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J Glowinski

Publications and source records attributed to J Glowinski.

At least 307 records · Page 17Linked to original sources
PubMed1984

A combined light and electron microscopic method for the visualization of the same in vitro neuron by radioautography and serial sections.

We report here on a technical improvement which makes it possible to study, at the ultrastructural level, a dopaminergic neuron which has been previously identified by light microscopy. Primary cultures of virtually pure mesencephalic neurons from mouse embryos were obtained. These cultures were kept for 6 days, then incubated with tritiated dopamine, fixed and embedded in Epon. The dopaminergic neurons were firstly visualized by radioautography directly through Epon blocks in toto by light microscopy. In a second step, ultrathin sections of the identified dopaminergic cells were prepared and the neurons observed at the electron microscopy level. The dopaminergic nature of these neurons was regularly checked by radioautographic control on some selected ultrathin sections.

Animals↗
PubMed1984

A new type of tachykinin binding site in the rat brain characterized by specific binding of a labeled eledoisin derivative.

A new ligand for investigating tachykinin-binding site subtypes was synthesized by coupling the 125I-Bolton and Hunter reagent to eledoisin (125I-BHE). Using a synaptosomal preparation (P2 fraction) of rat cerebral cortex, 125I-BHE was shown to bind with apparent high affinity (apparent Kd = 15.3 nM). When concentrations of up to 30 nM 125I-BHE were used, 125I-BHE binding was specific, saturable, reversible, and temperature-dependent. In contrast to [3H]dopamine, 125I-BHE was not taken up within synaptosomes by an ouabain-sensitive process. Eledoisin, kassinin, and substance P were examined for their ability to inhibit specific 125I-BHE binding to cortical synaptosomes. Eledoisin and kassinin were considerably more potent than substance P, in contrast to the order of potency observed for specific 125I-Bolton-Hunter substance P (125I-BHSP) binding. Specific 125I-BHE binding was highest in the cerebral cortex and hypothalamus; intermediate in the hippocampus, striatum, and thalamus; low in the mesencephalon, septum, and substantia nigra; and absent in the cerebellum. Comparison of these data with those previously obtained for 125I-BHSP binding to synaptosomes indicated that 125I-BHE-labeled binding sites differ markedly from those of 125I-BHSP-labeled binding sites. Therefore, tachykinin receptors other than substance P receptors seem to be present in the central nervous system.

Animals↗
PubMed1983

In vivo release of [3H]GABA in cat caudate nucleus and substantia nigra. I. Bilateral changes induced by a unilateral nigral application of muscimol.

Push-pull cannulae were implanted in both caudate nuclei and both substantiae nigrae (SN) of halothane-anesthetized cats and the release of [3H]GABA, continuously synthesized from [3H]glutamine, was measured in these structures during 4 h of superfusion. In some experiments, multi-unit neuronal activity was recorded at the tip of the nigral push-pull cannulae, using a bipolar electrode. Two hours after the onset of superfusion with [3H]glutamine, 10(-6)M of muscimol was added (for 1 h) in the superfusion medium delivered to one SN. This treatment increased locally the release of [3H]GABA and enhanced the neuronal activity of the nigral cells in the zona reticulata. An increased release of [3H]GABA was also observed in the contralateral SN, in association with an inhibition of the activity of the zona reticulata cells. The unilateral nigral application of muscimol also induced asymmetric changes in the release of [3H]GABA in both caudate nuclei, since [3H]GABA release was increased ipsilaterally and reduced on the contralateral side. The present findings are considered in relation to possible GABAergic neuronal populations affected by this local pharmacological treatment.

Animals↗
PubMed1983

Local and distal effects induced by unilateral striatal application of opiates in the absence or in the presence of naloxone on the release of dopamine in both caudate nuclei and substantiae nigrae of the cat.

