RESUMEN
Studies on the clinical course of familial ALS suggest that the duration of illness is relatively consistent for each mutation but variable among the different mutations. The authors analyzed the relative amount of mutant compared with normal SOD1 protein in the erythrocytes from 29 patients with ALS with 22 different mutations. Turnover of mutant SOD1 correlated with a shorter disease survival time.
Asunto(s)
Esclerosis Amiotrófica Lateral/genética , Esclerosis Amiotrófica Lateral/fisiopatología , Predisposición Genética a la Enfermedad/genética , Mutación/genética , Superóxido Dismutasa/genética , Adulto , Edad de Inicio , Anciano , Esclerosis Amiotrófica Lateral/metabolismo , Sistema Nervioso Central/metabolismo , Sistema Nervioso Central/patología , Sistema Nervioso Central/fisiopatología , Análisis Mutacional de ADN , Progresión de la Enfermedad , Femenino , Pruebas Genéticas , Humanos , Masculino , Persona de Mediana Edad , Neuronas Motoras/metabolismo , Neuronas Motoras/patología , Valor Predictivo de las Pruebas , Pronóstico , Superóxido Dismutasa/metabolismo , Superóxido Dismutasa-1 , Tasa de Supervivencia , Factores de TiempoRESUMEN
We investigated the expression pattern of versican, aggrecan, link protein and hyaluronan in the developing limb bud cartilage of the fetal mouse using in situ hybridization and/or immunohistochemistry. Versican mRNA and immunostaining were detected in the mesenchymal cell condensation of the future digital bone at E13. Versican mRNA expression rapidly disappeared from the tibial cartilage, as cartilage formation progressed during E13-15, but the immunostaining was gradually replaced by aggrecan immunostaining from the diaphysis. Immunostaining for both molecules thus had a 'nega-posi' pattern and consequently versican immunostaining was still detected at the epiphyseal end at E15. This result indicated that versican functions as a temporary framework in newly formed cartilage matrix. An aggrecan-positive region within the cartilage invariably had intense hyaluronan staining, whereas a versican-positive region also had affinity for hyaluronan within the cartilage, but not in the mesenchymal cell condensation. Therefore, the presence of versican aggregates was not confirmed in the developing limb bud cartilage. Furthermore, although link protein was more closely related with aggrecan than versican during limb bud cartilage formation, there was a discrepancy between the expression of aggrecan and link protein in tibial cartilage at E15. In particular, only a link protein-positive region was present in the marginal area of the metaphysis and the epiphysis at this stage. This finding may indicate a novel role for link protein.
Asunto(s)
Cartílago/embriología , Proteoglicanos Tipo Condroitín Sulfato/análisis , Proteínas de la Matriz Extracelular , Ácido Hialurónico/análisis , Esbozos de los Miembros/crecimiento & desarrollo , Proteínas/análisis , Proteoglicanos/análisis , Agrecanos , Animales , Biomarcadores/análisis , Cartílago/química , Proteoglicanos Tipo Condroitín Sulfato/genética , Edad Gestacional , Ácido Hialurónico/genética , Inmunohistoquímica/métodos , Hibridación in Situ/métodos , Lectinas Tipo C , Ratones , Ratones Endogámicos ICR , Proteínas/genética , Proteoglicanos/genética , ARN Mensajero/análisis , VersicanosAsunto(s)
Encefalitis Viral/complicaciones , Porfobilinógeno Sintasa/deficiencia , Porfirias Hepáticas/fisiopatología , Enfermedad Aguda , Adulto , Femenino , Hemo/biosíntesis , Humanos , Linaje , Fenotipo , Porfirias Hepáticas/complicaciones , Porfirias Hepáticas/genética , Convulsiones/tratamiento farmacológico , Convulsiones/etiologíaRESUMEN
We investigated the histochemical localisation of versican, aggrecan and hyaluronan in the developing condylar cartilage of the fetal rat mandible at d 15-17 of gestation. At d 15 of gestation, immunostaining for versican was detected in the anlage of the future condylar process (condylar anlage), although the staining intensity showed a considerable regional variation. At d 16 of gestation, a metachromatically stained matrix firstly appeared in the condylar anlage. Aggrecan, hyaluronan and versican were simultaneously detected in this newly formed condylar cartilage. At d 17 of gestation, immunostaining for versican became restricted to the perichondrium and was barely detected in the cartilage. Colocalisation of versican and aggrecan was also seen in the cranial base cartilage at d 14 of gestation. These results indicate that although versican is replaced by aggrecan during the transition from prechondrogenic tissue to cartilage, both molecules were temporally colocalised in the newly formed cartilage. A hyaluronan-rich, low-versican area was identified in the posterior end of the condylar anlage during d 15-17 of gestation. The existence of this area is a unique structural feature of the developing condylar cartilage.
