RESUMEN
The bone-derived hormone fibroblast growth factor-23 (FGF23) has recently received much attention due to its association with chronic kidney disease and cardiovascular disease progression. Extracellular sodium concentration ([Na+]) plays a significant role in bone metabolism. Hyponatremia (lower serum [Na+]) has recently been shown to be independently associated with FGF23 levels in patients with chronic systolic heart failure. However, nothing is known about the direct impact of [Na+] on FGF23 production. Here, we show that an elevated [Na+] (+20 mM) suppressed FGF23 formation, whereas low [Na+] (-20 mM) increased FGF23 synthesis in the osteoblast-like cell lines UMR-106 and MC3T3-E1. Similar bidirectional changes in FGF23 abundance were observed when osmolality was altered by mannitol but not by urea, suggesting a role of tonicity in FGF23 formation. Moreover, these changes in FGF23 were inversely proportional to the expression of NFAT5 (nuclear factor of activated T cells-5), a transcription factor responsible for tonicity-mediated cellular adaptations. Furthermore, arginine vasopressin, which is often responsible for hyponatremia, did not affect FGF23 production. Next, we performed a comprehensive and unbiased RNA-seq analysis of UMR-106 cells exposed to low versus high [Na+], which revealed several novel genes involved in cellular adaptation to altered tonicity. Additional analysis of cells with Crisp-Cas9-mediated NFAT5 deletion indicated that NFAT5 controls numerous genes associated with FGF23 synthesis, thereby confirming its role in [Na+]-mediated FGF23 regulation. In line with these in vitro observations, we found that hyponatremia patients have higher FGF23 levels. Our results suggest that [Na+] is a critical regulator of FGF23 synthesis.
Asunto(s)
Factor-23 de Crecimiento de Fibroblastos , Sodio , Humanos , Factor-23 de Crecimiento de Fibroblastos/genética , Factor-23 de Crecimiento de Fibroblastos/metabolismo , Hiponatremia/fisiopatología , Insuficiencia Renal Crónica/fisiopatología , Sodio/metabolismo , Sodio/farmacología , Línea Celular Tumoral , Línea Celular , Animales , Ratones , Ratones Endogámicos C57BL , Arginina Vasopresina/metabolismo , Osteoblastos/citología , Osteoblastos/efectos de los fármacos , Osteoblastos/metabolismo , Factores de Transcripción NFATC/genética , Factores de Transcripción NFATC/metabolismo , RatasRESUMEN
In mammalian reproduction, sexually active males seek female conspecifics, while estrous females try to approach males. This sex-specific response tendency is called sexual preference. In small rodents, sexual preference cues are mainly chemosensory signals, including pheromones. In this article, we review the physiological mechanisms involved in sexual preference for opposite-sex chemosensory signals in well-studied laboratory rodents, mice, rats, and hamsters of both sexes, especially an overview of peripheral sensory receptors, and hormonal and central regulation. In the hormonal regulation section, we discuss potential rodent brain bisexuality, as it includes neural substrates controlling both masculine and feminine sexual preferences, i.e., masculine preference for female odors and the opposite. In the central regulation section, we show the substantial circuit regulating sexual preference and also the influence of sexual experience that innate attractants activate in the brain reward system to establish the learned attractant. Finally, we review the regulation of sexual preference by neuropeptides, oxytocin, vasopressin, and kisspeptin. Through this review, we clarified the contradictions and deficiencies in our current knowledge on the neuroendocrine regulation of sexual preference and sought to present problems requiring further study.
