RESUMEN
BACKGROUND/AIMS: Down syndrome associated disorders are caused by a complex genetic context where trisomy 21 is a central component in relation to other changes involving epigenetic regulators and signaling molecules. This unique genetic context is responsible for the predisposition of people with Down syndrome to acute leukemia. Although, the research in this field has discovered some important pathogenic keys, the exact mechanism of this predisposition is not known. METHODS: In this study we applied functional enrichment analysis to evaluate the interactions between genes localized on chromosome 21, genes already identify as having a key role in acute leukemia of Down syndrome, miRNAs and signaling pathways implicated in cancer and cell development and found that miR-155 has a high impact in genes present on chromosome 21. Forward, we performed next generation sequencing on DNA samples from a cohort of patients diagnosed with acute leukemia of Down syndrome and in vitro functional assay using a CMK-86 cell line, transfected with either mimic or inhibitor of the microRNA-155-5p. RESULTS: Our results show that the epigenetic alteration of the TNF superfamily receptors in Down syndrome, which can be correlated to microRNA-155-5p aberrant activity, may play an important role in cell signaling and thus be linked to acute myeloid leukemia. CONCLUSION: Some genes, already shown to be mutated in AML-DS, are potential targets for miR-155. Our results show that the epigenetic alteration of the TNF superfamily receptors in Down syndrome may play an important role in cell signaling and thus be linked to acute myeloid leukemia.
Asunto(s)
Síndrome de Down/complicaciones , Epigénesis Genética , Regulación Leucémica de la Expresión Génica , Leucemia Mieloide Aguda/patología , Reacción Leucemoide/patología , MicroARNs/genética , Receptores del Factor de Necrosis Tumoral/genética , Diferenciación Celular , Estudios de Cohortes , Síndrome de Down/etiología , Síndrome de Down/genética , Síndrome de Down/metabolismo , Síndrome de Down/patología , Femenino , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Leucemia Mieloide Aguda/etiología , Leucemia Mieloide Aguda/metabolismo , Reacción Leucemoide/etiología , Reacción Leucemoide/metabolismo , Masculino , Receptores del Factor de Necrosis Tumoral/metabolismoRESUMEN
BACKGROUND: Improved immunization rates have reduced the incidence of vaccine-preventable diseases (VPDs) in advanced nations. Japan's unique vaccination system classifies vaccines into routine vaccines ostensibly required under the Preventive Vaccination Law and recommended but optional vaccines, although all vaccines are in fact voluntary. In Japan, low immunization rates, particularly for optional vaccines, have resulted in high rates of sequelae and death. The decision as to whether a child will receive a vaccine depends on the parents, who must obtain information, make inquiries, and make the required payment, the last of which is a major barrier. This randomized, controlled trial was conducted to evaluate the effectiveness of an immunization education program designed to meet mothers' needs. METHODS: This randomized controlled trial assigned pregnant women to intervention or control groups. The intervention was individual education sessions involving the children's fathers in shared decision-making on whether or not to immunize their child. A survey was conducted before and after the intervention. Data were analyzed using the intention-to-treat principle. RESULTS: Of 225 pregnant women, 175 (78%) participated and 171 replied to the post-survey. At age 3 months, intervention infants had higher self-reported immunization rates for hepatitis B virus vaccine (76% vs. 49%; P < 0.001) and rotavirus vaccine (84% vs. 68%; P = 0.019) than control group infants. The percentage of parents intending to vaccinate their infants was higher in the intervention group (77% vs. 52%; P < 0.01). Improvements in scores for basic knowledge (mean [SD]: 5.5 [3.6] vs. 3.0 [3.8], range: 10-30; P < 0.001), advanced knowledge (mean [SD]: 5.1 [2.4] vs. 2.8 [2.5], range: 5-15; P < 0.001), and health literacy regarding immunization (mean [SD]: 0.5 [0.8] vs. 0.2 [0.6], range: 1-5; P < 0.01) were higher in the intervention group. The rate of decision making by both parents (68% vs. 52%; P < 0.05) was higher in the intervention group. CONCLUSIONS: Our findings confirmed the program's effectiveness. The intervention improved immunization rates, the percentage of parents intending to vaccinate their infants and knowledge scores. Interventions which directly and indirectly involved fathers in shared decision-making on whether to immunize their child were effective, as were individualized interventions that provided parents with access to up-to-date information. TRIAL REGISTRATION: UMIN000012575 . Registered 14 December 2013 (The study was prospectively registered).