Halothane-anesthetized cats implanted with push-pull cannulae in both caudate nuclei (CN) and substantiae nigrae (SN) were used to study the effects of naloxone and various opiates when applied into the left CN on the release of newly synthetized tritiated dopamine (DA) from nerve terminals and dendrites of the two nigro-striatal dopaminergic pathways. In all cases, the drugs (naloxone, opiates alone or in the presence of naloxone) were applied for 30 min into the left CN. When applied alone, naloxone (10(-6) M) induced a delayed reduction in tritiated DA release both in the ipsilateral and contralateral CN. These effects were seen after removal of the drug from the superfusion fluid. Complementary experiments made with tritiated naloxone (10(-6) M) revealed that the contralateral effect on DA release was not due to a diffusion of the opiate antagonist from its application site. Locally, D-Ala2, Met-enkephalinamide (D-Ala2, Met-Enk, 10(-6) M) and the potent delta agonist Tyr-D-Ser-Gly-Phe-Leu-Thr (DSThr, 5 X 10(-8) M) induced a biphasic increase in tritiated DA release. The local changes in tritiated DA release evoked by morphine (10(-6) M) and mu agonists such as Tyr-D-Ala-Gly-NH-C6H13 (10(-8) M) and fentanyl (10(-8) M) differed from those of delta agonists and furthermore differed from each other. For instance, morphine induced a delayed increase in tritiated DA release whereas a biphasic increase followed by a delayed inhibition occurred with fentanyl. Among all the opiates tested D-Ala2-Met-Enk was the only one which elicited a distal effect, that is a reduction of tritiated DA release in the ipsilateral SN. Marked differences in these opiates' effects on tritiated DA release occurred both locally and in distal structures when opiates were applied simultaneously with naloxone (10(-6) M). Locally, the changes induced by mu agonists were particularly altered since during morphine's application with naloxone a reduction of tritiated DA release occurred. In addition, the opiate antagonist prevented the second increase and the delayed inhibition of tritiated DA release evoked by fentanyl (10(-8) M). Interestingly, the combined application of naloxone with either D-Ala2, Met-Enk (10(-6) M), DSThr (5 X 10(-8) M) or morphine (10(-6) M) resulted in the appearance of changes in tritiated DA release in contralateral structures. The most striking effect was seen with D-Ala2, Met-Enk which enhanced tritiated DA release in the contralateral CN and SN. These results are discussed in the light of the involvement of several types of opiate receptors and of the polysynaptic pathways responsible for the distal changes in dopaminergic transmission.

Animals↗
PubMed1983

Local cerebral glucose consumption in the rat. I. Effects of halothane anesthesia.

Rates of cerebral glucose utilization were measured by means of the autoradiographic 2-deoxy-D-(1-14C)glucose technique in normal rats under light halothane-anesthesia. Three types of region-specific metabolic alterations were elicited by inhalation of 0.5% halothane. The most striking effect observed was a significant increase of glucose consumption within the locus coeruleus, substantia nigra compacta and reticulata, interpeduncular nucleus, hippocampus, and fornix of the anesthetized animals in comparison to the corresponding brain areas of the conscious control rats. Halothane-anesthesia was also associated with significant metabolic depression in 21 (out of the 74 examined) discrete regions of the rat brain, distributed within the pons, cerebellum, diencephalon, and cortex, and was more prevalent in thalamus and neocortex. However, halothane failed to alter consistently the rates of glucose utilization in the rest of the rat brain areas investigated. The present findings suggest that halothane specifically alters the regional cerebral glucose utilization, with some limbic system components and the basal ganglia displaying increased metabolism, in contrast to the sensorimotor system which demonstrates significantly decreased metabolic activity.

Anesthesia, General↗
PubMed1983

Local cerebral glucose consumption in the rat. II. Effects of unilateral substantia nigra stimulation in conscious and in halothane-anesthetized animals.

