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Microscopic Immunocytochemistry (microscopic + immunocytochemistry)
Selected AbstractsSynapse-specific localization of vesicular glutamate transporters in the rat olfactory bulbEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 5 2007Marie-Madeleine Gabellec Abstract Vesicular glutamate transporters (VGLUTs) mediate the packaging of the excitatory neurotransmitter glutamate into synaptic vesicles. Three VGLUT subtypes have so far been identified, with distinct expression patterns in the adult brain. Here, we investigated the spatial distribution of the three VGLUTs in the rat olfactory bulb, a brain region containing a variety of glutamate synapses, both axodendritic and dendrodendritic. Using multilabelling confocal microscopy and electron microscopic immunocytochemistry, we showed that each VGLUT isoform has a highly selective localization in olfactory bulb synapses. VGLUT1 is present at dendrodendritic synapses established by the output neurones (mitral and tufted cells) with bulbar interneurones in the glomerular layer and external plexiform layer, as well as in axonal synapses of the granule cell layer. By contrast, VGLUT2 is strongly expressed in axon terminals of olfactory sensory neurones, which establish synapses with second-order neurones in the glomerular neuropil. VGLUT2 is also found in the outer part of the external plexiform layer and in the granule cell layer but colocalizes only partially with VGLUT1. Finally, we showed that VGLUT3 is exclusively located in the glomerular neuropil, where it colocalizes extensively with the vesicular inhibitory amino acid transporter vesicular GABA transporter, suggesting that it is associated with a subset of inhibitory synapses. Together, these observations extend previous findings on VGLUT distribution in the forebrain, and suggest that each VGLUT subtype has a specific function in the distinct features of axodendritic and dendrodendritic synapses that characterize the olfactory bulb circuit. [source] Cell type-dependent expression of HCN1 in the main olfactory bulbEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 2 2003Noémi B. Holderith Abstract In many brain regions, hyperpolarization-activated cationic currents (Ih) are involved in the generation of rhythmic activities, but the role of Ih in olfactory oscillations remains unclear. Knowledge of the cellular and subcellular distributions of hyperpolarization-activated and cyclic nucleotide-gated channel (HCN) subunits is necessary for understanding the role of Ih in olfactory network activities. Using light microscopic immunocytochemistry, we demonstrate strong HCN1 labelling of the glomerular layer and moderate staining of granule cell, internal and external plexiform layers of the rat main olfactory bulb. In the glomerular layer, among many unlabelled neurons, two distinct subpopulations of juxtaglomerular cells are labelled. Approximately 10% of the juxtaglomerular cells strongly express HCN1. These small diameter cells are immunoreactive for GABA and comprise a subpopulation of periglomerular cells. An additional subset of juxtaglomerular cells (, 1%) expresses low levels of HCN1. They are large in diameter, GABA immunonegative but immunopositive for vesicular glutamate transporter 2, characterizing them as external tufted cells. Quantitative immunogold localization revealed that the somatic plasma membranes of periglomerular cells contain approximately four times more HCN1 labelling than those of external tufted cells. Unlike in cortical pyramidal cells, immunogold density for HCN1 does not significantly differ in somatic and dendritic plasma membranes of external tufted cells, indicating that post-synaptic potentials arriving at proximal and distal dendrites are modulated by the same density of Ih. Our results demonstrate a cell type-dependent expression of HCN1 in the olfactory bulb and predict a differential contribution of distinct juxtaglomerular cell types to network oscillations. [source] The chondroitin sulphate proteoglycan brevican is upregulated by astrocytes after entorhinal cortex lesions in adult ratsEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 7 2000Niklas Thon Abstract The chondroitin sulphate proteoglycan brevican is one of the most abundant extracellular matrix molecules in the adult rat brain. It is primarily synthesized by astrocytes and is believed to influence astroglial motility during development and under certain pathological conditions. In order to study a potential role of brevican in the glial reaction after brain injury, its expression was analysed following entorhinal cortex lesion in rats (12 h, 1, 2, 4, 10, 14 and 28 days and 6 months post lesion). In situ hybridization and immunohistochemistry were employed to study brevican mRNA and protein, respectively, in the denervated outer molecular layer of the fascia dentata and at the lesion site. In both regions brevican mRNA was upregulated between 1 and 4 days post lesion. The combination of in situ hybridization with immunohistochemistry for glial fibrillary acidic protein demonstrated that many brevican mRNA-expressing cells are astrocytes. In the denervated zone of the fascia dentata, immunostaining for brevican was increased by 4 days, reached a maximum by 4 weeks and remained detectable up to 6 months post lesion. Electron microscopic immunocytochemistry showed that brevican is a component of the extracellular matrix compartment. At the lesion site a similar time course of brevican upregulation was observed. These data demonstrate that brevican is upregulated in areas of brain damage as well as in areas denervated by a lesion. They suggest a role of brevican in reactive gliosis and are compatible with the hypothesis that brevican is involved in the synaptic reorganization of denervated brain areas. [source] Distribution of P2X3 -immunoreactive fibers in hairy and glabrous skin of the ratTHE JOURNAL OF COMPARATIVE NEUROLOGY, Issue 6 2009Anna M.W. Taylor Abstract The skin is innervated by two populations of unmyelinated sensory fibers, the peptidergic and nonpeptidergic, which transmit nociceptive information to the central nervous system. The peptidergic population expresses neuropeptides such as substance P (SP) and calcitonin gene-related peptide (CGRP) and has both cutaneous and visceral targets. The nonpeptidergic population expresses the purinergic receptor P2X3, binds the isolectin B4 (IB4), and innervates mainly the epidermis. To date, the peptidergic nociceptor population in cutaneous tissue of the rat has been well characterized, whereas the nonpeptidergic innervation pattern has lacked an adequate description. To this aim, we used light microscopic immunocytochemistry to investigate the pattern of P2X3 -immunoreactive (-IR) fiber innervation of both hairy and glabrous skin from male Sprague-Dawley rats. Our results show extensive P2X3 -IR fibers throughout the upper and lower dermis. Thick bundles of P2X3 -IR fibers were found to run in parallel with the dermal-epidermal junction and projected multiple thin collateral axons that penetrated the epidermal layer, creating a dense network of innervation throughout the entire epidermis. The distribution of P2X3 -IR fibers in the epidermis was far more extensive than the distribution of CGRP-IR fibers. P2X3 -IR fibers also innervate hair follicles but were rarely found in close proximity to glands and blood vessels. The present results suggest a primary role for P2X3 -IR fibers in the detection of noxious stimuli in cutaneous tissue and provide an anatomical basis for future studies examining a possible functionally distinct role of nonpeptidergic nociceptors in the transmission of nociceptive signals. J. Comp. Neurol. 514:555,566, 2009. © 2009 Wiley-Liss, Inc. [source] | ||||||||||