Asunto(s)
Educación en Salud , Inmunización , Padres/educación , Adulto , Toma de Decisiones , Femenino , Conocimientos, Actitudes y Práctica en Salud , Humanos , Inmunización/psicología , Inmunización/estadística & datos numéricos , Lactante , Intención , Japón , Masculino , Padres/psicología , Periodo Posparto , Embarazo , Evaluación de Programas y Proyectos de Salud , Encuestas y CuestionariosRESUMEN
BACKGROUND: Naldemedine (S-297995) is a peripherally acting µ-opioid receptor antagonist developed as a once-daily oral drug for opioid-induced constipation (OIC) in adults with chronic noncancer or cancer pain. This study characterized the pharmacological effects of naldemedine in vitro and in vivo. METHODS: The binding affinity and antagonist activity of naldemedine against recombinant human µ-, δ-, and κ-opioid receptors were assayed in vitro. Pharmacologic effects of naldemedine were investigated using animal models of morphine-induced inhibition of small and large intestinal transit, castor oil-induced diarrhea, antinociception, and morphine withdrawal. KEY RESULTS: Naldemedine showed potent binding affinity and antagonist activities for recombinant human µ-, δ-, and κ-opioid receptors. Naldemedine significantly reduced opioid-induced inhibition of small intestinal transit (0.03-10 mg kg-1 ; P < 0.05) and large intestinal transit (0.3-1 µmol L-1 ; P < 0.05). Naldemedine (0.03-1 mg kg-1 ) pretreatment significantly reversed the inhibition of castor oil-induced diarrhea by subcutaneous morphine (P < 0.01). Naldemedine (1-30 mg kg-1 ) pretreatment (1 or 2 hours) did not alter the analgesic effects of morphine in a model measuring the latency of a rat to flick its tail following thermal stimulation. However, a significant delayed reduction of the analgesic effect of morphine was seen with higher doses of naldemedine (10-30 mg kg-1 ). Some centrally mediated and peripherally mediated withdrawal signs in morphine-dependent rats were seen with naldemedine doses ≥3 and ≥0.3 mg kg-1 , respectively. CONCLUSIONS & INFERENCES: Naldemedine displayed potent binding affinity to, and antagonistic activity against, µ-, δ-, and κ-opioid receptors. Naldemedine tempered OIC in vivo without compromising opioid analgesia.
Asunto(s)
Naltrexona/análogos & derivados , Antagonistas de Narcóticos/farmacología , Estreñimiento Inducido por Opioides , Animales , Motilidad Gastrointestinal/efectos de los fármacos , Humanos , Masculino , Naltrexona/farmacología , RatasRESUMEN
We investigated the mechanisms underlying the suppression of the rewarding effects of opioids using the femur bone cancer (FBC) mouse model. The rewarding and antinociceptive effects of subcutaneously administered morphine and oxycodone in the FBC model mice were assessed using the conditioned place preference test and the von-Frey test. In FBC mice, antinociceptive doses of morphine (30 mg/kg) and oxycodone (5 mg/kg) did not produce the rewarding effects but excessive doses of morphine (300 mg/kg) and oxycodone (100 mg/kg) did. Western blot analyses revealed a transient and significant increase in phosphorylated-extracellular regulated kinase (p-ERK) levels in ventral tegmental area (VTA) 5 min after the administration of morphine in sham-group. Interestingly, in FBC group, a regular dose of morphine did not increase p-ERK levels but a high dose of morphine caused an increase in p-ERK level 5 min after administration. The rewarding effects of a regular dose of and a high dose of morphine in the sham-operation and FBC model, respectively, were significantly inhibited by the MEK inhibitor. The suppression of p-ERK might result in resistance to these rewarding effects under the conditions of bone cancer.
Asunto(s)
Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Morfina/farmacología , Oxicodona/farmacología , Receptores Opioides mu/agonistas , Recompensa , Regulación hacia Arriba/efectos de los fármacos , Área Tegmental Ventral/efectos de los fármacos , Área Tegmental Ventral/enzimología , Analgésicos/farmacología , Animales , Butadienos/farmacología , Condicionamiento Psicológico/efectos de los fármacos , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Quinasas MAP Reguladas por Señal Extracelular/antagonistas & inhibidores , Ratones , Morfina/antagonistas & inhibidores , Nitrilos/farmacología , Oxicodona/antagonistas & inhibidores , Fosforilación/efectos de los fármacos , Ensayo de Unión Radioligante , Receptores Opioides mu/metabolismoRESUMEN
Although estrous cycle has been reported to influence antiociceptive effect of morphine in several pain conditions, its effect on cancer pain is not well established. We investigated the effect of estrogen on morphine antinociception using a bone cancer pain model and compared its potency with that of oxycodone. Female mice were ovariectomized (OVX) for preparation of a femur bone cancer pain (FBC) model. ß-estradiol was subcutaneously (s.c.) administered and antinociceptive effects of opioids was assessed using the von Frey monofilament test. Although morphine (5-20mg/kg, s.c.) did have significant antinociceptive effects in the FBC-OVX group, its effects in the FBC-OVX+ß-estradiol (OVX+E) group was limited. Oxycodone (1-5mg/kg, s.c.) exhibited significant effects in both groups. Expression changes in opioid-related genes (µ-, κ-, δ-opioid receptors, prodynorphin, proenkephalin, proopiomelanocortin) in the spinal and supraspinal sites were examined among the sham-OVX, sham-OVX+E, FBC-OVX, and FBC-OVX+E groups by in situ hybridization. These studies detected a significant increase in prodynorphin in the spinal dorsal horn of the FBC-OVX+E group. Spinal injection of a dynorphin-A antibody to FBC-OVX+E mice restored antinociception of morphine. In conclusion, we detected a differential effect of estrogen on morphine- and oxycodone-induced antinociception in a female FBC model. The effect of morphine was limited with estrogen exposure, which may be due to estrogen- and pain-mediated spinal expression of dynorphin-A.
