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J Rodrigo

Publications and source records attributed to J Rodrigo.

At least 73 records · Page 4Linked to original sources

Immunohistochemical distribution and electron microscopic subcellular localization of the proteasome in the rat CNS.

The proteasome multicatalytic proteinase (MCP) is a 20S complex that plays a major role in nonlysosomal pathways of intracellular protein degradation. A polyclonal antibody against rat liver MCP was used to investigate the distribution of MCP in the CNS of the rat and its subcellular localization within the neurons. As expected, MCP immunoreactivity (MCP-IR) was distributed ubiquitously in the rat CNS but not homogeneously. The most intensely stained neurons were the pyramidal cortical neurons of layer 5 and the motor neurons of the ventral horn in the spinal cord, which show an intense nuclear and cytoplasmatic MCP-IR and clearly stained processes. Additionally, some populations of large neurons in the mesencephalon and brainstem also displayed a moderate MCP-IR in their perikarya. The vast majority of neurons in the remaining structures did not show a strong cytoplasmatic MCP-IR, but their nuclei displayed an intense MCP-IR. The subcellular localization also was studied by immunoelectron microscopy. MCP-IR was intense in the neuronal nuclei, and significant staining also was found in the cytoplasm, dendritic, and axonic processes (including some myelinated axons) and in synaptic boutons, as illustrated in the cerebellar cortex. The distribution of MCP in the rat CNS and its subcellular localization are discussed in relation to (1) the distribution of calpain, the other major nonlysosomal cellular protease, and (2) the possible role of MCP in the degradation of regulatory proteins and key transcription factors that are essential in many neuronal responses.

Animals↗

A combined flap technique for earlobe reconstruction in one stage.

Many of the techniques available for earlobe reconstruction require that the rest of the ear is in continuity with retroauricular skin and they usually need two stages; in some cases a graft is also required. We present a method to create an earlobe in one stage and without grafts, illustrated by one case report. The technique is a combination of the flap techniques of Davis and Zenteno Alanis, using an extended retroauricular flap together with an anterior ear flap.

Adult↗

Nitric oxide-mediated beta 2-adrenoceptor relaxation is impaired in mesenteric veins from portal-hypertensive rats.

BACKGROUND & AIMS: beta-Adrenergic relaxation seems to be mediated by nitric oxide. The aim of this study was to evaluate changes induced by portal hypertension in beta 2-adrenergic vasorelaxation. METHODS: Isolated rat mesenteric veins were relaxed by salbutamol, and nerve-mediated vasocontractions were induced by electrical field stimulation. Responses were evaluated in the presence of NG-nitro-L-arginine methyl ester (L-NAME) or tetrodotoxin. Immunocytochemical techniques were used for localization of neuronal NO synthase. RESULTS: Salbutamol-induced relaxations were decreased in rings from portal-hypertensive animals. L-NAME reduced these relaxations, but its effects were more pronounced in sham-operated rats. Tetrodotoxin decreased the effect of salbutamol only in rings from sham-operated animals. Combination of L-NAME and tetrodotoxin did not exert a greater effect than either of these agents alone. Veins from portal-hypertensive animals were more sensitive to S-nitroso-N-acetyl penicillamine. L-NAME increased vasocontractions by electrical stimulation only in rings from sham-operated rats. Veins from portal-hypertensive animals exhibited a specific degeneration of NO-containing nerve endings. CONCLUSIONS: beta 2-Adrenergic relaxation is impaired in mesenteric veins from portal-hypertensive rats, possibly as a result of a defective neuronal release of NO.

Adrenergic beta-Agonists↗

Distribution of glyoxylate dehydrogenase activity in cortical and subcortical regions of the rat brain. A light microscopic histoenzymological study.

The distribution of the glyoxylate dehydrogenase (GLYO-DH) (Glyoxylate oxidoreductase) activity has been investigated in rat brain using the corresponding histochemical method. The results have demonstrated a positive histochemical reaction in neurons of several cortical and subcortical areas. These neurons are located in the same regions in which operate monoaminergic neurotransmitters. These facts suggest a possible interaction between glyoxylic acid and monoaminergic neurotransmission.

Aldehyde Oxidoreductases↗

Experimental model for local administration of nerve growth factor in microsurgical nerve reconnections.

