Biomedical subjects

I Colin

Publications and source records attributed to I Colin.

13 recordsLinked to original sources

PubMed2002

Neurokinin release in the rat nucleus of the solitary tract via NMDA and AMPA receptors.

Neurokinins (substance P, neurokinin A and neurokinin B) and the neurokinin receptors, the NK1 and NK3 receptors, are largely expressed in the nucleus of the solitary tract (NST) where they are involved in the central regulation of visceral function. Studying the mechanisms that control neurokinin release can provide valuable information concerning the control of autonomic functions subserved by the NST. Glutamate is the principal excitatory neurotransmitter in the NST and the main neurotransmitter of afferent vagal fibers. Neurokinins and glutamate may interact within the NST. In the present study, we have examined the contribution of the N-methyl-D-aspartate (NMDA) and alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) subtypes of glutamate receptors on the release of the endogenous neurokinins in the NST. We used internalization of the NK1 or NK3 receptor as an index of endogenous neurokinin release assessed by immunocytochemical visualization of the NK1 or NK3 receptor endocytosis. Experiments were performed in vitro using rat brainstem slices. A first series of experiments were done in order to validate our in vitro preparation. Application of substance P, neurokinin A or neurokinin B induced dose-dependent internalization of NK1 and NK3 receptor. This was blocked by the endocytosis inhibitor, phenylarzine oxide. The NK1 receptor antagonist SR140333 blocked internalization of NK1 receptor induced by the three neurokinins. In addition, the internalization NK1 or NK3 receptor was reversible. These results demonstrate that internalization and recycling mechanisms of NK1 or NK3 receptor were preserved in in vitro brainstem slices. Application of NMDA or AMPA induced internalization of NK1 receptor. This was blocked by the application of SR140333 suggesting that NK1 receptor internalization is due to the binding of endogenous neurokinin released under the effects of NMDA and AMPA. Application of NMDA or AMPA had no effect on NK3 receptor. Application of tetrodotoxin blocked NK1 receptor internalization induced by NMDA, demonstrating that the release of neurokinins is dependent of axon potential propagation. This result excludes the hypothesis of a release on neurokinins via pre-synaptic NMDA receptors located on neurokinin-containing axon terminals. NMDA or AMPA may directly induce neurokinin release in the NST by acting on receptors located on the cell bodies and dendrites of neurokinin-containing neurons. Release of neurokinins may also be the result of a general activation of neuron networks of the NST by NMDA or AMPA. To conclude, our results suggest that glutamate, through activation of post-synaptic NMDA and AMPA receptors, contributes to neurokinin signaling in the NST.

Animals↗

PubMed2000

Ontogenesis of CYP2C-dependent arachidonic acid metabolism in the human liver: relationship with sudden infant death syndrome.

A modification of the human monooxygenase system have been previously associated with the sudden infant death syndrome (SIDS): the hepatic CYP2C content was markedly enhanced and resulted from an activation of CYP2C gene transcription. To determine the possible consequence of the up-regulation of CYP2C in SIDS, we examined the metabolism of arachidonic acid (AA) an endogenous substrate of CYP2C involved in the physiologic regulation of vascular tone. The overall AA metabolism was extremely low during the fetal period and rose after birth to generate 14,15 epoxyeicosatrienoic acid (EET), 11,12 EET and the sum of 5,6 dihydroxyeicosatrienoic acid (diHETE)+omega/omega-1 hydroxy AA. In SIDS, the accumulation of CYP2C proteins was associated with a significant increase in the formation of 14,15 and 11,12 diHETE, which were shown to be supported by individually expressed CYP2C8 and 2C9 and HETE1 (presumably 15 HETE). This increase was markedly inhibited by addition of sulfaphenazole, a selective inhibitor of CYP2C9. So, we propose that the higher CYP2C content in SIDS stimulates the production of EETs and diHETEs and might have severe pathologic consequences in children.

8,11,14-Eicosatrienoic Acid↗

PubMed1998

Localization of components of glycinergic synapses during rat spinal cord development.

