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J P Denizot

Publications and source records attributed to J P Denizot.

At least 19 recordsLinked to original sources

Intermediate zone cells express calcium-permeable AMPA receptors and establish close contact with growing axons.

Recent studies have shown that cells in the intermediate zone (IZ) of the embryonic neocortex originate in the basal telencephalon and migrate tangentially in the cortical wall (;; ). We had previously observed growing cortical axons closely apposed to calbindin-positive, tangentially oriented cells in the IZ (), and it has been shown that neurites in the IZ express a glutamate transporter (). To test if glutamate released by corticofugal growth cones could influence the tangential IZ cells, we characterized the glutamate receptors expressed by IZ cells using patch-clamp techniques, histochemical labeling, and immunostaining on slices of embryonic mice forebrain. We show that tangential IZ cells express inwardly rectifying kainate responses, but not NMDA responses, and accumulate cobalt after AMPA receptor activation. We conclude that IZ cells express calcium-permeable AMPA receptors. This property correlates with our observation that the GluR2 subunit is not expressed in the IZ. AMPA receptors are activated by a millimolar concentration of glutamate. To know whether this high level of glutamate could occur at the surface of IZ cells, we examined contacts made by corticofugal growth cones and calbindin-positive IZ cells using electron microscopy. We show vesicle-containing neurites tightly apposed to calbindin-positive IZ cells over remarkably long length. This suggests that glutamate released by growing corticofugal axons could reach high concentrations close to AMPA receptors of tangential IZ cells and efficiently activate them to control the intracellular calcium in embryonic IZ cells.

Action Potentials↗

Synapse formation and spontaneous activity in rat brainstem neurons in primary culture.

The correlation between synaptogenesis and onset of spontaneous action potentials was assessed in rat brainstem cells up to 29 days in primary culture. Cells exhibited different stages of maturation followed by electron microscopy and patch clamp recordings. Terminal boutons with no preferential orientation of presynaptic vesicles appeared after 2 days in culture. After 5 days, preferential orientation of presynaptic vesicles and thickening of postsynaptic membranes were observed. The spontaneous discharge of action potentials, single or bursting, was observed after 7 days in vitro. This was followed by the expression of a 128-pS K(+) channel starting at 13 days in vitro. A 69-pS K(+) channel was also present throughout the duration of the cultures. These results suggest that spontaneous discharge of action potentials does not occur before synapses are formed and K(+) channel types develop differentially in brainstem neurons in vitro.

Action Potentials↗

Larval electroreceptors indicate a larval electric system in mormyrids.

The ontogeny of the electroreceptors of two species of mormyrids, Campylomormyrus cassaicus and Pollimyrus isidori, was studied. In young larvae (10 and 12 days old, respectively) ampullary organs, knollenorgans, promormyromasts and two types of tuberous organs (differing by their accessory cells) were found. These latter each possess a single sensory cell sitting on a platform of accessory cells. The platform is pierced by an unmyelinated nerve fibre. The promormyromast is composed of a single sensory cell surrounded by several accessory cells. The free apical area of the sensory cell and the accessory cells protrude into an intraepidermal cavity. The receptor cell is innervated by an unmyelinated nerve fibre ending in synaptic boutons. At the disappearance of the larval and appearance of the adult electric discharge the two types of tuberous organs degenerate, whereas the promormyromast differentiates into the typical mormyromast consisting of two types of sensory cells. The two types of tuberous organs and the promormyromast therefore are termed larval electroreceptors. Mormyrids are therefore the first group of electric fish possessing a complete larval electric system, comprising not only a larval electric organ, but also a larval electrosensory system.

Animals↗

Phosphorylated and non-phosphorylated neurofilament epitopes are co-distributed in the fish Mauthner axon.