Asunto(s)
Neoplasias Óseas/complicaciones , Estrógenos/farmacología , Fémur/efectos de los fármacos , Morfina/farmacología , Oxicodona/farmacología , Dolor/complicaciones , Dolor/tratamiento farmacológico , Analgésicos/farmacología , Analgésicos/uso terapéutico , Animales , Línea Celular Tumoral , Dinorfinas/genética , Ciclo Estral/efectos de los fármacos , Femenino , Regulación de la Expresión Génica/efectos de los fármacos , Masculino , Ratones , Morfina/uso terapéutico , Ovariectomía , Oxicodona/uso terapéutico , Dolor/genética , Dolor/fisiopatologíaRESUMEN
This study aimed to identify self-care strategies and assess physical and psychosocial factors associated with premenstrual distress among high school students. A cross-sectional survey of 217 adolescent girls aged 15 to 18 years was conducted in October 2009. Most (84.3 percent) had at least one or more symptoms of premenstrual distress. Premenstrual distress interfered with normal school activity in 51.2 percent. Most participants (57.1 percent) did not perform any self-care strategies for premenstrual distress. A hierarchical multiple linear regression analysis was conducted. Comprehension of one's own physical and mental states during premenstrual phases mediated the relationship between neuroticism and premenstrual distress. Activity restrictions due to menstrual distress mediated the relationship between the family's understanding of one's behavior during premenstrual phases and premenstrual distress. Findings suggest that, even if girls have neuroticism, it will be important to teach them to address the comprehension of one's own physical and mental states so that perceptions of both premenstruation and menstruation become more positive. Findings also suggest that the family's understanding was associated with alleviation of premenstrual distress. This study suggests the need for education to help adolescent girls and their families manage premenstrual distress and increase awareness of the benefit of managing its associated symptoms.
Asunto(s)
Pueblo Asiatico/psicología , Síndrome Premenstrual/psicología , Autocuidado , Estrés Psicológico/complicaciones , Estudiantes/psicología , Adolescente , Adulto , Pueblo Asiatico/estadística & datos numéricos , Concienciación , Estudios Transversales , Femenino , Educación en Salud , Conocimientos, Actitudes y Práctica en Salud , Humanos , Japón , Menstruación/psicología , Estudios Retrospectivos , Instituciones Académicas , Apoyo Social , Encuestas y CuestionariosRESUMEN
BACKGROUND AND PURPOSE: We demonstrated previously that oxycodone has potent antinociceptive effects at supraspinal sites. In this study, we investigated changes in neuronal function and antinociceptive mechanisms of oxycodone at ventrolateral periaqueductal gray (VLPAG) neurons, which are a major site of opioid action, in a femur bone cancer (FBC) model with bone cancer-related pain. EXPERIMENTAL APPROACH: We characterized the supraspinal antinociceptive profiles of oxycodone and morphine on mechanical hypersensitivity in the FBC model. Based on the disinhibition mechanism underlying supraspinal opioid antinociception, the effects of oxycodone and morphine on GABAA receptor-mediated inhibitory postsynaptic currents (IPSCs) in VLPAG neurons were evaluated in slices from the FBC model. KEY RESULTS: The supraspinal antinociceptive effects of oxycodone, but not morphine, were abolished by blocking G protein-gated inwardly rectifying potassium1 (Kir 3.1) channels. In slices from the FBC model, GABAergic synaptic transmission at VLPAG neurons was enhanced, as indicated by a leftward shift of the input-output relationship curve of evoked IPSCs, the increased paired-pulse facilitation and the enhancement of miniature IPSC frequency. Following treatment with oxycodone and morphine, IPSCs were reduced in the FBC model, and the inhibition of presynaptic GABA release by oxycodone, but not morphine was enhanced and dependent on Kir 3.1 channels. CONCLUSION AND IMPLICATIONS: Our results demonstrate that Kir 3.1 channels are important for supraspinal antinociception and presynaptic GABA release inhibition by oxycodone in the FBC model. Enhanced GABAergic synaptic transmission at VLPAG neurons in the FBC model is an important site of supraspinal antinociception by oxycodone via Kir 3.1 channel activation.
