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1.
J Colloid Interface Sci ; 321(1): 118-29, 2008 May 01.
Artículo en Inglés | MEDLINE | ID: mdl-18272170

RESUMEN

Wicking of liquids into porous media is of great importance to many applications. One example are propellant management devices (PMD) used in spacecraft tanks. PMDs are designed to ensure gas free delivery of propellant during all acceleration conditions of the flight. This might be achieved by a metallic weave which is wetted by the propellant and thus prevents gas from entering below a critical bubble point pressure. In the case of cryogenic or volatile liquids the weave may dry out and refilling of the structure becomes an important issue. In this study we analyze the wicking of different liquids into a dry Dutch twilled weave (DTW 200 x 1400) by combining experimental and analytical approaches. Experiments were performed under isothermal and terrestrial conditions to investigate the role of evaporation for the capillary rise. The standard wicking model from Lucas and Washburn is enhanced to account for evaporation and gravity effects, too. By comparing the experimental results with the enhanced wicking model we find good qualitative agreement. It is also noted that evaporation may have a major impact on the wicking process.

2.
Neuroscience ; 138(2): 561-73, 2006.
Artículo en Inglés | MEDLINE | ID: mdl-16364554

RESUMEN

Lumbar spinothalamic neurons in the lamina X of the L3-L4 spinal cord segment have been proposed to constitute the spinal ejaculation generator in male rats. Lumbar spinothalamic cells are immunoreactive for galanin and neurokinin-1 receptors. We previously showed that after injection of pseudorabies virus either in the bulbospongiosus muscle or in the prostate, retrogradely labeled cells in the L3-L4 segment also displayed galanin or neurokinin-1 receptor immunoreactivities, demonstrating a direct link between lumbar spinothalamic cells and two anatomical structures involved in the two phases of ejaculation i.e. the emission and the expulsion phases. In order to provide with a more precise anatomical support for the role of lumbar spinothalamic cells in controlling ejaculation, we injected simultaneously in male adult rats two strains of recombinant pseudorabies virus, expressing either beta-galactosidase (PRV-BaBlu) or green fluorescent protein (PRV-152) in the prostate and in the bulbospongiosus muscle, respectively. After 5 days, we performed multiple immunofluorescence experiments to detect PRV-BaBlu, PRV-152 and galanin or neurokinin-1 receptors in transverse sections of the L1-S1 segment. Double- and triple-labeled cells were counted using confocal laser scanning microscope. Double-labeled neurons with the two strains of pseudorabies virus were mainly found at the L3-L4 segment lateral to the central canal in lamina X and represented about 60% of the total number of pseudorabies virus-labeled neurons. All the double pseudorabies virus-labeled neurons also expressed lumbar spinothalamic and most of them neurokinin-1 receptor, identifying them as lumbar spinothalamic neurons. The convergence of retrograde labeling from prostate and bulbospongiosus muscle on the same lumbar spinothalamic cells strongly reinforce their role in the spinal control and coordination of the emission and expulsion of sperm.


Asunto(s)
Músculo Liso/inervación , Neuronas/fisiología , Próstata/inervación , Médula Espinal/fisiología , Animales , Colina O-Acetiltransferasa/análisis , Eyaculación , Genes Reporteros , Proteínas Fluorescentes Verdes/análisis , Proteínas Fluorescentes Verdes/genética , Herpesvirus Suido 1/genética , Vértebras Lumbares , Masculino , Ratas , Ratas Sprague-Dawley , beta-Galactosidasa/análisis , beta-Galactosidasa/genética
3.
Phys Rev E Stat Nonlin Soft Matter Phys ; 74(2 Pt 2): 026303, 2006 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-17025535

RESUMEN

The dynamic behavior of liquid metal drops submitted to a high-frequency magnetic field is investigated experimentally. The motivation for this study comes from the coating industry. In this industry, liquid metals showing a free surface held in a dome-type shape are evaporated by applying electromagnetic pressure. The Galinstan drops are placed on a curved glass plate. A ringlike inductor fed by an alternating electrical current generates the magnetic field. The surface contour of the drop is observed using a high-speed camera system. The data are analyzed by utilizing image processing methods. In the experiment, we vary the inductor current I and the drop volume V while the frequency is fixed at 20 kHz . Upon increasing the inductor current within the range 0I(C), these symmetric states become unstable to capillary waves. The critical current (critical electromagnetic Bond number) as well as the critical mode number, the critical frequency, and the amplitudes of the waves depend strongly on the volume (Bond number).

