RESUMO
The concept that postnatal health and development can be influenced by events that occur in utero originated from epidemiological studies in humans supported by numerous mechanistic (including epigenetic) studies in a variety of model species. Referred to as the 'developmental origins of health and disease' or 'DOHaD' hypothesis, the primary focus of large-animal studies until quite recently had been biomedical. Attention has since turned towards traits of commercial importance in farm animals. Herein we review the evidence that prenatal risk factors, including suboptimal parental nutrition, gestational stress, exposure to environmental chemicals and advanced breeding technologies, can determine traits such as postnatal growth, feed efficiency, milk yield, carcass composition, animal welfare and reproductive potential. We consider the role of epigenetic and cytoplasmic mechanisms of inheritance, and discuss implications for livestock production and future research endeavours. We conclude that although the concept is proven for several traits, issues relating to effect size, and hence commercial importance, remain. Studies have also invariably been conducted under controlled experimental conditions, frequently assessing single risk factors, thereby limiting their translational value for livestock production. We propose concerted international research efforts that consider multiple, concurrent stressors to better represent effects of contemporary animal production systems.
RESUMO
The physiological basis of seasonal breeding in the guinea fowl (Numida meleagris) still remains unknown, despite the socioeconomic importance of these birds, particularly in Ghana. A study involving a total of 50 local guinea cocks was conducted, and documented gross anatomical and histological differences in the reproductive organs of breeding and non-breeding male guinea fowls. The study also compared peripheral testosterone concentrations in breeding and non-breeding cocks. Seasonal differences in variables measured were determined using two-tailed t-test/Mann-Whitney U-test. All comparisons were made at 5% level of significance. Breeding males had significantly (P = 0.000) higher anatomical biometric parameters than their non-breeding counterparts. Also, breeding birds had thicker (P = 0.000) phalli than their non-breeding counterparts. Histologically, regressing testis was characterized by the presence of sloughed off cells and increased debris in the tubular lumen and within the excurrent duct system, collapsed tubules and reduction in tubular lumen. Germ and Sertoli cell populations and nuclear diameters and actual seminiferous tubular diameter and length in regressing testes were significantly (P = 0.000) lower than in active testes. Leydig cell nuclear diameters and populations were also significantly (P = 0.000) reduced. Relative volume of seminiferous tubules in the testis, testicular sperm production/mg testis and per testis and peripheral testosterone concentrations were all higher (P < 0.05) in breeding than non-breeding testis. The ducts in the epididymal region also saw significant (P < 0.05) reductions in luminal diameters in non-breeding birds. Significant regression in anatomical and histological structures of the guinea cock reproductive tract occurred during the non-breeding season, and lower peripheral testosterone concentrations may be responsible for this phenomenon.
Assuntos
Galliformes/metabolismo , Genitália Masculina/química , Estações do Ano , Testosterona/metabolismo , Animais , Galliformes/sangue , Gana , Masculino , Testosterona/sangueRESUMO
The study documented gross anatomical and histological differences in the reproductive organs of 28 breeding and non-breeding female guinea fowls. Peripheral progesterone and 17ß-oestradiol concentrations were also compared in breeding and non-breeding hens. In non-breeding females, all ovarian and oviducal gross anatomical features had significantly regressed. Histologically, some of the changes in a regressing oviduct include systematic changes in height and size of all epithelial cells in all regions of the duct, absence/sparse ciliation of portions of surface epithelium in the magnum, isthmian and uterine regions, general loss of cytoplasmic mass, reduction in size and degeneration of tubular glands. Mucosal folds in all regions of the oviduct except the infundibular lip were higher in breeding females. No difference was found between the two groups in plasma progesterone concentrations. Breeding females, however, had higher peripheral oestradiol concentrations than non-breeding females. About 2 h prior to oviposition, plasma oestradiol concentrations peaked at 2.4-fold (230 pg/ml) compared with baseline concentration and plasma progesterone concentrations by nearly 9-fold (5.29 ng/ml) of baseline. Significant regression and changes in the histological structure of the ovary and oviduct had occurred in non-breeding females, and lower peripheral oestrogen concentrations may be responsible for this phenomenon.