The sequence of events leading to the chemical matching of presynaptic neurotransmitters and postsynaptic transmitter receptors is investigated here in vivo for the spinal glycine receptor (GlyR) by using immunocytochemical methods. In the ventral horn of adult rat spinal cord, GlyRs are only present at glycinergic postsynaptic differentiations where they are stabilized by the associated protein gephyrin. With quantitative confocal microscopy, we found that gephyrin is detected before GlyRs at embryonic day (E)13-E14 and at E15, respectively, inside the cytoplasm and at plasmalemmal loci. Around the time of birth, the number of cell surface gephyrin-immunoreactive (-IR) spots exceeds that of GlyR. They first match 10 days after birth. The densities of postsynaptic gephyrin- and GlyR-IR were quantified between birth and the adult stage with post-embedding immunogold staining. Immunostaining for gephyrin and GlyR was not detected in the extrasynaptic membrane. The density of staining in postsynaptic membrane increased progressively with development. The inhibitory amino-acid content of the presynaptic terminal boutons opposed to gephyrin-IR sites was also analyzed. In the newborn, postnatal day 10, and adult, more than 90% of these boutons were immunostained for glycine. As seen with serial sections, 38% and 51.2% of the terminals also contained gamma-aminobutyric acid (GABA) in neonate and adult, respectively. These data indicate that around the time of birth, most glycine-containing boutons, some also containing GABA, are opposed to gephyrin-IR postsynaptic densities, whereas GlyRs are not present. Our results suggest that gephyrin determines subsynaptic loci on the plasma membrane where GlyR will subsequently accumulate.

Animals↗

PubMed1998

Antigoitrogenic effect of combined supplementation with dl-alpha-tocopherol, ascorbic acid and beta-carotene and of dl-alpha-tocopherol alone in the rat.

The effects of the vitamins dl-alpha-tocopherol, ascorbic acid and beta-carotene, free radical scavengers and lipid peroxidation inhibitors, were analyzed in male Wistar rats made goitrous by feeding a low iodine diet (< 20 micrograms iodine/kg) and perchlorate (1% in drinking water) for 4, 8, 16, and 32 days. Groups of control or goitrous rats received for at least 16 days before killing a diet containing 0.6% vitamin E (as dl-alpha-tocopherol acetate), 1.2% vitamin C (ascorbic acid) and 0.48% beta-carotene, either simultaneously (vitamin cocktail) or separately. This treatment led to a 5-fold increase of vitamin E in the thyroid gland, a 24-fold increase in the liver and a 3-fold increase in the plasma. In control rats, vitamin cocktail administration increased slightly the thyroid weight with little changes in thyroid function parameters. During iodine deficiency, administration of the vitamin cocktail or vitamin E alone reduced significantly the rate of increase in thyroid weight, and DNA and protein contents, as well as the proportion of [3H]thymidine labeled thyroid follicular cells, but not that of labeled endothelial cells. Plasma tri-iodothyronine, thyroxine, TSH levels, thyroid iodine content and concentration as well as relative volumes of glandular compartments were not modified. The proportion of necrotic cells rose from 0.5% in normal animals to about 2% after 16 days of goiter development. No significant protective effect of the vitamins was observed. These results suggest that these vitamins, particularly vitamin E, modulate one of the regulatory cascades involved in the control of thyroid follicular cell growth, without interfering with the proliferation of endothelial cells.

Animals↗

PubMed1996

Gephyrin accumulates at specific plasmalemma loci during neuronal maturation in vitro.

The distribution of glycine receptor (GlyR)-associated gephyrin has been investigated in rat spinal cord neurons maintained in vitro by means of immunocytochemical techniques. Gephyrin, which is crucial for the stabilization of postsynaptic GlyR microdomains, is present in mature neurons at postsynaptic differentiations. With immunofluorescence, discontinuous patches of gephyrin were detected within the neuronal soma of spinal cord neurons on the 1st day after plating. Subsequently, gephyrin was present at membrane areas that correspond to points of contact between cells or with the culture dish. By the 5th day, gephrin was mostly associated with the MAP2-positive somatodendritic compartment. With immunoelectron microscopy, gephyrin blobs detected at the earliest stages (1-3 days after plating) were found within the cytoplasm or associated with the plasma membrane. Asymmetrically immunostained intercellular contacts were only detected after 5 days, and gephyrin was found in association with clearly differentiated postsynaptic membranes at 7 days. At later stages, we observed gephyrin immunoreactivity only at some synapses. Our results suggest that gephyrin accumulates initially at the locus of cell-to-cell contacts involved in adhesion processes. These localizations may define hot spots for later accumulation of the GlyR and possibly other receptors.