Asunto(s)
Analgésicos Opioides/farmacología , Canales de Potasio Rectificados Internamente Asociados a la Proteína G/fisiología , Neuronas/efectos de los fármacos , Oxicodona/farmacología , Sustancia Gris Periacueductal/efectos de los fármacos , Ácido gamma-Aminobutírico/fisiología , Analgésicos Opioides/uso terapéutico , Animales , Neoplasias Óseas/tratamiento farmacológico , Neoplasias Óseas/fisiopatología , Línea Celular Tumoral , Canales de Potasio Rectificados Internamente Asociados a la Proteína G/antagonistas & inhibidores , Hiperalgesia/tratamiento farmacológico , Potenciales Postsinápticos Inhibidores/efectos de los fármacos , Masculino , Ratones Endogámicos C3H , Morfina/farmacología , Morfina/uso terapéutico , Neuronas/fisiología , Oxicodona/uso terapéutico , Dolor/tratamiento farmacológico , Dolor/fisiopatología , Sustancia Gris Periacueductal/fisiología , Transmisión Sináptica/efectos de los fármacosRESUMEN
In temperate zones, animals restrict breeding to specific seasons to maximize the survival of their offspring. Birds have evolved highly sophisticated mechanisms of seasonal regulation, and their testicular mass can change 100-fold within a few weeks. Recent studies on Japanese quail revealed that seasonal gonadal development is regulated by central thyroid hormone activation within the hypothalamus, depending on the photoperiodic changes. By contrast, the mechanisms underlying seasonal testicular regression remain unclear. Here we show the effects of short day and low temperature on testicular regression in quail. Low temperature stimulus accelerated short day-induced testicular regression by shutting down the hypothalamus-pituitary-gonadal axis and inducing meiotic arrest and germ cell apoptosis. Induction of T3 coincided with the climax of testicular regression. Temporal gene expression analysis over the course of apoptosis revealed the suppression of LH response genes and activation of T3 response genes involved in amphibian metamorphosis within the testis. Daily ip administration of T3 mimicked the effects of low temperature stimulus on germ cell apoptosis and testicular mass. Although type 2 deiodinase, a thyroid hormone-activating enzyme, in the brown adipose tissue generates circulating T3 under low-temperature conditions in mammals, there is no distinct brown adipose tissue in birds. In birds, type 2 deiodinase is induced by low temperature exclusively in the liver, which appears to be caused by increased food consumption. We conclude that birds use low temperature-induced circulating T3 not only for adaptive thermoregulation but also to trigger apoptosis to accelerate seasonal testicular regression.
Asunto(s)
Frío , Coturnix/fisiología , Fotoperiodo , Testículo/fisiología , Triyodotironina/sangre , Animales , Apoptosis , Ayuno/metabolismo , Regulación de la Expresión Génica , Yoduro Peroxidasa/metabolismo , Hígado/enzimología , Hormona Luteinizante/metabolismo , Masculino , Meiosis , Espermatozoides/fisiología , Yodotironina Deyodinasa Tipo IIRESUMEN
Thyroid-stimulating hormone (TSH; thyrotropin) is a glycoprotein secreted from the pituitary gland. Pars distalis-derived TSH (PD-TSH) stimulates the thyroid gland to produce thyroid hormones (THs), whereas pars tuberalis-derived TSH (PT-TSH) acts on the hypothalamus to regulate seasonal physiology and behavior. However, it had not been clear how these two TSHs avoid functional crosstalk. Here, we show that this regulation is mediated by tissue-specific glycosylation. Although PT-TSH is released into the circulation, it does not stimulate the thyroid gland. PD-TSH is known to have sulfated biantennary N-glycans, and sulfated TSH is rapidly metabolized in the liver. In contrast, PT-TSH has sialylated multibranched N-glycans; in the circulation, it forms the macro-TSH complex with immunoglobulin or albumin, resulting in the loss of its bioactivity. Glycosylation is fundamental to a wide range of biological processes. This report demonstrates its involvement in preventing functional crosstalk of signaling molecules in the body.
Asunto(s)
Especificidad de Órganos , Procesamiento Proteico-Postraduccional , Tirotropina/metabolismo , Albúminas/metabolismo , Animales , Glicosilación , Inmunoglobulina G/metabolismo , Ratones Endogámicos C57BL , Adenohipófisis/metabolismo , Tirotropina/sangre , Hormona Liberadora de Tirotropina/metabolismoRESUMEN
A system of self-sustained biological clocks controls the 24-h rhythms of behavioral and physiological processes such as the sleep-wake cycle. The circadian clock system is regulated by transcriptional and translational negative feedback loops of multiple clock genes. Polymorphisms in circadian clock genes have been associated with morningness-eveningness (diurnal) preference, familial advanced sleep phase type (ASPT), and delayed sleep phase type (DSPT). We genotyped single-nucleotide polymorphisms in circadian clock genes in 182 DSPT individuals, 67 free-running type (FRT) individuals, and 925 controls. The clock gene polymorphisms were tested for associations with diurnal preference and circadian rhythm sleep disorder (CRSD) phenotypes. The PER3 polymorphism (rs228697) was significantly associated with diurnal preference and the FRT phenotype. The minor allele of rs228697 was more prevalent in evening types than in morning types (sex-adjusted odds ratio (OR), 2.483, Bonferroni-corrected P = 0.012) and in FRT individuals compared with the controls (age- and sex-adjusted OR, 2.021, permutated P = 0.017). Our findings support the notion that PER3 polymorphisms could be a potential genetic marker for an individual's circadian and sleep phenotypes.