4.
Biochim Biophys Acta ; 1541(3): 150-60, 2001 Dec 19.
Artículo en Inglés | MEDLINE | ID: mdl-11755209

RESUMEN

Some biological properties of new bifunctional conjugates designed for drug targeting were evaluated through in vitro experiments. Eight peptidylcyclodextrin compounds were used, which correspond to modified beta- or gamma-cyclodextrin (CD) grafted on neuropeptide substance P (SP) or a shorter derivative (SP(4-11)). Using anti-SP and anti-CD antibodies as molecular probes, we showed that the main structural features of the two moieties of these adducts were preserved. Binding experiments, using CHO cells expressing the human SP-specific NK1 receptor, demonstrated the functionality of all peptidylcyclodextrin derivatives, which exhibited IC50 values in a 10(-9)-10(-7) M range. All compounds were able to induce a pharmacological response, triggering phosphatidylinositol turnover with EC50 values in the same range as the natural ligand. Moreover, autoradiography analysis of rat spinal corn sections proved that [125I]SP binding was dose-dependently displaced by one selected compound (a gamma-CD-SP), showing a similar affinity of this adduct for the rat neurokinin 1 receptor. Our observations demonstrate that these peptidylcyclodextrins efficiently target NK1 receptor-expressing cells.


Asunto(s)
Ciclodextrinas/farmacología , Sistemas de Liberación de Medicamentos , Receptores de Neuroquinina-1/efectos de los fármacos , Sustancia P/análogos & derivados , beta-Ciclodextrinas , gamma-Ciclodextrinas , Animales , Anticuerpos/inmunología , Autorradiografía , Unión Competitiva , Células CHO , Cricetinae , Ciclodextrinas/química , Ciclodextrinas/inmunología , Diseño de Fármacos , Estructura Molecular , Receptores de Neuroquinina-1/biosíntesis , Receptores de Neuroquinina-1/genética , Proteínas Recombinantes/biosíntesis , Sustancia P/química , Sustancia P/inmunología
5.
Neuroscience ; 134(4): 1325-41, 2005.
Artículo en Inglés | MEDLINE | ID: mdl-16054769

RESUMEN

Ejaculation requires the coordination of sympathetic, parasympathetic and somatic neural outputs. Timely occurrence of the emission and expulsion of sperm results from an interplay between spinal nuclei innervating the seminal tract and the sexual accessory glands including the prostate on the one hand, and on the other hand perineal striated muscles, particularly the bulbospongiosus muscle. A group of cells essential for ejaculation, located around the central canal and referred to as lumbar spinothalamic neurons have been recently identified. Lumbar spinothalamic neurons are immunoreactive for galanin and neurokinin-1 receptor. In order to investigate the anatomical relationships between lumbar spinothalamic neurons and both the prostate and the bulbospongiosus muscle, pseudorabies virus retrograde tracing technique was used combined with immunohistochemistry. Three to five days after pseudorabies virus injection in the bulbospongiosus muscle or the prostate in male rats, spinal cord sections were processed for double immunofluorescence against pseudorabies virus and galanin or neurokinin-1 receptor. Immunocytochemical experiments against pseudorabies virus and choline acetyltransferase were also performed to discriminate between motoneurons and preganglionic neurons, or interneurons. Spinal sections were examined with confocal laser scanning microscope. Three days after pseudorabies virus injection within the prostate and the bulbospongiosus muscle, sympathetic preganglionic neurons and motoneurons of the dorsomedial nucleus were retrogradely labeled, respectively. Five days after pseudorabies virus injection, transsynaptically labeled choline acetyltransferase-negative neurons were found mainly located in the medial gray surrounding the central canal from L1 to S1. At the L3-L4 level, most of transsynaptically labeled neurons were immunoreactive for galanin and to a lesser extent for neurokinin-1 receptor, strongly suggesting that they could be the lumbar spinothalamic cells. We have thus evidenced connections between these cells and motoneurons of the dorsomedial nucleus and both sympathetic and parasympathetic preganglionic neurons innervating the bulbospongiosus muscle and the prostate, respectively. These anatomical data reinforce the crucial role for lumbar spinothalamic cells in coordinating the spinal control of ejaculation.