Animals↗

PubMed1996

Expression of glycine receptor alpha subunits and gephyrin in cultured spinal neurons.

The inhibitory glycine receptor is a pentameric membrane protein composed of alpha and beta subunits. In the postsynaptic membrane, the glycine receptor and the copurifying peripheral membrane protein gephyrin are clustered underneath glycine-releasing nerve terminals. Here, we describe the expression of gephyrin and the neonatal and adult glycine receptor alpha subunit isoforms alpha1 and alpha2 during in vitro differentiation of rat spinal neurons. Analysis by immunoassays and the reverse transcriptase-polymerase chain reaction showed that gephyrin and alpha subunit mRNA and protein levels exhibited a marked increase from 1 to 5 days in vitro, i.e. prior to the formation of functional synaptic contacts. Using confocal and standard immunofluorescence, we determined the number of immunoreactive cells and the cellular localization of the alpha subunits and gephyrin. At 3 days in vitro, glycine receptor immunoreactivity revealed by the monoclonal antibody mAb4a was found in < 10% of cells and was mainly localized intracellularly; in contrast, gephyrin was detected in in vitro, gephyrin was essentially localized at the neuronal surface. At this stage, the number of glycine receptor-positive cells approached that of gephyrin-containing neurons (50%), and glycine receptor antigen was found both intracellularly and at the periphery of the cells. The antibody mAb2b, which binds exclusively to the alpha1 subunit, revealed aggregates at the surface of a few neurons. At 10 days in vitro, glycine receptor and gephyrin staining was localized in clusters at the periphery of the soma and the neurites. This quantitative analysis corroborates temporal differences in the cellular distribution of gephyrin and glycine receptor alpha subunits, the former being accumulated first at the neuronal surface.

Animals↗

PubMed1994

Effects of memory load on interhemispheric relay.

Experimentation with unilateral and bilateral tachistoscopic stimulation (the Dimond paradigm) increasingly suggests that interhemispheric cooperation (bilateral advantage) occurs or increases as a function of task complexity in general and memory load in particular. However, tachistoscopic experimentation with ipsilateral and contralateral field/hand relation conditions (the Poffenberger paradigm) has failed to provide any conclusive supporting evidence. The present investigation comprised a Sternberg "high speed memory scanning" task, modified as a go/no-go task, and formatted into the Poffenberger paradigm. Sets of items to be scanned (memory load) varied in size from one to four. A highly significant effect of load and a significant field/hand interaction were found, but the field/hand/load interaction did not reach significance. We concluded that the interhemispheric transfer time (ITT) metric drawn from this paradigm is not reliably sensitive to increasing memory load. However, our finding of significantly longer ITT in women than in men suggests that commissural anatomy and physiology may be sexually dimorphic.

Adult↗

PubMed1992

Long-term reduction of microalbuminuria after 3 years of angiotensin-converting enzyme inhibition by perindopril in hypertensive insulin-treated diabetic patients.