Asunto(s)
Células Quimiorreceptoras/fisiología , Vías Nerviosas/fisiología , Atractivos Sexuales/farmacología , Conducta Sexual Animal/fisiología , Animales , Células Quimiorreceptoras/efectos de los fármacos , Femenino , Masculino , Vías Nerviosas/efectos de los fármacos , Conducta Sexual Animal/efectos de los fármacosRESUMEN
Male sexual function in mammals is controlled by the brain neural circuits and the spinal cord centers located in the lamina X of the lumbar spinal cord (L3-L4). Recently, we reported that hypothalamic oxytocin neurons project to the lumbar spinal cord to activate the neurons located in the dorsal lamina X of the lumbar spinal cord (dXL) via oxytocin receptors, thereby facilitating male sexual activity. Sexual experiences can influence male sexual activity in rats. However, how this experience affects the brain-spinal cord neural circuits underlying male sexual activity remains unknown. Focusing on dXL neurons that are innervated by hypothalamic oxytocinergic neurons controlling male sexual function, we examined whether sexual experience affects such neural circuits. We found that >50% of dXL neurons were activated in the first ejaculation group and ~30% in the control and intromission groups in sexually naïve males. In contrast, in sexually experienced males, ~50% of dXL neurons were activated in both the intromission and ejaculation groups, compared to ~30% in the control group. Furthermore, sexual experience induced expressions of gastrin-releasing peptide and oxytocin receptors in the lumbar spinal cord. This is the first demonstration of the effects of sexual experience on molecular expressions in the neural circuits controlling male sexual activity in the spinal cord.
Asunto(s)
Eyaculación , Péptido Liberador de Gastrina/biosíntesis , Regulación de la Expresión Génica , Erección Peniana , Receptores de Oxitocina/biosíntesis , Médula Espinal/metabolismo , Animales , Femenino , Masculino , Ratas , Ratas Long-EvansRESUMEN
Exposure to endocrine-disrupting chemicals may adversely affect animals, particularly during development. Tris(1,3-dichloroisopropyl) phosphate (TDCIPP) is an organophosphate with anti-androgen function in vitro that is present in indoor dust at relatively high concentrations. In male rats, androgens are necessary for the development of reproductive organs, as well as the endocrine and central nervous systems. However, we currently do not know the exact effects of TDCIPP exposure through suckling on subsequent reproductive behavior in males. Here, we show that TDCIPP exposure (25-250 mg kg-1 via oral administration over 28 consecutive days post-birth) suppressed male sexual behavior and reduced testes size. These changes were dose-dependent and appeared first in adults rather than in juveniles. These results demonstrate that TDCIPP exposure led to normal body growth and appearance in juveniles, but disrupted the endocrine system and physiology in adults. Therefore, assays should be performed using adult animals to ensure accuracy, and to confirm the influence of chemical substances given during early mammalian life.
Asunto(s)
Disruptores Endocrinos/toxicidad , Conducta Sexual Animal/efectos de los fármacos , Animales , Relación Dosis-Respuesta a Droga , Masculino , Ratas , Ratas Wistar , Testosterona/sangreRESUMEN
Correct perinatal oestrogen levels are critical for sexual differentiation. For example, perinatal exposure to oestrogen causes masculinization and defeminization of the brain in female rats and also induces delayed effects after maturation characterized by early onset of abnormal oestrus cycling. However, the mechanisms underlying the above effects of oestrogen remain to be fully determined. 17α-ethinyloestradiol (EE), a common synthetic oestrogen widely used in oral contraceptives, binds specifically to oestrogen receptors. In this study, we demonstrated the effects of a single neonatal injection of high- or low-dose EE on reproductive behaviours. Female rats within 24 h after birth were subcutaneously injected with sesame oil, EE (0.02, 2 mg kg-1 ) and 17ß-oestradiol (E2 ) (20 mg kg-1 ). Between 11 and 15 weeks of age, sexual behaviour was tested twice in a paced mating situation. Latency to enter, lordosis and soliciting behaviour were recorded. Both high-dose EE- and E2 -treated females showed a significantly lower lordosis quotient, decreased soliciting behaviours, increased rejection and fighting numbers. Accessibility to males was also delayed by neonatal E2 exposure, although it was shortened by high-dose EE exposure. In contrast, low-dose EE-treated females did not exhibit impaired sexual behaviour. These results suggest that single neonatal exposure to a high dose of EE or E2 disturbs the normal development of the female brain, resulting in impaired sexual behaviours in a female-paced mating situation. Besides, the differences noted between high-dose EE- and E2 -treated females might be caused by different affinities of the oestrogen receptors, metabolic rates or mechanisms of action. Copyright © 2017 John Wiley & Sons, Ltd.
