RESUMO
Free-water imaging can predict and monitor dopamine system degeneration in people with Parkinson's disease. It can also enhance the sensitivity of traditional diffusion tensor imaging (DTI) metrics for indexing neurodegeneration. However, these tools are yet to be applied to investigate cholinergic system degeneration in Parkinson's disease, which involves both the pedunculopontine nucleus and cholinergic basal forebrain. Free-water imaging, free-water-corrected DTI and volumetry were used to extract structural metrics from the cholinergic basal forebrain and pedunculopontine nucleus in 99 people with Parkinson's disease and 46 age-matched controls. Cognitive ability was tracked over 4.5 years. Pearson's partial correlations revealed that free-water-corrected DTI metrics in the pedunculopontine nucleus were associated with performance on cognitive tasks that required participants to make rapid choices (behavioural flexibility). Volumetric, free-water content and DTI metrics in the cholinergic basal forebrain were elevated in a sub-group of people with Parkinson's disease with evidence of cognitive impairment, and linear mixed modelling revealed that these metrics were differently associated with current and future changes to cognition. Free water and free-water-corrected DTI can index cholinergic degeneration that could enable stratification of patients in clinical trials of cholinergic interventions for cognitive decline. In addition, degeneration of the pedunculopontine nucleus impairs behavioural flexibility in Parkinson's disease, which may explain this region's role in increased risk of falls.
Assuntos
Prosencéfalo Basal , Doença de Parkinson , Núcleo Tegmental Pedunculopontino , Humanos , Doença de Parkinson/complicações , Imagem de Tensor de Difusão , Prosencéfalo Basal/diagnóstico por imagem , Colinérgicos , Água , Neurônios ColinérgicosRESUMO
The ontogenetic development in teleost fish is sensitive to temperature, and the developmental rate has a direct relationship with the environmental temperature within a species' thermal tolerance limit. Temperature determines time to and survival at hatching. Yellow perch is a North American species of ecological and commercial importance, and its phenology is vulnerable to climate change. The embryonic development of yellow perch was comparable to closely related members of the family Percidae. Developmental progression was fastest at 18°C and slowest at 12°C, with medial progression at 15°C. Time to hatch and swim-up, feeding onset, and exogenous feeding phases were different across all incubation temperatures regardless of a gradual post-hatch warming of the 12 and 15°C groups to a common garden temperature of 18°C. Incubation temperature may lower the rate of survival to hatch at 15°C and had complex impacts on developmental abnormalities. Temperature had significant effects on the development rate, time of hatch, survival, and incidence of developmental abnormalities. Early ontogenetic thermal history in ectotherms is an important factor determining phenotypic variation. It will be important to link the thermally induced changes in development described here to the physiological and morphological differences and to link the developmental abnormalities to functional performance.
RESUMO
The exposure of ionizing radiation during early gestation often leads to deleterious and even lethal effects; however, few extensive studies have been conducted on late gestational exposures. This research examined the behavior al effects of C57Bl/6J mouse offspring exposed to low dose ionizing gamma irradiation during the equivalent third trimester. Pregnant dams were randomly assigned to sham or exposed groups to either low dose or sublethal dose radiation (50, 300, or 1000 mGy) at gestational day 15. Adult offspring underwent a behavioral and genetic analysis after being raised under normal murine housing conditions. Our results indicate very little change in the behavioral tasks measuring general anxiety, social anxiety, and stress-management in animals exposed prenatally across the low dose radiation conditions. Quantitative real-time polymerase chain reactions were conducted on the cerebral cortex, hippocampus, and cerebellum of each animal; results indicate some dysregulation in markers of DNA damage, synaptic activity, reactive oxygen species (ROS) regulation, and methylation pathways in the offspring. Together, our results provide evidence in the C57Bl/6J strain, that exposure to sublethal dose radiation (<1000 mGy) during the last period of gestation leads to no observable changes in behaviour when assessed as adults, although some changes in gene expression were observed for specific brain regions. These results indicate that the level of oxidative stress occurring during late gestation for this mouse strain is not sufficient for a change in the assessed behavioral phenotype, but results in some modest dysregulation of the genetic profile of the brain.