The energy metabolism of 74 anatomically discrete central nervous structures was investigated by means of the autoradiographic 2-deoxy-D-(1-14C)glucose method (14C-DG) in conscious awake, as well as in halothane-anesthetized rats, following unilateral substantia nigra (SN) electrical stimulation. All the basal ganglia structures displayed bilateral metabolic activation in conscious animals, with only the contralateral subthalamic nucleus being unaffected. In anesthetized rats only the SN reticulata, globus pallidus, and subthalamic nucleus were affected ipsilaterally whereas anesthesia masked the effects of SN stimulation in SN compacta and striatum ipsilaterally, as well as within all the above structures contralaterally. The entopeduncular nucleus was bilaterally activated no matter the state of consciousness. Unilateral SN stimulation also increased glucose utilization within several thalamic regions (ventromedial, ventrolateral, ventroanterior, intralaminar, ventrobasal, and mediodorsal nuclei), the habenular complex, a few mesencephalic, brainstem (locus coeruleus and dorsal raphe) and cerebellar structures, mostly bilaterally and independently of the state of consciousness. Some of the factors suggested to be responsible for the masking effects of halothane-anesthesia on the metabolic activations elicited by unilateral SN stimulation are the following: (1) absence of movements in anesthetized rats, (2) halothane-induced depression of polysynaptic pathways mostly mediated through the thalamus, and (3) the stimulatory effect of halothane-anesthesia itself on metabolic activity in both parts of the SN.

Anesthesia, General↗
PubMed1983

GABA in the thalamic motor nuclei modulates dopamine release from the two dopaminergic nigrostriatal pathways in the cat.

Halothane anesthetized cats were implanted with five push-pull cannulae and the release of (3H)dopamine (3H-DA), newly synthetized from (3H)tyrosine, was measured in both caudate nuclei (CN) and substantiae nigrae (SN). GABA was applied by means of one of the cannula in the ventralis medialis (VM) and the adjacent part of the ventralis lateralis (VL) nuclei of the thalamus. Autoradiographic studies performed with (14C)-GABA showed that the diffusion of the amino-acid in our experimental conditions was restricted to a 4.2 mm3 sphere at the tip of the push-pull cannula. Multi-unit activity was recorded both at the site of GABA injection and in the contralateral VM or VL nucleus. During and after a 30-min application of GABA (10(-5) M) in the VM-VL nuclei, neuronal firing was markedly enhanced locally and decreased in the homologous structures, while (3H)DA release increased in both CN and the contralateral SN. No effect was observed in the ipsilateral SN. These results show that thalamic motor nuclei could play a role in the regulation of the activity of the two dopaminergic nigrostriatal pathways and will be discussed in the light of previous data indicating a role of the thalamus in the bilateral regulation of dopaminergic transmission in the basal ganglia.

Animals↗
PubMed1983

Diencephalic and mesencephalic efferents of the medial prefrontal cortex in the rat: electrophysiological evidence for the existence of branched axons.

Efferents originating from the rat medial prefrontal cortex and projecting to diencephalic and mesencephalic structures were analyzed electrophysiologically. The antidromic activation technique was used in order to identify cortical neurons which innervate the medio-dorsal nucleus of the thalamus (MD), lateral habenula, ventro-medial mesencephalic tegmentum (VMT), substantia nigra (SN), superior colliculus (SC) and central gray (CG). Corresponding cortical neurons, found mainly in layers V and VI, were distributed in the rostro-caudal extension of the medial prefrontal cortex. Their conduction velocities were relatively slow and ranged between 0.4 and 5.5 m/s. The reciprocal collision test allowed us to determine whether a given cortical neuron innervated only one structure or whether it sent axon collaterals to other subcortical areas. Twelve patterns of axonal branching were established: MD-VMT, MD-SN, MD-SC, VMT-SN, VMT-SC, VMT-CG, SN-SC, habenula-VMT, habenula-CG, MD-SN-SC, VMT-SN-SC and VMT-habenula-CG. Finally, it was estimated that a large proportion of the prefrontal cells which project to a given subcortical structure also innervate one or two other subcortical areas. Results are discussed in the light of anatomical data and their functional significance.

Animals↗
PubMed1983

Properties of a 125I-substance P derivative binding to synaptosomes from various brain structures and the spinal cord of the rat.