We studied the long-term effects of the angiotensin-converting enzyme (ACE) inhibitor perindopril, administered for 36 months on glycemic control, creatinine clearance, and albuminuria in hypertensive insulin-treated diabetics. After 1 month treatment with placebo, 39 patients entered the study and received 4-8 mg perindopril/day. Within the first 3 months, diastolic blood pressure was normalized in 80% of the patients. From these, 23 were followed during a total of 3 years on perindopril therapy, and divided in three groups according to their initial urinary albumin excretion rate (AER): 11 had normal AER (less than 15 mg/24 hours), eight had microalbuminuria (AER 15-150 mg/24 hour), and four had AER greater than 150 mg/24 hours and had overt proteinuria. Long-term (3 years) diastolic blood pressure normalization (less than or equal to 90 mm Hg) was achieved throughout the study. Concomitant with blood pressure reduction, a long-term decrease in AER was observed in normo- and microalbuminuric patients. Macroproteinuria was unaffected by perindopril. Glycemic control and creatinine clearance remained stable during the whole study period. No major side effects were observed. We conclude that perindopril safely produces a long-term normalization of elevated blood pressure in hypertensive insulin-treated diabetics without affecting glycemic control. Blood pressure normalization is associated with long-term AER reduction in normo- and microalbuminuric patients.

Albuminuria↗

PubMed1992

Detection and identification of endothelin-1 immunoreactivity in rat and porcine thyroid follicular cells.

Endothelin-1 immunoreactivity (irET-1) was observed in rat and porcine thyroid glands. Using a radioimmunoassay for endothelin-1, the mean concentration in extracts of rat and porcine thyroid glands were 0.75 pg/mg +/- 0.03 (n = 4) and 1.5 pg/mg +/- 0.2 (n = 8) (mean +/- SE) respectively. Gel-filtration and reverse-phase HPLC showed that ir ET-1 eluted in a position identical to synthetic endothelin-1. In addition, immunohistochemical study showed that irET-1 is located within epithelial follicular cells. No immunostaining was seen in parafollicular C-cells nor in parathyroid.

Animals↗

PubMed1990

Urinary proteins and red blood cell membrane negative charges in diabetes mellitus.

The nature and origin of proteinuria in diabetes mellitus have been investigated by measuring the urinary excretion of seven specific proteins of low (beta 2-microglobin, retinol-binding protein) or high molecular weight (albumin, transferrin, hemopexin and IgG). Using the Alcian Blue binding test, we also measured negative charges on red blood cell (RBC) membrane which according to recent studies might mirror the glomerular polyanion charge. A group of 190 diabetics was examined, including 90 patients with type I diabetes, 23 type II diabetics treated with diet and/or hypoglycaemic agents and 77 longstanding type II diabetics requiring insulin therapy. With the exception of beta 2-microglobulin all proteins measured were excreted in the urine of diabetics in significantly higher amounts than in controls. The assay of transferrin proved the most sensitive (58% positive) followed by albumin (49%), IgG (34%), hemopexin (28%) and retinol-binding protein (26%). Practically the same ranking was obtained when only type I diabetics were considered. RBC membrane negative charges were diminished in diabetics and negatively correlated with the urinary excretion of albumin (r = -0.61, n = 190). RBC charges were also negatively correlated with other urinary proteins of high molecular mass (r between - 0.5 and - 0.2) but presented no relation with urinary beta 2-microglobulin or retinol-binding protein. The loss of RBC charges in diabetics most likely reflects the concomitant depletion of the glomerular polyanion responsible for the increased glomerular leakage of high molecular mass plasma proteins. The preferential increase in transferrin excretion together with the progressive rise in the urinary excretion of IgG lead us to postulate that the loss of glomerular polyanion in diabetes is accompanied, from the early stage, by a progressive decrease in the size-selectivity of the glomerular filter. The urinary excretion of retinol-binding protein was weakly correlated with albuminuria (r = 0.26, n = 186). Eight % of diabetics showed an elevation of urinary retinol-binding protein without evidence of microalbuminuria, which clearly demonstrates that a proximal tubular impairment can occur independently of the glomerular alterations in the course of diabetic nephropathy.

Adult↗

PubMed1990

Correlated morphological and functional study of isolated rat thyroid follicles in suspension culture.

