RESUMO
Hydrogen sulfide (H2S) is the first inorganic compound identified as both a substrate for mitochondrial oxidative phosphorylation and a transmitter in mammalian cells. H2S seems to mediate effects that are correlated with those of nitric oxide (NO) by a reciprocal regulation. Moreover, H2S is consumed by mitochondrial oxidation mediated by sulfide-quinone reductase-like protein (SQRDL)-the vertebrate homolog of sulfide-quinone oxidoreductase (SQR). There is evidence that SQR plays an essential role in regulating H2S levels in fission yeast. To start understanding the role of SQRDL in the mammalian metabolism of H2S, we examine rat tissues. Our results show that SQRDL protein is present in all tissues tested, albeit restricted to specific mitochondrial populations at the cellular level. We demonstrate a developmental regulation of Sqrdl transcription in the kidney, where SQRDL protein is detectable in glomerular podocytes and in tubular cells of the renal medulla. We also show that Sqrdl transcription in T cells is responsive to external H2S. Taken together, our results suggest that Sqrdl transcription is adaptively regulated, probably to meet the need of H2S oxidation. Thus far, SQRDL has only been studied in a limited set of tissues. The present report demonstrates the presence and specific localization of SQRDL in various mammalian tissues.
Assuntos
Mamíferos/metabolismo , Mitocôndrias/enzimologia , Quinona Redutases/metabolismo , Homologia de Sequência de Aminoácidos , Envelhecimento/metabolismo , Animais , Linhagem Celular , Humanos , Imuno-Histoquímica , Glomérulos Renais/efeitos dos fármacos , Glomérulos Renais/enzimologia , Medula Renal/efeitos dos fármacos , Medula Renal/enzimologia , Leucócitos/efeitos dos fármacos , Leucócitos/enzimologia , Masculino , Camundongos , Proteínas dos Microfilamentos/metabolismo , Mitocôndrias/efeitos dos fármacos , Podócitos/citologia , Podócitos/efeitos dos fármacos , Podócitos/enzimologia , Quinona Redutases/genética , Ratos , Espermatozoides/efeitos dos fármacos , Espermatozoides/enzimologia , Sulfetos/farmacologia , Transcrição Gênica/efeitos dos fármacosRESUMO
Hydrogen sulfide (H2S) can be consumed by both invertebrates and vertebrates as an inorganic substrate. The pathway metabolizing H2S probably involves three mitochondrial enzymes, one of which is sulfide-quinone oxidoreductase (SQR), known as sulfide-quinone reductase-like protein (SQRDL) in vertebrates. Evidence from fission yeast suggests that SQR might have a role in regulating sulfide levels in the cell. Regulation might be essential for H2S to act as a gaseous transmitter (gasotransmitter). The brain is an organ with high activity of gasotransmitters, like nitric oxide (NO) and H2S, which are known to affect synaptic transmission. In this study, we provide evidence that SQRDL is expressed in the mammalian brain. Real-time polymerase chain reaction (PCR) showed an increase in the number of Sqrdl transcripts in the brain with increasing age. Cellular fractionation and subsequent analysis by Western blotting indicated that the protein is located in mitochondria, which is the site of sulfide consumption in the cell. With an immunohistochemical approach, we demonstrated that the SQRDL protein is expressed in neurons, oligodendrocytes, and endothelial cells. Taken together, our data suggest that brain tissue harbors the machinery required for local regulation of sulfide levels.
Assuntos
Química Encefálica/fisiologia , Encéfalo/enzimologia , Mitocôndrias/enzimologia , Quinona Redutases/análise , Quinona Redutases/biossíntese , Animais , Western Blotting , Imuno-Histoquímica , Imunoprecipitação , Ratos , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , VertebradosRESUMO
UNLABELLED: Leber congenital amaurosis (LCA, MIM 204001) is a clinically and genetically heterogeneous retinal disorder characterized by severe visual loss from birth, nystagmus, poor pupillary reflexes, retinal pigmentary or atrophic changes, and a markedly diminished electroretinogram (ERG). PURPOSE: To examine 100 consecutive patients with LCA in order to assess the relative burden of the three known genes involved in LCA, namely retinal guanylyl cyclase (GUCY2D), retinal pigment epithelium protein ( RPE65), and the cone-rod homeobox (CRX), and to define their clinical correlates. METHODS: Mutational analysis and detailed clinical examinations were performed in patients diagnosed with LCA at the Johns Hopkins Center for Hereditary Eye Diseases and the Montreal Children's Hospital. RESULTS: Mutations were identified in 11% of our patients: GUCY2D mutations accounted for 6%, while RPE65 and CRX gene mutations accounted for 3% and 2%, respectively. The clinical presentation was variable; however, the visual evolution in patients with mutations in GUCY2D and CRX remained stable, while individuals with mutations in the RPE65 gene showed progressive visual loss. CONCLUSIONS: This study suggests that molecular diagnosis of Leber congenital amaurosis could provide important information concerning prognosis and course of treatment.