The neurofilament (NF) polypeptides of the fish giant Mauthner cell axons (MAs) and their degree of phosphorylation were investigated in the adult by means of immunoblot and immunohistochemical staining. Fasciculus longitudinalis medialis (FLM) axons of much smaller caliber, among which MAs run in the ventral spinal cord, were also analyzed in two teleost fish belonging to continuously growing species. To detect NF polypeptide subunits, commercially available monoclonal antibodies against mammalian NF-L (68 kDa), NF-M (160 kDa), phosphorylated (P) and non-phosphorylated (nP) NF-H (200 kDa) epitopes were used. These antibodies labelled bands of the identical molecular weight in fish cervical spinal cord total protein immunoblots. A peculiar NF composition of the MAs was observed, following immunohistochemical staining i.e. a low NF-M expression and a codistribution of P and (nP)-NF-H epitopes. Moreover, on the basis of immunoperoxidase staining in ultrathin longitudinal MA sections, we suggest that P NF-H are arranged in bundles, whilst (nP)-NF-H are likely to be free in the axoplasm. By contrast, FLM axons were found reactive with antibodies only against P NF-H. These results confirm that in carp and trout, NF have epitopes cross-reacting with monoclonal antibodies directed against the mammalian NF subunits. Furthermore, as regards the co-distribution of phosphorylated and non-phosphorylated NF-H epitopes in the M-cell axons, these might be considered as not yet completely mature axons taking into account that carp and trout belong to continuously growing species.

Animals↗

Projection neurons of the mormyrid electrosensory lateral line lobe: morphology, immunohistochemistry, and synaptology.

This paper describes the morphological, immunohistochemical, and synaptic properties of projection neurons in the highly laminated medial and dorsolateral zones of the mormyrid electrosensory lateral line lobe (ELL). These structures are involved in active electrolocation, i.e., the detection and localization of objects in the nearby environment of the fish on the basis of changes in the reafferent electrosensory signal generated by the animal's own electric organ discharge. Electrosensory, corollary electromotor command-associated signals (corollary discharges), and a variety of other inputs are integrated within the ELL microcircuit. The organization of ELL projection neurons is analyzed at the light and electron microscopic levels based on Golgi impregnations, intracellular labeling, neuroanatomical tracer techniques, and gamma-aminobutyric acid (GABA), gamma-aminobutyric acid decarboxylase (GAD), and glutamate immunohistochemistry. Two main types of ELL projection neurons have been distinguished in mormyrids: large ganglionic (LG) and large fusiform (LF) cells. LG cells have a multipolar cell body (average diameter 13 microns) in the ganglionic layer, whereas LF cells have a fusiform cell body (on average, about 10 x 20 microns) in the granular layer. Apart from the location and shape of their soma, the morphological properties of these cell types are largely similar. They are glutamaterigic and project to the midbrain torus semicircularis, where their axon terminals make axodendritic synaptic contacts in the lateral nucleus. They have 6-12 apical dendrites in the molecular layer, with about 10,000 spines contacted by GABA-negative terminals and about 3,000 GABA-positive contacts on the smooth dendritic surface between the spines. Their somata and short, smooth basal dendrites, which arborize in the plexiform layer (LG cells) or in the granular layer (LF cells), are densely covered with GABA-positive, inhibitory terminals. Correlation with physiological data suggests that LG cells are I units, which are inhibited by stimulation of the center of their receptive fields, and LF cells are E units, excited by electric stimulation of the receptive field center. Comparison with the projection neurons of the ELL of gymnotiform fish, which constitute another group of active electrolocating teleosts, shows some striking differences, emphasizing the independent development of the ELL in both groups of teleosts.

Animals↗

Interneurons of the ganglionic layer in the mormyrid electrosensory lateral line lobe: morphology, immunohistochemistry, and synaptology.