Asunto(s)
Estrógenos/toxicidad , Etinilestradiol/toxicidad , Conducta Sexual Animal/efectos de los fármacos , Animales , Animales Recién Nacidos , Anticonceptivos Orales , Relación Dosis-Respuesta a Droga , Estrógenos/administración & dosificación , Etinilestradiol/administración & dosificación , Femenino , Inyecciones Subcutáneas , Masculino , Ratas WistarRESUMEN
In the rat, induction of maternal behavior depends on the parity of the female. For example, nulliparous (NP) females need longer exposure to pups than multiparous (MP) or lactating (L) females to exhibit similar maternal behavior. In this study, we investigated the role of brain oxytocin in the approaching behavior of these female rats. Olfactory preferences for pup odors were examined for 8 consecutive days. Each preference test was followed by direct overnight exposure to pups. On the 8th day, MP and L, but not NP females showed robust pup-odor preferences. After the behavioral test, half of the females were exposed to pups for 2 h, whereas the other half were not. The females were then sacrificed to analyze brain oxytocin (OXT) and vasopressin (AVP) activities by cFos immunohistochemistry and to quantify their receptor mRNA expression using real-time PCR. In the paraventricular nucleus (PVN), the percentage of cFos-positive OXT neurons was significantly larger in MP and L females than in NP females after pup exposure. No significant differences were found in cFos expression in OXT neurons of the supraoptic nucleus (SON) or in AVP neurons of either the PVN or SON. Expression of OXT receptor mRNA in the medial preoptic area and amygdala of the control groups was also higher in MP females than in NP females. Finally, we demonstrated that infusion of OXT into the lateral ventricle of NP females promoted preferences for pup odors. These results indicate that puerperal and parental experiences enhance the responsiveness of OXT neurons in the PVN to pup stimuli and establish olfactory preferences for these odors in a parity-dependent manner.
Asunto(s)
Odorantes , Oxitocina/fisiología , Animales , Conducta Animal , Encéfalo/metabolismo , Femenino , Humanos , Inmunohistoquímica , Lactancia , Conducta Materna , Neuronas/metabolismo , Núcleo Hipotalámico Paraventricular/metabolismo , Embarazo , Preñez , Proteínas Proto-Oncogénicas c-fos/fisiología , ARN Mensajero/metabolismo , Ratas , Ratas Long-Evans , Olfato , Núcleo Supraóptico/metabolismo , Vasopresinas/fisiologíaRESUMEN
Arginine vasopressin (AVP) and oxytocin (OT) are well-known as neuropeptides that regulate various social behaviors in mammals. However, little is known about their role in mouse female sexual behavior. Thus, we investigated the role of AVP (v1a and v1b) and OT receptors on female sexual behavior. First, we devised a new apparatus, the bilevel chamber, to accurately observe female mouse sexual behavior. This apparatus allowed for a more precisely measurement of lordosis as receptivity and rejection-like behavior (newly defined in this study), a reversed expression of proceptivity. To address our research question, we evaluated female sexual behavior in mice lacking v1a (aKO), v1b (bKO), both v1a and v1b (dKO), and OT (OTRKO) receptors. aKO females showed decreased rejection-like behavior but a normal level of lordosis, whereas bKO females showed almost no lordosis and no change in rejection-like behavior. In addition, dKO females showed normal lordosis levels, suggesting that the v1b receptor promotes lordosis, but not necessarily, while the v1a receptor latently suppresses it. In contrast, although OTRKO did not influence lordosis, it significantly increased rejection-like behavior. In summary, the present results demonstrated that the v1a receptor inhibits proceptivity and receptivity, whereas the v1b and OT receptors facilitate receptivity and proceptivity, respectively.