Asunto(s)
Cartílago/química , Cartílago/embriología , Proteínas de la Matriz Extracelular , Glicosaminoglicanos/análisis , Mandíbula/embriología , Agrecanos , Animales , Proteoglicanos Tipo Condroitín Sulfato/análisis , Edad Gestacional , Histocitoquímica/métodos , Ácido Hialurónico/análisis , Lectinas Tipo C , Proteoglicanos/análisis , Ratas , Ratas Sprague-Dawley , VersicanosRESUMEN
Transgenic mice carrying familial amyotrophic lateral sclerosis (FALS)-linked mutant Cu/Zn superoxide dismutase (SOD1) genes such as G93A (G93A-mice) and G85R (G85R-mice) genes develop limb paresis. Introduction of human wild type SOD1 (hWT-SOD1) gene, which does not cause motor impairment by itself, into different FALS mice resulted in different effects on their clinical courses, from no effect in G85R-mice to acceleration of disease progression in G93A-mice. However, the molecular mechanism which causes the observed difference, has not been clarified. We hypothesized that the difference might be caused by the stability of mutant SOD1 proteins. Using a combination of mass spectrometry and enzyme-linked immunosorbent assay, we found that the concentration of G93A-SOD1 protein was markedly elevated in tissues of transgenic mice carrying both G93A- and hWT-SOD1 genes (G93A/hWT-mice) compared to that in G93A-mice, and also found that the concentration of G93A-SOD1 protein had a close relation to the disease duration. The concentration of metallothionein-I/II in the spinal cord, reflecting the degree of copper-mediated oxidative stress, was highest in G93A/hWT-mice, second in G93A-mice, and normal in the mice carrying hWT-SOD1 gene. These results indicated that the increase of G93A-SOD1 protein was responsible for the increase of oxidative stress and disease acceleration in G93A/hWT-mice. We speculate that coexpression of hWT-SOD1 protein is deleterious to transgenic mice carrying a stable mutant such as G93A-SOD1, because this mutant protein is stabilized by hWT-SOD1 protein, but not to transgenic mice carrying an unstable mutant such as G85R-SOD1, because this mutant protein is not stabilized by hWT-SOD1.