This is the second paper in a series that describes the morphology, immunohistochemistry, and synaptology of the mormyrid electrosensory lateral line lobe (ELL). The ELL is a highly laminated cerebellum-like structure in the rhombencephalon that subserves an active electric sense: Objects in the nearby environment of the fish are detected on the basis of changes in the reafferent electrosensory signals that are generated by the animal's own electric organ discharge. The present paper describes interneurons in the superficial (molecular, ganglionic, and plexiform) layers of the ELL cortex that were analyzed in the light and electron microscopes after Golgi impregnation, intracellular labeling, neuroanatomical tracing, and gamma-aminobutyric acid (GABA) immunohistochemistry. The most numerous interneurons in the ganglionic layer are GABAergic medium-sized ganglionic (MG) cells and small ganglionic (SG) cells. MG cells have 10-20 spiny apical dendrites in the molecular layer, a cell body of 10-12 microns diameter in the ganglionic layer, a single basal dendrite that gives rise to fine, beaded, axon-like branches in either the plexiform layer (MG1 subtype) or the deeper granular layer (MG2 subtype), and an axon that terminates in the plexiform layer. Their apical dendritic tree has 12,000-22,000 spines that are contacted by GABA-negative terminals, and it receives, 1,250-2,500 GABA-positive contacts on the smooth dendritic surface between the spines. The average ratio of GABA-negative to GABA-positive contacts on the interneuron apical dendrites (14:1) is significantly higher than that for the efferent projection cells that have been described previously (Grant et al. [1996] J. Comp. Neurol., this issue). The somata and basal dendrites of MG cells receive a low to moderate density of GABAergic synaptic input, and their axons make GABAergic synaptic contacts with the somata and cell bodies of MG as well as with large ganglionic (LG) cells. SG cells probably represent immature, growing MG cells. Other interneurons in the superficial ELL layers include GABAergic stellate cells in the molecular layer, two types of non-GABAergic cells with smooth dendrites in the deep molecular layer that are named thick-smooth dendrite cells and deep molecular layer cells, and horizontal cells that are encountered particularly in the plexiform layer. Comparison with the ELL of waveform gymnotiform fish, which is another group of active electrolocating teleosts that has been investigated thoroughly, shows striking differences. In these fish, no GABAergic interneurons are found in the ganglionic (pyramidal) layer of the ELL, and GABA-negative interneurons with smooth dendrites in the molecular layer also seem to be lacking. At present, the phylogenetic origin of the described superficial interneurons in the mormyrid ELL is uncertain.

Animals↗

Characterization and sleep deprivation-induced expression modulation of dendrin, a novel dendritic protein in rat brain neurons.

We report on the characterization of the novel rat brain protein dendrin which is encoded by the brain-specific transcript 464. On immunoblots, two protein variants (81 kD, 89 kD) were identified in cytosolic and membraneous protein fractions. The variants are most abundant in the hippocampus, notably in apical dendrites of CA1 pyramidal cells. Dendritic and perikaryal immunolabelling is apparent in neurons of the cerebral cortex, dentate gyrus, subiculum, amygdala, and preoptic areas. In cortical and hippocampal dendrites, electron-dense immunoreaction is associated with the endoplasmic reticulum, the plasma membrane, and spine heads. An association of dendrin with polyribosomes and the presence of its mRNA in dendrites both provide evidence for dendritic mRNA translation. In the rat forebrain, dendrin expression is altered after an extended period of wakefulness. Twenty-four-hour sleep deprivation decreases the mRNA and protein concentrations of both variants in subcortical forebrain plus midbrain areas by 24 +/- 11% (P < 0.05) and 40 +/- 14% (P < 0.1), respectively, as measured relative to beta-actin mRNA and neural actin. In the cerebral cortex and hippocampus, the relative mRNA level remains unchanged whereas the cortical protein concentration is reduced by 42 +/- 10% (P < 0.05). Thus, dendrin belongs to a new class of dendritic proteins whose expression is differentially modulated by prolonged behavioral activity.

Animals↗

Neurogranin is locally concentrated in rat cortical and hippocampal neurons.