Asunto(s)
Proteínas CLOCK/genética , Relojes Circadianos/genética , Ritmo Circadiano/genética , Proteínas Circadianas Period/genética , Trastornos del Sueño del Ritmo Circadiano/genética , Factores de Transcripción ARNTL/genética , Adulto , Alelos , Quinasa de la Caseína I/genética , Proteínas de Ciclo Celular/genética , Relojes Circadianos/fisiología , Ritmo Circadiano/fisiología , Criptocromos/genética , Femenino , Frecuencia de los Genes/genética , Marcadores Genéticos/genética , Predisposición Genética a la Enfermedad , Humanos , Péptidos y Proteínas de Señalización Intracelular/genética , Masculino , Polimorfismo de Nucleótido Simple , Sueño/fisiologíaRESUMEN
Although norepinephrine transporter (NET) inhibition has an additional effect on µ-opioid receptor (MOR)-mediated anti-nociception in inflammatory and neuropathic pain, its effect on cancer pain is not well characterized. We investigated the additional effect of NET inhibition on MOR activation using a mouse femur bone cancer (FBC) pain model by comparing the anti-nociceptive effect of the dual-acting opioids tramadol and tapentadol and the clinically used MOR-targeted opioids oxycodone and morphine. The anti-nociceptive effects of subcutaneously administered opioids were assessed using the von-Frey filament test. Oxycodone (1 - 10 mg/kg) and morphine (5 - 50 mg/kg) dose-dependently exhibited potent anti-nociceptive effects, whereas tramadol (10 - 56 mg/kg) and tapentadol (10 - 30 mg/kg) exhibited partial effects. Rota-rod analyses of tapentadol at a higher dose (> 30 mg/kg) showed a significant decrease in motor coordination, which was partially recovered by pretreatment with MOR or α(1)-adrenoceptor antagonists. The partial anti-nociceptive effect of tapentadol (30 mg/kg) was completely suppressed by a MOR antagonist, but not by α(1)- or α(2)-adrenoceptor antagonists, suggesting that neither α(1)-adrenoceptor- nor α(2)-adrenoceptor-mediated pathways are involved in anti-nociception in the FBC model. We conclude that addition of NET inhibition does not contribute to MOR-mediated anti-nociception in bone cancer pain.
Asunto(s)
Analgésicos Opioides/uso terapéutico , Neoplasias Óseas/complicaciones , Proteínas de Transporte de Noradrenalina a través de la Membrana Plasmática/antagonistas & inhibidores , Dolor/tratamiento farmacológico , Dolor/etiología , Receptores Opioides mu/metabolismo , Analgésicos Opioides/administración & dosificación , Analgésicos Opioides/farmacología , Animales , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Masculino , Ratones Endogámicos C3H , Morfina/administración & dosificación , Morfina/farmacología , Trasplante de Neoplasias , Oxicodona/administración & dosificación , Oxicodona/farmacología , Oxicodona/uso terapéutico , Fenoles/administración & dosificación , Fenoles/farmacología , Fenoles/uso terapéutico , Ratas Sprague-Dawley , Receptores Adrenérgicos alfa 1/fisiología , Receptores Adrenérgicos alfa 2/fisiología , Receptores Opioides mu/fisiología , Tapentadol , Tramadol/administración & dosificación , Tramadol/farmacología , Tramadol/uso terapéutico , Células Tumorales CultivadasRESUMEN
BACKGROUND AND PURPOSE: Oxycodone and morphine are µ-opioid receptor agonists prescribed to control moderate-to-severe pain. Previous studies suggested that these opioids exhibit different analgesic profiles. We hypothesized that distinct mechanisms mediate the differential effects of these two opioids and investigated the role of G protein-gated inwardly rectifying potassium (K(IR)3 also known as GIRK) channels in their antinociceptive effects. EXPERIMENTAL APPROACH: Opioid-induced antinociceptive effects were assessed in mice, using the tail-flick test, by i.c.v. and intrathecal (i.t.) administration of morphine and oxycodone, alone and following inhibition of K(IR)3.1 channels with tertiapin-Q (30 pmol per mouse, i.c.v. and i.t.) and K(IR)3.1-specific siRNA. The antinociceptive effects of oxycodone and morphine were also examined after tertiapin-Q administration in the mouse femur bone cancer and neuropathic pain models. KEY RESULTS: The antinociceptive effects of oxycodone, after both i.c.v. and i.t. administrations, were markedly attenuated by K(IR)3.1 channel inhibition. In contrast, the antinociceptive effects of i.c.v. morphine were unaffected, whereas those induced by i.t. morphine were attenuated, by K(IR)3.1 channel inhibition. In the two chronic pain models, the antinociceptive effects of s.c. oxycodone, but not morphine, were inhibited by supraspinal administration of tertiapin-Q. CONCLUSION AND IMPLICATIONS: These results demonstrate that K(IR)3.1 channels play a primary role in the antinociceptive effects of oxycodone, but not those of morphine, at supraspinal sites and suggest that supraspinal K(IR)3.1 channels are responsible for the unique analgesic profile of oxycodone.