Asunto(s)
Esclerosis Amiotrófica Lateral/enzimología , Esclerosis Amiotrófica Lateral/genética , Regulación Enzimológica de la Expresión Génica/fisiología , Mutación/fisiología , Médula Espinal/enzimología , Superóxido Dismutasa/genética , Superóxido Dismutasa/metabolismo , Animales , Femenino , Radicales Libres/metabolismo , Humanos , Masculino , Metalotioneína/genética , Metalotioneína/metabolismo , Ratones , Ratones Mutantes Neurológicos , Ratones Transgénicos , Estrés Oxidativo/genética , Médula Espinal/fisiopatología , Superóxido Dismutasa-1 , Regulación hacia Arriba/fisiologíaRESUMEN
The hydrophilicity of polar and apolar domains of various amphiphiles was systematically estimated for their homologues and analogues by measuring the molar adiabatic compressibility of an aqueous solution at infinite dilution. The homologues of protic alkyl H(CH(2))(n)-, perfluoroalkyl F(CF(2))(n)-, and alkylphenyl H(CH(2))(n)(C(6)H(5))- groups (n=0-10) were chosen to represent apolar hydrophobic domains. The polar hydrophilic domains tested were -SO(4)Na, -SO(3)Na, -COONH(4), -N(CH(3))(3)Br, N(C(m)H(2m+1))(4)Br (m=1-5), and -NH(CH(2))(n)SO(3) (n=3, 4) groups. Also tested were the tetraphenyl ionic compounds (C(6)H(5))(4)MX (M=B/X=Na, M=P/X=Cl, M=As/X=Cl) to study the effect of the ionic sign of the core atom across the tetraphenyl apolar shell, the polyethylene glycols H(OCH(2)CH(2))(m)OH (m=1-4) to study the role of apolar -CH(2)- units in the hydrophilic oxyethylene group, and the zwitterionic dimethylaminoalkylsulfonate (CH(3))(2)NH(CH(2))(n)SO(3) homologues to study the effect of intramolecular salt formation on the hydrophilicity of the zwitterion. The adiabatic compressibility of the solution was calculated from measurement of the sound velocity and density of solutions. The introduction of laboratory automation and the numerical control of the system improved the accuracies and efficiencies of the measurements a great deal. The range of the temperature scan was 0-40 degrees C with an effective accuracy of +/-0.001 degrees C and the concentration was automatically scanned down to far below the cmc of the surfactant. The hydrophilicity of various polar and apolar substances was estimated as the decrease of molar adiabatic compressibility of the aqueous solution with increased concentration of their homologues and analogues. The hydrophobic hydration of nonpolar substances was found to be very small at room temperature and was barely detected above 40 degrees C; however, it became large as the temperature was lowered and attained a maximum at 0 degrees C. The cationic charge of quaternary ammonium N(+)(C(n)H(2n+1))(4) was found to enhance the hydrophobic hydration of methylene groups located at a distance of 4 to 6 Å from the core nitrogen atom, while the terminal negative charge of the anionic surfactant R-SO(4)(-), R-SO(3)(-), or R-COO(-) was found to decrease the hydrophobic hydration of -CH(2)- units within the same range. The hydrophilicity of quaternary ammonium and the tetraphenyl ions should be synergistically given by both hydrophobic and ionic hydrations. The hydrophilicity of the perfluoromethylene unit -CF(2)- was found to have a value comparable to that of the protic methylene unit -CH(2)-. The hydrophobic hydration seems to offer a good measure of the hydrophilicity of apolar substances; however, it does not necessarily represent the "hydrophobicity" of the apolar segment when the "surface activity" of the amphiphile is concerned. Copyright 2000 Academic Press.
RESUMEN
The nucleotide sequence of 10 isolates of human parvovirus B19 (B19) were determined and compared throughout 96.3% of the open reading frames (4145 nucleotides from nt. 509-4653). In the 4145 nucleotides, 122 mutation sites were found, of which 24 were accompanied by amino acid displacement. Furthermore, the polymorphism of the amino acids was seen in about 110 bases near the carboxy terminal of the non-structural protein, ranging from nt. 2011 to 2123, where four amino acid mutation points were found to exist. Based on the amino acid polymorphism of these four mutation sites in this area, 10 isolates of the B19 parvovirus could be divided into 4 subtypes (subtypes A, B, C, and D). The frequency of isolation of the subtypes depended on the time and location of collection of the B19 viremic blood specimens.
