0000000000079592
AUTHOR
A. Molowny
Endocytosis in cultured neurons is altered by chronic alcohol exposure.
Endocytosis is required for many cellular pivotal processes, including membrane recycling, nutrient uptake, and signal transduction. This complex process is particularly relevant in polarized cells, such as neurons. Previous studies have demonstrated that alcohol alters intracellular traffic, including endocytosis, in several cell types. However, information on the effect of chronic alcohol exposure on this process in neurons is scarce. As an approach, we investigated the effect of alcohol exposure on the internalization of two widely used endocytic markers, albumin and transferrin, in developing hippocampal neurons in primary culture. The effect of this treatment on the levels of several r…
PSA-NCAM immunocytochemistry in the cerebral cortex and other telencephalic areas of the lizard Podarcis hispanica: differential expression during medial cortex neuronal regeneration.
The lizard medial cortex, a region homologous to the mammalian dentate gyrus, shows postnatal neurogenesis and the surprising ability to replace its neurons after being lesioned specifically with the neurotoxin 3-acetylpyridine. As the polysialylated form of the neural cell adhesion molecule (PSA-NCAM) is expressed during neuronal migration and differentiation, we have studied its distribution in adult lizards and also during the lesion-regeneration process. In the medial cortex of control animals, many labeled fusiform somata, presumably corresponding to migratory neuroblasts, appeared in the inner plexiform layer. There were also scattered immunoreactive granule neurons in the cell layer.…
The lizard cerebral cortex as a model to study neuronal regeneration
The medial cerebral cortex of lizards, an area homologous to the hippocampal fascia dentata, shows delayed postnatal neurogenesis, i.e., cells in the medial cortex ependyma proliferate and give rise to immature neurons, which migrate to the cell layer. There, recruited neurons differentiate and give rise to zinc containing axons directed to the rest of cortical areas, thus resulting in a continuous growth of the medial cortex and its zinc-enriched axonal projection. This happens along the lizard life span, even in adult lizards, thus allowing one of their most important characteristics: neuronal regeneration. Experiments in our laboratory have shown that chemical lesion of the medial cortex…
Early histological maturation in the hippocampus of the guinea pig.
The vesicular zinc-rich synaptic systems of the principal neurons of the hippocampus are well developed in newborn guinea pigs, a precocial species. In addition, alvear and fimbrial myelinated fibers as well as significant inhibitory interneurons (i.e. somatostatin, parvalbumin and opioid immunoreactive hippocampal interneurons) are also well developed. On the contrary, neither vesicular zinc synapses nor myelinated fibers nor the above mentioned immunoreactive interneurons are detectable in newborn specimens of other related altricial species such as rats or rabbits. These data suggest that early maturation of a highly integrative center related to cognitive map building such as the hippoc…
Ontogeny of somatostatin immunoreactive neurons in the medial cerebral cortex and other cortical areas of the lizardPodarcis hispanica
The ontogeny of somatostatin immunoreactive interneurons in the cerebral cortex of the lizard Podarcis hispanica has been studied in histological series of embryos, perinatal specimens, and adults. Somatostatin immunoreactive interneurons appear in the early stages of lizard cerebral cortex ontogeny, their number increases during embryonary development, reaches a peak in early postnatal life, and decreases in adult lizards. The first somatostatin immunoreactive somata in the lizard forebrain appeared on E36, and they were located in non cortical areas. Then, on E39 and later, somatostatin immunoreactive neurons were seen in the lizard cortex in a rostral-to-caudal spatial gradient, which pa…
Ethanol reduces zincosome formation in cultured astrocytes.
Aims Zinc is an ion that participates in basic cellular and tissular functions. Zinc deficiency is present in many physiological and health problems affecting most body organs, including the brain. Among the circumstances involved in zinc deficiency, ethanol consumption is probably one of the most frequent. A dietary zinc supplement has been proposed as possibly being an efficient method to palliate zinc deficiency. Astrocytes form part of the hematoencephalic barrier, and they are apparently implicated in the homeostasis of the neuronal medium. In this work, we analyze the effect of ethanol on extracellular zinc management by rat astrocytes in culture. Methods Intracellular levels of 'free…
Capture of extracellular zinc ions by astrocytes.