Assuntos
Efeitos Tardios da Exposição Pré-Natal , Humanos , Feminino , Gravidez , Animais , Camundongos , Efeitos Tardios da Exposição Pré-Natal/genética , Camundongos Endogâmicos C57BL , Radiação Ionizante , Raios gama , Ansiedade/etiologia , Comportamento AnimalRESUMO
Long-term temperature shifts associated with seasonal variability are common in temperate regions. However, these natural shifts could place significant strain on thermal stress responses of fishes when combined with mean increases in water temperatures predicted by climate change models. We examined the relationship between thermal acclimation, basal expression of heat shock protein (hsp) genes and the activation of the heat shock response (HSR) in lake whitefish (LWF; Coregonus clupeaformis), a cold water species of cultural and commercial significance. Juveniles were acclimated to either 6, 12, or 18°C water for several months prior to the quantification of hsp mRNA levels in the presence or absence of acute heat shock (HS). Acclimation to 18°C increased basal mRNA levels of hsp70 and hsp47, but not hsc70 or hsp90ß in gill, liver and white muscle, while 6°C acclimation had no effect on basal hsp transcription. Fish in all acclimation groups were capable of eliciting a robust HSR following acute HS, as indicated by the upregulation of hsp70 and hsp47. An increase of only 2°C above the 18°C acclimation temperature was required to trigger these transcriptional changes, suggesting that the HSR may be frequently initiated in LWF populations living at mildly elevated temperatures. Collectively, these expression profiles show that environmental temperature influences both basal hsp levels and the HSR in LWF, and indicate that these fish may have a greater physiological and ecological susceptibility to elevated temperatures than to cooler temperatures.
Assuntos
Proteínas de Choque Térmico HSP47/genética , Proteínas de Choque Térmico HSP70/genética , Resposta ao Choque Térmico/genética , Salmonidae/genética , Aclimatação , Animais , Mudança Climática , Expressão Gênica , Lagos , RNA Mensageiro/genética , Temperatura , Regulação para Cima/genéticaRESUMO
BACKGROUND: Gait disturbance is an early, disabling feature of Parkinson's disease (PD) that is typically refractory to dopaminergic medication. The cortical cholinergic system, originating in the nucleus basalis of Meynert of the basal forebrain, has been implicated. However, it is not known if degeneration in this region relates to a worsening of disease-specific gait impairment. OBJECTIVE: To evaluate associations between sub-regional cholinergic basal forebrain volumes and longitudinal progression of gait impairment in PD. METHODS: 99 PD participants and 47 control participants completed gait assessments via an instrumented walkway during 2 minutes of continuous walking, at baseline and for up to 3 years, from which 16 spatiotemporal characteristics were derived. Sub-regional cholinergic basal forebrain volumes were measured at baseline via MRI and a regional map derived from post-mortem histology. Univariate analyses evaluated cross-sectional associations between sub-regional volumes and gait. Linear mixed-effects models assessed whether volumes predicted longitudinal gait changes. RESULTS: There were no cross-sectional, age-independent relationships between sub-regional volumes and gait. However, nucleus basalis of Meynert volumes predicted longitudinal gait changes unique to PD. Specifically, smaller nucleus basalis of Meynert volume predicted increasing step time variability (P = 0.019) and shortening swing time (P = 0.015); smaller posterior nucleus portions predicted shortening step length (P = 0.007) and increasing step time variability (P = 0.041). CONCLUSIONS: This is the first study to demonstrate that degeneration of the cortical cholinergic system predicts longitudinal progression of gait impairments in PD. Measures of this degeneration may therefore provide a novel biomarker for identifying future mobility loss and falls. © 2020 The Authors. Movement Disorders published by Wiley Periodicals LLC on behalf of International Parkinson and Movement Disorder Society.
Assuntos
Prosencéfalo Basal , Doença de Parkinson , Prosencéfalo Basal/diagnóstico por imagem , Colinérgicos , Estudos Transversais , Marcha , Humanos , Doença de Parkinson/complicaçõesRESUMO
Characterizing the thermal preference of fish is important in conservation, environmental and evolutionary physiology and can be determined using a shuttle box system. Initial tank acclimation and trial lengths are important considerations in experimental design, yet systematic studies of these factors are missing. Three different behavioral assay experimental designs were tested to determine the effect of tank acclimation and trial length (hours of tank acclimation:behavioral trial: 12:12, 0:12, 2:2) on the temperature preference of juvenile lake whitefish (Coregonus clupeaformis), using a shuttle box. Average temperature preferences for the 12 h:12 h, 0 h:12 h, 2 h:2 h experimental designs were 16.10±1.07°C, 16.02±1.56°C and 16.12±1.59°C respectively, with no significant differences between experimental designs (P=0.9337). Ultimately, length of acclimation time and trial length had no significant effect on thermal preference.