Asunto(s)
Parvovirus B19 Humano/clasificación , Polimorfismo Genético , Proteínas no Estructurales Virales/genética , Secuencia de Aminoácidos , Sustitución de Aminoácidos , Secuencia de Bases , Análisis Mutacional de ADN , ADN Viral/sangre , Genotipo , Humanos , Sistemas de Lectura Abierta , Parvovirus B19 Humano/genética , Parvovirus B19 Humano/aislamiento & purificación , Reacción en Cadena de la Polimerasa , Polimorfismo Conformacional Retorcido-SimpleRESUMEN
A 22-year-old man had choreatic movements in upper limbs, neck and trunk for over twelve years which were associated with dystonia in lower limbs upon initiating voluntary movements. The choreatic movement lasted for a few seconds and the dystonia lasted for a few minutes. He also had high serum CK levels and hypertrophic calf muscles. His muscle strength and deep tendon reflexes were normal. His choreatic movements fulfill the criteria for paroxysmal kinesigenic choreoathetosis (PKC). However, it was unclear what the symptom of dystonia was due to. From a muscle biopsy and DNA analysis, he was diagnosed as having Becker muscular dystrophy. Administration of anticonvulsant improved the dystonia as well as the choreatic movement, which showed that the dystonia was a symptom of PKC. Coincidence of choreatic movements and dystonias which had different lasting time in a patient of PKC was atypical and had not previously reported.
Asunto(s)
Corea/complicaciones , Distrofia Muscular de Duchenne/complicaciones , Adulto , Anticonvulsivantes/uso terapéutico , Carbamazepina/uso terapéutico , Creatina Quinasa/sangre , Humanos , Hipertrofia/complicaciones , Pierna , Masculino , Músculo Esquelético/patología , Resultado del TratamientoRESUMEN
Using a video game format, this study examined the effects of time pressure (TP) on behavioral and electrocortical indices. The behavioral results were consistent with previous time pressure research in that TP reduced time to perform a task and increases behavioral errors. In addition, electroencephalogram (EEG) measures showed distinctive patterns associated with TP in the theta, mu, and gamma bands along the midline. Site specific changes in the success vs. failure trials were also seen in midline theta at Fz, gamma at Fz, and mu at Cz. Right parietal alpha also differentiated TP and success vs. failure trials. In specific TP (1) increased frontal midline theta activity and (2) increased gamma at midline (frontal, central, and partietal) and in right frontal areas. The results of these findings are discussed in terms of the formation of specific neurocognitive strategies as evidenced by the topographic distribution of task-related modulation of the EEG within certain frequency bands. It is suggested that the effect of TP on visuomotor performance is mediated by adopting either task-relevant or task-irrelevant neurocognitive strategies as evidenced by successful or failed trials, respectively. Whether these strategies are formulated prior to performance or appear spontaneously during task performance remains unclear and is awaiting further experimentation.
Asunto(s)
Corteza Cerebral/fisiología , Desempeño Psicomotor/fisiología , Adulto , Ritmo alfa , Femenino , Humanos , Masculino , Tiempo de Reacción/fisiología , Ritmo TetaRESUMEN
The aim of this study was to investigate the developmental characteristics of the mandibular condyle in sequential phases at the gene level using in situ hybridisation. At d 14.5 of gestation, although no expression of type II collagen mRNA was observed, aggrecan mRNA was detected with type I collagen mRNA in the posterior region of the mesenchymal cell aggregation continuous with the ossifying mandibular bone anlage prior to chondrogenesis. At d 15.0 of gestation, the first cartilaginous tissue appeared at the posterior edge of the ossifying mandibular bone anlage. The primarily formed chondrocytes in the cartilage matrix had already shown the appearance of hypertrophy and expressed types I, II and X collagens and aggrecan mRNAs simultaneously. At d 16.0 of gestation, the condylar cartilage increased in size due to accumulation of hypertrophic chondrocytes characterised by the expression of type X collagen mRNA, whereas the expression of type I collagen mRNA had been reduced in the hypertrophic chondrocytes and was confined to the periosteal osteogenic cells surrounding the cartilaginous tissue. At d 18.0 of gestation before birth, cartilage-characteristic gene expression had been reduced in the chondrocytes of the lower half of the hypertrophic cell layer. The present findings demonstrate that the initial chondrogenesis for the mandibular condyle starts continuous with the posterior edge of the mandibular periosteum and that chondroprogenitor cells for the condylar cartilage rapidly differentiate into hypertrophic chondrocytes. Further, it is indicated that sequential rapid changes and reductions of each mRNA might be closely related to the construction of the temporal mandibular ramus in the fetal stage.