Assuntos
Cegueira/genética , Proteínas do Olho/genética , Guanilato Ciclase/genética , Proteínas de Homeodomínio/genética , Mutação/genética , Atrofias Ópticas Hereditárias/genética , Proteínas/genética , Transativadores/genética , Adulto , Cegueira/congênito , Cegueira/diagnóstico , Proteínas de Transporte , Criança , Pré-Escolar , Análise Mutacional de DNA , Feminino , Seguimentos , Genótipo , Humanos , Lactente , Masculino , Atrofias Ópticas Hereditárias/diagnóstico , Linhagem , Fenótipo , Reação em Cadeia da Polimerase , Polimorfismo Conformacional de Fita Simples , cis-trans-IsomerasesRESUMO
PURPOSE: Leber congenital amaurosis (LCA) has been mapped to chromosome 17p13.1. From the candidate genes mapped to this region, thus far, only Retinal Guanylate Cyclase (RetGC), has been found to have pathogenic LCA mutations, in families from North African origin. However, early reports, demonstrated eight LCA families linked to 17p13.1, but only four of them showed mutations in RetGC. Mapped in proximity to this locus is the candidate gene Pigment Epithelium Derived actor (PEDF), a factor implicated in photoreceptor differentiation and neuronal survival. Our purpose in this study was to identify mutations and polymorphisms in the PEDF gene in LCA patients of diverse ethnic origin. METHODS: Automated genotyping with four 17p13.1 markers flanking the PEDF gene was performed to assess homozygosity and PCR-SSCP combined with direct sequencing was used to detect mutations in the PEDF gene in 17 LCA patients. RESULTS: Homozygosity of markers D17S796 and D17S804 was found and four new intragenic basepair alterations were discovered: a Met72Thr polymorphism in exon 3 (T331C), a Thr130Thr polymorphism in exon 4 (T506C), a G to A transition in intron 5 (nine base pairs upstream from splice acceptor site), and a Tyr321Tyr polymorphism in exon 7 (C1079T) were detected. CONCLUSIONS: We report the discovery of four new polymorphic alterations in the PEDF gene in LCA patients and exclude by RFLP analysis the PEDF gene as a common cause of Leber congenital amaurosis. These single nucleotide polymorphisms will aid in future linkage analysis of complex multifactorial diseases involving retinal and RPE dysfunctions.
Assuntos
Fatores de Crescimento Neural , Atrofias Ópticas Hereditárias/genética , Proteínas/genética , Serpinas/genética , Substituição de Aminoácidos , Cromossomos Humanos Par 17 , Etnicidade/genética , Proteínas do Olho/genética , Feminino , Humanos , Masculino , Mutação , Linhagem , Reação em Cadeia da Polimerase , Polimorfismo Genético , Polimorfismo Conformacional de Fita SimplesRESUMO
A free-ranging group of Saimiri sciureus was studied in a semi-natural forest habitat in eastern Amazonia, where behaviour patterns were broadly similar to those recorded for the species in the wild. According to focal-animal samples, the monkeys spent the vast majority of their time foraging and feeding, in particular for arthropod prey, which contributed almost half of identified food items. The predation of a small-bodied bat was also observed, although the study animals did not appear to forage systematically for chiropterans in the manner recorded for Saimiri oerstedi.