Asunto(s)
Ratones Noqueados , Receptores de Oxitocina , Receptores de Vasopresinas , Conducta Sexual Animal , Animales , Femenino , Receptores de Vasopresinas/metabolismo , Receptores de Vasopresinas/genética , Receptores de Oxitocina/metabolismo , Receptores de Oxitocina/genética , Conducta Sexual Animal/fisiología , Ratones , Masculino , Oxitocina/metabolismo , Ratones Endogámicos C57BL , Arginina Vasopresina/metabolismoRESUMEN
Mutual attachment between mother and pup is important to enable the mother to care for her pup and for the pup to receive care from its mother. Pups eventually leave their mothers, which is also very important to their growth. The mechanism of preference by which pups transfer attachment from their mother to others remains unknown. In this study, we assessed mother/novel dam preferences and examined the brain regions associated with the regulation of this preference in C57BL/6 mice pups. We found that C57BL/6 mice pups had variety in their mother/novel dam preferences at 16 days old. This variety was not related to the sex of the pups, their weight, or the litter size. In order to clarify the brain mechanisms responsible for this variety, we examined the relationship between mother/novel dam preference and neuronal activation induced by contact with the mother. We found that pups exhibiting novel dam preference had higher neural activity in the anterior cingulate cortex (ACC) and bed nucleus of the stria terminalis (BNST) when exposed to their mother. These results suggest that ACC and/or BNST neural activity may be associated with mother/novel dam preferences in infant mice.
Asunto(s)
Conducta Materna , Madres , Animales , Encéfalo , Femenino , Humanos , Conducta Materna/fisiología , Ratones , Ratones Endogámicos C57BL , NeuronasRESUMEN
Both volatile and nonvolatile molecules are involved in chemosensory communication in rodents. Volatile odors from physically inaccessible estrous females induced increased numbers of c-Fos-positive cells in the preoptic area (POA) and in the cortical nucleus of the amygdala (CoA) of male rats. The numbers of c-Fos-positive cells in the medial nucleus of the amygdala (MeA) increased in response to the nonvolatile odors of bedding soiled with the excreta of estrous females. In an alternate choice paradigm, male rats carrying ibotenic acid lesions in either the MeA or the CoA--or a combination of both--distinguished the odors of estrous females from those of males, although the time spent sniffing the stimuli was diminished. Males with POA lesions showed complete loss of this capability. Males carrying either of the lesions did not detect differences between estrous and anestrous females or between intact and orchidectomized males. Lesions in the POA or MeA severely impaired male sexual behavior, whereas a CoA lesion had no effects. Thus, c-Fos-positive cells in the CoA might be involved in chemosensory transmission relevant to certain social contexts, but not in the execution of male sexual behavior. The POA is indispensable for both olfactory preferences and sexual behavior. The residual olfactory preference in males with MeA or CoA lesions or the combination of both could reflect an additional route for chemosensory transmission from the main olfactory bulb to the POA.
Asunto(s)
Amígdala del Cerebelo/fisiología , Conducta de Elección/fisiología , Vías Olfatorias/fisiología , Área Preóptica/fisiología , Conducta Sexual Animal/fisiología , Análisis de Varianza , Animales , Ciclo Estral , Femenino , Inmunohistoquímica , Masculino , Odorantes , Bulbo Olfatorio/fisiología , Proteínas Proto-Oncogénicas c-fos/metabolismo , Ratas , Ratas Long-EvansRESUMEN
Sexually experienced male rats show an olfactory preference for estrous female odor compared to male odor. Notably, they also prefer castrated male over gonadally intact male odor. This study examined the role of elevated circulating gonadotropins and gonadotropin-releasing hormone (GnRH) induced by disinhibition of the hypothalamus-pituitary-gonad axis in the castration-induced attractiveness of male rats. Experiment I compared preference of sexually experienced males between odors of castrated males injected with a GnRH antagonist (Cast+Ant), castrated males injected with saline (Cast), and gonadally intact males (Sham). These olfactory preference tests revealed that probe males significantly preferred Cast over Sham and Cast+Ant odors but had no preference between Sham and Cast+Ant odors. Experiment II used hypophysectomy (HPx) to remove the gonadotropin source in place of pharmacologically antagonizing GnRH. Similarly, the probe males preferred Cast (high both GnRH and gonadotropins) over HPx (high GnRH but no gonadotropin) odors, but also preferred HPx than Sham (low both GnRH and gonadotropins) odors. We then examined the effects of exogenous gonadotropins, human chorionic gonadotropin (hCG) and equine chorionic gonadotropin (eCG), to reveal which gonadotropins critically affect on the attractiveness of HPx males. The olfactory preference tests revealed that injecting eCG, but not hCG, significantly increased the probe males` preference compared to HPx male odor, suggesting that the attractiveness is augmented via follicle stimulating hormone (FSH) receptor rather than luteinizing hormone receptor. These results indicate that sexually active male rats show olfactory preference for castrated male rats due to elevated GnRH and FSH blood levels produced by a lack of androgens.