Assuntos
Aclimatação , Salmonidae , Animais , Evolução Biológica , TemperaturaRESUMO
BACKGROUND: The invasive benthic round goby (Neogobius melanostomus) is the most successful temperate invasive fish and has spread in aquatic ecosystems on both sides of the Atlantic. Invasive species constitute powerful in situ experimental systems to study fast adaptation and directional selection on short ecological timescales and present promising case studies to understand factors involved the impressive ability of some species to colonize novel environments. We seize the unique opportunity presented by the round goby invasion to study genomic substrates potentially involved in colonization success. RESULTS: We report a highly contiguous long-read-based genome and analyze gene families that we hypothesize to relate to the ability of these fish to deal with novel environments. The analyses provide novel insights from the large evolutionary scale to the small species-specific scale. We describe expansions in specific cytochrome P450 enzymes, a remarkably diverse innate immune system, an ancient duplication in red light vision accompanied by red skin fluorescence, evolutionary patterns of epigenetic regulators, and the presence of osmoregulatory genes that may have contributed to the round goby's capacity to invade cold and salty waters. A recurring theme across all analyzed gene families is gene expansions. CONCLUSIONS: The expanded innate immune system of round goby may potentially contribute to its ability to colonize novel areas. Since other gene families also feature copy number expansions in the round goby, and since other Gobiidae also feature fascinating environmental adaptations and are excellent colonizers, further long-read genome approaches across the goby family may reveal whether gene copy number expansions are more generally related to the ability to conquer new habitats in Gobiidae or in fish.
Assuntos
Peixes/fisiologia , Genoma , Espécies Introduzidas , Características de História de Vida , Animais , Feminino , Peixes/genética , MasculinoRESUMO
We examined the impact of repeated thermal stress on the heat shock response (HSR) of thermally sensitive lake whitefish (Coregonus clupeaformis) embryos. Our treatments were designed to mimic temperature fluctuations in the vicinity of industrial thermal effluents. Embryos were either maintained at control temperatures (3 oC) or exposed to a repeated thermal stress (TS) of 3 or 6 oC above control temperature every 3 or 6 days throughout embryonic development. At 82 days post-fertilisation, repeated TS treatments were stopped and embryos received either a high level TS of 12, 15, or 18 oC above ambient temperature for 1 or 4 h, or no additional TS. These treatments were carried out after a 6 h recovery from the last repeated TS. Embryos in the no repeated TS group responded, as expected, with increases in hsp70 mRNA in response to 12, 15 and 18 oC high-level TS. However, exposure to repeated TS of 3 or 6 °C every 6 days also resulted in a significant upregulation of hsp70 mRNA relative to the controls. Importantly, these repeated TS events and the associated elevations in hsp70 attenuated the upregulation of hsp70 in response to a 1 h, high-level TS of 12 oC above ambient, but not to either longer (4 h) or higher (15 or 18 oC) TS events. Conversely, hsp90α mRNA levels were not consistently elevated in the no repeated TS groups exposed to high-level TS. In some instances, hsp90α levels appeared to decrease in embryos exposed to repeated TS followed by a high-level TS. The observed attenuation of the HSR in lake whitefish embryos demonstrates that embryos of this species have plasticity in their HSR and repeated TS may protect against high-level TS, but the response differs based on repeated TS treatment, high-level TS temperature and duration, and the gene of interest.
Assuntos
Resposta ao Choque Térmico , Salmonidae/metabolismo , Animais , Proteínas de Peixes/genética , Proteínas de Peixes/metabolismo , Proteínas de Choque Térmico HSP70/genética , Proteínas de Choque Térmico HSP70/metabolismo , Salmonidae/embriologiaRESUMO
A laboratory flume was constructed to examine substrate effects on aquatic development. The flume was designed as a once-through system with a submerged cobble-filled corebox. Lake whitefish (Coregonus clupeaformis) embryos and temperature probes were deployed at multiple sites within the cobble and in the open water channel. Embryos were incubated in the flume for two different experimental periods: one to examine substrate impacts during natural lake cooling (37 days: 5 December 2016 to 10 January 2017) and the second to investigate substrate effects while administering a twice weekly 1 h heat shock (51 days: 11 January to 2 March 2017). During incubation, no significant difference was found in the average temperature between locations; however, temperatures were more stable within the cobble. Following both incubation periods, embryos retrieved from the cobble were significantly smaller in both dry mass and body length by up to 20%. These results demonstrate differences between embryos submerged in a cobble substrate and in the open water column, highlighting the need to consider the physical influences from the incubation environment when assessing development effects as part of any scientific study or environmental assessment.