The rat protein kinase C substrate neurogranin has a granular distribution in cortical and hippocampal neurons. We demonstrate that in these cells, granular labelling corresponds to a local concentration of neurogranin-immunoreactivity at both the membranes of mitochondria and trans-Golgi vesicles and soma-proximal dendritic shaft and spinal head structures. Our findings suggest that the function of neurogranin could be affected by protein assembly at these cellular sites.

Animals↗

The GTPase Rab3a is associated with large dense core vesicles in bovine chromaffin cells and rat PC12 cells.

Small GTPases of the rab family control intracellular vesicle traffic in eukaryotic cells. Although the molecular mechanisms underlying the activity of the Rab proteins have not been elucidated yet, it is known that the function of these proteins is dependent on their precise subcellular localization. It has been suggested that Rab3a, which is mainly expressed in neural and endocrine cells, might regulate exocytosis. Recently, direct experimental evidence supporting this hypothesis has been obtained. Consistent with such a role for Rab3a in regulated exocytosis was the previously reported specific association of Rab3a with synaptic vesicles and with secretory granules in adrenal chromaffin cells. Since the latter result, based on subcellular fractionation, has been controversial, we have re-investigated the subcellular localization of this GTP-binding protein by using a combination of morphological techniques. Bovine chromaffin cells were labelled with an affinity-purified polyclonal anti-Rab3a antibody and analyzed by confocal microcopy. Rab3a was found to colocalize partially with dopamine beta-hydroxylase, a chromaffin granule marker. In agreement with this observation, immunoelectron microscopy revealed a specific staining of chromaffin granules. In addition to large dense core vesicles, some small vesicles were labelled. To eliminate the possibility that the staining was due to a Rab3a-related protein, we investigated by immunoelectron microscopy the localization of an epitope-tagged Rab3a expressed in rat PC12 cells. Secretory granules were specifically labelled, whereas clear microvesicles were not. These results provide further evidence supporting a specific association of the GTPase Rab3a with large dense core secretory vesicles.

Adrenal Medulla↗

Ultrastructural distribution of calcium in cutaneous electroreceptor organs of teleost fish.

The calcium distribution in the ampullary electroreceptor and the type B electroreceptor organ (gymnarchomast) of Gymnarchus niloticus (Glymnarchidae) and in the tuberous organ of Apteronotus leptorhynchus (gymnotidae) was studied. Endogenous calcium appeared as electron-dense precipitates when the cutaneous organs were pre-fixed with phosphate-buffered glutaraldehyde and postfixed with osmium tetroxide plus potassium bichromate. Calcium precipitates were localized in both intracellular compartments of sensory cells, and afferent nerve fibers. In contrast to sensory cells, small amounts of calcium precipitates were found in the cytoplasm of accessory cells. In sensory cells, electron-dense deposits were apparent mainly in synaptic vesicles near synaptic ribbons, inside vacuoles of the endoplasmic reticulum, and between the layers of the nuclear membrane. Very few deposits were found in mitochondria. Precipitates were also observed within the axons of afferent nerves and between the layers of the myelin sheath. The synaptic cleft was devoid of calcium. Calcium deposits have a specific cellular distribution in electroreceptor organs of teleost fish.

Afferent Pathways↗

[Post-embryonic differentiation of a cutaneous electro-receptor].

In order to decrease the rate of postembryonic development of electroreceptor organs, excisions of epidermis and deafferentations were carried out in the gymnotid fish Eigenmannia virescens. Twenty-five days later, the epidermis showed electroreceptor organs without innervation. Some of these at the beginning of their development consisted of masses of identical cells, whereas others showed presumed sensory cells whose cytoplasm contained rudimentary synaptic structures. The epidermis also showed differentiated tuberous organs with a low number of sensory cells. In all these organs, radioactive thymidine was fixed in the nuclei of the platform accessory cells. Thirty-five-40 days after surgery, tuberous organs were identical to the functional organs, and thymidine was detected in the nuclei of the cavity accessory cells. These results show that the gymnotid electroreceptor organs can develop before any nervous contact occurs, and suggest that they might originate from epidermal cells.