Asunto(s)
Hormona Folículo Estimulante , Hormona Luteinizante , Animales , Femenino , Hormona Liberadora de Gonadotropina/metabolismo , Gónadas , Caballos , Hipotálamo/metabolismo , Masculino , Odorantes , Hipófisis/metabolismo , RatasRESUMEN
Among the intact male rats, a subpopulation has been found to show little or no sexual behavior, even after experiencing several mating sessions. This study investigated whether sexually sluggish (SS) males show behavioral differences from normal copulatory (NC) males, other than those concerning sexual behavior. The olfactory preference of males was measured through the time spent displaying nose-poking behavior directed at sexually active males and estrous females for odor exploration in a three-chamber apparatus. Both the NC and SS males showed a significant preference for the odor of estrous females compared with that of male odors. However, SS males spent significantly less time nose-poking estrous females than NC males. The food-finding test was performed after overnight fasting. Our findings showed that all the NC males found the buried pellet within 5 min, whereas over 60% of the SS males failed to find it. The males were also tested for their ability to find a buried bag containing soiled bedding from estrous female cages. The bag was found by 80% of NC males, but only by 20% of SS males. Our results suggest that SS and NC male rats differ not only in sexual behavior but also in other functions such as olfaction.
Asunto(s)
Copulación/fisiología , Odorantes , Ratas Long-Evans/psicología , Conducta Sexual Animal/fisiología , Olfato/fisiología , Animales , Ciclo Estral , Femenino , Humanos , MasculinoRESUMEN
Oxytocin (OT) has been demonstrated to be involved in various social behaviors in mammals. However, OT gene knockout (OTKO) mice can conceive and deliver successfully, though females cannot rear their pups because of lack of lactation. Here, we investigated the sociosexual behavior of both sexes in two experimental setups: olfactory preference for sexual partner's odor and direct social interaction in an enriched condition. In the preference test, mice were given a choice of two airborne odors derived from intact male and receptive female mice, or from intact or castrated male mice. Wild-type (WT) mice significantly preferred opposite-sex odors, whereas OTKO mice showed vigorous but equivalent exploration to all stimuli. In social interactions in the enriched condition, no difference in sexual behavior was found between WT and OTKO males. In contrast, WT female initiated sexual behavior at the second week test, while OTKO females required 4 weeks to receive successful mounts. Neuronal activation by odor stimulation was compared between WT and OTKO mice. The numbers of cFos-immunoreactive cells increased in the medial amygdala and the preoptic area after exposure to opposite-sex odors in WT mice, whereas the increase was suppressed in OTKO mice. We conclude that OT plays an important role in the regulation of olfactory-related social behavior in both male and female mice. The influence of OT was greater in female mice, especially during social interactions involving the acquisition of sexual experience.
Asunto(s)
Oxitocina/fisiología , Conducta Sexual Animal/fisiología , Amígdala del Cerebelo/metabolismo , Animales , Cognición , Complejo Nuclear Corticomedial , Femenino , Técnicas de Inactivación de Genes , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Oxitocina/deficiencia , Área Preóptica/metabolismo , Proteínas Proto-Oncogénicas c-fos/metabolismo , Conducta SocialRESUMEN
Although the involvement of two types of vasopressin (AVP) receptors, v1a and v1b, in neural regulation of social behavior is well documented in rodents, there is no report on combined actions of them in regulation of social behavior. In this study, we investigated behavioral differences between wild-type (WT) and v1a and v1b double knockout (dKO) mice. For this, we measured olfactory preference, sexual behavior with receptive females (four weekly tests) in an enriched large observation cage, and anxiety-like behaviors. No difference between WT and dKO mice was found in olfactory preferences for estrous female odor to male odor. Over all four mating tests, the number of mounts and pursuits after receptive females was significantly greater in dKO mice than in WT mice. In the elevated plus maze and the open field test, dKO mice showed lower anxiety-like behavior than WT mice. Finally, we measured approach behavior to several types of objects, figurines, and caged anestrous or estrous females placed in the open field apparatus. The only difference observed was that dKO mice spent longer in the vicinity of estrous females than did WT mice. These findings suggest that vasopressin receptors are involved in the regulation of sociosexual behavior, presumably partly mediated by emotional responses, in male mice.
