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1.
Mol Cell ; 75(6): 1161-1177.e11, 2019 09 19.
Artigo em Inglês | MEDLINE | ID: mdl-31421980

RESUMO

Genes are transcribed in a discontinuous pattern referred to as RNA bursting, but the mechanisms regulating this process are unclear. Although many physiological signals, including glucocorticoid hormones, are pulsatile, the effects of transient stimulation on bursting are unknown. Here we characterize RNA synthesis from single-copy glucocorticoid receptor (GR)-regulated transcription sites (TSs) under pulsed (ultradian) and constant hormone stimulation. In contrast to constant stimulation, pulsed stimulation induces restricted bursting centered around the hormonal pulse. Moreover, we demonstrate that transcription factor (TF) nuclear mobility determines burst duration, whereas its bound fraction determines burst frequency. Using 3D tracking of TSs, we directly correlate TF binding and RNA synthesis at a specific promoter. Finally, we uncover a striking co-bursting pattern between TSs located at proximal and distal positions in the nucleus. Together, our data reveal a dynamic interplay between TF mobility and RNA bursting that is responsive to stimuli strength, type, modality, and duration.


Assuntos
Glucocorticoides/farmacologia , Regiões Promotoras Genéticas , RNA/biossíntese , Receptores de Glucocorticoides/metabolismo , Sítio de Iniciação de Transcrição , Transcrição Gênica/efeitos dos fármacos , Animais , Camundongos , RNA/genética
2.
Trends Biochem Sci ; 46(5): 417-428, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33309324

RESUMO

To identify the appropriate times for growth and development, organisms must sense and process information about the availability of nutrients, energy status, and environmental cues. For sessile eukaryotes such as plants, integrating such information can be critical in life or death decisions. For nearly 30 years, the conserved phosphatidylinositol 3-kinase-related protein kinases (PIKKs) target of rapamycin (TOR) has been established as a central hub for integrating external and internal metabolic cues. Despite the functional conservation across eukaryotes, the TOR complex has evolved specific functional and mechanistic features in plants. Here, we present recent findings on the plant TOR complex that highlight the conserved and unique nature of this critical growth regulator and its role in multiple aspects of plant life.


Assuntos
Sirolimo , Serina-Treonina Quinases TOR , Plantas/metabolismo , Transdução de Sinais , Serina-Treonina Quinases TOR/metabolismo
3.
J Pineal Res ; 76(5): e13001, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-39092800

RESUMO

This study explores the 24-h rhythmic cycle of protein O-GlcNAcylation within the brain and highlights its crucial role in regulating the circadian cycle and neuronal function based on zebrafish as an animal model. In our experiments, disruption of the circadian rhythm, achieved through inversion of the light-dark cycle or daytime melatonin treatment, not only impaired the rhythmic changes of O-GlcNAcylation along with altering expression patterns of O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA) in zebrafish brain but also significantly impeded learning and memory function. In particular, circadian disruption affected rhythmic expression of protein O-GlcNAcylation and OGT in the nuclear fraction. Notably, the circadian cycle induces rhythmic alterations in O-GlcNAcylation of H2B histone protein that correspond to changes in H3 trimethylation. Disruption of the cycle interfered with these periodic histone code alterations. Pharmacological inhibition of OGT with OSMI-1 disrupted the wake-sleep patterns of zebrafish without affecting expression of circadian rhythm-regulating genes. OSMI-1 inhibited the expression of c-fos, bdnf, and calm1, key genes associated with brain function and synaptic plasticity, and decreased the binding of O-GlcNAcylated H2B and OGT to promoter regions of these genes. The collective findings support the potential involvement of circadian cycling of the O-GlcNAc histone code in regulating synaptic plasticity and brain function. Overall, data from this study provide evidence that protein O-GlcNAcylation serves as a pivotal posttranslational mechanism integrating circadian signals and neuronal function to regulate rhythmic physiology.