Asunto(s)
Eyaculación/fisiología , Neuronas/citología , Próstata/inervación , Receptores de Galanina/metabolismo , Receptores de Neuroquinina-1/metabolismo , Animales , Colina O-Acetiltransferasa/metabolismo , Inmunohistoquímica , Región Lumbosacra , Masculino , Microscopía Confocal , Músculo Esquelético/inervación , Vías Nerviosas/citología , Vías Nerviosas/metabolismo , Neuronas/metabolismo , Ratas , Ratas Sprague-Dawley , Médula Espinal/citología
6.
Eur J Cell Biol ; 57(2): 273-84, 1992 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-1324839

RESUMEN

Four different rat monoclonal antibodies were produced against delta opioid receptor using an antiidiotypic approach in which antibodies directed against the opioid agonist DADLE were used as immunogen. In the first step, seven hybridomas were selected on the basis of their ability to inhibit the DADLE-anti-DADLE antibody interaction. After purification from ascitic fluids, these monoclonal antibodies were characterized. Four antiidiotypic antibodies, named 5, 11, 16, and 51, directed toward different epitopes, recognized the delta opioid receptor: (i) they bound directly to the NG108-15 cells, (ii) they inhibited the [3H]DADLE binding on the NG108-15 cells, (iii) they immunoprecipitated a 52,500 dalton protein present on the surface of the NG108-15 cells. The four monoclonal antiidiotypic anti-opioid receptor antibodies were used to immunocytologically detect the opioid receptors under light and electron microscopy in the rat spinal cord. The regional distribution of the immunoreactivity corresponded to layers known to be rich delta opioid receptor subtype. Moreover, at the ultrastructural level, the labeling was located mainly on plasma membranes, especially on non-synaptic zones. Our results show that monoclonal antiidiotypic antibodies constitute a valuable tool for visualizing cell surface receptors.


Asunto(s)
Anticuerpos Antiidiotipos/inmunología , Anticuerpos Monoclonales/inmunología , Receptores Opioides/inmunología , Animales , Anticuerpos Antiidiotipos/aislamiento & purificación , Anticuerpos Monoclonales/aislamiento & purificación , Línea Celular , Células Híbridas , Técnicas para Inmunoenzimas , Inmunoglobulina G/inmunología , Masculino , Ratones , Ratones Desnudos , Microscopía Electrónica , Pruebas de Precipitina , Conejos , Ratas , Receptores Opioides delta
7.
J Comp Neurol ; 433(2): 183-92, 2001 Apr 30.
Artículo en Inglés | MEDLINE | ID: mdl-11283958

RESUMEN

Tachykinins are involved in pain transmission at the spinal level. In frog, at least four tachykinins [TK] have been isolated from the brain, but their organization in the dorsal horn of the spinal cord is still poorly known. We have reexamined TK distribution by immunocytochemistry using an antibody recognizing the sequence common to all tachykinins in the spinal cord and dorsal root ganglia of the green frog Rana esculenta. A dense tachykinin-like immunoreactivity (TK-LI) was observed in the dorsolateral fasciculus or Lissauer's tract running ventromedial to the entry of the dorsal root and in numerous small and medium-sized dorsal root ganglion cells showing a primary afferent origin for part of TK-LI of the dorsal horn. The observation of numerous cell bodies in the dorsal horn, in addition, suggested a local or propriospinal origin. One group of cells was localized at the entrance of the Lissauer's tract TK-LI fibers into the dorsal horn, and another group was localized in the upper dorsal horn, a region with a low density of TK-LI fibers. It was suggested that the latter group may correspond to neurokinin B. Electron microscopic examination of the Lissauer's tract showed numerous immunoreactive axons, some located at the center of glomerular-like arrangements, suggesting that the information brought by these fibers may be transmitted and most probably modulated before their entry in the dorsal horn. In conclusion, the functional organization of tachykinins in the frog spinal cord seems to be similar to that of mammals, albeit with a different morphological organization.


Asunto(s)
Rana esculenta/metabolismo , Médula Espinal/metabolismo , Taquicininas/metabolismo , Animales , Ganglios Espinales/citología , Ganglios Espinales/metabolismo , Ganglios Espinales/ultraestructura , Inmunohistoquímica , Masculino , Microscopía Electrónica , Neuronas/metabolismo , Neuronas/ultraestructura , Médula Espinal/citología , Médula Espinal/ultraestructura , Distribución Tisular
8.
J Neuroimmunol ; 81(1-2): 211-24, 1998 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-9521624

RESUMEN

Site-directed antibodies against synthetic related dermorphin peptides were previously produced and characterized. One of them, which specifically recognizes the crucial 'opioid message' (the N-terminal part of the dermorphin molecule (i.e. Tyr-D-Ala-Phe-Gly) was selected in order to detect and locate endogenous dermorphin-like molecules in rat, mouse and guinea pig tissues. Dermorphin-like peptides were found to be present in tissues known to contain peptides such as neurons in the central nervous system, nerve fibers in the gut and B and T immune cells. With all the tissues assayed, the HPLC profile obtained on the immunoreactive material showed the same main peak eluted at a retention time of 32 +/- 1 min. The results of biochemical experiments in which enzymatic treatments were performed on the dermorphin-like immunoreactivity indicate the immunoreactivity is a peptide resistant to aminopeptidase hydrolysis. This finding suggests the presence of a residue conferring resistance to proteolytic processes of this kind, which is likely to be a D-amino acid residue.