Assuntos
Embrião não Mamífero/fisiologia , Desenvolvimento Embrionário/fisiologia , Salmonidae/embriologia , Animais , Meio Ambiente , Salmonidae/fisiologia , TemperaturaRESUMO
Cytochrome P450 (CYPs) enzymes are critical for the metabolism of exogenous and endogenous compounds. In mammals, the CYP3s are arguably the most important xenobiotic metabolizing enzymes and are all contained within the CYP3A subfamily. In fish, CYP3s include CYP3A and multiple subfamilies unique to the teleost lineage. The goal of this study was to provide insight on the regulation of genes in the CYP3C subfamily. Zebrafish, which have 4 CYP3C genes, were exposed to 17ß-estradiol (E2; 0.001-10⯵M) or ß-naphthoflavone (ßNF; 0.005-1⯵M), prototypical ligands of the estrogen receptor (ER) and the aryl hydrocarbon receptor (AhR), respectively. Gene expression was measured in the liver, intestine and gonads using quantitative PCR. CYP1A and vitellogenin (VTG) gene expression were used as positive controls for AhR and ER regulation, respectively. Exposure to ßNF resulted in the dose-dependant induction of CYP1A and CYP3C genes in the female intestine but not in the liver. E2 exposure resulted in the induction of all CYP3Cs in the male intestine and in the female liver. VTG was induced in both female and male livers. CYP3C3 and CYP3C4 were induced in the testis; CYP3C1 and CYP3C4 were slightly induced in the ovary. The time-course of gene induction was investigated in the liver and intestine after exposure to ßNF (0.5⯵M) and E2 (0.1⯵M). Inducible genes were up-regulated within 12â¯h after exposure. These data support a role for the AhR and ER in the regulation of CYP3Cs. Overall, the induction of CYP3Cs by AhR and ER ligands is different from mammalian CYP3A and may suggest a functional role for CYP3Cs that involves planar aromatic hydrocarbons and steroids.
Assuntos
Sistema Enzimático do Citocromo P-450/genética , Receptores de Hidrocarboneto Arílico/metabolismo , Receptores de Estrogênio/metabolismo , Peixe-Zebra/genética , Animais , Estradiol/farmacologia , Estrogênios/farmacologia , Feminino , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/metabolismo , Fígado/efeitos dos fármacos , Fígado/metabolismo , Masculino , Ovário/efeitos dos fármacos , Ovário/metabolismo , Testículo/efeitos dos fármacos , Testículo/metabolismo , Vitelogeninas/genética , Peixe-Zebra/metabolismo , beta-Naftoflavona/farmacologiaRESUMO
Chronic low-concentration chemical exposures may have both direct health outcomes on adults and indirect effects on their offspring. Using zebrafish, we examined the impacts of chronic, low-concentration carbamazepine (CBZ) exposure on a suite of male reproductive endpoints in the parents and four generations of offspring reared in clean water. CBZ is one of the most frequently detected pharmaceutical residues in water, is a histone deacetylase inhibitor in mammals, and is reported to lower androgens in mammals and fish. Exposure of adult zebrafish to 10 µg/L CBZ for 6 weeks decreased reproductive output, courtship and aggressive behaviors, 11-ketotestosterone (11KT), and sperm morphology but did not impact milt volume or sperm swimming speed. Pairwise breeding generated lineages of offspring with both parents exposed and two lineages where only one parent was exposed; the control lineage had unexposed parents. Reproductive output and male reproductive indices were assessed in F1-F4 offspring to determine whether parental CBZ exposure had transgenerational impacts. The offspring of CBZ-exposed males had lower 11KT, reproductive output, altered courtship, aggression, and sperm morphology compared to the lineage from unexposed parents. Our results indicate that parental carbamazepine exposure history impacts the unexposed progeny up to the F4 generations and that paternal, but not maternal, exposure is most important for the reproductive health of male offspring.
