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Biomedical subjects

J F Pujol

Publications and source records attributed to J F Pujol.

At least 109 records · Page 6Linked to original sources
PubMed1981

Radioautographic evidence for an innervation of the subcommissural organ by GABA-containing nerve fibres.

Light-and electron-microscopic radioautographic studies were carried out in rats after intraventricular injection of [3H]GABAergic elements in the subcommissural organ (SCO). This amino acid is specifically accumulated in SCO ependymocytes and numerous nerve terminals and fibres. Some labelled terminals contain clear round vesicles and large granular vesicles and sometimes display synaptic contacts on the SCO ependymocytes. They disappear after 5,7-dihydroxytryptamine (5,7-DHT) treatment or lesions of mesencephalic raphé nuclei. They are morphologically and pharmacologically similar to the serotonin-containing terminals of the basal SCO. The other labelled terminals contain exclusively clear round vesicles, are dispersed throughout the SCO and survive after the 5,7-DHT treatment or raphé nuclei lesions; some of them are in synaptic contact with neuronal elements of the basal SCO. In both SCO ependymocytes and fibres or terminals, [3H]GABA is accumulated according to the pharmacological criteria of a GABA neuronal uptake. Under the same experimental conditions [3H[glutamine and beta[3H]alanine fail to label the SCO: [3H] glutamate produces a very light labelling and [3H] serotonin is accumulated only in nerve profiles of the basal SCO. These results, and the presence of a glutamate decarboxylase activity in the SCO, suggest the existence of GABA-synthesizing elements in the SCO and could indicate the possible involvement of GABA in the secretory activity of the SCO.

5,7-Dihydroxytryptamine↗
PubMed1981

Importance of fixation in immunohistochemistry: use of formaldehyde solutions at variable pH for the localization of tyrosine hydroxylase.

Adequate fixative in immunohistochemistry requires not only a rapid and total immobilization of the antigen, but also a sufficient preservation of its immunoreactivity and maintenance of its accessibility to the immunochemical reagents for localization. Thus, the optimal fixation condition for a specific antigen necessitates a compromise between these opposing variables and can be determined by the preparation of a series of tissues with a progressively increasing degree of fixation. Unless the results of localization using such a series is available, one must be satisfied with adequate but less than optimal results. In the present study, this principle is demonstrated using the localization of tyrosine hydroxylase in the dopaminergic system with formaldehyde as the fixative. The rate and degree of fixation with formaldehyde was shown to be highly pH dependent. By perfusing the tissue with formaldehyde at pH 6.5 (where the rate of fixation is extremely slow) it is possible to rapidly distribute the fixative homogeneously into the tissue. By suddenly changing to a formaldehyde perfusate of higher pH, the cross-linking reaction is rapidly increased. This two-step fixation procedure provides a means of obtaining a rapid and uniform immobilization of the antigen, so that its translocation can be avoided. The final degree of fixation is controlled by the duration and pH of the second fixative solution. The results obtained by increasing the pH of the second solution demonstrated that complete fixation of tyrosine hydroxylase in the dopaminergic system with formaldehyde maybe obtained using a very basic formaldehyde solution (pH 11) while still retaining immunoreactivity of the enzyme. The localization that was achieved at lower pH appeared adequate until it was compared to the results obtained by perfusion at pH 11 in the second step.

Animals↗
PubMed1981

Functional recuperation of the serotoninergic innervation in the rat locus coeruleus.

1. 5,6-dihydroxytryptamine (5,6-DHT) or a lesion of the raphe centralis superior (RCS) cause significant decreases in the serotonin (5-HT) content and significant increases in the tyrosine hydroxylase activity in the locus coeruleus (LC) of the rat. This suggests that noradrenaline (NA) synthesis is controlled by serotonin-containing neurons in the raphe system via their terminals in the LC. 2. Radioautography after intraventricular infusion of tritiated serotonin (3H-5-HT) and biochemical determinations of endogenous 5-HT content showed an almost complete disappearance of serotoninergic axonal varicosities and content in the LC region 10-15 days after intraventricular administration of 75 micrograms of 5,6-DHT. Two to 4 months after neurotoxin administration, 5-HT fibers had regrown in the LC but, contrary to the normal innervation pattern, the majority of them invaded the medial most portion of the nucleus and the adjacent subependymal region. The LC region regained almost all of its endogenous 5-HT content in the same time period. 3. Functional recuperation of these 5-HT fibers was demonstrated by the fact that the RCS had, after regeneration, the same functional control on NA synthesis as in the normal animal.

5,6-Dihydroxytryptamine↗
PubMed1981

Is the retrograde axonal transport of 3H-5-HT a specific process of serotoninergic neurons?