Assuntos
Comportamento Alimentar , Comportamento Predatório , Saimiri/psicologia , Animais , Animais Selvagens , Artrópodes , Brasil , Quirópteros , Frutas , Insetos , Locomoção , Atividade Motora , Plantas Comestíveis , Comportamento SocialRESUMO
Four groups of male Wistar rats showing disrupted inhibitory avoidance conditioning due to striatal lesions were studied. Three groups received striatal, cortical, or ventral mesencephalic brain grafts and the fourth group remained as a lesioned control. Sixty days postgraft the animals were retrained in an inhibitory avoidance task. The striatal-grafted animals were the only group that significantly improved in the ability to acquire the inhibitory avoidance task. Acetylcholinesterase histochemistry revealed positive patches of cells in the striatal grafts. Cortical grafts showed less reactivity, without patches. Immunocytochemical analyses for tyrosine hydroxylase revealed positive cell reactivity in the mesencephalic grafts and few positive fibers were detected in the border between the striatal grafts and the host tissue. These results demonstrate that striatal but not cortical or mesencephalic brain grafts can promote the restoration of the ability to acquire an inhibitory avoidance task and suggest that the acetylcholine tissue content is involved in the behavioral recovery.
Assuntos
Aprendizagem da Esquiva/fisiologia , Transplante de Tecido Encefálico/fisiologia , Córtex Cerebral/transplante , Corpo Estriado/transplante , Transplante de Tecido Fetal/fisiologia , Mesencéfalo/transplante , Regeneração Nervosa/fisiologia , Inibição Neural/fisiologia , Acetilcolina/fisiologia , Animais , Mapeamento Encefálico , Córtex Cerebral/fisiologia , Corpo Estriado/fisiologia , Masculino , Rememoração Mental/fisiologia , Mesencéfalo/fisiologia , Neurônios/fisiologia , Ratos , Ratos WistarRESUMO
Four groups of male Wistar rats showing disrupted inhibitory avoidance conditioning due to striatal lesions received either striatal or ventral mesencephalic brain grafts. Two additional non-lesioned groups were used as controls. Half of the groups was retrained in an inhibitory avoidance task at fifteen days postgraft and the other half at sixty days postgraft. Those animals receiving striatal grafts significantly improved their ability to acquire the inhibitory avoidance task at fifteen and sixty days postgraft, as opposed to those receiving mesencephalic grafts, which did not show behavioral recovery. Choline acetyltransferase and glutamate decarboxylase activities, as well as dopamine content, were measured in the grafted tissue. Striatal grafts showed levels of choline acetyltransferase activity similar to the control group. Moreover, a positive correlation was found between the choline acetyltransferase activity and the behavioral recovery. In contrast, both glutamate decarboxylase activity and dopamine levels were significantly lower in striatal and in mesencephalic grafts, as compared to the controls. These results show that striatal but not mesencephalic grafts can promote the restoration of the ability to acquire an inhibitory avoidance task even at early stages (15 days) of the development of the grafts. The results also suggest that acetylcholine plays an important role in behavioral recovery.
Assuntos
Aprendizagem da Esquiva/fisiologia , Transplante de Tecido Encefálico/fisiologia , Colina O-Acetiltransferase/fisiologia , Transplante de Tecido Fetal/fisiologia , Neostriado/fisiologia , Neostriado/transplante , Animais , Dopamina/metabolismo , Glutamato Descarboxilase/metabolismo , Masculino , Mesencéfalo/transplante , Neostriado/enzimologia , Ratos , Ratos WistarRESUMO
We have previously shown that hypothalamic fetal brain grafts induced recovery of sexual behavior in medial preoptic area (MPOA)-lesioned male rats. In the present series of experiments, male rats with completely abolished sexual behavior by MPOA lesions received either hypothalamic or frontal cortical fetal grafts. The animals that received hypothalamic grafts showed a gradual recovery of sexual behavior. In contrast, those animals who received cortical grafts did not recover sexual behavior during the 15 weeks after the graft. In addition, to evaluate the connectivity of the grafted tissue with the host brain, a retrograde tracer, fluorogold, was injected in the dorsal tegmental area. Fluorogold-labeled cells were found in the hypothalamic, but not in the cortical grafts. These results suggest that specificity of the grafted tissue and connectivity between brain grafts and host tissue are necessary for the recovery of male sexual behavior in MPOA-lesioned rats.