Assuntos
Poluentes Químicos da Água , Peixe-Zebra , Animais , Carbamazepina , Feminino , Humanos , Masculino , Exposição Paterna , ReproduçãoRESUMO
Lake whitefish (Coregonus clupeaformis) utilize overwintering embryonic development (up to 180â¯days), and such stenothermic, cold-water embryos may be particularly susceptible to thermal shifts. We incubated whitefish embryos in temperature treatments that were constant temperature (2.0⯱â¯0.1⯰C, 5.0⯱â¯0.1⯰C, and 8.0⯱â¯0.1⯰C; mean⯱â¯SD) or variable temperature (VT, meanâ¯=â¯5.0⯱â¯0.3⯰C). In the VT, a daily 2⯰C temperature change followed a continuous pattern throughout development: 2-4-6-8-6-4-2⯰C. Hatchling survival proportion from fertilization to hatch was significantly impacted by incubation temperature (Pâ¯<â¯0.001): 2⯰C (0.88⯱â¯0.01) and 5⯰C (0.91⯱â¯0.01) showed higher survival than both the VT (0.83⯱â¯0.02) and 8⯰C groups (0.15⯱â¯0.06), which were statistically distinct from each other. Time to hatch (dpf) was significantly different across all treatments (Pâ¯<â¯0.001): 8⯰C (68⯱â¯2â¯dpf), VT (111⯱â¯4â¯dpf), 5⯰C (116⯱â¯4â¯dpf), 2⯰C (170⯱â¯3â¯dpf). Likewise, hatchling yolk-free dry mass (mg) and total body length (mm) were significantly different across all treatments (Pâ¯<â¯0.001): 8⯰C (0.66⯱â¯0.08â¯mg; 11.1⯱â¯0.08â¯mm), VT (0.97⯱â¯0.06â¯mg; 11.7⯱â¯0.05â¯mm), 5⯰C (1.07⯱â¯0.03â¯mg; 12.0⯱â¯0.02â¯mm), 2⯰C (1.36⯱â¯0.04â¯mg; 12.8⯱â¯0.05â¯mm). Oxygen consumption rate (VÌo2) was significantly affected by the interaction between treatment and measurement temperature (Pâ¯<â¯0.001). Hatchling VT whitefish showed mean VÌo2 that was higher compared to the 2⯰C group measured at 2⯰C, and lower compared to the 2⯰C and 5⯰C group measured at 8⯰C. This study demonstrates that the VT incubation treatment produced fewer (increased mortality), smaller embryos that hatched earlier than 2⯰C and 5⯰C embryos. The plasticity of VÌo2 for this stenothermic-incubating fish species under variable incubation conditions reveals a metabolic cost to cycling thermal incubation conditions.
Assuntos
Salmonidae/fisiologia , Animais , Embrião não Mamífero/fisiologia , Desenvolvimento Embrionário , Consumo de Oxigênio/fisiologia , Salmonidae/embriologia , Salmonidae/crescimento & desenvolvimento , Salmonidae/metabolismoRESUMO
The Epstein-Barr virus (EBV) nuclear antigen 1 (EBNA1) is a sequence-specific DNA-binding protein that plays an essential role in viral episome replication and segregation, by recruiting the cellular complex of DNA replication onto the origin (oriP) and by tethering the viral DNA onto the mitotic chromosomes. Whereas the mechanisms of viral DNA replication are well documented, those involved in tethering EBNA1 to the cellular chromatin are far from being understood. Here, we have identified regulator of chromosome condensation 1 (RCC1) as a novel cellular partner for EBNA1. RCC1 is the major nuclear guanine nucleotide exchange factor for the small GTPase Ran enzyme. RCC1, associated with chromatin, is involved in the formation of RanGTP gradients critical for nucleo-cytoplasmic transport, mitotic spindle formation and nuclear envelope reassembly following mitosis. Using several approaches, we have demonstrated a direct interaction between these two proteins and found that the EBNA1 domains responsible for EBNA1 tethering to the mitotic chromosomes are also involved in the interaction with RCC1. The use of an EBNA1 peptide array confirmed the interaction of RCC1 with these regions and also the importance of the N-terminal region of RCC1 in this interaction. Finally, using confocal microscopy and Förster resonance energy transfer analysis to follow the dynamics of interaction between the two proteins throughout the cell cycle, we have demonstrated that EBNA1 and RCC1 closely associate on the chromosomes during metaphase, suggesting an essential role for the interaction during this phase, perhaps in tethering EBNA1 to mitotic chromosomes.