Asunto(s)
Ansiedad/metabolismo , Receptores de Vasopresinas/metabolismo , Conducta Sexual Animal/fisiología , Animales , Conducta de Elección/fisiología , Ciclo Estral , Conducta Exploratoria/fisiología , Femenino , Masculino , Ratones Endogámicos C57BL , Ratones Noqueados , Percepción Olfatoria/fisiología , Receptores de Vasopresinas/genéticaRESUMEN
The central part of the medial preoptic nucleus (MPNc) is associated with sexual arousal induction in male rats. However, it is largely unclear how males are sexually aroused and achieve their first copulation. We previously reported that more MPNc neurons activate during the first copulation than the second copulation. In this study, to explore the molecules responsible for sexual arousal induction, we performed DNA microarray of the MPNc in sexually naive males and males after they copulated for their first and second times. We then performed quantitative PCR analyses to validate the results of the DNA microarray. Six genes were identified. Their expression increased following copulation and was higher in males after they copulated for the first time than after the second time. The genes encode transcription factors (Fos, Nfil3, and Nr4a3), a serine/threonine kinase (Sik1), an antioxidant protein (Srxn1), and a neuropeptide precursor VGF (Vgf), which may be the candidate genes responsible for sexual arousal induction. We examined the effects of Vgf knockdown in the MPNc on sexual partner preference and sexual behavior in sexually inexperienced and experienced males to determine the role of VGF in sexual arousal induction. A preference for estrous female rats was reinforced, and the latency of mount and intromission became short after sexually inexperienced males copulated for the first time. However, Vgf knockdown disrupted these phenomena. Vgf knockdown did not have any significant effect in sexually experienced males. VGF-derived neuropeptides presumably serve as an effector molecule to increase sexual activity following sexual arousal induction.
Asunto(s)
Nivel de Alerta/genética , Neuropéptidos/fisiología , Área Preóptica/metabolismo , Conducta Sexual Animal/fisiología , Animales , Copulación/fisiología , Femenino , Perfilación de la Expresión Génica , Técnicas de Silenciamiento del Gen , Masculino , Neuropéptidos/metabolismo , Análisis de Secuencia por Matrices de Oligonucleótidos , Ratas , Ratas Transgénicas , Ratas Wistar , Factores SexualesRESUMEN
In situ hybridization detected a transient, sex-specific transcription of somatostatin gene in the central part of the rat medial preoptic nucleus, which coincides with the sexually dimorphic nucleus of the preoptic area (SDN-POA), during, but not after, the establishment of sex difference. On postnatal d 1 (day of birth), somatostatin mRNA was detected in the SDN-POA of both sexes. On d 8 through 35, the area of somatostatin mRNA-positive cells was significantly larger in males than in females. In males the area attained its maximum size on d 15 and diminished gradually thereafter. In females the area did not change in size during this period. On d 60 expression of somatostatin mRNA was low and not different between sexes. Throughout the observed period, Nissl staining and calbindin immunohistochemistry enabled visualization of the typical SDN-POA in the same region. As with Nissl staining and calbindin immunohistochemistry, somatostatin mRNA hybridization on d 15 revealed a reversal of the sexual dimorphism in the size of the SDN-POA in males that had been neonatally orchidectomized or females given estrogen as pups, showing that sex steroid milieu during the organizational period determines the area occupied by somatostatin mRNA-positive cells. Sex-specific, transient transcription of the somatostatin gene may causally relate to the estrogen-dependent organization of the SDN-POA.