An experimental model for local administration of neuronotrophic substances at the site of peripheral nerve lesion is presented. The model consists of a subcutaneously located silicone reservoir and a connecting tube with its distal end fixed in the proximity of the severed and repaired nerve. The results of the preliminary tests of the model are presented. Sixty Sprague-Dawley rats were divided into two groups: control (saline-treated) (n = 30) and NGF-treated (n = 30). After axotomy of the sciatic nerve, an epineurial repair is performed. NGF or saline is injected daily into the subcutaneous reservoir during the first 3 weeks after axotomy and a single dose in the fourth week. The regenerated nerve observed in the NGF-treated group after four weeks of treatment presents a greater percentage of myelinated axons, thicker myeline sheaths, and more mature endoneurial layers. This experimental model provides a reliable and quantitative way to deliver neuronotrophic substances in site and at different administration rates.

Animals↗

Chemical heterogeneity in adult rat cerebellar Purkinje cells as revealed by zebrin I and low-affinity nerve growth factor receptor immunocytochemical expression following injury.

Cerebellar Purkinje cells in rat express low-affinity nerve growth factor receptor during development, but rarely in normal adult animals. However, after either mechanical injury or colchicine treatment during adulthood, these cells re-express low-affinity nerve growth factor receptor-immunoreactive protein. Two Purkinje cell subpopulations were defined in normal adult cerebellum by the presence or the absence of zebrin I antigen. Nevertheless, it remains an open question as to whether low-affinity nerve growth factor receptor-immunoreactive protein can be expressed by all damaged Purkinje cells, independent of their location and their staining with antibodies against intrinsic molecular markers that reveal Purkinje cell heterogeneity, such as zebrin I. In this study, a serial-section immunocytochemical mapping of the expression zebrin I and low-affinity nerve growth factor receptor, using specific monoclonal antibodies, we carried out in colchicine-treated rats. After mechanical damage of the cerebellar cortex, co-localization of these antigens at the cellular level was also analysed in thin adjacent sections, and by using a combined immunocytochemical staining method in individual sections. The findings revealed the existence of three sub-sets of Purkinje cells: (1) two complementary groups distinctly immunoreactive to one antibody, but not to the other and (2) a third group that contained double-labelled cells. In contrast, co-expression of both antigens was never observed following mechanical lesions. The seemingly independent response to mechanical injury of Purkinje cells located in different zebrin-defined compartments, indicates that particular subpopulations of Purkinje cells may respond differentially to traumatic injury.

Animals↗

Localization of nitric oxide synthase in the adult rat brain.

The distribution of the immunoreactivity to nitric oxide synthase has been examined from rostral to caudal areas of the rat central nervous system using light microscopy. Endogenous nitric oxide synthase was located using a specific polyclonal antiserum, produced against affinity purified nitric oxide synthase from whole rat brain, following the avidin-biotin peroxidase procedure. Immunoreactive cell bodies and processes showed a widespread distribution in the brain. In the telencephalon, immunoreactive structures were distributed in all areas of the cerebral cortex, the ventral endopiriform nucleus and claustrum, the main and accessory olfactory bulb, the anterior and posterior olfactory nuclei, the precommisural hippocampus, the taenia tecta, the nucleus accumbens, the stria terminalis, the caudate putamen, the olfactory tubercle and islands of Calleja, septum, globus pallidus and substantia innominata, hippocampus and amygdala. In the diencephalon, the immunoreactivity was largely found in both the hypothalamus and thalamus. In the hypothalamus, immunoreactive cell bodies were characteristically located in the perivascular-neurosecretory systems and mamillary bodies. In addition, immunoreactive nerve fibres were detected in the median eminence of the infundibular stem. The mesencephalon showed nitric oxide synthase immunoreactivity in the ventral tegmental area, the interpeduncular nucleus, the rostral linear nucleus of the raphe and the dorsal raphe nucleus. Immunoreactive structures were also found in the nuclei of the central grey, the peripeduncular nucleus and substantia nigra pars lateralis, the geniculate nucleus and in the superior and inferior colliculi. The pons displayed immunoreactive structures principally in the pedunculopontine and laterodorsal tegmental nuclei, the ventral tegmental nucleus, the reticulotegmental pontine nucleus, the parabrachial nucleus and locus coeruleus. In the medulla oblongata, immunoreactive neurons and processes were detected in the principal sensory trigeminal nucleus, the trapezoid body, the raphe magnus, the pontine reticular nuclei, the supragenual nucleus, the prepositus hypoglossal nucleus, the medial and spinal vestibular nuclei, the dorsal cochlear nucleus, the medullary reticular field, the nucleus of the solitary tract, the gracile and cuneate nuclei, the dorsal nucleus of the vagus nerve and the oral, interpolar and caudal parts of the spinal trigeminal nucleus. In the cerebellum, the stellate and basket cells showed immunoreactivity, which was also seen in the basket terminal fibres of the Purkinje cell layer. Isolated immunoreactive Purkinje cells were found in the vermis and parafloccular regions of the cerebellum. In the granular layer of the cerebellum, the granular cells and glomeruli were also immunoreactive. Numerous positive varicose nerve fibres and occasional neurons were also found in the lateral and interposed cerebellar nuclei.(ABSTRACT TRUNCATED AT 400 WORDS)