The specificity of the retrograde axonal transport of 3H-serotonin (3H-5-HT) was radioautographically studied in the afferents to the olfactory bulb (O.B.). Injections of 3H-5-HT of different concentrations (10(-2), 10(-3), 10(-4) and 10(-5) M) were performed into the O.B. of catron pretreated rats. Following injection of 3H-5-HT (10(-2) M), a cytoplasmic perikaryal labeling was observed in the bulk of afferents to the O.B. (aminergic and non-aminergic neurons). When lower concentrations of 3H-5-HT (10(-5) M) were injected into the O.B., the retrograde labeling was only seen in the raphe dorsalis (RD) serotoninergic perikarya. The specificity of the uptake-retrograde transport of 3H-5-HT seems to depend on the selectivity of uptake by nerve terminals.

Afferent Pathways↗
PubMed1981

In vivo measurements by differential pulse voltammetry of extra-cellular 5-hydroxyindoleacetic acid in the rat brain.

The use of differential pulse voltammetry, performed with electrochemically treated carbon fiber electrodes, enables us to detect in vitro or in vivo, in the striatum of anaesthetized rats, an oxidation peak (3) at a potential of + 300 mV. Electrolytic or 5-7-dihydroxytryptamine lesions of the medial forebrain bundle are followed by a decrease of 59 and 62% respectively of this peak. Biochemical measurements are significantly correlated with the measured peak (3) and decreases. Thus, peak (3) increases obtained after injection of L-tryptophan and/or Reserpine, as well as the decreases observed after injection of Clorgyline or 3-hydroxybenzylhydrazine, confirm that peak (3) is dependent upon 5-hydroxyindoleacetic acid concentration. The detection of a peak (3) in the cerebrospinal fluid and its increase after injection of Probenecid reinforce this conclusion.

5,7-Dihydroxytryptamine↗
PubMed1981

[Possible alteration of adrenergic neurons in the mesolimbic system of psychotics (author's transl)].

The activity of Phenylethanolamine-N-Methyltransferase (PNMT) was assayed in 12 brain structures in a group of 8 chronic psychotics, a sub-group of 5 schizophrenics and a group of 7 normal controls. It was found significant decreased in substantia perforata posterior and nucleus accumbens. These two structures belong to the meso-limbic system. These data, therefore, suggest an inhibition of PNMT and/or an alteration of putative adrenergic neurons in the meso-limbic system, in schizophrenia.

Humans↗
PubMed1980

[Micromethod of measuring the 3 brain isoenzymes of enolase].

The brain of the vertebrates contains three isozymic forms of the glycolytic enzyme enolase, two of which are neuron specific. These proteins could therefore be utilized as biochemical markers for various studies of neurons. We have devised a separation procedure which allows the measurement of the three enolase forms from minute amounts of tissue (50 to 100 microgram total protein). We show that this method can be applied to studies of brain microstructures, in vivo and in vitro (clonal cell lines) neural differentiation, and in measurements in biological fluids.

Animals↗
PubMed1980

[Activities of enzymes synthesizing catecholamines in areas of the brain stem in sudden infant death].

In victims of Sudden Infant Death Syndrome, the activity of phenylethanolamine-N-methyltransferase was found to decrease in the nucleus retroambigu, the Kolliker-Fuse nucleus and the C 2 area of medulla, whereas a decrease in the dopamine-beta-hydroxylase activity was found only in the C 2 area. These results suggest a hypoactivity of adrenergic neurons in respiratory and cardiac centres in the Sudden Infant Death Syndrome.

Adrenergic Fibers↗
PubMed1980

Retrograde axonal transport following injection of [3H]serotonin in the olfactory bulb. I. Biochemical study.

A retrograde axonal transport from the serotonergic nerve terminals in the olfactory bulb (OB) to their parent cell bodies in the midbrain raphe nuclei has been demonstrated after stereotaxic injection of [2H]5-HT into the OB of rats pretreated with a monoamine oxidase (MAO) inhibitor: at various time intervals thereafter (4-92 h) there was a preferential accumulation of radioactivity mainly in the raphe dorsalis nucleus (RDN). Maximal accumulation occurred at 24 h. Of this radioactivity, 30-50% was recovered as 5-HT. The accumulation was estimated to take place at two rates: a fast one (48 mm/day) and a slower one (16 mm/day). Under the same experimental conditions there was no clear evidence for a retrograde accumulation of [3H]norepinephrine in the RDN. A passive diffusion mechanism could be excluded since the diffuson of tracer towards the cerebrospinal fluid was prevented by prior mechanical obstruction of the olfactory diverticle of the lateral ventricle. Furthermore, colchicine strongly reduced (by 80%) the radioactive accumulatin in the RDN. Destruction of serotonergic nerve terminals by 5,6-dihydroxytryptamine or inhibiton of 5-HT uptake by fluoxetine decreased this retrograde accumulation whereas destruction of catecholaminergic nerve terminals by 6-hydroxydopamine was without effect. Pretreatment with reserpine decreased the amount of radioactivity transported to the RDN by 40%. In the absence of MAO inhibition pretreatment, animals still presentd 35% of the tracer transported to the RDN. Intrabulbar injection of MAO inhibitor did not affect the accumulation rates when compared with animals which received the inhibitor by the intraperitoneal route. In conclusion, the retrograde axonal transport following [3H]5-HT injection in the serotonergic RDN-OB system occurs via an active process which depends on a colchicine-sensitive mechanism and is partially linked to a reserp ine-sensitive structure. During its transport, the amine seems to be relatively protected from metabolic inactivation.