Assuntos
Proteínas de Ciclo Celular/metabolismo , Antígenos Nucleares do Vírus Epstein-Barr/metabolismo , Fatores de Troca do Nucleotídeo Guanina/metabolismo , Mitose , Proteínas Nucleares/metabolismo , Domínios e Motivos de Interação entre Proteínas , Motivos de Aminoácidos , Proteínas de Ciclo Celular/química , Proteínas de Ciclo Celular/genética , Cromatina/metabolismo , Cromossomos Humanos/metabolismo , Antígenos Nucleares do Vírus Epstein-Barr/química , Antígenos Nucleares do Vírus Epstein-Barr/genética , Transferência Ressonante de Energia de Fluorescência , Fatores de Troca do Nucleotídeo Guanina/química , Fatores de Troca do Nucleotídeo Guanina/genética , Células HEK293 , Células HeLa , Humanos , Metáfase , Microscopia Confocal , Proteínas Nucleares/química , Proteínas Nucleares/genética , Análise Serial de Proteínas , Mapeamento de Interação de Proteínas , Fuso Acromático/metabolismoRESUMO
Lipids serve as energy sources, structural components, and signaling molecules during fish embryonic development, and utilization of lipids may vary with temperature. Embryonic energy utilization under different temperatures is an important area of research in light of the changing global climate. Therefore, we examined percent lipid content and fatty acid profiles of lake whitefish (Coregonus clupeaformis) throughout embryonic development at three incubation temperatures. We sampled fertilized eggs and embryos at gastrulation, eyed and fin flutter stages following chronic incubation at temperatures of 1.8, 4.9 and 8.0°C. Hatchlings were also sampled following incubation at temperatures of 3.3, 4.9 and 8.0°C. Fertilized eggs had an initial high percentage of dry mass composed of lipid (percent lipid content; ~29%) consisting of ~20% saturated fatty acids (SFA), ~32% monounsaturated fatty acids (MUFA), ~44% polyunsaturated fatty acids (PUFA), and 4% unidentified. The most abundant fatty acids were 16:0, 16:1, 18:1(n-9c), 20:4(n-6), 20:5(n-3) and 22:6(n-3). This lipid profile matches that of other cold-water fish species. Percent lipid content increased during embryonic development, suggesting protein or other yolk components were preferentially used for energy. Total percentage of MUFA decreased during development, which indicated MUFA were the primary lipid catabolized for energy during embryonic development. Total percentage of PUFA increased during development, driven largely by an increase in 22:6(n-3). Temperature did not influence percent lipid content or percent MUFA at any development stage, and had inconsistent effects on percent SFA and percent PUFA during development. Thus, lake whitefish embryos appear to be highly adapted to low temperatures, and do not alter lipids in response to temperature within their natural incubation conditions.
Assuntos
Embrião não Mamífero/metabolismo , Desenvolvimento Embrionário , Ácidos Graxos/metabolismo , Metabolismo dos Lipídeos , Salmonidae/fisiologia , Zigoto/metabolismo , Animais , Temperatura Baixa/efeitos adversos , Metabolismo Energético , Feminino , Ionização de Chama/veterinária , Gastrulação , Lagos , Masculino , Ontário , Salmonidae/embriologia , Salmonidae/crescimento & desenvolvimentoRESUMO
Fluctuating incubation temperatures may have significant effects on fish embryogenesis; yet most laboratory-based studies use constant temperatures. For species that experience large, natural seasonal temperature changes during embryogenesis, such as lake whitefish (Coregonus clupeaformis), seasonal temperature regimes are likely optimal for development. Anthropogenic activities can increase average and/or variability of natural incubation temperatures over large (e.g. through climate change) or smaller (e.g. thermal effluent discharge) geographic scales. To investigate this, we incubated lake whitefish embryos under constant (2, 5, or 8°C) and fluctuating temperature regimes. Fluctuating temperature regimes had a base temperature of 2°C with: 1) seasonal temperature changes that modeled natural declines/inclines; 2) tri-weekly +3°C, 1h temperature spikes; or 3) both seasonal temperature changes and temperature spikes. We compared mortality to hatch, morphometrics, and heart rate at three developmental stages. Mortality rate was similar for embryos incubated at constant 2°C, constant 5°C, or with seasonal temperatures, but was significantly greater at constant 8°C. Embryos incubated constantly at >2°C had reduced body growth and yolk consumption compared to embryos incubated with seasonal temperature changes. When measured at the common base temperature of 2°C, embryos incubated at constant 2°C had lower heart rates than embryos incubated with both seasonal temperature changes and temperature spikes. Our study suggests that incubating lake whitefish embryos with constant temperatures may significantly alter development, growth, and heart rate compared to incubating with seasonal temperature changes, emphasizing the need to include seasonal temperature changes in laboratory-based studies.