Amino Acid Oxidoreductases↗

Subcellular localization of the inositol 1,4,5-trisphosphate receptor, P400, in the vestibular complex and dorsal cochlear nucleus of the rat.

The subcellular localization of the inositol 1,4,5-trisphosphate receptor protein, P400, was studied in the vestibular complex, an area to which Purkinje cells project, as well as in neurons of the dorsal cochlear nucleus and in ectopic Purkinje cells of adult rat brain. The receptor was demonstrated by electron microscopical immunocytochemistry using the avidin-biotin peroxidase complex procedure, with the monoclonal antibody 4C11 raised against mouse cerebellar inositol 1,4,5-trisphosphate receptor protein. Immunoreactivity was found in preterminal fibres and terminal boutons in the nuclei of the vestibular complex, generally associated with the subsurface systems and stacks or fragments of smooth endoplasmic reticulum. Ectopic Purkinje cells and cartwheel cells of the dorsal cochlear nucleus also displayed immunoreactivity, but this was much less intense in the latter. The results of the present study suggest that this receptor protein, involved in the release of Ca2+, is located in sites that enable it to influence the synthesis, transport and release of neurotransmitters.

Animals↗

C-PON immunoreactive neurons in the neostriatum of the hedgehog (Erinaceus europaeus): a correlated light- and electron-microscopic study.

The present study provides light- and electron-microscopic immunocytochemical data on the presence of neurons that are immunoreactive to the C-terminal flanking peptide of neuropeptide Y, C-PON, in the neostriatum of the hedgehog (Erinaceus europaeus). Positive neurons have mostly fusiform or round perikarya from which two to four poorly branched processes arise. Immunostained fibers and puncta are also evenly distributed throughout the neostriatum. Ultrastructurally, each neuron exhibits a deeply invaginated nucleus surrounded by abundant cytoplasm with a well-developed rough endoplasmic reticulum and Golgi apparatus. Positive neurons receive symmetric and asymmetric synapses from unlabeled terminals. The results of this study can be correlated with previous findings, as the C-PON-positive neurons of the hedgehog resemble medium-sized neostriatal neurons that are known to be local circuit neurons exhibiting C-PON in the rat. Thus, a high degree of C-PON neuronal system phylogenetic conservation and function can be postulated for the neostriatum of mammals.

Animals↗

Presence of calcitonin gene-related peptide in intraepithelial nerve fibers and motor end-plates of the cat esophagus: a light and electron microscopic study.

The morphology and distribution of the motor end-plates in the striated muscle and the terminal nerve fibers in the epithelium of the wall of the esophagus, which contain calcitonin gene-related peptide, were studied by light and electron microscopic immunocytochemistry, Varicose immunoreactive nerve fibers arising from the subepithelial plexus were seen to penetrate into the epithelium where they ended in terminal boutons. These nerve fibers lost their Schwann cells just at the point of penetration into the epithelium. Characteristically, the epithelial cells of the spinous layer showed prominent tonofilaments in the part of the cytoplasm in contact with the immunoreactive nerve varicosities, but membrane specializations between these structures were not observed. In the striated muscle of the esophageal wall there were small, elliptical, immunoreactive motor end-plates, which contained a small number of axonal clear vesicles and mitochondria. They were associated with relatively short and rarely branched junctional folds, reduced postjunctional surfaces and few organelles in the underlying sarcoplasm, features characteristic of the neuromuscular junctions of slow-fatiguing red muscle fibers. The two types of immunoreactive nerve endings, epithelial and muscular, presumably participate in afferent and efferent limbs respectively of the neural control of esophageal motility. The relationship between immunoreactive nerve terminals and epithelial cells in the spinous layer exhibiting prominent tonofilaments allowed us to speculate about the existence of two different patterns of reception to sensory stimuli. The intraepithelial fibers that end in the middle layer of the epithelium could be related to mechanoreceptor reflexes, while those that end in the upper layer may be related to thermoreceptor reflexes or facilitate information about the chemical and other characteristics of foods.