Animals↗
PubMed1980

Retrograde axonal transport following injection of [3H]-serotonin into the olfactory bulb. II. Radioautographic study.

Radioautography was used to study the intraneuronal distribution of [3H]-serotonin (5-HT) and/or its derivatives selectively taken up by the olfactory bulb (OB) serotonergic terminals and subsequently transported to their parent cell bodies in the midbrain raphe nuclei. This was done 24 h after injection of [3H]5-HT into the main OB of rats either pretreated or not with monoamine oxidase (MAO) inhibitor. A prior mechanical obstruction of the rostral ventricular cavities prevented diffusion of the tracer towards cerebrospinal fluid. Heavily labelled nerve cell bodies were found mainly in the ipsilateral raphe dorsalis nucleus (RDN) and to a lesser extent in the raphe centralis nucleus. The radioautographic reaction often extended to dendritic processes while sparing the nucleus. A diffuse reaction was also observed but limited to the raphe area. The supraependymal 5-HT fibers were found to be free of labelling. Neither local destruction of catecholaminergic terminals with 6-OHDA, nor absence of MAO inhibition, impaired this radioautographic pattern, while destruction of serotonergic terminals with 5,6-dihydroxytryptamine in OB resulted in the disappearance of labelled axonal varicosities and neurons in the OB and the RDN respectively. At the electron microscopy level, labelled cell bodies in the RDN were medium-sized (12-15 micrometers). Silver grains were localized mainly on mitochondria and, to a lesser extent, on lysosomes and endoplasmic reticulum but spared the nucleus and the nucleolus. Silver grains were also found near the nuclear membrane and outside the neuronal membrane. The observation of heavy metal impregnated thick sections confirmed the preferential localization of silver grains on mitochondria with or without inhibition of MAO. These results could account for the subcellular compartments involved in the retrograde axonal transport of [3H]5-HT and its subsequent degradation and/or dendritic release.

Animals↗
PubMed1980

In vivo electrochemical detection of catechols in the neostriatum of anaesthetized rats: dopamine or DOPAC?

Electroanalytical techniques for the in vivo measurement of neurotransmitters in brain tissue have been applied especially to the catecholamines, which are easily oxidizable. Measurements are, however, complicated by the presence of ascorbic acid (AA) in brain tissue. Lane et al. have been able to circumvent this problem, at least in part, by the application of differential pulse voltametry (DPV) to a surface-modified platinum electrode, obtaining distinct oxidation current peaks in recordings from the rat neostriatum which are attributed to AA and to dopamine (DA), respectively, but which are also unstable. We have recently described a new type of electrode, consisting of a pyrolytic carbon fibre 8 micrometers thick and 0.5 mm long. We now report that the DPV method used in conjunction with an electrochemical treatment of this electrode yields stable and reproducible peaks in which catecholamines and AA are resolved from each other. Moreover, pharmacological investigations suggest that the catecholamine peak measured in vivo in the rat neostriatum should be attributed to 3, 4-dihydroxyphenylacetic acid (DOPAC), suggesting that our technique may be a useful means of following dopaminergic activity in vivo.

3,4-Dihydroxyphenylacetic Acid↗
PubMed1980

Alteration of tyrosine hydroxylase activity in the locus coeruleus after administration of p-chlorophenylalanine.

The time course of the variations in tyrosine hydroxylase activity (THA) and serotonin (5-HT) content were measured in the rat locus coeruleus after parachlorophenylalanine (PCPA) administration. Highly significant decreases in the 5-HT content in LC were found 24-48 h after PCPA treatment (300 mg/kg daily). An increase in THA (in the LC) was found to be significant 4 days after 2 successive injections of PCPA and after 4 successive injections of the drug THA gradually increased, reaching a maximum around 4 days after the last injection. This maximum increase in THA was greatly reduced when 5-HTP was simultaneously administered with PCPA. These results join others which suggest that a serotonin-mediated mechanism could be one of the processes controlling noradrenaline metabolism in the locus coeruleus.

Animals↗
PubMed1980

Genetic studies of daily variations of first-step enzymes of monoamines metabolism in the brain of inbred strains of mice and hybrids. I. Daily variations of tryptophan hydroxylase activity in the nuclei raphe dorsalis, raphe centralis and in the striatum.

A daily variation of tryptophan hydroxylase (TrH) activity was observed in the raphe dorsalis (RD), raphe centralis (RC) and striatum (St) of 3 inbred strains of mice (BALB/c, C57BL/6, C57BR) and of the reciprocal hybrids obtained from Balb/c and C57BL6. Significant differences of the characteristics of these rhythms have been found in the same strain between different structures and for the same structure between different strains. In RD and RC hybridization led to less defined daily variations which, in the striatum, remained well synchronized and could be controlled by a dominant genetic mechanism part. These results help to discuss evidence for selective control mechanisms of regulation responsible for daily variation of TrH in the 5-HT cell bodies and terminals and their relative independence.

Animals↗