Assuntos
Ritmo Circadiano , N-Acetilglucosaminiltransferases , Peixe-Zebra , Animais , Peixe-Zebra/metabolismo , Ritmo Circadiano/fisiologia , N-Acetilglucosaminiltransferases/metabolismo , N-Acetilglucosaminiltransferases/genética , Cognição/fisiologia , Proteínas de Peixe-Zebra/metabolismo , Proteínas de Peixe-Zebra/genética , Luz , Encéfalo/metabolismo
4.
J Exp Biol ; 226(17)2023 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-37408509

RESUMO

Gymnotiformes are nocturnal fishes inhabiting the root mats of floating plants. They use their electric organ discharge (EOD) to explore the environment and to communicate. Here, we show and describe tonic and phasic sensory-electromotor responses to light distinct from indirect effects depending on the light-induced endogenous circadian rhythm. In the dark, principally during the night, inter-EOD interval histograms are bimodal: the main peak corresponds to the basal rate and a secondary peak corresponds to high-frequency bouts. Light causes a twofold tonic but opposing effect on the EOD histogram: (i) decreasing the main mode and (ii) blocking the high-frequency bouts and consequently increasing the main peak at the expense of removal of the secondary one. Additionally, light evokes phasic responses whose amplitude increases with intensity but whose slow time course and poor adaptation differentiate from the so-called novelty responses evoked by abrupt changes in sensory stimuli of other modalities. We confirmed that Gymnotus omarorum tends to escape from light, suggesting that these phasic responses are probably part of a global 'light-avoidance response'. We interpret the data within an ecological context. Fish rest under the shade of aquatic plants during the day and light spots due to the sun's relative movement alert the fish to hide in shady zones to avoid macroptic predators and facilitate tracking the movement of floating plant islands by wind and/or water currents.


Assuntos
Peixe Elétrico , Gimnotiformes , Animais , Órgão Elétrico/fisiologia , Gimnotiformes/fisiologia , Movimento , Peixe Elétrico/fisiologia
5.
Pediatr Nephrol ; 38(5): 1427-1438, 2023 05.
Artigo em Inglês | MEDLINE | ID: mdl-36427087

RESUMO

BACKGROUND: Nocturnal enuresis (NE) is a multifactorial and complex condition. One less understood factor in its pathophysiology is the enuretic inability to wake up when the bladder is full (impaired arousal). OBJECTIVE: We aimed to investigate the relationship between sleep and NE in children and adolescents. METHODS: A systematic review was performed following the PRISMA guidelines, and the electronic databases MEDLINE (via PubMed) and SCOPUS were searched until March 2022. Eligibility criteria were studies that recruited patients aged five-17 years with a diagnosis of NE according to the International Child Continence Society (ICCS), Diagnostic and Statistical Manual of Mental Disorders-Fifth Edition (DSM-5), or International Classification Criteria of Sleep Disorders-Third edition (ICSD-3) who had their sleep assessed using validated questionnaires and/or polysomnography. The tool used to analyze the risk of bias in the included studies was the risk of bias in non-randomized studies of exposure. RESULTS: Of 1582 citations screened, nine were included, giving 1685 participants, 581 with NE. All studies were observational and half had a low risk of bias. Four studies evaluated sleep by questionnaires only; two used questionnaires and polysomnography; two used only polysomnography, and one used sleep logs and actigraphy. Sleep questionnaires showed that children with enuresis had more sleep problems than controls, especially parasomnias, breathing disorders, and daytime sleepiness. Among the polysomnography parameters, the sleep stage architecture and periodic limb movements during sleep had conflicting data between the two studies. LIMITATIONS: The studies evaluated sleep through heterogeneous tools. They used different questionnaires; even those considered by polysomnography did not record the same channels. CONCLUSION: It seems that enuretic children and adolescents sleep differently from those who are non-enuretic. More studies are needed to clarify the best way to assess sleep and better understand this relationship. The review protocol was registered with PROSPERO, CRD42021266338. There was no funding.


Assuntos
Enurese Noturna , Incontinência Urinária , Humanos , Criança , Adolescente , Enurese Noturna/epidemiologia , Sono , Polissonografia , Bexiga Urinária
6.
Proc Natl Acad Sci U S A ; 117(1): 668-676, 2020 01 07.
Artigo em Inglês | MEDLINE | ID: mdl-31848247

RESUMO

The glymphatic system functions in the removal of potentially harmful metabolites and proteins from the brain. Dynamic, contrast-enhanced MRI was used in fully awake rats to follow the redistribution of intraventricular contrast agent entrained to the light-dark cycle and its hypothetical relationship to the sleep-waking cycle, blood flow, and brain temperature in specific brain areas. Brain areas involved in circadian timing and sleep-wake rhythms showed the lowest redistribution of contrast agent during the light phase or time of inactivity and sleep in rats. Global brain redistribution of contrast agent was heterogeneous. The redistribution was highest along the dorsal cerebrum and lowest in the midbrain/pons and along the ventral surface of the brain. This heterogeneous redistribution of contrast agent paralleled the gradients and regional variations in brain temperatures reported in the literature for awake animals. Three-dimensional quantitative ultrashort time-to-echo contrast-enhanced imaging was used to reconstruct small, medium, and large arteries and veins in the rat brain and revealed areas of lowest redistribution overlapped with this macrovasculature. This study raises new questions and theoretical considerations of the impact of the light-dark cycle, brain temperature, and blood flow on the function of the glymphatic system.