Animals↗

Insulin-like growth factor I-immunoreactive peptide in adult human cerebellar Purkinje cells: co-localization with low-affinity nerve growth factor receptor.

It has been proposed that Insulin-like growth factor I is involved in the development, growth and maintenance of the central nervous system possibly interacting with other trophic factors. High levels of insulin-like growth factor I have been detected in the cerebellum during development and adulthood suggesting a specific role for insulin-like growth factor I in this brain area. While there is ever increasing data regarding the cell types containing endogenous insulin-like growth factor I in the rat brain, no information on the human brain is yet available. In the present study we sought to analyse the precise location of insulin-like growth factor I peptide in the adult human cerebellum using a specific antiserum against recombinant human insulin-like growth factor I. After immunocytochemistry, numerous Purkinje cells exhibited intense positive staining occupying the cell soma, dendrites and dendritic spines as well as axons. Occasionally, immunoreactive Purkinje cell axons were arciform and exhibited bulbous dilatations along their proximal length. Putative recurrent collaterals of Purkinje cell axons were also insulin-like growth factor I reactive. Double-staining immunocytochemistry in the same sections consistently showed, as expected, co-expression of insulin-like growth factor I and calbindin, although a few calbindin containing Purkinje cells lacked insulin-like growth factor I immunostaining suggesting there are insulin-like growth factor I positive Purkinje cell subsets in the human cerebellum. In addition, co-expression of insulin-like growth factor I and low-affinity nerve growth factor receptor-immunoreactive protein was found in a subpopulation of insulin-like growth factor I positive Purkinje cells. The results of this study prove the presence of insulin-like growth factor I immunoreactivity in a Purkinje cell subpopulation of the adult human cerebellum suggesting that insulin-like growth factor I may participate in paracrine or autocrine regulatory systems in the adult human brain.

Adult↗

Distribution of the inositol 1,4,5-trisphosphate receptor, P400, in adult rat brain.

The distribution of the inositol 1,4,5-trisphosphate receptor protein, P400, was investigated in adult rat brain by immunocytochemistry with the monoclonal antibody 4C11 raised against mouse cerebellar inositol 1,4,5-trisphosphate receptor protein. Immunoreactive neuronal cell bodies were detected in the cerebral cortex, the claustrum, the endopiriform nucleus, the corpus callosum, the anterior olfactory nuclei, the olfactory tubercle, the nucleus accumbens, the lateral septum, the bed nucleus of the stria terminalis, the hippocampal formation, the dentate gyrus, the caudate-putamen, the fundus striatum, the amygdaloid complex, the thalamus, the caudolateral part of the hypothalamus, the supramammillary nuclei, the substantia nigra, the pedunculopontine tegmental nucleus, the ventrotegmental area, the Purkinje cells in the cerebellum, the dorsal cochlear nucleus, the subnucleus oralis and caudalis of trigeminal nerve, and the dorsal horn of the spinal cord. Immunoreactive fibres were found in the medial forebrain bundle, the globus pallidus, the stria terminalis, the pyramidal tract, the spinal tract of trigeminal nerve, and the ventral horn of spinal cord. Nerve fibres forming a dense plexus ending in terminal-like boutons were detected in relation to nonimmunoreactive neurons of the dentate, interpositus, and fastigial nuclei of the cerebellum and around neurons of the vestibular nuclei. This receptor protein binds a specific second messenger, inositol 1,4,5-trisphosphate, which produces a mobilization of intracellular Ca2+ and a modulation of transmitter release.

Animals↗

Presence of C-flanking peptide of neuropeptide Y(C-PON)-immunoreactive neurons in the olfactory cortex of the hedgehog (Erinaceus europaeus).

Although the presence of the neuropeptide C-terminal flanking peptide of neuropeptide Y, C-PON, has been described in the central nervous system (CNS) of mammals, to date there is no information related with its involvement in brain functions. An analysis of the location of C-PON in specific neuronal circuits of known anatomy and physiological action should provide light on its physiological role. The presence, distribution and morphology of C-PON-containing neurons in the olfactory cortex of the hedgehog was studied by immunocytochemistry. Immunoreactive neurons to C-PON were widely distributed in the three layers of the olfactory cortex of this primitive mammal. These neurons were medium sized and showed two or three immunostained, poorly branched, dendrites. In some positive neurons, a fine, beaded axon-like process was also immunostained. Although direct evidence of a physiological function of C-PON in the olfactory cortex of the hedgehog cannot be accurately stated from our findings, the morphology of C-PON neurons and their distribution in the deep cortical layers, where the majority of pyramidal neurons are located, suggest that this neuropeptide may play a role in the intrinsic neuronal circuitry of the relatively well-developed hedgehog paleocortex. A regulatory vascular role of some peptide-immunoreactive neurons can be inferred since occasional C-PON-positive neurons have been located near blood vessels.