Synaptic zinc ions released during synaptic transmission interact with pre- and postsynaptic neuroreceptors, thus modulating neurotransmission. It is likely that they have to be efficiently cleared from the extracellular milieu to assure subsequent synaptic events. Both neurons and glia are assumed to participate in this clearance by mechanisms that are not fully understood. In this study, electron microscopic zinc cytochemistry has shown zinc-electrondense particles associated with hippocampal astrocytic membranes frequently found accumulated in stacked lamellae. In cultured astrocytes, the use of zinc fluorochromes and endocytic markers allowed the simultaneous imaging of the capture of e…
Cytochemical techniques for zinc and heavy metals localization in nerve cells
Zinc is one of the most abundant oligoelements in the living cell. It appears tightly bound to metallothioneins, loosely bound to some metalloproteins and nucleic acids, or even as free ion. Small amounts of zinc ions (in the nanomolar range) regulate a plentitude of enzymatic proteins, receptors, and transcription factors; thus, cells need accurate homeostasis of zinc ions. Some neurons have developed mechanisms to accumulate zinc in specific membrane compartments ("vesicular zinc") which can be revealed using histochemical techniques. This article is a short report on the different direct-indirect experimental approaches for zinc and heavy metal detection in neurons. Substances giving a b…
Photoperiod-temperature and neuroblast proliferation-migration in the adult lizard cortex.
The lizard medial cortex (a zone homologous to the mammalian fascia dentata) shows delayed postnatal neurogenesis throughout the lifetime of these animals. Experimental lesioning of this area is followed by neuronal regeneration, a unique phenomenon in the adult amniote telencephalon. The differential effects of temperature and photoperiod on postnatal neurogenetic activity were studied using tritiated thymidine pulses and posterior autoradiography as well as proliferating cell nuclear antigen (PCNA) immunostaining. Long (summer) photoperiods increased the number of proliferating neuroblasts in the ependymal neuroepithelium. Cold (winter) temperature prevented migration of the newly generat…
Ultrastructure of putative migrating cells in the cerebral cortex of Lacerta galloti.
Cells considered to be migratory in the cerebral cortex of adult lizards are ultrastructurally of two types. Nuclei in the first type have highly dispersed chromatin, creating a spongy appearance, whereas in the second type the chromatin is irregularly clumped. Both types of cells are closely associated with processes of radial ependymal glia cells, which perhaps orient their migratory pathways. Cells with spongy chromatin show an increase in cytoplasmic organelles and progressive chromatin condensation as they travel from the ependymal layer to the granular layer. Possibly these cells account for the neuronal increase that takes place in the granular layer during postnatal life. Cells with…
CRMP-4 expression in the adult cerebral cortex and other telencephalic areas of the lizard Podarcis hispanica.
The control of neuritogenesis is crucial for the development, maturation and regeneration of the nervous system. The collapsin response-mediated protein 4 (CRMP-4) is a member of a family of proteins that are involved in neuronal differentiation and axonal outgrowth. In rodents, this protein is expressed in recently generated neurons such as some granule neurons of the dentate gyrus, as well as in certain differentiated neurons undergoing neurite outgrowth or synaptogenesis during adulthood. Since CRMP-4 protein appears to be highly conserved throughout the evolutionary scale, we have used immunocytochemistry to study its distribution in the lizard cerebral cortex. We have found pronounced …
Delayed postnatal neurogenesis in the cerebral cortex of lizards
Labelled cells were consistently observed in the medial cortex of the lizard brain after i.p. injections of tritiated thymidine (5 microCi/g b. wt.), 1, 7, 18 or 28 days of survival and posterior autoradiographic evaluation. In 3 groups of specimens (postnatal, young and adult) of the species Podarcis hispanica, after one day of survival, labelled cells were located in the ependymal cell layer underlying the medial cortex. After intermediate survival times (7, 18 days), labelled cells were found in 3 zones: the ependymal layer, the inner plexiform layer and the granular layer. After one month of survival, most labelled cells were observed in the granular layer. In the granular layer, these …
Zinc-positive presynaptic boutons of the rabbit hippocampus during early postnatal development
Abstract The evolution of vesicular zinc-containing boutons in the developing rabbit hippocampus has been studied during early postnatal life using the selenite–Danscher histochemical method. By P3, large immature mossy fiber boutons with labeled synaptic vesicles were seen in the hilus of the dentate gyrus and in the stratum lucidum of the CA3–CA4 hippocampal areas. After P5, smaller boutons with labeled vesicles were identified in the stratum oriens and stratum radiatum of all hippocampal areas, and even transiently in the stratum lacunosum of P6 animals. Vesicular zinc-containing boutons increased in number and underwent ultrastructural maturation; light microscope densitometric–volumetr…
Neurons of the dentate molecular layer in the rabbit hippocampus.