Assuntos
Estradiol/sangue , Galliformes/fisiologia , Genitália Feminina/anatomia & histologia , Progesterona/sangue , Estações do Ano , Animais , Feminino , Genitália Feminina/fisiologia , ReproduçãoRESUMO
The rhythm of life on earth is shaped by seasonal changes in the environment. Plants and animals show profound annual cycles in physiology, health, morphology, behaviour and demography in response to environmental cues. Seasonal biology impacts ecosystems and agriculture, with consequences for humans and biodiversity. Human populations show robust annual rhythms in health and well-being, and the birth month can have lasting effects that persist throughout life. This review emphasizes the need for a better understanding of seasonal biology against the backdrop of its rapidly progressing disruption through climate change, human lifestyles and other anthropogenic impact. Climate change is modifying annual rhythms to which numerous organisms have adapted, with potential consequences for industries relating to health, ecosystems and food security. Disconcertingly, human lifestyles under artificial conditions of eternal summer provide the most extreme example for disconnect from natural seasons, making humans vulnerable to increased morbidity and mortality. In this review, we introduce scenarios of seasonal disruption, highlight key aspects of seasonal biology and summarize from biomedical, anthropological, veterinary, agricultural and environmental perspectives the recent evidence for seasonal desynchronization between environmental factors and internal rhythms. Because annual rhythms are pervasive across biological systems, they provide a common framework for trans-disciplinary research.
Assuntos
Ecossistema , Abastecimento de Alimentos , Periodicidade , Estações do Ano , Agricultura , Animais , Biodiversidade , Mudança Climática , Humanos , PlantasRESUMO
The elderly are at much higher risk for developing pneumonia than younger individuals. Pneumonia is a leading cause of death and is the third most common reason for hospitalization in the elderly. One reason that elderly people may be more susceptible to pneumonia is a breakdown in the lung's first line of defense, mucociliary clearance. Cilia beat in a coordinated manner to propel out invading microorganisms and particles. Ciliary beat frequency (CBF) is known to slow with aging, however, little is known about the mechanism(s) involved. We compared the CBF in BALB/c and C57BL/6 mice aged 2, 12, and 24 mo and found that CBF diminishes with age. Cilia in the mice at age 12 and 24 mo retained their ability to be stimulated by the ß2 agonist procaterol. To help determine the mechanism of ciliary slowing, we measured protein kinase C alpha and epsilon (PKCα and PKCε) activity. There were no activity differences in PKCα between the mice aged 2, 12, or 24 mo. However, we demonstrated a significantly higher PKCε activity in the mice at 12 and 24 mo than the in the mice 2 mo of age. The increase in activity is likely due to a nearly threefold increase in PKCε protein in the lung during aging. To strengthen the connection between activation of PKCε and ciliary slowing, we treated tracheas of mice at 2 mo with the PKCε agonist 8-[2-(2-pentylcyclopropylmethyl)-cyclopropyl]-octanoic acid (DCP-LA). We noted a similar decrease in baseline CBF, and the cilia remained sensitive to stimulation with ß2 agonists. The mechanisms for the slowing of baseline CBF have not been previously determined. In this mouse model of aging we were able to show that decreases in CBF are related to an increase in PKCε activity.
Assuntos
Envelhecimento/fisiologia , Pulmão/fisiopatologia , Depuração Mucociliar/fisiologia , Pneumonia/enzimologia , Proteína Quinase C-épsilon/metabolismo , Agonistas de Receptores Adrenérgicos beta 2/farmacologia , Fatores Etários , Animais , Caprilatos/farmacologia , Cílios/enzimologia , Cílios/fisiologia , Células Epiteliais/fisiologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Modelos Animais , Procaterol/farmacologia , Proteína Quinase C-alfa/metabolismoRESUMO
Prenatal exposure to endogenous or exogenous androgens alters the development of the female reproductive tract. Although lesions in ovaries and external genitalia of androgenized female sheep have been reported, lesions of the tubular genitalia have not. Testosterone propionate (TP) or dihydrotestosterone (DHT) was administered by intramuscular injection twice weekly to 32 ewes from 30 to 90 days of pregnancy. The ewes lambed normally. The reproductive tracts from 24 treated and 13 control postpubertal female offspring were examined at 10 months of age. The ovaries, oviducts, and uteri were grossly and histologically normal in both TP- and DHT-exposed sheep. However, in the DHT-treated sheep, the uterus connected to a misshapen, saccular vagina that opened into the urethra; in the TP-treated sheep, it ended in a blind sac. In both TP- and DHT-treated sheep, the urethra was approximately 5 times longer than that of control sheep, and it resembled a male urethra with bilateral male accessory genital glands. The urethra terminated in a fully developed penis in both TP- and DHT-treated sheep, and a scrotal sac was present (without testes). These results show that prenatal exposure of female sheep to exogenous androgens results in masculinization of the tubular and external genitalia.