Asunto(s)
Cartílago/embriología , Colágeno/genética , Proteínas de la Matriz Extracelular , Cóndilo Mandibular/embriología , Proteoglicanos/genética , ARN Mensajero/análisis , Agrecanos , Animales , Cartílago/metabolismo , Desarrollo Embrionario y Fetal , Hibridación in Situ , Lectinas Tipo C , Cóndilo Mandibular/metabolismo , Ratones , Ratones Endogámicos C57BLRESUMEN
BACKGROUND: It has been 10 years since the implementation in Japan of donor blood screening for human T-cell lymphotropic virus type I (HTLV-I). This report reviews the effectiveness of screening in preventing transmission of HTLV-I through blood transfusion and the current status of patients with confirmed seroconversion due to transfusions given before the implementation of screening. STUDY DESIGN AND METHODS: Patients who received blood at Kyushu University Hospital from 1990 to 1997 were followed. Serum samples were collected before transfusion and 60 days or more after transfusion. Seroconversion was determined by a second-generation particle agglutination test. Confirmation tests were an immunofluorescence assay, enzyme-linked immunosorbent assay, and immunoblotting. Confirmed seroconverted patients were followed by a search of hospital records. RESULTS: Seroconversion was found in one of 4672 transfused patients, but the donor was identified and confirmed to be negative for anti-HTLV-I and virus genome by nested polymerase chain reaction. A total of 23,323 red cell concentrates and 17,237 platelet concentrates were transfused to these 4672 patients. Therefore, the anti-HTLV-I prevalence in blood for transfusion after screening was estimated at 1 in 45,560 (0.0022%; the upper 95% CI was 0.0080%). One hundred two seroconverted patients who were transfused before donor screening for HTLV-I were followed. One patient developed HTLV-I-associated myelopathy, diagnosed 18 weeks after seroconversion, and another patient developed uveitis 1 month after seroconversion. No patients developed adult T-cell lymphoma, and the survival rate of seroconverted patients was 92.5 percent 15 years after transfusion. CONCLUSION: This study confirmed that the present donor screening program for HTLV-I by the new particle agglutination test can almost completely prevent virus transmission by transfusion. Complications of HTLV-I transmission were at lower rates than expected.
Asunto(s)
Donantes de Sangre , Infecciones por HTLV-I/diagnóstico , Infecciones por HTLV-I/transmisión , Tamizaje Masivo , Reacción a la Transfusión , Adolescente , Adulto , Anciano , Pruebas de Aglutinación , Niño , Preescolar , Femenino , Infecciones por HTLV-I/fisiopatología , Humanos , Lactante , Masculino , Persona de Mediana Edad , Paraparesia Espástica Tropical/etiología , Pruebas Serológicas , Análisis de Supervivencia , Uveítis/virologíaRESUMEN
The nature of adsorbed cationic amphiphiles at the mica/solution interface was studied by XPS and contact angle measurements. The elemental analyses of freshly cleaved mica surfaces by XPS showed that the potassium atoms on the surface lattice of mica are not necessarily distributed equally to each surface on cleavage. The adsorbed cationic amphiphile molecules remaining on mica surfaces after rinsing with distilled water were found to be anchored to the surface by ion-exchange, replacing surface potassium and/or other cations. The ratio of adsorbed cationic amphiphile molecules with single alkyl chains to the maximum potassium ions on mica surface was estimated to be twice as large as that of amphiphiles having two alkyl chains. The contact angle of water drops placed on the adsorbed surface showed a gradual decrease with the elapse of time due to the dissolution of adsorbed surfactant into the water drop; however, the decrease was not observed for those mica surfaces when aged for more than 3 days in the adsorption bath. The anchoring of adsorbed molecules by ion-exchange was found to occur extremely slowly, however; the anchored molecules may not easily be desorbed when rinsed with deionized water. The time dependent anchoring of adsorbed molecules was studied in terms of adsorption time, alkyl chain length, and concentration of cationic surfactant. Copyright 1999 Academic Press.