Asunto(s)
Encéfalo/efectos de los fármacos , Dolor Crónico/prevención & control , Canales de Potasio Rectificados Internamente Asociados a la Proteína G/efectos de los fármacos , Morfina/farmacología , Narcóticos/farmacología , Neuralgia/prevención & control , Nocicepción/efectos de los fármacos , Oxicodona/farmacología , Animales , Encéfalo/metabolismo , Dolor Crónico/genética , Dolor Crónico/metabolismo , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Canales de Potasio Rectificados Internamente Asociados a la Proteína G/genética , Canales de Potasio Rectificados Internamente Asociados a la Proteína G/metabolismo , Inyecciones Intraventriculares , Inyecciones Espinales , Masculino , Ratones , Ratones Endogámicos C3H , Ratones Endogámicos C57BL , Morfina/administración & dosificación , Narcóticos/administración & dosificación , Neuralgia/genética , Neuralgia/metabolismo , Oxicodona/administración & dosificación , Dimensión del Dolor , Bloqueadores de los Canales de Potasio/farmacología , Interferencia de ARN , ARN Interferente Pequeño/metabolismo , Xenopus laevisRESUMEN
The pars tuberalis of the pituitary gland is the regulatory hub for seasonal reproduction in birds and mammals. Although fish also exhibit robust seasonal responses, they do not possess an anatomically distinct pars tuberalis. Here we report that the saccus vasculosus of fish is a seasonal sensor. We observe expression of key genes regulating seasonal reproduction and rhodopsin family genes in the saccus vasculosus of masu salmon. Immunohistochemical studies demonstrate that all of these genes are expressed in the coronet cells of the saccus vasculosus, suggesting the existence of a photoperiodic signalling pathway from light input to neuroendocrine output. In addition, isolated saccus vasculosus has the capacity to respond to photoperiodic signals, and its removal abolishes photoperiodic response of the gonad. Although the physiological role of the saccus vasculosus has been a mystery for several centuries, our findings indicate that the saccus vasculosus acts as a sensor of seasonal changes in day length in fish.
Asunto(s)
Estructuras Animales/fisiología , Peces/anatomía & histología , Peces/fisiología , Fotoperiodo , Estaciones del Año , Estructuras Animales/citología , Estructuras Animales/ultraestructura , Animales , Autorradiografía , Encéfalo/metabolismo , Células Cultivadas , Peces/genética , Regulación de la Expresión Génica , Masculino , Familia de Multigenes , Reproducción/genética , Rodopsina/genética , Rodopsina/metabolismo , Transducción de Señal/genética , Testículo/crecimiento & desarrollo , Testículo/metabolismoRESUMEN
Because of the presence of sperm-storage tubules (SST) in the utero-vaginal junction (UVJ) in the oviduct, once ejaculated sperm have entered the female reproductive tract, they can survive for a prolonged time in domestic birds, although the specific mechanisms involved in the sperm uptake into, maintenance within, and controlled release from the SST remain to be elucidated. In this report, we provide evidence that progesterone triggers the release of the resident sperm from the SST in the UVJ. The ultrastructural observation of the SST indicated that the resident sperm are released from the SST around 20 h after oviposition. When laying birds were injected with progesterone, most of the sperm were released from the SST within 1 h of injection. In situ hybridization analyses demonstrated the presence of the transcripts of membrane progestin receptor α in the UVJ, and the translated proteins were detected in the UVJ extracts by Western blotting. Moreover, the number of secretory granules in the SST epithelial cells fluctuates during the ovulatory cycle, and the progesterone administration mimics this phenomena. A binding assay using [(3)H]-progesterone indicated the presence of a high affinity, limited capacity, saturable and single binding site for [(3)H]-progesterone in the membrane fraction of the UVJ, and this receptor did not interact with the synthetic antiprogestin RU486. These results demonstrated for the first time that the progesterone stimulates the release of the resident sperm from the SST and that the release of the sperm might occur via membrane progestin receptor α-mediating signal transduction.