Assuntos
Androgênios/farmacologia , Genitália Feminina/efeitos dos fármacos , Efeitos Tardios da Exposição Pré-Natal/veterinária , Ovinos/metabolismo , Virilismo/veterinária , Androgênios/metabolismo , Animais , Di-Hidrotestosterona/metabolismo , Di-Hidrotestosterona/farmacologia , Feminino , Genitália Feminina/crescimento & desenvolvimento , Técnicas Histológicas/veterinária , Gravidez , Efeitos Tardios da Exposição Pré-Natal/metabolismo , Propionato de Testosterona/metabolismo , Propionato de Testosterona/farmacologia , Virilismo/metabolismoRESUMO
During the acute phase (1 wk of symptoms or less) of infectious mononucleosis (IM), 70--80% of circulating Epstein-Barr virus nuclear antigen (EBNA)-positive cells have differentiated toward plasma cells. Thus the characteristics of the infected cells in the majority of IM patients during early disease are indistinguishable from EBNA-positive tumor cells of a previously reported child who developed lymphoma during IM. IgA and IgG were the most frequent and IgM the least frequent immunoglobulin isotypes detected in EBNA-positive cells. In acute disease EBNA was present in 5.5--20% of T cell-depleted blood lymphocytes but in the 2nd or 3rd wk of illness the number of EBNA-positive cells sharply decreased to 0.4--1.4%. At the same time the fraction of antigen-positive cells containing cytoplasmic immunoglobulins also diminished, suggesting either that differentiation of infected cells was altered during the disease or that nondifferentiated antigen-positive cells had a survival advantage. Both the high proportion of plasmacytic EBNA-positive cells seen during acute disease and the apparent loss of differentiation by these cells later in disease may be regulated by host immunologic factors. Immunoglobulin-producing EBNA-positive cells may be the source of heterophile antibodies and other seemingly inappropriate antibodies usually found in serum during IM; however, increased numbers of noninfected plasma cells were present in some patients and may also be a potential source of these unusual antibodies.
Assuntos
Linfócitos B/imunologia , Mononucleose Infecciosa/imunologia , Plasmócitos/citologia , Doença Aguda , Antígenos Virais , Diferenciação Celular , Citoplasma/imunologia , Imunofluorescência , Herpesvirus Humano 4/imunologia , Imunoglobulina M/biossínteseRESUMO
Chronic gonadotropin-releasing hormone agonist (GnRHa) treatment is effective for the medical suppression of the hypothalamic-pituitary-gonadal axis in situations like central precocious puberty and gender dysphoria. However, its administration during the peripubertal period could influence normal brain development and function because GnRH receptors are expressed in brain regions that regulate emotions, cognition, motivation and memory. This study used an ovine model to determine whether chronic peripubertal GnRHa-treatment affected the developmental shift from preference of familiarity to novelty. Experimental groups included Controls and GnRHa-treated rams. To differentiate between effects of altered GnRH signaling and those associated with the loss of sex steroids, a group was also included that received testosterone replacement as well as GnRHa (GnRHaâ¯+â¯T). Preference for a novel versus familiar object was assessed during 5-min social isolation at 8, 28 and 46 weeks of age. Approach behavior was measured as interactions with and time spent near the objects, whereas avoidance behavior was measured by time spent in the entrance zone and attempts to escape the arena via the entry point. Emotional reactivity was measured by the number of vocalizations, escape attempts and urinations. As Control and GnRHa-treated rams aged, their approach behaviors showed a shift from preference for familiarity (8 weeks) to novelty (46 weeks). In contrast, relative to the Controls the GnRHaâ¯+â¯T rams exhibited more approach behaviors towards both objects, at 28 and 46 weeks of age and preferred familiarity at 46 weeks of age. Vocalisation rate was increased in GnRHa treated rams in late puberty (28 weeks) compared to both Control and GnRHaâ¯+â¯T rams but this effect was not seen in young adulthood (46 weeks). These results suggest that the specific suppression of testosterone during a developmental window in late puberty may reduce emotional reactivity and hamper learning a flexible adjustment to environmental change. The results also suggest that disruption of either endogenous testosterone signalling or a synergistic action between GnRH and testosterone signalling, may delay maturation of cognitive processes (e.g. information processing) that affects the motivation of rams to approach and avoid objects.
Assuntos
Comportamento Exploratório/efeitos dos fármacos , Gosserrelina/farmacologia , Maturidade Sexual/fisiologia , Animais , Hormônios Esteroides Gonadais/farmacologia , Hormônio Liberador de Gonadotropina/agonistas , Hormônio Liberador de Gonadotropina/fisiologia , Masculino , Reconhecimento Psicológico/efeitos dos fármacos , Caracteres Sexuais , Carneiro Doméstico/fisiologia , Testículo/efeitos dos fármacos , Testosterona/farmacologiaRESUMO
Entrainment of circadian rhythms (CR) to the light dark cycle has been well described under controlled, experimental conditions. However, studies in rodents have reported that rhythms in the laboratory are not always reproduced under field conditions. The aim of this study was to characterise the CR of sheep maintained under conditions of standard UK farm animal husbandry and to investigate the effects of environmental challenges presented by season, weaning and changes in housing on CR. Male sheep (nâ¯=â¯9) were kept at pasture, or group housed in barns, under natural photoperiod for one year. CR in locomotor activity were monitored using accelerometry, and 24â¯h patterns in plasma cortisol and melatonin were measured every 4â¯h by ELISA. CR was measured before and after weaning, in summer and winter, and at pasture and by barn housing. Cosinor analysis revealed high amplitude, diurnal rhythms in locomotor activity that were disrupted by weaning and by barn housing. Rhythms in winter showed an interrupted night time activity pattern, but only when the sheep were kept at pasture. Cortisol and melatonin secretion followed typical circadian patterns in winter and summer. The CR of the sheep under the field conditions of this study were strikingly robust under basal conditions, but easily disrupted by environmental challenges. Interrupted patterns of activity during the long nights of wintertime, not previously reported for sheep kept in experimental conditions were recorded. Based on these findings, we propose that animals require exposure to more complex environments than the laboratory in order to exhibit their true circadian phenotype.