RESUMEN
PURPOSE: To prepare poly(lactide-co-glycolide)(PLGA) microspheres containing recombinant hepatitis B core antigen (HBcAg; Mw = 3,600,000) by a w/o/w emulsion/solvent evaporation method and evaluate the possibility of this system as a potent long-acting carrier for hepatitis B core antigen in mice. METHODS: Various additives had been incorporated in the internal aqueous phase during the process of microencapsulating HBcAg, HBcAg antigenicity in the medium extracted from the prepared microspheres were measured by ELISA. Shape confirmation of the HBcAg antigen was performed by a sucrose gradient velocity centrifugal technique. For in vivo study, prepared microspheres were administered subcutaneously to Balb/C mice, and the serum IgG level was determined by ELISA. RESULTS: The inactivation of HBcAg by methylene chloride was dramatically reduced by the addition of gelatin (4-8% (w/v)) to the internal aqueous phase during the preparation. Further improvement of the loading efficiency to almost 61% resulted with cooling (4 degrees C). The prepared microspheres (4.27 microm+/-1.23 microm) containing 0.15% HBcAg displayed burst release (50-60% within 2 days). In subcutaneous inoculation, the adjuvant effect of PLGA microspheres was almost the same as that of the complete Freund's adjuvant. Whereas oral inoculation using the microspheres was not effective. CONCLUSIONS: The pH of the added gelatin seemed to be the key to the stabilization of HBcAg from various stability tests and CD spectrum study. Finally, the possibility of using this system as a potent long-acting hepatitis B vaccine was demonstrated.
Asunto(s)
Composición de Medicamentos , Antígenos del Núcleo de la Hepatitis B/administración & dosificación , Vacunas contra Hepatitis B/administración & dosificación , Animales , Estabilidad de Medicamentos , Femenino , Ratones , Ratones Endogámicos BALB C , Microesferas , Peso MolecularRESUMEN
Leukemia inhibitory factor (LIF) is a cytokine that acts on a wide range of cell types in vitro, but knowledge of its physiological role is limited. High levels of LIF protein have been selectively detected in the thymus throughout postnatal development. LIF-deficient mice have shown impaired thymic T cell maturation, suggesting the possibility that T cells require LIF for maturation. We have used highly specific antibodies raised against native rat LIF to inhibit LIF function during a defined and restricted period of thymic T cell maturation (first postnatal week). Surprisingly, we observed increased T cell activation in the LIF-deprived wild-type rat. The increased T cell response is retained even 4 weeks after anti-LIF treatment when the level of LIF in the thymic microenvironment has returned to normal. Our results are in contrast to findings with LIF knockout mice, where decreased T cell activation was observed. These observations suggest that LIF may have alternative effects on various phases of T cell development and that LIF may be involved in the restriction of the T cell repertoire during maturation occurring in the first postnatal week.
Asunto(s)
Anticuerpos Bloqueadores/inmunología , Inhibidores de Crecimiento/fisiología , Interleucina-6 , Activación de Linfocitos , Linfocinas/fisiología , Linfocitos T/inmunología , Animales , Citometría de Flujo , Inhibidores de Crecimiento/deficiencia , Inhibidores de Crecimiento/inmunología , Factor Inhibidor de Leucemia , Leucopoyesis , Linfocinas/deficiencia , Linfocinas/inmunología , Masculino , Ratones , Ratones Noqueados , Ratas , Ratas Endogámicas Lew , Timo/citologíaRESUMEN
For the fully hydrated multilamellar stack of dimyristoylphosphatidylcholine (DMPC) fluid membranes containing hydrophobic peptide gramicidin A (GrA), the membrane thickness and the bilayer-bilayer separation (i.e., water layer thickness) were determined by measuring small-angle X-ray scattering and the density of aqueous suspensions of DMPC-GrA mixtures. When the molar ratio of GrA to DMPC was 0.04, the membrane thickness decreased by 2-3 A by the incorporation of GrA molecules into DMPC bilayers, whereas the water layer thickness increased by 3-4 A. As the cause of the increment of water layer thickness, two possibilities were considered; (1) attractive van der Waals force acting between the bilayer membranes weakened by the decrease of membrane thickness, and (2) repulsive undulation force enhanced by the incorporation of GrA which may stabilize the gauche conformers of the lipid acyl chains.