Assuntos
Ritmo Circadiano/fisiologia , Sistema Glinfático/diagnóstico por imagem , Fotoperíodo , Vigília/fisiologia , Animais , Temperatura Corporal/fisiologia , Circulação Cerebrovascular/fisiologia , Meios de Contraste/administração & dosagem , Sistema Glinfático/fisiologia , Imageamento Tridimensional , Imageamento por Ressonância Magnética , Masculino , Modelos Animais , Ratos , Sono/fisiologia
7.
Genomics ; 114(2): 110283, 2022 03.
Artigo em Inglês | MEDLINE | ID: mdl-35143886

RESUMO

The liver plays a principal role in avian migration. Here, we characterised the liver transcriptome of a long-distance migrant, the Northern Wheatear (Oenanthe oenanthe), sampled at different migratory stages, looking for molecular processes linked with adaptations to migration. The analysis of the differentially expressed genes suggested changes in the periods of the circadian rhythm, variation in the proportion of cells in G1/S cell-cycle stages and the putative polyploidization of this cell population. This may explain the dramatic increment in the liver's metabolic capacities towards migration. Additionally, genes involved in anti-oxidative stress, detoxification and innate immune responses, lipid metabolism, inflammation and angiogenesis were regulated. Lipophagy and lipid catabolism were active at all migratory stages and increased towards the fattening and fat periods, explaining the relevance of lipolysis in controlling steatosis and maintaining liver health. Our study clears the way for future functional studies regarding long-distance avian migration.


Assuntos
Migração Animal , Aves Canoras , Migração Animal/fisiologia , Animais , Fígado , Aves Canoras/genética , Transcriptoma
8.
Int J Mol Sci ; 24(3)2023 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-36769150

RESUMO

The aim of the experiment was to test the effect of an elevated level of glucocorticoids on the mouse hippocampal transcriptome after 12 h of treatment with corticosterone that was administered during an active phase of the circadian cycle. Additionally, we also tested the circadian changes in gene expression and the decay time of transcriptomic response to corticosterone. Gene expression was analyzed using microarrays. Obtained results show that transcriptomic responses to glucocorticoids are heterogeneous in terms of the decay time with some genes displaying persistent changes in expression during 9 h of rest. We have also found a considerable overlap between genes regulated by corticosterone and genes implicated previously in stress response. The examples of such genes are Acer2, Agt, Apod, Aqp4, Etnppl, Fabp7, Fam107a, Fjx1, Fmo2, Galnt15, Gjc2, Heph, Hes5, Htra1, Jdp2, Kif5a, Lfng, Lrg1, Mgp, Mt1, Pglyrp1, Pla2g3, Plin4, Pllp, Ptgds, Ptn, Slc2a1, Slco1c1, Sult1a1, Thbd and Txnip. This indicates that the applied model is a useful tool for the investigation of mechanisms underlying the stress response.


Assuntos
Corticosterona , Glucocorticoides , Camundongos , Animais , Corticosterona/farmacologia , Corticosterona/metabolismo , Glucocorticoides/metabolismo , Hipocampo/metabolismo , Perfilação da Expressão Gênica , Transcriptoma
9.
Int J Mol Sci ; 23(21)2022 Oct 27.
Artigo em Inglês | MEDLINE | ID: mdl-36361798

RESUMO

Dinoflagellates bioluminescence mechanism depends upon a luciferin-luciferase reaction that promotes blue light emission (480 nm) in specialized luminogenic organelles called scintillons. The scintillons contain luciferin, luciferase and, in some cases, a luciferin-binding protein (LBP), which prevents luciferin from non-enzymatic oxidation in vivo. Even though dinoflagellate bioluminescence has been studied since the 1950s, there is still a lack of mechanistic understanding on whether the light emission process involves a peroxidic intermediate or not. Still, bioassays employing luminous dinoflagellates, usually from Gonyaulax or Pyrocystis genus, can be used to assess the toxicity of metals or organic compounds. In these dinoflagellates, the response to toxicity is observed as a change in luminescence, which is linked to cellular respiration. As a result, these changes can be used to calculate a percentage of light inhibition that correlates directly with toxicity. This current approach, which lies in between fast bacterial assays and more complex toxicity tests involving vertebrates and invertebrates, can provide a valuable tool for detecting certain pollutants, e.g., metals, in marine sediment and seawater. Thus, the present review focuses on how the dinoflagellates bioluminescence can be applied to evaluate the risks caused by contaminants in the marine environment.