Assuntos
Ritmo Circadiano/fisiologia , Abrigo para Animais , Melatonina/sangue , Atividade Motora/fisiologia , Fotoperíodo , Desmame , Acelerometria , Animais , Hidrocortisona/sangue , Masculino , Estações do Ano , OvinosRESUMO
There is increasing evidence that sera from HIV-1-infected individuals contain antibodies that enhance infection by HIV-1 in vitro. Previous work has demonstrated that complement receptors on T lymphoid cells and Fc receptors for IgG (Fc gamma R) on monocytic cells are required for enhanced infection by antibody-complexed HIV-1. Characterization of such infection-enhancing antibodies is essential because immunogenic epitopes which induce enhancing antibodies should be excluded from HIV-1 vaccines. This study was conducted to identify enhancing antibodies involved in Fc R-mediated enhancement of HIV-1 infection employing IgG human monoclonal antibodies (HMAbs) reactive against gp120 of HIV-1, which were produced by B cell lines derived from an HIV-1-infected individual. A potent neutralizing HMAb N70-1.5e did not enhance infection by HIV-1 (IIIB and MN strains), whereas HMAb N70-2.3a mediated enhancement of HIV-1 infection, but had little neutralizing activity. A competition radio immunoassay demonstrated that the two antibodies bind to distinct epitopes. These results indicated that enhancing and neutralizing antibodies can be induced by different epitopes on gp120, suggesting the potential for development of safe vaccines against HIV-1 by exclusion of immunogenic epitopes for enhancing antibodies. We made attempts to identify the epitope on gp120 that is recognized by the enhancing antibody N70-2.3a by using recombinant HIV-1 proteins and found that the antibody binds to a conformational site of nonvariable sequences in the carboxyl half (aa 272-509) of gp120.
Assuntos
Anticorpos Monoclonais/imunologia , Anticorpos Anti-HIV/imunologia , Proteína gp120 do Envelope de HIV/imunologia , HIV-1/imunologia , Animais , Ligação Competitiva , Epitopos/imunologia , Infecções por HIV/imunologia , Infecções por HIV/microbiologia , HIV-1/fisiologia , Humanos , Imunoglobulina G/imunologia , Testes de Neutralização , Coelhos , Radioimunoensaio , Receptores Fc/imunologia , Replicação ViralRESUMO
Galanin is a small neuropeptide that mediates its effects via three receptor isoforms: galanin receptor-1, galanin receptor-2 and galanin receptor-3 (Gal-R1, Gal-R2 and Gal-R3). Galanin is thought to be an important intermediate in signalling in the hypothalamic-pituitary-gonadal axis and has been widely detected in the ovine hypothalamus. The expression of galanin and Gal-R1 has been reported to fluctuate during the reproductive cycle. Although the distribution of Gal-R1 has been determined in the ovine hypothalamus, the distribution of Gal-R2 was hitherto unknown. Using immunohistological and immunofluorescence techniques, we have mapped the distribution of Gal-R2 in the ovine hypothalamus, collected during the follicular phase of the oestrous cycle and examined colocalisation of Gal-R2 with oestrogen receptor alpha (ERalpha) and gonadotrophin-releasing hormone (GnRH). Gal-R2 was expressed in several regions of the hypothalamus (supraoptic nucleus, paraventricular nucleus, ventromedial nucleus, arcuate nucleus) but not as widely expressed as Gal-R1. Areas of Gal-R2 expression overlapped with those reported for Gal-R1. We observed that, in certain defined regions of the hypothalamus, up to 50% of neurones that express Gal-R2 also express ERalpha. No neurones coexpressed Gal-R2 and GnRH. Thus, we conclude that, in follicular phase animals, this receptor plays little or no role in direct intermediary signal transmission in GnRH-mediated control of the reproductive cycle.