Asunto(s)
Oviductos/fisiología , Progesterona/fisiología , Espermatozoides/metabolismo , Animales , Coturnix , Femenino , Masculino , Ovulación , Receptores de Progesterona/fisiologíaRESUMEN
The avian perivitelline layer, an extracellular matrix homologous to the zona pellucida (ZP) of mammalian oocytes, is composed mainly by zona pellucida gene family glycoproteins. Our previous studies in Japanese quail have demonstrated that the matrix's components, ZP3 and ZPD, are synthesized in ovarian granulosa cells. Another component, ZP1, is synthesized in the liver. Recently, we demonstrated that another minor constituent, ZP2 is produced in the oocytes of the immature follicles. In the present study, we report the isolation of complementary DNA encoding quail ZP4 and its expression and origin in the female birds. By ribonuclease protection assay and in situ hybridization, we demonstrated that ZP4 transcripts were transcribed in the oocytes of small white follicles. The expression level of ZP4 decreased dramatically during follicular development, and the highest expression was observed in the small white follicles. Western blot analysis using the specific antibody against ZP4 indicated that the immunoreactive 58.2 kDa protein was present in the lysates of the small white follicles. These results demonstrate for the first time that the avian ZP4 is expressed in the oocyte, and that the expression pattern of the gene is similar to that of ZP2.
Asunto(s)
Coturnix/metabolismo , Proteínas del Huevo/análisis , Glicoproteínas de Membrana/análisis , Oocitos/química , Receptores de Superficie Celular/análisis , Animales , Western Blotting , Coturnix/genética , ADN Complementario/análisis , Proteínas del Huevo/genética , Proteínas del Huevo/inmunología , Electroforesis en Gel de Poliacrilamida , Femenino , Hibridación in Situ , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/inmunología , ARN Mensajero/análisis , Receptores de Superficie Celular/genética , Receptores de Superficie Celular/inmunología , Transcripción Genética , Glicoproteínas de la Zona PelúcidaRESUMEN
To investigate the genetic characteristics of the ancient populations of Hokkaido, northern Japan, polymorphisms of the ABO blood group gene were analyzed for 17 Jomon/Epi-Jomon specimens and 15 Okhotsk specimens using amplified product-length polymorphism and restriction fragment length polymorphism analyses. Five ABO alleles were identified from the Jomon/ Epi-Jomon and Okhotsk people. Allele frequencies of the Jomon/Epi-Jomon and Okhotsk people were compared with those of the modern Asian, European and Oceanic populations. The genetic relationships inferred from principal component analyses indicated that both Jomon/Epi-Jomon and Okhotsk people are included in the same group as modern Asian populations. However, the genetic characteristics of these ancient populations in Hokkaido were significantly different from each other, which is in agreement with the conclusions from mitochondrial DNA and ABCC11 gene analyses that were previously reported.
Asunto(s)
Sistema del Grupo Sanguíneo ABO/genética , Pueblo Asiatico/genética , Etnicidad/genética , Frecuencia de los Genes , Polimorfismo Genético , Alelos , Arqueología , Historia Antigua , Humanos , Japón , Análisis de Componente PrincipalRESUMEN
It has been known for many decades that nonmammalian vertebrates detect light by deep brain photoreceptors that lie outside the retina and pineal organ to regulate seasonal cycle of reproduction. However, the identity of these photoreceptors has so far remained unclear. Here we report that Opsin 5 is a deep brain photoreceptive molecule in the quail brain. Expression analysis of members of the opsin superfamily identified as Opsin 5 (OPN5; also known as Gpr136, Neuropsin, PGR12, and TMEM13) mRNA in the paraventricular organ (PVO), an area long believed to be capable of phototransduction. Immunohistochemistry identified Opsin 5 in neurons that contact the cerebrospinal fluid in the PVO, as well as fibers extending to the external zone of the median eminence adjacent to the pars tuberalis of the pituitary gland, which translates photoperiodic information into neuroendocrine responses. Heterologous expression of Opsin 5 in Xenopus oocytes resulted in light-dependent activation of membrane currents, the action spectrum of which showed peak sensitivity (lambda(max)) at approximately 420 nm. We also found that short-wavelength light, i.e., between UV-B and blue light, induced photoperiodic responses in eye-patched, pinealectomized quail. Thus, Opsin 5 appears to be one of the deep brain photoreceptive molecules that regulates seasonal reproduction in birds.