Asunto(s)
Estrógenos/farmacología , Área Preóptica/efectos de los fármacos , Área Preóptica/metabolismo , Somatostatina/genética , Transcripción Genética , Animales , Animales Recién Nacidos , Calbindinas , Femenino , Masculino , Embarazo , ARN Mensajero/metabolismo , Ratas , Ratas Endogámicas , Ratas Sprague-Dawley , Proteína G de Unión al Calcio S100/metabolismo , Caracteres Sexuales , Somatostatina/metabolismo , Factores de TiempoRESUMEN
The DNA extracted from a high-temperature environment in which micro-organisms are living will be a good source for the isolation of thermostable enzymes. Using a metagenomic approach, we aimed to isolate thermostable ß-xylosidases that will be exploited for biofuel production from lignocellulosic biomass. DNA samples obtained from the soil near a spout of a hot spring (70°C, pH7.2) were subjected to sequencing, which generated a total of 84.2 Gbp with 967,925 contigs of >500 bp in length. Similarity search for ß-xylosidase in the contigs revealed the presence of 168 candidate sequences, each of which may have arisen from more than one gene. Individual genes were amplified by PCR using sequence-specific primers. The resultant DNA fragments were cloned and introduced into Escherichia coli BL21 Star(DE3). Consequently, 269 proteins were successfully expressed in the E. coli cells and then examined for ß-xylosidase activity. A total of 82 proteins exhibited ß-xylosidase activity at 50°C, six of which retained the activity even at 90°C. Out of the six, three proteins were originated from a single candidate sequence, AR19M-311. An amino acid sequence comparison suggested the amino acid residues that appeared to be crucial for thermal stability of the enzymes.
Asunto(s)
Escherichia coli/enzimología , Manantiales de Aguas Termales/microbiología , Metagenómica/métodos , Microbiología del Suelo , Xilosidasas/genética , Xilosidasas/aislamiento & purificación , Secuencia de Aminoácidos , Sustitución de Aminoácidos , ADN Bacteriano/genética , Escherichia coli/genética , Regulación Bacteriana de la Expresión Génica , Manantiales de Aguas Termales/análisis , Filogenia , Homología de Secuencia , Xilosidasas/metabolismoRESUMEN
We previously reported that social isolation promotes parental care in sexually naïve male mice. This effect was blocked by exposure to chemosensory and auditory social signals derived from males in an adjacent compartment. In the present study, we examined whether the chemosensory signals detected in the vomeronasal organ (VNO) are involved in parental behavior by using mice deficient for a VNO-specific ion channel (Trpc2-/-) and thus impaired in VNO-input signaling. We housed virgin homozygous Trpc2-/- and heterozygous Trpc2± males for 3weeks during puberty (5-8weeks old) alone or in groups of 3-5 males. At 8weeks of age, the mice were placed with three pups in an observation cage and tested for parental behavior. The Trpc2-/- males housed under isolated conditions spent significantly longer in the vicinity of pups than did the Trpc2-/- males than had been group housed, whereas no isolation effect was observed in heterozygous Trpc2± males. Both Trpc2 knockout and isolation housing significantly increased the time males spent licking pups and crouching (arched back posture over pups to enable nursing), whereas only isolation housing increased the incidence of retrieval behavior. These results demonstrated that social signals transmitted not only through the VNO but also from other modalities, independent of each other, suppress the expression of parental behavior during puberty in sexually naïve males.
Asunto(s)
Conducta Paterna/fisiología , Aislamiento Social/psicología , Órgano Vomeronasal/fisiopatología , Análisis de Varianza , Animales , Animales Recién Nacidos , Femenino , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Estadísticas no Paramétricas , Canales Catiónicos TRPC/deficiencia , Canales Catiónicos TRPC/genéticaRESUMEN
Copulatory behavior has been reported to raise the pain threshold in male rats. In this study, we examined the effect of copulatory behavior with or without ejaculation on pain threshold measured by electrical shock via an electrode attached to the tail. It was demonstrated that ejaculation is not necessary to raise the pain threshold in male rats. In addition, we examined whether oxytocin, a hypothalamic neuropeptide, was involved in copulation-induced hypoalgesia. Sexually experienced males were subjected to stereotaxic implantation of a guide cannula targeting the lateral ventricle. After the recovery period, half of the males were intracerebroventricularly treated with an oxytocin antagonist (OTA, 100ng d(CH2)51,Tyr(Me)2,Thr4, Orn8,Tyr-NH29]-vasotocin/1µL saline) and the remaining half were administered saline without anesthesia. Fifteen minutes later, half of each group were given sexual behavior with receptive females. We found no effect of OTA on sexual activity. Immediately after ejaculation, pain threshold was measured. While raised pain threshold was observed after sexual behavior in saline-treated males, no change in pain threshold was found in OTA-treated males even after copulation. The results suggest that central oxytocin mediates copulation-induced hypoalgesia in male rats.