Asunto(s)
Aminopeptidasas/farmacología , Subgrupos Linfocitarios/química , Proteínas del Tejido Nervioso/análisis , Neuronas/química , Oligopéptidos/análisis , Animales , Química Encefálica , Cromatografía Líquida de Alta Presión , Colon/química , Colon/citología , Sistema Digestivo/química , Endopeptidasas/metabolismo , Cobayas , Técnicas para Inmunoenzimas , Masculino , Metionil Aminopeptidasas , Ratones , Ratones Endogámicos BALB C , Proteínas del Tejido Nervioso/química , Proteínas del Tejido Nervioso/metabolismo , Neuroinmunomodulación , Neuropéptidos/análisis , Oligopéptidos/química , Oligopéptidos/metabolismo , Péptidos Opioides , Especificidad de Órganos , Fragmentos de Péptidos/síntesis química , Fragmentos de Péptidos/inmunología , Nervios Periféricos/química , Hipófisis/química , Radioinmunoensayo , Ratas , Ratas Sprague-Dawley , Ratas Wistar , Especificidad de la Especie , Bazo/química
9.
J Neuroimmunol ; 76(1-2): 1-9, 1997 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-9184627

RESUMEN

Monoclonal antibodies to the binding site of the NK1 receptor for the neuropeptide substance P were produced in mice using the complementary or antisense peptide methodology. Among several anti-peptide monoclonal antibodies, we selected the mAb12 antibody which specifically crossreacted, through its paratope, with a binding site present on membranes from rat parotid gland cells, with an affinity close to 2 x 10(-7) M and with membranes from CHO cells expressing human brain NK1 receptors. Immunocytochemical investigations using mAb12 revealed immunostaining whose distribution in the dorsal horns of rat spinal cord fits well with the known location of NK1 receptors. In both biochemical and immunocytochemical experiments, the competition occurring between the antibody and substance P, or a substance P-protein conjugate, indicates that mAb12 recognizes a membrane epitope located at or near the substance P binding domain on the NK1 receptor. Immunization of mice with mAb12 led to the production of specific anti-substance P antibodies, again suggesting that mAb12 shares common structural features with the neuropeptide. This monoclonal antibody can now be used in further biochemical or cytochemical characterizations of NK1 receptors. Owing to its fine specificity, mAb12 could also serve as a molecular model for designing peptides, possibly displaying pharmacological properties in the various processes in which substance P is involved, e.g. immunomodulation, inflammation or chronic pain.


Asunto(s)
Anticuerpos Monoclonales/inmunología , Receptores de Neuroquinina-1/análisis , Sustancia P/inmunología , Animales , Sitios de Unión , Células CHO , Cricetinae , Femenino , Humanos , Inmunización , Inmunohistoquímica , Masculino , Ratones , Ratas , Ratas Wistar , Receptores de Neuroquinina-1/inmunología
10.
Neuroscience ; 110(4): 755-64, 2002.
Artículo en Inglés | MEDLINE | ID: mdl-11934482

RESUMEN

Several lines of evidence show that endogenous and exogenous cannabinoids modulate pain transmission at the spinal level through specific cannabinoid-1 (CB1) receptors. Since anatomical data concerning spinal CB1 receptors are rather contradictory, we studied the cellular and subcellular localizations of the CB1 receptors by immunocytochemistry. Results show a dual pre- and postsynaptic localization of CB1 receptors. Presynaptic receptors are evidenced by the labeling of (1) heterogeneous dorsal root ganglion neurons and (2) axons of Lissauer's tract. Postsynaptic receptors are shown by the labeling of numerous interneurons in the outer part of lamina II. Double immunolabelings show that lamina II outer CB1 neurons, probably islet cells, may also contain GABA or nitric oxide synthase. Numerous CB1-containing neurons in lamina X are also immunostained with anti-nitric oxide synthase (NOS) antibody. Under the electron microscope, CB1 immunoreactivity is exclusively localized postsynaptically in both somatic and dendritic compartments. The absence of labeling on primary afferent axon terminals is discussed and compared to the absence of labeling on terminals or vesicle-containing dendrites of islet cells, where a presynaptic localization was expected according to data of the literature.