Rat thyroid follicles were isolated by collagenase digestion and cultured in suspension on agarose for 1-12 days with 0-0.1-1 mU/ml thyrotropin (TSH). After a 4 h exposure to Na125I they were processed for light and electron microscopy, autoradiography and biochemical analysis. Follicular 125I accumulation (A) and organification (PBI) were measured. Thyroglobulin (Tg) content of follicles and 125I-labelled amino acids in Tg were analyzed by high-performance liquid chromatography (HPLC). Without TSH, follicular lumina and cell polarity persisted. From day 3, the rough endoplasmic reticulum (RER) and ribosomes disappeared while autophagic vacuoles appeared: 125I accumulation and PBI were significantly reduced. From day 6, ultrastructural cell dedifferentiation occurred. At day 12, autoradiographic labelling was found over very few lumina; half of the 125I accumulated was still organified. With 1 mU TSH, follicles formed aggregates with narrow densely labelled lumina lined by tall cells. The RER was well developed up to day 12. 125I accumulation, PBI and iodothyronine (T3, T4) formation in Tg remained significantly higher than in follicles cultured without TSH, showing a transient decrease at days 6 and 9. Monoiodotyrosine/diiodotyrosine (MIT/DIT) and T3/T4 ratios in Tg were not modified, suggesting the persistence in the follicles of a significant iodine pool available for iodination. With 0.1 mU TSH, alterations of cell morphology and reduction of functional properties occurred later than without TSH. In the presence of TSH, morphological signs of new follicle formation were seen. These data demonstrate that closed follicles keep their follicular structure up to 12 days of culture, even without TSH. However, TSH is necessary to maintain iodine accumulation and organification.

Amino Acids↗

PubMed1989

Are adrenergic neurons subject to a serotoninergic influence in the nucleus tractus solitarii? A morphological and biochemical study in the rat.

The possible relationships between adrenaline-synthesizing neurons and serotoninergic afferent fibers in the nucleus tractus solitarii of the rat were investigated both morphologically and biochemically. Adrenergic elements (cell bodies, dendrites and nerve endings) were detected simultaneously with serotoninergic axonal varicosities in the same electron-microscopic sections by means of combined phenylethanolamine-N-methyltransferase immunocytochemistry and [3H]serotonin-uptake radioautography. Among some 500 serotoninergic varicosities scanned in the areas of significant overlap between the 2 types of labeling, only 3 were directly apposed to an adrenergic process, identified as a dendrite in each case. No synaptic membrane differentiations were seen at these occasional sites of contact. Destruction of the serotonin input by 5,7-dihydroxytryptamine had no significant effect on the tyrosine hydroxylase dopamine-beta-hydroxylase and phenylethanolamine-N-methyltransferase enzymatic activities in the C2 adrenergic region, but induced 22% and 38% increases of tyrosine hydroxylase and dopamine-beta-hydroxylase activities, respectively, in the neighboring A2 noradrenergic area. Taken together, these results suggest that serotoninergic and adrenergic neurons do not significantly interact in the nucleus tractus solitarii; this implies that the possible catecholaminergic relays for the action of serotonin in autonomic regulation at this level could consist of noradrenergic neurons rather than of their adrenergic counterparts.

Animals↗

PubMed1986

Direct toxic effect of iodide in excess on iodine-deficient thyroid glands: epithelial necrosis and inflammation associated with lipofuscin accumulation.

Involution of thyroid hyperplasia (induced by a low iodine diet and a goitrogen, propylthiouracil, PTU) was obtained in mice by administering a high or a moderate dose of iodide (HID or MID, respectively). In HID involuting glands, vasoconstriction was observed after 12 hr whereas necrosis and inflammation were very abundant as early as after 6 hr and maximal after 48 hr. They were not prevented by papaverine by which vasoconstriction was inhibited, but were inhibited by the continuation of PTU by which iodide oxidation and organification were inhibited. Lipofuscin inclusions in thyroid and inflammatory cells were always associated with necrosis. On the contrary, when involution was induced by MID or by HID + triiodothyronine (T3), or by T3 alone, neither necrosis nor inflammation occurred and apoptosis was the only mode of cell deletion. No lipofuscin inclusion occurred. Our results demonstrate that iodide in excess, after being oxidized or organified, is directly toxic for iodine-deficient thyroid cells. The presence of lipofuscin suggests that its toxicity is mediated by lipid peroxidation, a consequence of production of free radicals in excess.

Animals↗