Assuntos
Hormônio Liberador de Gonadotropina/metabolismo , Hipotálamo/metabolismo , Neurônios/metabolismo , Receptor Tipo 2 de Galanina/metabolismo , Animais , Receptor alfa de Estrogênio/metabolismo , Receptor alfa de Estrogênio/fisiologia , Ciclo Estral/fisiologia , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Hormônio Liberador de Gonadotropina/biossíntese , Hipotálamo/citologia , Imuno-Histoquímica , Receptor Tipo 2 de Galanina/biossíntese , OvinosRESUMO
Chronic gonadotropin-releasing hormone agonist (GnRHa) administration is used where suppression of hypothalamic-pituitary-gonadal axis activity is beneficial, such as steroid-dependent cancers, early onset gender dysphoria, central precocious puberty and as a reversible contraceptive in veterinary medicine. GnRH receptors, however, are expressed outside the reproductive axis, e.g. brain areas such as the hippocampus which is crucial for learning and memory processes. Previous work, using an ovine model, has demonstrated that long-term spatial memory is reduced in adult rams (45 weeks of age), following peripubertal blockade of GnRH signaling (GnRHa: goserelin acetate), and this was independent of the associated loss of gonadal steroid signaling. The current study investigated whether this effect is reversed after discontinuation of GnRHa-treatment. The results demonstrate that peripubertal GnRHa-treatment suppressed reproductive function in rams, which was restored after cessation of GnRHa-treatment at 44 weeks of age, as indicated by similar testes size (relative to body weight) in both GnRHa-Recovery and Control rams at 81 weeks of age. Rams in which GnRHa-treatment was discontinued (GnRHa-Recovery) had comparable spatial maze traverse times to Controls, during spatial orientation and learning assessments at 85 and 99 weeks of age. Former GnRHa-treatment altered how quickly the rams progressed beyond a specific point in the spatial maze at 83 and 99 weeks of age, and the direction of this effect depended on gonadal steroid exposure, i.e. GnRHa-Recovery rams progressed quicker during breeding season and slower during non-breeding season, compared to Controls. The long-term spatial memory performance of GnRHa-Recovery rams remained reduced (P<0.05, 1.5-fold slower) after discontinuation of GnRHa, compared to Controls. This result suggests that the time at which puberty normally occurs may represent a critical period of hippocampal plasticity. Perturbing normal hippocampal formation in this peripubertal period may also have long lasting effects on other brain areas and aspects of cognitive function.
Assuntos
Hormônio Liberador de Gonadotropina/agonistas , Gosserrelina/farmacologia , Orientação Espacial/efeitos dos fármacos , Aprendizagem Espacial/efeitos dos fármacos , Memória Espacial/efeitos dos fármacos , Animais , Peso Corporal/efeitos dos fármacos , Masculino , Tamanho do Órgão/efeitos dos fármacos , Maturidade Sexual/efeitos dos fármacos , Maturidade Sexual/fisiologia , Ovinos , Testículo/efeitos dos fármacos , Testículo/crescimento & desenvolvimentoRESUMO
Chronic gonadotropin-releasing hormone agonist (GnRHa) is used therapeutically to block activity within the reproductive axis through down-regulation of GnRH receptors within the pituitary gland. GnRH receptors are also expressed in non-reproductive tissues, including areas of the brain such as the hippocampus and amygdala. The impact of long-term GnRHa-treatment on hippocampus-dependent cognitive functions, such as spatial orientation, learning and memory, is not well studied, particularly when treatment encompasses a critical window of development such as puberty. The current study used an ovine model to assess spatial maze performance and memory of rams that were untreated (Controls), had both GnRH and testosterone signaling blocked (GnRHa-treated), or specifically had GnRH signaling blocked (GnRHa-treated with testosterone replacement) during the peripubertal period (8, 27 and 41 weeks of age). The results demonstrate that emotional reactivity during spatial tasks was compromised by the blockade of gonadal steroid signaling, as seen by the restorative effects of testosterone replacement, while traverse times remained unchanged during assessment of spatial orientation and learning. The blockade of GnRH signaling alone was associated with impaired retention of long-term spatial memory and this effect was not restored with the replacement of testosterone signaling. These results indicate that GnRH signaling is involved in the retention and recollection of spatial information, potentially via alterations to spatial reference memory, and that therapeutic medical treatments using chronic GnRHa may have effects on this aspect of cognitive function.