Using crude synaptosomal fractions (P2 fractions) and 125I-Bolton and Hunter substance P (125I-BHSP) as a ligand, the characteristics of specific binding sites were examined in various brain structures and in the spinal cord (dorsal and ventral parts) of the rat. Scatchard plots revealed the occurrence of a single class of binding sites in the various structures studied with comparable Kd values (from 0.46 to 1.10 nmol/l in the brain and 0.51, 0.56 nmol/l in the spinal cord dorsal and ventral parts respectively) and of marked differences in the number of binding sites (Bmax) (septum greater than striatum greater than hippocampus, hypothalamus greater than mesencephalon greater than cerebral cortex and dorsal part of the spinal cord greater than ventral part). In the brain no correlation was found between the number of 125I-BHSP binding sites and the amount of substance P levels (substance P-like immunoreactivity) in synaptosomes, particularly in the hippocampus and the substantia nigra since the former structure was characterized by its low substance P content and its high number of binding sites and the reverse was observed in the substantia nigra. The ability of several C- and N-terminal fragments of substance P and of tachykinins to compete with 125I-BHSP binding to synaptosomes from the hippocampus, the hypothalamus and the dorsal part of the spinal cord was then determined. Results obtained were closely similar from one structure to another and comparable to those previously reported using whole brain synaptosomes.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1983

Biogenic amines and adenosine-sensitive adenylate cyclases in primary cultures of striatal neurons.

Primary cultures of virtually pure striatal neurons from 16-day-old mouse embryos can be obtained using a serum-free chemically defined medium. Membranes prepared from these cells contain dopamine, beta-adrenergic, serotonin and adenosine sensitive adenylate cyclases. The pharmacological properties of the dopamine receptors are similar to those found for D1 receptors in adults except for the apparent affinities for agonists which were 5-10 times higher in fetal neurons. Beta-adrenergic receptors of striatal and cerebellar fetal neurons are of the beta 1-subtype as indicated by their identical affinity for adrenaline and noradrenaline and by their homogeneous, high affinity for practolol (Ki = 1.3 X 10(-6)M). Adenosine and serotonin sensitive adenylate cyclases present classical characteristics. An extensive study of the additive effects of the 4 neurotransmitter-sensitive adenylate cyclases indicates that: (1) part of the neurons bear more than one type of biogenic amine receptors; (2) the serotonin receptors are always associated with adenosine receptors on the same neurons; (3) adenosine- and dopamine-sensitive adenylate cyclases are additive. From this it can be concluded that as far as their adenylate cyclases-linked amine receptors are concerned, a maximal number of 15 types of neurons are present in these striatal cell cultures.

Adenosine↗
PubMed1983

Effects of unilateral electrical stimulation of various thalamic nuclei on the release of dopamine from dendrites and nerve terminals of neurons of the two nigrostriatal dopaminergic pathways.

The role of several motor and intralaminar thalamic nuclei in the regulation of dopamine release from terminals and dendrites of the nigrostriatal dopaminergic neurons was investigated in halothane-anaesthetized cats. For this purpose, the effects of the unilateral electrical stimulation of various thalamic nuclei on the release of newly synthesized [3H]dopamine were simultaneously determined in both substantiae nigrae and caudate nuclei using the push-pull cannula method. The electrical stimulation of the motor nuclei was the only one to induce asymmetric changes in the four structures since [3H]dopamine release was enhanced in the ipsilateral caudate nucleus and reduced in the contralateral structure while opposite responses were observed in the corresponding substantiae nigrae. A reduction of [3H]dopamine release occurred in the four structures or only in the contralateral substantia nigra and caudate nucleus following the stimulation of the parafascicularis nucleus and the adjacent posterior part of the nucleus centrum medianum or of the nucleus centralis lateralis and the adjacent paralaminar part of the nucleus medialis dorsalis, respectively. The stimulation of the anterior part of the nucleus centrum medianum, which in contrast to other thalamic nuclei examined, receives few nigral inputs, selectively enhanced [3H]dopamine release in the contralateral substantia nigra. No significant changes in [3H]dopamine release were seen either in the substantiae nigrae or in the caudate nuclei following the stimulation of midline thalamic nuclei. These results indicate that the motor and intralaminar thalamic nuclei exert multiple and selective influences on the release of dopamine from terminals and/or dendrites of the dopaminergic neurons. They also further support a role of thalamic nuclei in the transfer of information from one substantia nigra to the contralateral dopaminergic neurons. The possible involvement of connections between paired thalamic nuclei was underlined by the observations of evoked potentials in contralateral homologous nuclei following unilateral stimulation of motor, or some intralaminar, nuclei. The present report provides new insights on the mechanisms contributing to the reciprocal and/or bilateral regulations of nigrostriatal dopaminergic pathways.