Assuntos
Embrião não Mamífero/fisiologia , Desenvolvimento Embrionário , Salmonidae/embriologia , Estresse Fisiológico , Termotolerância , Animais , Aquicultura , Fertilização in vitro/veterinária , Great Lakes Region , Frequência Cardíaca , Temperatura Alta/efeitos adversos , Lagos , Ontário , Distribuição Aleatória , Salmonidae/crescimento & desenvolvimento , Salmonidae/fisiologia , Estações do Ano , Análise de Sobrevida , Saco Vitelino/embriologia , Saco Vitelino/fisiologiaRESUMO
We present a method to characterize variable thermal regimes in terms of an equivalent or effective temperature. Our method is based on a first order exponential transformation of a time series of temperatures to yield an exponentially-weighted mean temperature characteristic of the regime and independent of any particular species or end point. The resulting effective temperature or exponential mean, Te¯, offers an improved method for summarizing mean temperature where biological response scales exponentially to temperature. The exponential mean allows growth under varying thermal regimes to be predicted using constant temperature models and offers a compact descriptor communicating the growth capacity of variable thermal regimes. The method combines mathematical simplicity with translatability to different Q10 values without recourse to the underlining time series data. It also provides a quantitative baseline that improves on mean temperature by incorporating the effect of Jensen's inequality and it remains applicable at near zero temperatures where thermal sums lack accuracy.
Assuntos
Modelos Biológicos , Perciformes/crescimento & desenvolvimento , Salmonidae/crescimento & desenvolvimento , Algoritmos , Animais , Mudança Climática , Simulação por Computador , Ecossistema , TemperaturaRESUMO
Lake whitefish (Coregonus clupeaformis) embryos were exposed to thermal stress (TS) at different developmental stages to determine when the heat shock response (HSR) can be initiated and if it is altered by exposure to repeated TS. First, embryos were subject to one of three different TS temperatures (6, 9, or 12°C above control) at 4 points in development (21, 38, 60 and 70 days post-fertilisation (dpf)) for 2h followed by a 2h recovery to understand the ontogeny of the HSR. A second experiment explored the effects of repeated TS on the HSR in embryos from 15 to 75 dpf. Embryos were subjected to one of two TS regimes; +6°C TS for 1h every 6 days or +9°C TS for 1h every 6 days. Following a 2h recovery, a subset of embryos was sampled. Our results show that embryos could initiate a HSR via upregulation of heat shock protein 70 (hsp70) mRNA at all developmental ages studied, but that this response varied with age and was only observed with a TS of +9 or +12°C. In comparison, when embryos received multiple TS treatments, hsp70 was not induced in response to the 1h TS and 2h recovery, and a downregulation was observed at 39 dpf. Downregulation of hsp47 and hsp90α mRNA was also observed in early age embryos. Collectively, these data suggest that embryos are capable of initiating a HSR at early age and throughout embryogenesis, but that repeated TS can alter the HSR, and may result in either reduced responsiveness or a downregulation of inducible hsps. Our findings warrant further investigation into both the short- and long-term effects of repeated TS on lake whitefish development.
Assuntos
Resposta ao Choque Térmico , Salmonidae/embriologia , Animais , Regulação para Baixo , Embrião não Mamífero/embriologia , Embrião não Mamífero/fisiologia , Proteínas de Peixes/genética , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Choque Térmico HSP47/genética , Proteínas de Choque Térmico HSP70/genética , Proteínas de Choque Térmico HSP90/genética , Temperatura Alta , RNA Mensageiro/genética , Salmonidae/fisiologia , Regulação para CimaRESUMO
At present, the zebrafish embryo is increasingly used as an alternative animal model to screen for developmental toxicity after exposure to xenobiotics. Since zebrafish embryos depend on their own drug-metabolizing capacity, knowledge of their intrinsic biotransformation is pivotal in order to correctly interpret the outcome of teratogenicity assays. Therefore, the aim of this in vitro study was to assess the activity of cytochrome P450 (CYP)-a group of drug-metabolizing enzymes-in microsomes from whole zebrafish embryos (ZEM) of 5, 24, 48, 72, 96 and 120 h post-fertilization (hpf) by means of a mammalian CYP substrate, i.e., benzyloxy-methyl-resorufin (BOMR). The same CYP activity assays were performed in adult zebrafish liver microsomes (ZLM) to serve as a reference for the embryos. In addition, activity assays with the human CYP3A4-specific Luciferin isopropyl acetal (Luciferin-IPA) as well as inhibition studies with ketoconazole and CYP3cide were carried out to identify CYP activity in ZLM. In the present study, biotransformation of BOMR was detected at 72 and 96 hpf; however, metabolite formation was low compared with ZLM. Furthermore, Luciferin-IPA was not metabolized by the zebrafish. In conclusion, the capacity of intrinsic biotransformation in zebrafish embryos appears to be lacking during a major part of organogenesis.