Assuntos
Hormônio Liberador de Gonadotropina/agonistas , Hormônio Liberador de Gonadotropina/farmacologia , Orientação Espacial/efeitos dos fármacos , Transdução de Sinais/fisiologia , Aprendizagem Espacial/efeitos dos fármacos , Memória Espacial/efeitos dos fármacos , Testosterona/farmacologia , Fatores Etários , Animais , Masculino , OvinosRESUMO
It has been postulated that synaptic potentiation during waking is offset by a homoeostatic reduction in net synaptic strength during sleep. However, molecular mechanisms to support such a process are lacking. Here we demonstrate that deficiencies in the RNA-editing gene Adar increase sleep due to synaptic dysfunction in glutamatergic neurons in Drosophila. Specifically, the vesicular glutamate transporter is upregulated, leading to over-activation of NMDA receptors, and the reserve pool of glutamatergic synaptic vesicles is selectively expanded in Adar mutants. Collectively these changes lead to sustained neurotransmitter release under conditions that would otherwise result in synaptic depression. We propose that a shift in the balance from synaptic depression towards synaptic potentiation in sleep-promoting neurons underlies the increased sleep pressure of Adar-deficient animals. Our findings provide a plausible molecular mechanism linking sleep and synaptic plasticity.
Assuntos
Adenosina Desaminase/metabolismo , Proteínas de Drosophila/metabolismo , Drosophila/enzimologia , Drosophila/fisiologia , Ácido Glutâmico/metabolismo , Plasticidade Neuronal , Adenosina Desaminase/genética , Animais , Drosophila/genética , Proteínas de Drosophila/genética , Feminino , Masculino , Neurônios/metabolismo , Neurotransmissores/metabolismo , Edição de RNA , Sono , Vesículas Sinápticas/metabolismoRESUMO
Elevated oestradiol concentrations during the follicular phase stimulate a surge in gonadotropin-releasing hormone (GnRH) and luteinising hormone (LH) concentrations, which leads to ovulation. Progesterone can block the oestradiol-induced GnRH/LH surge, but the mechanism that is involved is unclear. We examined the effect of progesterone on oestradiol-induced activation of cells within the ovine hypothalamus/preoptic area (POA) to determine: (i) in which regions progesterone acts to block the GnRH/LH surge and (ii) whether progesterone directly or indirectly prevents activation of oestradiol-responsive cells. Cellular activation was assessed by measuring the number of cells that expressed Fos (an immediate early gene). Exposure to increased oestradiol concentrations in the absence of progesterone (which normally stimulates a LH surge) did not cause any region-specific changes in hypothalamic Fos expression during the activation stage of the LH surge-induction process (Experiment 1). The same treatment significantly increased cellular activation within the POA, lateral septum (LS), and arcuate nucleus at the time of surge onset (Experiment 2). Concurrent exposure to increased oestradiol and progesterone concentrations during the activation stage of the surge-induction process (which normally blocks the LH surge) was associated with significantly reduced cellular activation within the ventromedial hypothalamus and anterior hypothalamic area, relative to the positive controls (oestradiol increment alone) and arcuate nucleus relative to the negative controls (no increment in oestradiol) during the activation stage (Experiment 1). At the time of surge onset (Experiment 2), exposure to progesterone during the activation period prevented the oestradiol-induced increase in cellular activation that occurred in the POA, LS and arcuate nucleus of the positive controls. These results demonstrated that oestradiol and progesterone induced differential region- and time-specific effects on cellular activation within the regions of the ovine brain that generate the preovulatory GnRH/LH surge. Moreover, the lack of cellular activation within the POA, LS and arcuate nucleus at the time of surge onset in animals exposed to progesterone during the activation stage is consistent with the hypothesis that progesterone can block the preovulatory surge by direct inhibition of oestradiol-induced cellular activation in these areas.
Assuntos
Estradiol/metabolismo , Ciclo Estral/sangue , Hormônio Liberador de Gonadotropina/sangue , Hipotálamo/metabolismo , Hormônio Luteinizante/sangue , Progesterona/fisiologia , Animais , Núcleo Arqueado do Hipotálamo/citologia , Núcleo Arqueado do Hipotálamo/metabolismo , Regulação para Baixo , Feminino , Hipotálamo/citologia , Sistemas Neurossecretores/fisiologia , Área Pré-Óptica/citologia , Área Pré-Óptica/metabolismo , Proteínas Proto-Oncogênicas c-fos/metabolismo , Septo do Cérebro/citologia , Septo do Cérebro/metabolismo , OvinosRESUMO
Dredging and associated screening at a dredge site in the southern North Sea (Area 408) is associated with areas of well-sorted fine sand that extend for up to 3 km to the south-east of the dredged area and overlay sediments with a more variable particle size composition. This well-sorted fine sand may reflect deposition and transport of material mobilised by the dredging and screening processes at the dredge site. Multivariate analysis of the benthic community structure suggests that marine aggregate dredging, at the level of intensity employed in the study area prior to sample collection, has had a limited impact on benthic community composition compared with that reported from studies elsewhere. This is ascribed to the likely rapid rates of recolonisation by the mobile opportunistic polychaetes and crustaceans that dominate the macrofauna of the sandy gravel deposits at this particular dredge site. Analysis of variance showed, however, that significant differences existed between the sample treatments in terms of species evenness (Pielou's J). Dredged samples were found to have the lowest mean species evenness (0.71) when compared to controls (0.77). The present study highlights the inherent difficulties in the application of general impact/recovery predictions to dredged sites with varying environmental characteristics.