Animals↗

Formation of new sensory cells in deafferented tuberous organs of the gymnotid fish Eigenmannia virescens.

The cutaneous electroreceptor "tuberous organs" of the lateral line system of Eigenmannia virescens were studied at light and electron microscopic levels with immunohistochemical and autoradiographic techniques after sectioning of the posterior branch of the lateral line nerve. After deafferentation total degeneration of the sensory cells was observed. The accessory cells of the basal platform, however, remain intact and undergo a process of differentiation. The cytoplasms and nuclei of these cells increase in volume, and the nuclei incorporate tritiated thymidine. In control tuberous organs with intact innervation, tritiated thymidine is absorbed by the nuclei of the elongated epidermal cells surrounding the sensory cavity. The newly formed sensory cells, but not those of the intact organs, are substance P immunoreactive. They have synaptic bars surrounded by vesicles and their free membrane surface is covered with microvilli. The new sensory cells are fully differentiated 35 days after the lateral line was cut. These results demonstrate that in the tuberous organs of E. virescens new sensory cells are formed in the absence of an innervation.

Afferent Pathways↗

[Development, during ontogenetic development of gymnotids, of substance p expression in tuberous organs (electroreceptors)].

The evolution of the neuropeptidic expression of Substance P has been investigated with immunohistochemistry in the cutaneous electroreceptors, tuberous organs, during ontogenetic development of Apteronotus leptorhynchus. In the present data, antiSP antiserum has been applied to serial sections of Apteronotus leptorhynchus larvae obtained from several egg layings. Larvae were taken during development from hatching up to one hundred days old. SP immunoreactivity appeared just after hatching, in the epidermal zones which give rise to cutaneous sense organs. Four days after hatching, the tuberous organs are differentiated and immunoreactivity was observed in these organs, in both sensory cells and accessory cells. From day 30 after hatching, there was a regular decrease in the number of tuberous organs showing labelled accessory cells, and one hundred days later only 8% of tuberous organs had immunoreactive accessory cells. The adult accessory cells were no longer labelled with anti-SP antiserum. The results showed that in Apteronotus leptorhynchus, the epidermal structures which give rise to the cutaneous sensory organs were immunoreactive at a very early stage of development; this suggests that SP could have an effect upon their differentiation.

Animals↗

Substance P immunoreactivity of rat brain stem neurons in primary culture.

Substance P (SP)-ergic neurons from 16/17 day-embryonic rat brain stem in primary culture were identified by immunocytochemistry using biotinylated avidin and phosphatase alkaline methods with affinity-purified anti-SP antibodies. An average of 84% of neurons contained SP from day 9 to day 21 after plating. These in vitro data show that SP-containing neurons develop in our culture conditions. SP may act as a maturation factor as well as a neurotransmitter.

Animals↗

The olfactoretinalis system = terminal nerve?

Immunohistochemistry revealed a distinct terminal nerve (nT) from the olfactoretinalis system (ORS) in several gymnotid species. The ganglion (NOR) of the ORS is a cluster of FMRF-amide immunoreactive neurones located on the olfactory nerve and bulb. The NOR projects to the olfactory bulb and optic nerve but not to the olfactory epithelium. In contrast, the nT is a small substance P or GnRH immunoreactive fibre bundle which originates from a ganglion located rostral to the olfactory epithelium, courses caudally with the olfactory nerve and terminates as glomeruli in the olfactory bulb. Peripherally the ganglion projects to the epithelium of the anterior nostril. Thus, the terminal nerve appears to be a cranial nerve, clearly distinct from the ORS which consequently should no longer be considered as the terminal nerve.