Asunto(s)
Vías Aferentes/metabolismo , Ganglios Espinales/metabolismo , Nociceptores/metabolismo , Dolor/metabolismo , Células del Asta Posterior/metabolismo , Terminales Presinápticos/metabolismo , Receptores de Droga/metabolismo , Vías Aferentes/ultraestructura , Animales , Cannabinoides/metabolismo , Ganglios Espinales/ultraestructura , Inmunohistoquímica , Interneuronas/metabolismo , Interneuronas/ultraestructura , Masculino , Microscopía Electrónica , Óxido Nítrico/metabolismo , Nociceptores/ultraestructura , Dolor/fisiopatología , Células del Asta Posterior/ultraestructura , Terminales Presinápticos/ultraestructura , Ratas , Ratas Wistar , Receptores de Cannabinoides , Receptores de Droga/ultraestructura , Transmisión Sináptica/fisiología , Ácido gamma-Aminobutírico/metabolismo
11.
Neuroscience ; 92(2): 729-43, 1999.
Artículo en Inglés | MEDLINE | ID: mdl-10408621

RESUMEN

Rxt1, a member of the Na+/Cl- orphan transporter family, exhibits numerous features suggesting a role as plasma membrane transporter. Despite numerous attempts, its substrate has not yet been identified, although immunocytochemical studies have shown that Rxt1 distribution generally matches that of glutamate or GABA. In order to further characterize Rxt1, its detailed immunocytochemical distribution in the rat spinal cord and dorsal root ganglia was studied at both light microscope and ultrastructural levels. The widespread distribution of Rxt1 in spinal cord and ganglia cannot be correlated with any known classical or peptidergic transmitter. Rxt1 is expressed in a subpopulation of glutamatergic primary afferent fibers, in large and medium-sized ganglion cells, while small glutamate cells exhibit generally no Rxt1-like immunoreactivity. In the spinal cord, Rxt1-immunoreactive cell body distribution is quite ubiquitous since Rxt1 is expressed in all laminae in various neuronal types like interneurons, some projection neurons and motoneurons. Some of these neurons are cholinergic. At the electron microscope level, the peroxidase labeling was never localized to the plasma membrane, but rather associated with different organelles including the outer membrane of small synaptic vesicles and large granular vesicles. This localization resembles that of vesicular transporters detected with the same method and suggests that Rxt1, in contrast to other Na+/Cl- transporters, is expressed on vesicles. This was confirmed using a pre-embedding silver-intensified colloidal gold method. Indeed, most gold particles appeared to be localized into the axoplasm on synaptic vesicle accumulations; only few gold particles were observed close to the plasma membrane. These results suggest that Rxt1, despite its molecular characteristics predicting a plasma membrane localization, might be a vesicular transporter.


Asunto(s)
Proteínas Portadoras/análisis , Ganglios Espinales/química , Proteínas de Transporte de Membrana , Proteínas del Tejido Nervioso/análisis , Médula Espinal/química , Simportadores , Vesículas Sinápticas/química , Animales , Inmunohistoquímica , Masculino , Ratas , Ratas Wistar , Simportadores del Cloruro de Sodio
12.
J Histochem Cytochem ; 36(11): 1397-401, 1988 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-2844889

RESUMEN

Anti-idiotypic antibodies may serve as valuable probes for cytological identification of peptide receptors in the CNS. We have previously described the preparation of anti-substance P (SP) anti-idiotypic antibodies (anti-Id Ab) and have shown that they recognize SP receptors. These anti-Id Ab can be used in cytology to label SP receptors in CNS. We chose rat cervical spinal cord as a model because SP is present in large amounts in the dorsal and ventral horns, where it is implicated in pain and in motor function, respectively. After application of an indirect immunoperoxidase technique to tissue sections from perfused animals, immunolabeling was seen in the two superficial layers of the dorsal horn, the area surrounding the central canal, extending along the white matter in lamina VII, and in part of the ventral horn. This localization is in accordance with the classical distribution of SP receptors as seen by autoradiography with labeled SP. In the light of control experiments, as well as of biochemical and pharmacological arguments, we discuss the specificity of the immunolabeling. We conclude that anti-Id Ab recognize NK-P receptors, although crossreaction with NK-A or NK-B receptors cannot be totally ruled out.