Assuntos
Ecossistema , Monitoramento Ambiental/estatística & dados numéricos , Poluição Ambiental/análise , Sedimentos Geológicos/análise , Invertebrados/fisiologia , Eliminação de Resíduos , Análise de Variância , Animais , Análise Multivariada , Mar do Norte , Tamanho da Partícula , Dinâmica PopulacionalRESUMO
We used a human monoclonal antibody (MAb; 15e) to identify an antibody-dependent cell-mediated cytotoxicity (ADCC) epitope on HIV-1 gp120. 15e has been shown to recognize a conformation-dependent epitope on gp120 which is important in both CD4 binding and neutralizing of HIV-1 infection. 15e binds to gp120 of HIV-1IIIB but not HIV-1RF. Using a standard ADCC assay, 15e was found to mediate ADCC against cells infected with HIV-1IIIB but not HIV-1RF. 15e did not mediate ADCC against cells with recombinant gp120 bound to surface CD4, indicating that 15e does not mediate innocent bystander ADCC against uninfected CD4 cells. To better define the 15e epitope, we performed ADCC against target cells infected with a vaccinia vector which expresses processed HIV-1IIIB gp160 from which the third variable region was deleted (amino acids, 312-328). MAb 15e efficiently mediated ADCC against cells expressing this altered form of gp120, indicating that this region is not contributing to the conformational epitope defined by 15e. 15e defines an important epitope in the human immune response to HIV-1 infection. Antibodies with 15e-like activity may be useful in immunoprophylaxis or immunotherapy of HIV-1 infection.
Assuntos
Anticorpos Monoclonais , Citotoxicidade Celular Dependente de Anticorpos , Anticorpos Anti-HIV , Proteína gp120 do Envelope de HIV/imunologia , HIV-1/imunologia , Sítios de Ligação , Linhagem Celular , Epitopos , Produtos do Gene env/imunologia , Proteína gp160 do Envelope de HIV , Humanos , Precursores de Proteínas/imunologiaRESUMO
Exposure of the female ovine fetus to exogenous androgens during early gestation permanently masculinizes the reproductive anatomy, physiology, and behavior of the adult ewe. In utero testosterone exposure has been shown to act centrally on the GnRH neuronal network to alter the response to both the stimulatory and inhibitory actions of estrogen. It is currently unknown whether fetal androgens alter other mechanisms that are critical for the regulation of GnRH release and, specifically, other important regulatory steroid feedback loops. Three studies were performed on gonadectomized postpubertal sheep to determine whether the inhibitory actions of progesterone on episodic LH release are also sex-specific and engendered by early in utero exposure to testosterone. In each study, the pulsatile pattern of LH release was determined both before and after the sc implantation of a progesterone releasing CIDR device. The studies involved 7 female, 7 male, and 12 androgenized female sheep (T60 (n = 7) and T30 (n = 5) groups; 200 mg testosterone propionate/week im to the mother for 60 or 30 days, respectively, from day 30-90 or 60-90 of pregnancy). The first two studies were performed in the anestrous season in the presence (Exp 1) or absence (Exp 2) of a low circulating concentration of estradiol. Exp 3 was carried out in the breeding season in the absence of exogenous estrogen. In all three studies progesterone inhibited LH pulse frequency only in the females. Progesterone had no action on mean LH concentrations or the frequency or amplitude of LH pulses in the males or either group of androgenized ewes. We conclude that the inhibition of episodic LH release by progesterone is sexually differentiated in the sheep, males being less responsive than females to steroid negative feedback. Further, these sex differences are a consequence of in utero exposure to androgens for a period as short as 30 days between days 60 and 90 of a 147-day pregnancy.