Animals↗

Immunohistochemical localization of the inositol 1,4,5-trisphosphate receptor in the human nervous system.

A monoclonal antibody raised against the mouse cerebellar inositol trisphosphate receptor was used to study the immunohistochemical localization of this protein in the human central nervous system. As in the brain of rodents, strong immunoreactivity was found in dendrites, axon and cell bodies of Purkinje cells, as well as in nerve endings in the cerebellar and vestibular nuclei. Cerebellar efferent fibres were the only positive structures demonstrated in the brainstem and no immunostaining could be detected in the spinal cord or dorsal root ganglia. By contrast, numerous immunoreactive neurons were present in several telencephalic and diencephalic structures, including the brain cortex, hippocampus, basal ganglia, basal forebrain, amygdala and thalamus. Immunostaining of these brain neurons was weaker than that found in Purkinje cells and was evident in cell bodies and dendrites. Thus, the human brain contains a molecule cross-reacting with the mouse inositol trisphosphate receptor protein that is expressed in a pattern similar to that found in rodents. These findings can be of great importance for understanding the function of this protein in normal brain and its modifications in neuropathological disorders.

Aged↗

Distribution of insulin-like growth factor-I receptor mRNA in rat brain. Regulation in the hypothalamo-neurohypophysial system.

The distribution and regulation of mRNA for the IGF-I receptor (IGF-I-R) in the adult rat brain were studied by in-situ hybridization with a 35S-labelled cRNA probe. The pituitary gland showed a strong hybridization signal in the pars tuberalis (the surface of the median eminence), pars distalis and pars intermedia. Within the brain, a strong hybridization signal was found in the circumventricular organs, olfactory bulb, hippocampus, cerebellum and hypothalamus. IGF-I-R mRNA was consistently found in cell bodies of the hypothalamo-neurohypophysial system. Six days of intermittent salt-loading resulted in an increase in IGF-I-R gene expression in the supraoptic nucleus. The increase in IGF-I-R mRNA was accompanied by a high expression of c-Fos immunoreactivity in the same cells. The presence and regulation of IGF-I-R mRNA in the hypothalamus suggest that IGF-I may affect the local plasticity or modulation of activated magnocellular neurones by an autocrine or paracrine action through specific receptors in the hypothalamo-neurohypophysial system.

Animals↗

Subcellular localization of insulin-like growth factor I (IGF-I) in Purkinje cells of the adult rat: an immunocytochemical study.

Insulin-like growth factor I (IFG-I) was localized by light and electron microscopic immunocytochemistry in the adult rat cerebellum. Neurons showing the same cytological characteristics and pattern of synaptic input as Purkinje cells were IGF-I immunoreactive. In these neurons, intense electron-dense reaction product was consistently located in the rough endoplasmic reticulum and, occasionally, in multivesicular bodies and associated with the nuclear envelope. Thus, the results of this study suggest synthesis IGF-I in Purkinje cells and furthermore provide anatomical support for IGF-I participation in paracrine or autocrine regulatory systems in the brain.

Animals↗

Immunocytochemical localization of insulin-like growth factor I in the hypothalamo-hypophyseal system of the adult rat.

Insulin-like growth factor I (IGF-I) is shown to be involved in the regulation of pituitary hormones. High IGF-I concentrations were detected in hypothalamus and pituitary during adulthood. This study was undertaken to analyze the cellular distribution of IGF-I in the hypothalamo-hypophyseal system of the adult rat using immunocytochemical procedures. IGF-I was found to be widely distributed throughout the hypothalamus; it was present in the magnocellular neurons of the supraoptic, paraventricular, and accessory nuclei. Moreover, nerve fibres and puncta containing immunoreactive IGF-I were localized in the median eminence and the posterior lobe of the pituitary. These results support possible IGF-I neuromodulatory or neurohormonal action in the hypothalamus on pituitary hormone regulation.

Animals↗