Assuntos
Citocromo P-450 CYP3A/metabolismo , Sondas Moleculares/metabolismo , Peixe-Zebra/embriologia , Peixe-Zebra/metabolismo , Animais , Biotransformação/efeitos dos fármacos , Embrião não Mamífero/metabolismo , Desenvolvimento Embrionário/efeitos dos fármacos , Feminino , Luciferina de Vaga-Lumes/metabolismo , Humanos , Cetoconazol/farmacologia , Microssomos Hepáticos/efeitos dos fármacos , Microssomos Hepáticos/metabolismo , Oxazinas/metabolismo , Recombinação Genética/genética , Especificidade por Substrato/efeitos dos fármacosRESUMO
We investigated the effects of thermal stress on embryonic (fin flutter, vitelline circulation stage) and young of the year (YOY) juvenile lake whitefish by characterizing the kinetics of the heat shock response (HSR). Lake whitefish were subjected to one of three different heat shock (HS) temperatures (3, 6, or 9 °C above control) for six different lengths of time (0.25, 0.50, 1, 2, 3, or 4h) followed by a 2h recovery period at the control temperature of 2 °C or 14 °C for embryos and YOY juveniles, respectively. The duration of the HSR was examined by allowing the fish to recover for 1, 2, 4, 8, 12, 16, 24, 36, or 48 h following a 2h HS. In embryos, at the fin flutter stage, only hsp70 mRNA levels were upregulated in response to the various HS treatments. By comparison, all three typically inducible hsps, hsp90α, hsp70 and hsp47, were upregulated in the YOY juveniles. In both instances the HSR was long lasting, but much more so in embryos where hsp70 mRNA levels continued to increase for 48 h after a 2h HS and remained significantly higher than untreated controls. Collectively our data indicate that both embryo and YOY juvenile lake whitefish have a robust HSR which permits them to survive a 4h, 9 °C HS. Moreover, both life history stages are capable of triggering a HSR following a moderate 3 °C HS which is likely an important protective mechanism against environmental stressors during embryogenesis and early life history stages of lake whitefish.
Assuntos
Desenvolvimento Embrionário/genética , Peixes/genética , Resposta ao Choque Térmico/genética , Animais , Proteínas de Choque Térmico HSP70/genética , Lagos , RNA Mensageiro/genética , Temperatura , Regulação para Cima/genéticaRESUMO
Increasing incubation temperatures, caused by global climate change or thermal effluent from industrial processes, may influence embryonic development of fish. This study investigates the cumulative effects of increased incubation temperature and repeated heat shocks on developing Lake Whitefish (Coregonus clupeaformis) embryos. We studied the effects of three constant incubation temperatures (2°C, 5°C or 8°C water) and weekly, 1-h heat shocks (+3°C) on hatching time, survival and morphology of embryos, as these endpoints may be particularly susceptible to temperature changes. The constant temperatures represent the predicted magnitude of elevated water temperatures from climate change and industrial thermal plumes. Time to the pre-hatch stage decreased as constant incubation temperature increased (148d at 2°C, 92d at 5°C, 50d at 8°C), but weekly heat shocks did not affect time to hatch. Mean survival rates and embryo morphometrics were compared at specific developmental time-points (blastopore, eyed, fin flutter and pre-hatch) across all treatments. Constant incubation temperatures or +3°C heat-shock exposures did not significantly alter cumulative survival percentage (~50% cumulative survival to pre-hatch stage). Constant warm incubation temperatures did result in differences in morphology in pre-hatch stage embryos. 8°C and 5°C embryos were significantly smaller and had larger yolks than 2°C embryos, but heat-shocked embryos did not differ from their respective constant temperature treatment groups. Elevated incubation temperatures may adversely alter Lake Whitefish embryo size at hatch, but weekly 1-h heat shocks did not affect size or survival at hatch. These results suggest that intermittent bouts of warm water effluent (e.g., variable industrial emissions) are less likely to negatively affect Lake Whitefish embryonic development than warmer constant incubation temperatures that may occur due to climate change.