Asunto(s)
Dimiristoilfosfatidilcolina/química , Gramicidina/química , Lípidos de la Membrana/química , Membranas Artificiales , Secuencia de Aminoácidos , Rastreo Diferencial de Calorimetría , Membrana Dobles de Lípidos/química , Fluidez de la Membrana , Datos de Secuencia Molecular , Dispersión de Radiación , Temperatura , Agua/química , Rayos XRESUMEN
Cholinergic neuronal differentiation factor/leukemia inhibitory factor (CDF/LIF) is a multifunctional cytokine that affects neurons as well as many other cell types. Toward elucidating its neural functions in vivo, we previously investigated the distribution of CDF/LIF binding sites with iodinated native CDF/LIF in embryonic to postnatal day 0 (P0) rats. In the present study, we have extended our examination to postnatal ages and find that specific CDF/LIF binding sites are present at defined developmental stages in additional brain regions not previously exhibiting binding by P0. High levels of binding are detected in all P7 sensory and autonomic ganglia examined, but only in restricted postnatal central nervous system structures. Cranial motor and mesencephalic trigeminal neurons maintain high levels throughout, while binding to spinal motor neurons, which decreases to low levels at P0, reappears by P14 and increases with age. Most other structures, which show detectable binding by P0, exhibit higher levels at postnatal ages, including the red, deep, ventral cochlear, trapezoid, superior olivary, vestibular, ventral tegmental, and ventral posterior thalamic nuclei as well as the glomerular layer of the olfactory bulb. High levels are also detected in several structures for the first time after P0, including the cerebellar cortex (molecular and Purkinje cell layers), lateral reticular nucleus of the medulla and reticular formation, as well as the reticulotegmental, medial geniculate, solitary (rostral, dorsomedial, and commissural regions), medial septal, lateral mammillary, and lateral habenular nuclei. These results not only identify regions of potential CDF/LIF-responsive neurons and glia throughout development but suggest new CDF/LIF roles in the nervous system.
Asunto(s)
Sitios de Unión/efectos de los fármacos , Fibras Colinérgicas/metabolismo , Citocinas/farmacología , Inhibidores de Crecimiento/farmacología , Interleucina-6 , Linfocinas/farmacología , Sistema Nervioso/efectos de los fármacos , Animales , Autorradiografía , Factor Inhibidor de Leucemia , Sistema Nervioso/crecimiento & desarrollo , RatasRESUMEN
To define the physiological role of leukemia inhibitory factor (LIF), it is essential to localize sites of LIF synthesis in vivo. We generated polyclonal antibodies specific for native rat LIF, and developed a two-site immunoassay to detect 10 pg LIF/ml. Using this immunoassay, we determined LIF content of 18 organs, CNS regions, and ganglia throughout postnatal development of rats. High levels of LIF protein (1.0-11.0 ng/g tissue) are present in relatively few tissues: the uterus at late proestrus to estrus and on day 5 of pregnancy, ovary at estrus to early metestrus-1, footpads during early postnatal development and thymus throughout. Intermediate levels (0.5-1.0 ng) are detected in the gut, skin, skeletal muscle, pancreas and lung at one or more postnatal ages. Low levels (0.1-0.5 ng) are observed in most other non-nervous and nervous tissues. LIF protein levels do not completely correspond to reported LIF mRNA levels.