Animals↗

[Extra-bulbar primary olfactory projection in teleost fishes].

Immunohistochemical investigations with anti-substance P antiserum demonstrate the existence of an extensive extrabulbar primary olfactory projection in several gymnotid teleost fish. This projection, never described before, originates in particular primary olfactory bundles which enter with the olfactory nerve into the olfactory bulb. While the bulk of the olfactory fibers end with glomeruli in the glomerular layer of the olfactory bulb, two particular bundles penetrate into the telencephalon and end, without forming glomeruli, in several telencephalic and diencephalic regions. A few fibers run as far as to the hypothalamus. In the light of these findings, the general notion that the primary olfactory projection is limited to the olfactory bulb and forms only glomeruli-like terminals, should be reconsidered.

Animals↗

A well defined spinocerebellar system in the weakly electric teleost fish Gnathonemus petersii. A tracing and immuno-histochemical study.

Long ascending fiber systems were investigated in the spinal cord of a teleost fish, Gnathonemus petersii. Concomitant results of Fink-Heimer degeneration tracing as well as CaBP28K immunohistochemical labelling demonstrate the existence of a well defined direct pathway from the very lowest spinal level to the caudal lobe of the cerebellum. HRP retrograde labelling shows that this pathway originates in a cellular column located in the most ventral part of the lateral column next to the lateral extremity of the ventral horn. From each spinal segment, the large axons of these cells gather and form a strip shaped tract at the periphery of the lateral column immediately dorsal to the cell column from which they originate. The spinal course of these fibers is ipsilateral; they give off a large number of collaterals to the lateral reticular nucleus. Bypassing the trigeminal motor nucleus, the lateral column tract courses dorsally to the paratrigeminal command associated nucleus between the lateral lemniscus and the nucleus preeminentialis and with a ventro-dorsally oriented large loop, turns in the caudal direction and penetrates into the cerebellar caudal lobe. Running caudally in the dorsal granular layer of the caudal lobe, it shifts more and more medially and crosses the midline whilst decussating with the contralateral tract on the dorsal margin of the molecular layer of the caudal lobe. Finally, the tract splits off and terminates throughout the granular layer of the caudal lobe. The main characteristics of this pathway are similar to those of the ventral spinocerebellar tract of higher vertebrates; it conveys information from all spinal levels directly to the contralateral cerebellum. However, it does not seem to receive direct synaptic input from the periphery, since projection of the dorsal root fibers appears to be limited to the dorsal ipsilateral half of the spinal cord. The appearance of such a pathway in a teleost fish is probably related to the existence of a well developed proprioceptive system in this species.

Animals↗

Immunohistochemical demonstration of calbindin-D 28K (CABP28K) in the spinal cord motoneurons of teleost fish.

The distribution and localization of the calcium-binding protein, calbindin-D 28K (CaBP28K), in the spinal cord motoneurons of larvae of the teleost fish, Apteronotus leptorhynchus (Gymnotidae) and Pollimyrus isidori (Mormyridae), and in the adult goldfish, Carassius auratus (Cyprinidae), were determined by means of immunohistochemistry. Sections of whole larvae and goldfish spinal cord were reacted with a polyclonal antibody to rat renal CaBP28K. CaBP28K was located by the PAP technique (Sternberger). It was found in the soma, dendrites, axons and axon terminals of spinal motoneurons but not in those of electromotoneurons of Apteronotus leptorhynchus, whereas it occurred in both motoneurons and electromotoneurons of the larval electric organ of Pollimyrus isidori. In these species CaBP28K was also present in the electromotoneuron axon terminals that make synaptic contacts with the pedicles of the electrocytes. In adult Carassius auratus, CaBP28K was found in the soma, dendrites and axons of certain spinal motoneurons. The results indicate that, in teleosts, the motoneurons containing CaBP28K may represent a well-defined population within the spinal cord; the role of this protein in these cells remains to be determined.

Animals↗