Asunto(s)
Anticuerpos Antiidiotipos , Idiotipos de Inmunoglobulinas/inmunología , Inmunohistoquímica , Receptores de Neurotransmisores/análisis , Médula Espinal/análisis , Animales , Técnicas para Inmunoenzimas , Ratas , Receptores de Neuroquinina-1 , Receptores de Neurotransmisores/inmunología , Distribución Tisular
13.
J Chem Neuroanat ; 2(6): 335-47, 1989.
Artículo en Inglés | MEDLINE | ID: mdl-2610948

RESUMEN

Calcitonin gene-related peptide (CGRP) in the dorsal horn of the rat spinal cord was assumed until now to be principally of primary afferent origin. It is shown here, on the basis of both light and electron microscopic immunocytochemical evidence, that some cell bodies of the dorsal horn and lateral spinal nucleus (LSn) of the rat cervical spinal cord contain a CGRP-like immunoreactivity. At the light microscopic level, immunoreactive cell bodies were observed in animals pretreated with colchicine injected intraventricularly, CGRP-like cell bodies were morphologically heterogeneous and distributed in the three superficial layers of the dorsal horn. They were very rare in lamina I and more numerous in laminae II and III. A group of immunoreactive cell bodies was also observed in the LSn. Using electron microscopic techniques, a few immunoreactive cell bodies were observed even in control animals. In addition, relatively numerous immunoreactive dendrites were observed in lamina II. The specificity of the reaction and the physiological implications of the results are discussed.


Asunto(s)
Péptido Relacionado con Gen de Calcitonina/fisiología , Neuronas/ultraestructura , Médula Espinal/citología , Animales , Péptido Relacionado con Gen de Calcitonina/inmunología , Colchicina , Inmunohistoquímica , Inyecciones Intraventriculares , Masculino , Microscopía Electrónica , Neuronas/metabolismo , Ratas , Ratas Endogámicas
14.
J Chem Neuroanat ; 24(3): 153-62, 2002 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-12297261

RESUMEN

The role of cannabinoids in spinal analgesia has so far been investigated in mammals and the interactions between cannabinoid receptors and markers involved in nociception have been described in the rat spinal cord. An endocannabinoid system is well developed also in the amphibian brain. However, the anatomical substrates of pain modulation have been scarcely investigated in anamniotes, neither is there reference to such a role for cannabinoids in lower vertebrates. In the present paper we employed multiple cytochemical approaches to study the distribution of CB1 cannabinoid receptors and their morphofunctional relationships with some nociception markers (i.e. Substance P, nitric oxide synthase, GABA and mu opioid receptors) in the spinal cord of the anuran amphibian Xenopus laevis. We found a co-distribution of CB1 receptors with the aforementioned signaling molecules, as well as a more limited cellular co-localization, in the dorsal and central fields of the spinal cord. These regions correspond to the mammalian laminae I-IV and X, respectively, areas strongly involved in spinal analgesia. Comparison of these results with those previously obtained in the mammalian spinal cord, reveals a number of similarities between the two systems and suggests that cannabinoids might participate in the control of pain sensitivity also in the amphibian spinal cord.


Asunto(s)
Cannabinoides/metabolismo , Dolor/metabolismo , Receptores de Droga/análisis , Médula Espinal/química , Animales , Moduladores de Receptores de Cannabinoides , Cannabinoides/biosíntesis , Femenino , Receptores de Cannabinoides , Receptores de Droga/biosíntesis , Médula Espinal/metabolismo , Sustancia P/análisis , Sustancia P/biosíntesis , Xenopus laevis
15.
J Chem Neuroanat ; 9(1): 65-77, 1995 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-8527039

RESUMEN

A monoclonal antibody directed against a peptide (PS5) specified by RNA complementary to the mRNA coding for substance P (SP), was used to label SP receptors in the rat spinal cord as demonstrated by light and electron microscopy. An immunocytochemical method (avidin-biotin-peroxidase) was used on vibratome sections from rats perfused with paraformaldehyde. Immunoreactivity was observed principally in the two superficial layers of the dorsal horn, in lamina X and the region of motoneurons. The labeling was absent when the antibody was preincubated with the complementary peptide (PS5) used as immunogen. Competition between the anti-complementary peptide antibody and different ligands was tested by preincubation of tissue sections with the ligand in the presence of peptidase inhibitors before addition of the antibody. A specific agonist (SP) or antagonist (spantide, RP 67580) at 10(-6)M led to total absence of labeling. These results indicate that under our experimental conditions, the anti-complementary peptide antibody recognizes a SP binding site in the rat spinal cord. Electron microscopic study of the two superficial laminae of the dorsal horn showed that immunolabeling was mainly localized extracellularly at apposing neuronal plasma membranes. It was mostly associated with axodendritic or axosomatic appositions. Occasionally labeling was observed between two axon terminals. In all cases, these appositions were non-junctional. Generally, neuronal processes involved in these appositions did not contain large granular vesicles. These observations suggest that SP may act in a diffuse, nonsynaptic manner probably on targets distant from SP release sites.


