axonal sizes. A few millimetres proximal and distal to the sleeve, the nerve tissue and the axonal microtubular content were close to normal, i.e. the effects of drugs were local. Present results suggest that the local turnover of amino acids in the axon is necessary to maintain the integrity of microtubule and neurofilament proteins. We propose that the Schwann cell down-regulates the axonal cytomatrix.
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    CALIBERS AND MICROTUBULES OF NERVE-FIBERS - DIFFERENTIAL EFFECT OF UNDERNUTRITION IN DEVELOPING AND ADULT-RATS
    (1990) FAUNDEZ, V; CORDERO, ME; ROSSO, P; ALVAREZ, J
    Sural nerves of 9-week-old rats undernourished since birth, and of adult rats food-restricted for 27 and 48 days, were studied to explore the effect of severe undernutrition on the caliber and microtubules of axons in growing and non-growing animals. In 9-week-old undernourished rats, the number and caliber of myelinated fibers were normal while the cross-sectional area of non-medullated fibers was 29% smaller than controls. By contrast, in adult undernourished rats the cross-sectional area of myelinated fibers was affected sooner and to a greater extent (-28%) than that of non-medullated fibers (-23%). Regardless of age, in both controls and in undernourished rats non-medullatedfibers of equal caliber had similar microtubular content. The same was found in 3-.mu.m myelinated axons. These findings indicate that food restriction affects proportionately caliber and microtubules of axons. It is proposed that the anatomy of the axon is in a dynamic equilibrium and that microtubules participate in the specification of the axonal caliber.
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    CIPROFIBRATE, A CARCINOGENIC PEROXISOME PROLIFERATOR, INCREASES THE PHOSPHORYLATION OF EPIDERMAL-GROWTH-FACTOR RECEPTOR IN ISOLATED RAT HEPATOCYTES
    (1993) ORELLANA, A; HOLUIGUE, L; HIDALGO, PC; FAUNDEZ, V; GONZALEZ, A; BRONFMAN, M
    Ciprofibrate, a hypolipidaemic drug with carcinogenic and peroxisome-proliferation effects in rat liver, was found to increase the phosphorylation of epidermal-growth-factor receptor in P-32-labeled isolated rat hepatocytes. This effect was suppressed by protein-kinase-C inhibitors, and was accompanied by an almost complete inhibition of the receptor autophosphorylation normally induced by its ligand. However, in vitro experiments showed that protein-kinase-C phosphorylation of purified epidermal-growth-factor receptor was activated by ciprofibroyl-CoA, the acyl-CoA derivative of the drug, but not by the unmodified drug. Neither compound affected the ligand induction of epidermal-growth-factor-receptor autophosphorylation in isolated liver membranes. These results suggest that metabolically produced ciprofibroyl-CoA in liver cells would activate protein-kinase-C and produce changes in epidermal-growth-factor-receptor function.
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    EPIDERMAL GROWTH-FACTOR RECEPTOR IN SYNAPTIC FRACTIONS OF THE RAT CENTRAL-NERVOUS-SYSTEM
    (1992) FAUNDEZ, V; KRAUSS, R; HOLUIGUE, L; GARRIDO, J; GONZALEZ, A
    Functional relationships between epidermal growth factor (EGF) and neural tissues have of late attracted increasing interest. However, in spite of reported EGF effects on neurons, the expression of the EGF receptor (EGF-R) has not yet been unambiguously demonstrated in these cells. This 170-kDa protein bears an intracellular tyrosine kinase domain in which activity is ligand-dependent. We give definitive evidence here for its presence in neonatal and adult rat neurons showing also, for the first time, its binding and functional tyrosine kinase activities in the synaptic region. Immunohistochemistry using a polyclonal antibody prepared against the receptor purified from rat liver showed positive staining localized exclusively to neurons without regionalization to any particular brain zone. Binding studies made in Percoll-obtained synaptosomes revealed specific high affinity I-125-EGF binding sites (K(d), 1.42 x 10(-10) +/- 0.58 M) accounting for 17% of total binding and a great majority of low affinity (K(d), 2.55 x 10(-9) +/- 0.35 M) binding sites. Higher binding capacity was found in synaptosomal fractions obtained from newborn rats. The identity of the synaptosomal EGF binding activity with the 170-kDa EGF-R protein was demonstrated by cross-linking experiments. Furthermore, EGF-Affi-Prep affinity chromatography adsorbs a 170-kDa protein with EGF-R immunoreactivity from whole homogenates of adult rat brain. Phosphorylation assays made in freeze-thawed or intact synaptosomes showed EGF-induced tyrosine phosphorylation in the range of 170-, 126-150-, 124-, 113-, 98-, and 70-kDa proteins including the EGF-R. Thus, the EGF-R/EGF regulatory system could have a role in synaptic function that remains to be explored.
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    MICROTUBULES AND CALIBERS IN DEVELOPING AXONS
    (1986) FAUNDEZ, V; ALVAREZ, J
    The caliber and microtubular content of growing axons were assessed with the electron microscope in the sural nerve of the rat from birth until the age of 63 days (young adult). The caliber of both nonmedullated and myelinated fibers increased throughout the period of observation. At birth, nonmedullated fibers smaller than 0.2 .mu.m2 represented 69% of the population; at day 63 less than 7% were that small. Irrespective of the age of the rat, the number of microtubule profiles of nonmedullated fibers increased with the cross-sectional area of the axon although their packing decreased. In myelinated fibers of a given caliber, the packing of microtubules increased with time, and by day 63 the density had reached its final value. In nonmedullated fibers of a given caliber, a similar trend was observed after day 31; i.e., fibers showed a small but consistent increase in density. However, before that, and in contrast to myelinated fibers, nonmedullated fibers of defined calibers exhibited a transient increase in the microtubular density. Notwithstanding, during the history of an individual fiber the packing of its microtubules may decrease continuously until stabilizing, because the developing fiber is increasing its caliber and hence decreasing its microtubular density. In the 5-day-old rats, the caliber spectra of myelinated and nonmedullated axons overlapped in the 0.49-0.83 .mu.m2 range and their microtubular densities were similar. We conclude that the microtubular content of developing axons correlates with caliber, an architectural feature, and is largely independent of growth and of its myelinated condition. We propose that the cytoskeletal rather than the transport function commands the organization of axonal microtubules.