Assuntos
Transplante de Tecido Encefálico/fisiologia , Córtex Cerebral/transplante , Hipotálamo/transplante , Área Pré-Óptica/fisiologia , Comportamento Sexual Animal/fisiologia , Animais , Córtex Cerebral/fisiologia , Estradiol/farmacologia , Feminino , Transplante de Tecido Fetal/fisiologia , Hipotálamo/fisiologia , Masculino , Especificidade de Órgãos , Progesterona/farmacologia , Ratos , Ratos Wistar , Comportamento Sexual Animal/efeitos dos fármacosRESUMO
We recently showed that fetal brain transplants produced a significant recovery in the ability of gustatory neocortex-lesioned rats to learn a conditioned taste aversion. In this report we assessed the capability of gustatory neocortex fetal brain transplants to produce behavioral recovery at different times. Four groups of male Wistar rats showing disrupted taste aversions due to gustatory neocortex lesions were studied. The lesioned animals received fetal cortical grafts, obtained from 16-day-old fetuses, and were retrained in the behavioral procedure after 15, 30, 45, or 60 days postgraft. Behavioral results showed a very good functional recuperation at 60 days, slight recovery at 45 and 30 days, and a poor recovery at 15 days postgraft. Results with HRP histochemistry revealed that at 30, 45, and 60 days postgrafting there were increased connections with the ventromedial nucleus of the thalamus and with the amygdala. At 15 days postgrafting there was an absence of HRP-labeled cells. In addition, behavioral recovery was correlated with increased acetylcholinesterase activity, detected histochemically, and with morphological neuronal maturation, revealed by Golgi staining. These results suggest that morphological maturity and reconnectivity between grafts and host tissue are important for behavioral recovery in gustatory neocortex-lesioned rats.
Assuntos
Aprendizagem da Esquiva/fisiologia , Córtex Cerebral/fisiologia , Condicionamento Clássico/fisiologia , Regeneração Nervosa/fisiologia , Plasticidade Neuronal/fisiologia , Paladar/fisiologia , Acetilcolinesterase/biossíntese , Animais , Transplante de Tecido Encefálico/fisiologia , Córtex Cerebral/embriologia , Córtex Cerebral/transplante , Cloretos/toxicidade , Transplante de Tecido Fetal/fisiologia , Lítio/toxicidade , Cloreto de Lítio , Neurônios/fisiologia , Ratos , Ratos EndogâmicosRESUMO
An immunohistochemical study of opioid peptides in the hypophysis of the axolotl, Ambystoma mexicanum, was carried out with antisera against leu-enkephalin, beta-endorphin, met-enkephalin, and dynorphin A (1-8). We found leu-enkephalin immunoreactivity in some fibers of the neural lobe and the median eminence. In contrast to previous reports on mammals and other vertebrates, we found leu-enkephalin immunoreactivity in many cells scattered throughout the anterior lobe. As in other vertebrates, the beta-endorphin immunoreactivity was present in all the cells of the intermediate lobe and in a few cells of the anterior lobe. Met-enkephalin and dynorphin A (1-8) immunoreactivities were only present in the neural lobe and the median eminence. The presence of leu-enkephalin and beta-endorphin in the anterior lobe suggests that these peptides could be acting as hormones released from the hypophysis of the unmetamorphosed amphibian.
Assuntos
Endorfinas/análise , Encefalinas/análise , Adeno-Hipófise/citologia , Ambystoma mexicanum , Animais , Anticorpos , Reações Cruzadas , Dinorfinas/análise , Encefalina Leucina/análise , Encefalina Metionina/análise , Imunofluorescência , Imuno-Histoquímica , Fibras Nervosas/ultraestrutura , Adeno-Hipófise/anatomia & histologia , beta-Endorfina/análiseRESUMO
Male rats received bilateral lesions within the medial preoptic area which completely abolished sexual behavior. Hypothalamic fetal brain transplants gradually restored sexual behavior to prelesion levels by the 6th week after the transplant. Immunocytochemical analyses revealed tyrosine hydroxylase immunoreactivity neurons within the transplanted tissue. These results demonstrate that fetal brain transplants can restore an innate complex behavior in which no spontaneous recovery is observed.