Asunto(s)
Receptores de Neuroquinina-1/análisis , Médula Espinal/química , Secuencia de Aminoácidos , Animales , Anticuerpos Monoclonales , Membrana Celular/química , Membrana Celular/ultraestructura , Inmunohistoquímica , Indoles/farmacología , Isoindoles , Masculino , Ratones , Microscopía Electrónica , Datos de Secuencia Molecular , Antagonistas del Receptor de Neuroquinina-1 , ARN Complementario , Ratas , Ratas Wistar , Receptores de Neuroquinina-1/agonistas , Médula Espinal/ultraestructura , Sustancia P/análogos & derivados , Sustancia P/inmunología , Sustancia P/metabolismo , Sustancia P/farmacología
16.
J Chem Neuroanat ; 7(3): 159-70, 1994 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-7848572

RESUMEN

The ultrastructural localization of delta-opioid receptors was studied using monoclonal anti-idiotypic antibody prepared with an anti-D-Ala2-D-Leu5-enkephalin. Immunocytochemical techniques were used on vibratome sections from rats perfused with paraformaldehyde. A high density of immunoreactivity was observed in the dorsal horn of the spinal cord, particularly the two superficial layers, the dorsolateral funiculus and the area surrounding the central canal. The labelling was absent when the antibody was preincubated with the immunogen. Competition between the anti-idiotypic antibody and different ligands, delta or mu, was controlled by preincubation of tissue sections with the ligand in the presence of peptidase inhibitors for 3-4 h before addition of the anti-idiotypic antibody. Enkephalin, dermenkephalin and naltrindole induced disappearance of the labelling at 10(-9) M while dermorphin or dermorphin Lys7 were ineffective at the same concentration. Lamina II of the dorsal horn was studied by electron microscopy. The immunolabelling was mainly localized on cell membranes at appositions between the two neurons. About one third were localized between an axon terminal and a dendrite, the same proportion of labellings were between two axon terminals. Labelling was occasionally observed at appositions between a glomerular terminal and a dendrite or a terminal or at axoglial appositions. Axosomatic localizations were rare. The presynaptic localization of the labelling is in favor of a presynaptic mechanism of action for delta-opioids in the spinal cord, providing that these receptors are functional. delta-Opioid peptides probably act non-synaptically since receptors were never localized on synaptic differentiations.


Asunto(s)
Anticuerpos Antiidiotipos , Receptores Opioides delta/análisis , Médula Espinal/química , Animales , Anticuerpos Monoclonales , Especificidad de Anticuerpos , Inmunohistoquímica , Microscopía Electrónica , Ratas , Ratas Wistar , Receptores Opioides delta/ultraestructura
17.
Neuroreport ; 12(17): 3689-92, 2001 Dec 04.
Artículo en Inglés | MEDLINE | ID: mdl-11726775

RESUMEN

Cannabinoids and opioids interact in the control of nociception at the spinal level. Likely, several mechanisms are involved, with one of them being co-localization of cannabinoid and opioid receptors. In order to validate this hypothesis, a double labeling study of CB1 cannabinoid receptors and mu-opioid receptors in the dorsal horn of the rat spinal cord was performed. A strong co-localization of CB1 and mu-opioid receptors was observed in lamina II interneurons at the ultrastructural level. The physiological consequences of the co-localization are discussed.


Asunto(s)
Nociceptores/metabolismo , Dolor/metabolismo , Células del Asta Posterior/metabolismo , Receptores de Droga/metabolismo , Receptores Opioides mu/metabolismo , Membranas Sinápticas/metabolismo , Transmisión Sináptica/fisiología , Animales , Cannabinoides/metabolismo , Dendritas/metabolismo , Dendritas/ultraestructura , Inmunohistoquímica , Interneuronas/metabolismo , Interneuronas/ultraestructura , Masculino , Microscopía Electrónica , Nociceptores/ultraestructura , Péptidos Opioides/metabolismo , Orgánulos/metabolismo , Orgánulos/ultraestructura , Dolor/fisiopatología , Células del Asta Posterior/ultraestructura , Terminales Presinápticos/metabolismo , Terminales Presinápticos/ultraestructura , Ratas , Ratas Wistar , Receptores de Cannabinoides , Membranas Sinápticas/ultraestructura
18.
Neurosci Res ; 5(4): 353-60, 1988 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-3374865

RESUMEN

The distribution of methionine-enkephalin-like cell bodies in the hypothalamus of the cat was studied using an indirect immunoperoxidase technique. Previously, intraventricular or intratissular injections of colchicine were carried out. Met-enkephalin-like neurons were located in all hypothalamic nuclei. The densest clusters of immunoreactive perikarya were observed in the nucleus arcuatus, median eminence, regio praeoptica and perifornical region, whereas the nucleus supraopticus and hypothalamus anterior had the lowest density. In other hypothalamic nuclei the density of methionine-enkephalin (Met-enkephalin) cell bodies was intermediate.