Animals↗
PubMed1983

Effects of nigral application of muscimol on release of [3H] gamma-aminobutyrate and on multiunit activity in various cat thalamic nuclei.

The release of [3H] gamma-aminobutyrate (GABA) neosynthesized from [3H]glutamine was estimated in one substantia nigra and in the ipsilateral thalamus of halothane-anesthetized cats by perfusing a [3H]glutamine-enriched physiological medium through a push-pull cannula implanted in the two structures under investigation. After two hours of superfusion, muscimol (10(-6)M) was delivered through the nigral push-pull cannula for 50-60 min and local- and distal-evoked changes of [3H]GABA release were analyzed. In some experiments, changes of global neuronal activity induced by muscimol application were recorded in different thalamic nuclei, using a bipolar electrode. In a few of the above experiments, biochemical and electrophysiological determinations were simultaneously performed in the substantia nigra and the thalamus. The nigral application of muscimol (10(-6)M) induced locally an activation of the substantia nigra reticulata cells, as well as an increase in release of [3H]GABA. Distally, in the thalamus, two types of biochemical and electrophysiological responses were observed according to the localization of the tip of the push-pull cannula or the electrode. (1) An increased release of [3H]GABA and a depression of the global multi-unit cellular activity were obtained in the ventralis medialis-ventralis lateralis, the centralis lateralis and the paracentralis nuclei. These effects could reflect an activation of the GABAergic nigrothalamic neurons projecting to these different thalamic nuclei. (2) In contrast, in the medialis dorsalis paralamellar zone adjacent to the intralaminar nuclei of the thalamus, a decrease of [3H]GABA release and an activation of the multi-unit activity were obtained. These latter results may suggest either a polysynaptic response or the non-GABAergic nature of the nigrothalamic neurons afferent to the medialis dorsalis paralamellar zone.

Animals↗
PubMed1983

GABA in the intralaminar thalamic nuclei modulates dopamine release from the two dopaminergic nigro-striatal pathways in the cat.

Halothane-anaesthetized cats implanted with several push-pull cannulae were used to study the effects of the unilateral application of GABA (10(-5) M, 30 min) into thalamic intralaminar nuclei on the in vivo release of (3H)-dopamine [3H)-DA) newly synthesized from (3H)-tyrosine in both caudate nuclei (CN) and substantiae nigrae (SN). GABA applied into the left centralis lateralis nucleus (CL) elicited symmetric changes in the two CN, (3H)-DA release being initially reduced and thereafter markedly increased. On the contrary, an asymmetric pattern of responses was observed in the two CN during and after GABA application into the left centrum medianum parafascicular complex (CM/PF) since (3H)-DA release was decreased ipsilaterally and enhanced contralaterally. Changes in (3H)-DA release intervening in the two SN appeared to be triggered by processes independant from those operating in the CN. Hence, an immediate increase in (3H)-DA release with a time course distinct from that observed in other structures occurred in the contralateral SN after the application of GABA either into the left CL or the left CM/Pf. Furthermore in both cases, a biphasic response (decrease followed by an increase) similar to that intervening in the CN following GABA application into the CL was seen in the ipsilateral SN. Autoradiographic studies performed with (14C)-GABA (10(-5) M) revealed that the amino-acid did not spread out from the site of application. During and after GABA application (10(-5) M, 30 min) into the CL or the CM/Pf neuronal firing was markedly enhanced not only locally but also into respective contralateral homologous structures. These results further confirm the important and specific roles of distinct thalamic nuclei in the bilateral regulation of DA release from dendrites and nerve terminals of the nigro-striatal dopaminergic neurons.

Animals↗