Assuntos
Feto/efeitos dos fármacos , Hormônio Liberador de Gonadotropina/metabolismo , Neurônios/efeitos dos fármacos , Progesterona/farmacologia , Testosterona/farmacologia , Animais , Implantes de Medicamento , Estradiol/sangue , Feminino , Idade Gestacional , Hormônio Luteinizante/metabolismo , Masculino , Neurônios/fisiologia , Periodicidade , Gravidez , Caracteres Sexuais , Ovinos , Testosterona/administração & dosagemRESUMO
The preovulatory LH surge in the ewe is stimulated by the massive and sustained release of GnRH into the pituitary portal vessels. This study has examined the temporal relationship between changes in LH secretion and GnRH messenger RNA (mRNA) expression at the time of the estradiol-induced LH surge. Ovariectomized Clun Forest ewes were treated with exogenous progesterone and estradiol (E) to mimic estrous cycle concentrations of these gonadal steroids and to induce the LH surge. Ewes were killed at five time points relative to the time of onset of the LH surge: pre-E, before E insertion (n = 6); presurge, after E insertion and 8-10 h before surge onset (n = 5); ascending limb, 2-6 h after surge onset (n = 5); midpeak, 9-12 h after surge onset (n = 5); and postsurge, 21-27 h after surge onset (n = 5). Control animals (n = 5/group), which received no E, were killed at identical time intervals alongside the E-treated ewes. Coronal sections containing the diagonal band of Broca through to the anterior hypothalamus were processed for cellular in situ hybridization using an 35S-labeled oligonucleotide probe complementary to ovine GnRH. No changes were found in the number of GnRH mRNA-expressing cells detected in the rostral preoptic area or the medial septum in either gonadal steroid-treated or control ewes. In contrast, cellular GnRH mRNA expression (as assessed by silver grain density) decreased significantly (P < 0.05) between presurge and ascending limb groups within both the rostral preoptic area (0.64 +/- 0.06 vs. 0.43 +/- 0.05 silver grain density/microm2) and medial septum cells (1.08 +/- 0.09 vs. 0.77 +/- 0.07). No significant changes were detected in control ewes. These results show that the estradiol-induced LH surge in the ewe is associated with a decrease in GnRH mRNA expression that occurs in advance of the onset of the GnRH surge. This suggests that neural mechanisms controlling GnRH biosynthesis may be distinct from those regulating GnRH secretion.
Assuntos
Estradiol/farmacologia , Hormônio Liberador de Gonadotropina/genética , Hormônio Luteinizante/sangue , RNA Mensageiro/análise , Animais , Feminino , Hormônio Liberador de Gonadotropina/análise , Hormônio Liberador de Gonadotropina/biossíntese , OvinosRESUMO
The preovulatory GnRH/LH surge in the ewe is stimulated by a rise in the circulating estradiol concentration that occurs in conjunction with preovulatory ovarian follicle development. In the presence of high levels of progesterone, such as during the luteal phase of the estrous/menstrual cycle, the stimulatory effects of elevated estradiol on GnRH/LH secretion are blocked. Recent work in the ewe has shown that a relatively short period of estradiol exposure can stimulate a GnRH/LH surge that begins after estrogenic support has been removed. This result suggests that surge generation is characterized by an estradiol-dependent period (during which the signal is read) and an estradiol-independent period (during which a cascade of neuronal events transmits the stimulatory signal to the GnRH neurosecretory system, which releases a surge of GnRH). In this series of studies, we addressed the hypothesis that progesterone can block transmission of the stimulatory estradiol signal after it has been read. Nine ovariectomized ewes were run through repeated artificial estrous cycles by sequential addition and removal of exogenous steroids. In study one, ewes received three treatments in a randomized cross-over design. Exposure to a follicular phase estradiol concentration for 10 h (positive control treatment) stimulated an LH surge in all ewes, as determined in hourly jugular blood samples. Maintenance of luteal phase progesterone concentrations throughout the artificial follicular phase (2 x CIDR-G devices, negative control) blocked the stimulatory effects of a 10-h estradiol signal, and no ewes that received this treatment expressed an LH surge. In the experimental group, exposure to luteal phase levels of progesterone, during the period after the surge generating system had been activated by estradiol, blocked the LH surge in six of nine ewes. This result demonstrates that progesterone can block the surge, even when applied after the surge-generating system has been activated and, therefore, that it inhibits either the transmission of the estradiol signal and/or the release of the GnRH/LH surge. In study 2, we assessed whether sensitivity to the inhibitory effects of progesterone was confined to a specific stage of the transmission of the estradiol signal. Eight ewes were exposed to four treatments, over successive artificial estrous cycles. Positive and negative controls were similar to those described in Study 1, except the duration of the stimulatory estradiol signal was reduced to 8 h. The two experimental groups consisted of an EARLY P (progesterone) treatment, in which progesterone was given from hours 8-13 after estradiol insertion (immediately after estradiol removal), and a LATE P treatment, in which progesterone was given from hours 13-18 (immediately before LH surge secretion). As expected, LH surges were stimulated and blocked, in response to the positive and negative controls, respectively. Whereas the EARLY P treatment blocked the LH surge in seven of eight ewes, the LATE P treatment was only successful in inhibiting a surge in one of eight animals. This result demonstrates that progesterone can block the estradiol-induced surge-generating signal soon after the onset of signal transmission (immediately after estradiol removal) but not during the later stages of signal transmission (at the time of GnRH/LH surge onset).