Asunto(s)
Encefalina Metionina/análisis , Hipotálamo/citología , Neuronas/análisis , Animales , Gatos , Hipotálamo/análisis , Inmunohistoquímica
19.
Neuropeptides ; 17(4): 177-85, 1990 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-2093150

RESUMEN

This study investigated the nature of the cholecystokinin-like immunoreactivity (CCKLI) in the dorsal horn of the rat spinal cord. Indeed, using the same antiserum, no variation of radioimmunoassayable CCKLI in the dorsal horn of rats treated neonatally with capsaicin could be seen, whereas the density of immunoreactive CCKLI fibres was greatly reduced in immunocytochemistry (IC). By comparing the technical conditions used in IC and radioimmunoassay (RIA), it could be concluded that the contradictory results obtained by the two techniques were very unlikely due to the fixation of the tissues, but rather to the concentration of the antiserum. First, CCKLI levels in extracts from fresh or 4% paraformaldehyde fixed tissues were not significantly different. Second, glomerular terminals, which are typical terminals of the fine primary afferents were immunoreactive at the surface of the block but unlabelled in its depth, where the concentration of antiserum might gradually reach a value closer to that used in radioimmunoassay. High pressure liquid chromatography analyses of acidic extracts of the dorsal part of the lumbar spinal cord of control rats revealed the presence, besides two major peaks coeluting with sulphated and non-sulphated CCK8, of two minor peaks '1' and '2' accounting for 23% of total CCKLI. Capsaicin treatment resulted in the disappearance of peak 2 and a marked reduction in the proportion of CCKLI in peak 1. Reported data suggested that the CCKLI material present in capsaicin-sensitive neurons, accounting for only about 10% of radioimmunoassayable CCKLI but for a much higher proportion of that detected in IC, was distinct from both genuine CCK and CGRP.


Asunto(s)
Colecistoquinina/análisis , Médula Espinal/química , Animales , Animales Recién Nacidos , Capsaicina/farmacología , Cromatografía Líquida de Alta Presión , Fijadores , Inmunohistoquímica/métodos , Masculino , Radioinmunoensayo/métodos , Ratas , Ratas Endogámicas
20.
Neuropeptides ; 37(4): 211-9, 2003 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-12906839

RESUMEN

Although chronic inflammatory pain is known to be associated with hypersensitivity to mu opioid receptor agonists, no evidence for changes in the expression and/or characteristics of central mu opioid receptors has yet been reported in relevant models of this type of pain. In the present study, both immunohistochemical and autoradiographic approaches were used to address this question in polyarthritic rats, on the 4th week after intradermal injection of complete Freund's adjuvant, when inflammatory pain was at its maximum. Immunohistochemical labeling with specific anti-mu opioid receptor antibodies and autoradiographic labeling with [3H]DAMGO showed an upregulation of mu opioid receptors in the dorsal root ganglia but no changes in the density of these receptors in the dorsal horn at the level of L4-L6 segments in polyarthritic compared to age-paired control rats. On the other hand, autoradiographic quantification of the concentration-dependent increase in [35S]GTP-gamma-S binding by the mu-opioid receptor agonist DAMGO did not show any significant differences within the lumbar dorsal horn between polyarthritic and control rats. These data indicate that chronic inflammatory pain caused by polyarthritis was associated with an increased expression of mu-opioid receptors in dorsal root ganglion sensory neurones that did not result in an increased spinal density of these receptors, in spite of their well established axonal transport in the central portion of primary afferent fibres to the dorsal horn. In contrast, axonal transport of mu-opioid receptors in the peripheral portion of these fibres probably accounts for the increased receptor density in inflamed tissues already reported in the literature.


Asunto(s)
Artritis Experimental/metabolismo , Proteínas de Unión al GTP/metabolismo , Ganglios Espinales/metabolismo , Receptores Opioides mu/metabolismo , Médula Espinal/metabolismo , Analgésicos Opioides/metabolismo , Analgésicos Opioides/farmacología , Animales , Autorradiografía , Enfermedad Crónica , Encefalina Ala(2)-MeFe(4)-Gli(5)/metabolismo , Encefalina Ala(2)-MeFe(4)-Gli(5)/farmacología , Guanosina 5'-O-(3-Tiotrifosfato)/metabolismo , Guanosina 5'-O-(3-Tiotrifosfato)/farmacología , Vértebras Lumbares , Masculino , Dolor/metabolismo , Ratas , Ratas Sprague-Dawley , Radioisótopos de Azufre , Tritio
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