Asunto(s)
Proteínas Aviares/fisiología , Encéfalo/fisiología , Coturnix/fisiología , Proteínas del Tejido Nervioso/fisiología , Opsinas/fisiología , Células Fotorreceptoras de Vertebrados/fisiología , Secuencia de Aminoácidos , Animales , Proteínas Aviares/genética , Encéfalo/anatomía & histología , Coturnix/anatomía & histología , Coturnix/genética , Femenino , Técnicas In Vitro , Masculino , Eminencia Media/anatomía & histología , Eminencia Media/fisiología , Modelos Neurológicos , Datos de Secuencia Molecular , Proteínas del Tejido Nervioso/genética , Vías Nerviosas/anatomía & histología , Vías Nerviosas/fisiología , Oocitos/metabolismo , Opsinas/genética , Núcleo Hipotalámico Paraventricular/anatomía & histología , Núcleo Hipotalámico Paraventricular/fisiología , Estimulación Luminosa , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Testículo/crecimiento & desarrollo , Xenopus laevisRESUMEN
The avian perivitelline layer (PL), a vestment homologous to the zona pellucida (ZP) of mammalian oocytes, is composed of at least three glycoproteins. Our previous studies have demonstrated that the matrix's components, ZP3 and ZPD, are synthesized in ovarian granulosa cells. Another component, ZP1, is synthesized in the liver and is transported to the ovary by blood circulation. In this study, we report the isolation of cDNA encoding quail ZP2 and its expression in the female bird. By RNase protection assay and in situ hybridization, we demonstrate that ZP2 transcripts are restricted to the oocytes of small white follicles (SWF). The expression level of ZP2 decreased dramatically during follicular development, and the highest expression was observed in the SWF. Western blot and immunohistochemical analyses using the specific antibody against ZP2 indicate that the 80 kDa protein is the authentic ZP2, and the immunoreactive ZP2 protein is also present in the oocytes. Moreover, ultrastructural analysis demonstrated that the immunoreactive ZP2 localizes to the zona radiata, the perivitelline space, and the oocyte cytoplasm in the SWF. By means of western blot analysis and immunofluorescence microscopy, we detected a possible interaction of the recombinant ZP2 with ZP3 and that this interaction might lead to the formation of amorphous structure on the cell surface. These results demonstrate for the first time that the avian ZP gene is expressed in the oocyte, and that the ZP2 protein in the oocyte might play a role for the PL formation in the immature follicles of the ovary.
Asunto(s)
Coturnix/metabolismo , Proteínas del Huevo/metabolismo , Glicoproteínas de Membrana/metabolismo , Oocitos/metabolismo , Receptores de Superficie Celular/metabolismo , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Western Blotting , Células CHO , Clonación Molecular , Coturnix/genética , Cricetinae , Cricetulus , Proteínas del Huevo/genética , Femenino , Regulación del Desarrollo de la Expresión Génica , Inmunohistoquímica , Hibridación in Situ , Glicoproteínas de Membrana/genética , Microscopía Fluorescente , Datos de Secuencia Molecular , Peso Molecular , Oocitos/ultraestructura , ARN Mensajero/metabolismo , Receptores de Superficie Celular/genética , Proteínas Recombinantes/metabolismo , Transfección , Glicoproteínas de la Zona PelúcidaRESUMEN
Human earwax is classified into wet and dry types, which are determined by a single-nucleotide polymorphism in the adenosine triphosphate-binding cassette, sub-family C11 (ABCC11) gene locus. To investigate the allele frequencies of the ABCC11 locus within ancient populations on the Northern Japanese island of Hokkaido, amplified product-length polymorphisms were analyzed for 50 specimens of the Okhotsk people and 35 specimens of the Jomon and Epi-Jomon people excavated from various archaeological sites of Hokkaido. Of these specimens, 31 Okhotsk and 19 Jomon/Epi-Jomon samples were genotyped successfully. Frequencies of the wet-type allele in the Jomon/Epi-Jomon people, considered a major ancestor of the Ainu, were higher than those of other Northeastern Asian populations, including the modern Ainu. By contrast, in the Okhotsk people, believed to originate from East Siberia, frequencies of the dry-type allele were relatively higher than those in the Ainu and Jomon/Epi-Jomon people. These results suggest that gene flow from the Northeastern Asian Continent to descendants of the Jomon/Epi-Jomon people of Hokkaido through the Okhotsk people occurred, resulting in the establishment of the Ainu.
Asunto(s)
Transportadoras de Casetes de Unión a ATP/genética , Pueblo Asiatico/genética , Cerumen/metabolismo , Fósiles , Frecuencia de los Genes , Alelos , Arqueología , Genotipo , Geografía , Humanos , Japón , FenotipoRESUMEN
Photoperiodism is an adaptation mechanism that enables animals to predict seasonal changes in the environment. Japanese quail is the best model organism for studying photoperiodism. Although the recent availability of chicken genome sequences has permitted the expansion from single gene to genome-wide transcriptional analysis in this organism, the photoperiodic response of the domestic chicken is less robust than that of the quail. Therefore, in the present study, we examined the photoperiodic response of the red jungle fowl (Gallus gallus), a predecessor of the domestic chicken, to test whether this animal could be developed as an ideal model for studying the molecular mechanisms of seasonal reproduction. When red jungle fowls were transferred from short-day- to long-day conditions, gonadal development and an increase in plasma LH concentration were observed. Furthermore, rapid induction of thyrotropin beta subunit, a master regulator of photoperiodism, was observed at 16 h after dawn on the first long day. In addition, the long-day condition induced the expression of type 2 deiodinase, the key output gene of photoperiodism. These results were consistent with the results obtained in quail and suggest that the red jungle fowl could be an ideal model animal for the genome-wide transcriptional analysis of photoperiodism.