The molecular layer of the dentate gyrus appears as the main entrance gate for information into the hippocampus, i.e., where the perforant path axons from the entorhinal cortex synapse onto the spines and dendrites of granule cells. A few dispersed neuronal somata appear intermingled in between and probably control the flow of information in this area. In rabbits, the number of neurons in the molecular layer increases in the first week of postnatal life and then stabilizes to appear permanent and heterogeneous over the individuals' life span, including old animals. By means of Golgi impregnations, NADPH histochemistry, immunocytochemical stainings and intracellular labelings (lucifer yellow…
Postnatal neurogenesis in the medial cortex of the tropical lizard Tropidurus hispidus.
Young, adult and presumed old specimens of the tropical lizard Tropidurus hispidus, living in an almost steady warm habitat, have been the subjects of a 5-bromodeoxiuridine immunocytochemical study to label proliferating brain cells. All animals showed abundant 5-bromodeoxiuridine-labeled nuclei in the ependyma of their telencephalic lateral ventricles, with these being especially abundant in the medial cortex ependyma. Surprisingly, adult animals displayed higher numbers of labeled nuclei when compared with those of young specimens. In a second experiment, in order to check the evolution of ependymal-labeled nuclei, adult specimens were allowed 4 h or 2, 4, 7, 15 or 30 days of survival aft…
Transitory disappearance of microglia during the regeneration of the lizard medial cortex
In normal lizards, microglial cells populate the medial cortex (a zone homologous to the hippocampal fascia dentata), with a preferential distribution along the border between the granular cell layer and the plexiform layers. Intraperitoneal injection of the neurotoxin 3-acetylpyridine (3AP) induces a selective lesion in the medial cortex with a rapid degeneration of the granular layer and its zinc-enriched axonal projection. Within 6-8 weeks, the granular layer is, however, re- populated by a new set of neurons generated in the subjacent ependyma and the cell debris is removed. The aim of this study was to determine to what extent microglia were involved in the scavenging processes during …
Timm-staining intensity is correlated with the density of Timm-positive presynaptic structures in the cerebral cortex of lizards
In cortical areas of the lizard, Podarcis hispanica, Timm staining reveals a distinct pattern of lamination. At the electron-microscope level, virtually all of the reaction product is located in the synaptic vesicles of Timm-positive boutons. Using linear-regression analysis, the area density of Timm-positive bouton profiles as well as the numerical and volume density of stained vesicles were found to be closely correlated with the light-microscopic densitometric values obtained for each Timm-positive cortical zone. We discuss the possibility of estimating stereological electron-microscopic data parameters from densitometric measurements at the light-microscope level.
Reactive neurogenesis during regeneration of the lesioned medial cerebral cortex of lizards
Abstract This study reports that lesion of the adult lizard medial cortex (lizard hipocampal fascia dentata) induces a short period of intensive neurogenesis which we have termed reactive neurogenesis; a cell proliferation event that occurs in the subjacent ependyma. Specific lesion of the medial cortex was achieved by intraperitoneal injection of the neurotoxin 3-acetylpyridine and proliferating cells were detected using tritiated thymidine or 5-bromodeoxiuridine pulse labelling. After lesion, granule neurons in the lizard medial cortex cell layer appeared pyknotic and died; they were then removed and progressively replaced by a set of new neurons. These neurons were mostly generated from …
Imaging synaptic zinc release in living nervous tissue
Zinc enriched neurons have a pool of synaptic vesicles which contain free or loosely-bound zinc ions. The movement of the vesicular zinc ions into the synaptic clefts has been previously studied by microdialysis, fluorescence postmortem staining for zinc and radioactive zinc isotope. In this study the zinc fluorescence probe N-6-metoxy-p-toluensulfonamide quinoline (TSQ) has been applied as a tracer of synaptic release of zinc ions. This fluorochrome permeates cell membranes and when exposed to living brain slices gives rise to a staining pattern similar to that seen with autometallography. In the living brain slices, fluorescence emission persists after exposure to calcium saturated ethyle…
Zinc accumulation in the telencephalon of lizards.
The zinc concentration in the brains of two species of lizard was determined by atomic-absorption spectrophotometry. The zinc concentration was found to be highest in the telencephalon of Lacerta galloti (21.1 micrograms/g fresh weight) and Podarcis hispanica (16.77 +/- 0.8 micrograms/g) while the mesencephalon and brain stem exhibited lower zinc concentrations, i.e., 7.0 micrograms/g in Lacerta galloti and 6.08 +/- 0.4 micrograms/g in Podarcis hispanica. This high telencephalic concentration of zinc is paralleled by intense and well-defined Timm reactivity used for demonstrating the presence of zinc-containing boutons at the light-microscope level. Volumetric-densitometric studies of these…