Asunto(s)
Copulación , Oxitocina/fisiología , Dolor/fisiopatología , Animales , Eyaculación , Femenino , Inyecciones Intraventriculares , Masculino , Umbral del Dolor , Ratas Long-Evans , Receptores de Oxitocina/antagonistas & inhibidores , Vasotocina/análogos & derivados , Vasotocina/farmacologíaRESUMEN
Although perinatal exposure of female rats to estrogenic compounds produces irreversible changes in brain function, it is still unclear how the amount and timing of exposure to those substances affect learning function, or if exposure alters estrogen receptor α (ERα) expression in the hippocampus and cortex. In adult female rats, we investigated the effects of neonatal exposure to a model estrogenic compound, ethinyl estradiol (EE), on passive avoidance learning and ERα expression. Female Wistar-Imamichi rats were subcutaneously injected with oil, 0.02 mg/kg EE, 2 mg/kg EE, or 20 mg/kg 17ß-estradiol within 24 h after birth. All females were tested for passive avoidance learning at the age of 6 weeks. Neonatal 0.02 mg/kg EE administration significantly disrupted passive avoidance compared with oil treatment in gonadally intact females. In a second experiment, another set of experimental females, treated as described above, was ovariectomized under pentobarbital anesthesia at 10 weeks of age. At 15-17 weeks of age, half of each group received a subcutaneous injection of 5 µg estradiol benzoate a day before the passive avoidance learning test. Passive avoidance learning behavior was impaired by the 0.02 mg/kg EE dose, but notably only in the estradiol benzoate-injected group. At 17-19 weeks of age, hippocampal and cortical samples were collected from rats with or without the 5 µg estradiol benzoate injection, and western blots used to determine ERα expression. A significant decrease in ERα expression was observed in the hippocampus of the estradiol-injected, neonatal EE-treated females. The results demonstrated that exposure to EE immediately after birth decreased learning ability in adult female rats, and that this may be at least partly mediated by the decreased expression of ERα in the hippocampus.
Asunto(s)
Reacción de Prevención/efectos de los fármacos , Corteza Cerebral/efectos de los fármacos , Receptor alfa de Estrógeno/antagonistas & inhibidores , Estrógenos/farmacología , Etinilestradiol/farmacología , Hipocampo/efectos de los fármacos , Animales , Animales Recién Nacidos , Corteza Cerebral/fisiología , Estradiol/análogos & derivados , Estradiol/farmacología , Receptor alfa de Estrógeno/genética , Receptor alfa de Estrógeno/metabolismo , Femenino , Expresión Génica/efectos de los fármacos , Hipocampo/fisiología , Inyecciones Subcutáneas , Ovariectomía , Ovario/fisiología , Ovario/cirugía , Ratas , Ratas WistarRESUMEN
Partner preference and paced mating tests were accomplished in ovariectomized female rats following bilateral radiofrequency lesions in the medial amygdala. Open field behavior and passive avoidance learning were also examined to investigate the underlying behavioral mechanism. Partner preference was determined in a chamber located between castrated and sexually active males. Airborne olfactory cues were presented to the female through small holes on the partition. The lesion diminished preference for the odor of sexually active males over that of castrated males, even after injection with a high-dose of estrogen. On the other hand, in a paced mating test the lesioned females without estrogen treatment showed a significantly shorter latency for entering the male's compartment in a two-compartment apparatus, which allowed the females, but not the males, to cross the barrier through a narrow opening at the bottom. However, an administration of estrogen and progesterone reduced the effect. The lesion had no effect on emotionality or exploratory behavior in an open field test, but it impaired passive avoidance learning capability. We suggest that a male poses an inherent threat to a female. The seemingly incompatible results of partner preference and paced mating tests can be compromised if the male is inherently aversive to the female; this emotional response can be removed by the medial amygdala lesion.