Assuntos
Dinoflagellida , Animais , Dinoflagellida/metabolismo , Luciferases/metabolismo , Luminescência , Bioensaio , Sedimentos Geológicos , Medições Luminescentes
11.
Plant Cell Environ ; 42(6): 1897-1912, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30673142

RESUMO

Solar irradiance and air temperature are characterized by dramatic circadian fluctuations and are known to significantly modulate fruit composition. To date, it remains unclear whether the abrupt, yet predictive, diurnal changes in radiation and temperature prompt direct metabolic turn-over in the fruit. We assessed the role of fruit insolation, air temperature, and source-tissue CO2 assimilation in the diurnal compositional changes in ripening grape berries. This was performed by comparing the diurnal changes in metabolite profiles of berries positioned such that they experienced (a) contrasting diurnal solar irradiance patterns, and (b) similar irradiance but contrasting diurnal CO2 assimilation patterns of adjacent leaves. Grape carbon levels increased during the morning and decreased thereafter. Sucrose levels decreased throughout the day and were correlated with air temperature, but not with the diurnal pattern of leaf CO2 assimilation. Tight correlation between sucrose and glucose-6-phosphate indicated the involvement of photorespiration/glycolysis in sucrose depletion. Amino acids, polyamines, and phenylpropanoids fluctuated diurnally, and were highly responsive to the diurnal insolation pattern of the fruit. Our results fill the knowledge gap regarding the circadian pattern of source-sink assimilate-translocation in grapevine. In addition, they suggest that short-term direct solar exposure of the fruit impacts both its diurnal and nocturnal metabolism.


Assuntos
Frutas/anatomia & histologia , Frutas/metabolismo , Metaboloma , Vitis/metabolismo , Aminoácidos/metabolismo , Carbono/metabolismo , Dióxido de Carbono , Israel , Folhas de Planta/metabolismo , Sacarose/metabolismo , Temperatura
12.
J Eukaryot Microbiol ; 66(2): 254-266, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30027647

RESUMO

Two genes of the RACK1 homolog from the photosynthetic dinoflagellate Symbiodinium microadriaticum ssp. microadriaticum (SmicRACK1), termed SmicRACK1A and SmicRACK1B, were found tandemly arrayed and displayed a single synonymous substitution (T/C) encoding threonine. They included two exons of 942 bp each, encoding 313 amino acids with seven WD-40 repeats and two PKC-binding motifs. The protein theoretical mass and pI were 34,200 Da and 5.9, respectively. SmicRACK1 showed maximum identities with RACK1 homologs at the amino acid and nucleotide level, respectively, of 92 and 84% with S. minutum, and phylogenetic analysis revealed clustered related RACK1 sequences from the marine dinoflagellates S. minutum, Heterocapsa triquetra, Karenia brevis, and Alexandrium tamarense. Interestingly, light-dependent regulatory elements were found both within the 282 bp SmicRACK1A promotor sequence, and within an intergenic sequence of 359 nucleotides that separated both genes, which strongly suggest light-related functions. This was further supported by mRNA accumulation analysis, which fluctuated along the light and dark phases of the growth cycle showing maximum specific peaks under either condition. Finally, qRT-PCR analysis revealed differential SmicRACK1 mRNA accumulation with maxima at 6 and 20 d of culture. Our SmicRACK1 characterization suggests roles in active growth and proliferation, as well as light/dark cycle regulation in S. microadriaticum.


Assuntos
Dinoflagellida/genética , Expressão Gênica , Proteínas de Protozoários/genética , RNA Mensageiro/genética , Receptores de Quinase C Ativada/genética , Proteínas de Algas/química , Proteínas de Algas/genética , Proteínas de Algas/metabolismo , Sequência de Aminoácidos , Sequência de Bases , Dinoflagellida/metabolismo , Filogenia , Proteínas de Protozoários/química , Proteínas de Protozoários/metabolismo , RNA Mensageiro/metabolismo , Receptores de Quinase C Ativada/química , Receptores de Quinase C Ativada/metabolismo
13.
Int J Biometeorol ; 63(11): 1487-1496, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31147777

RESUMO

Long-term assessments of thermal responses of housed Jersey cows raised in tropical conditions were performed to investigate the effect of climate environment on their physiological performance and thermal equilibrium. Twelve Jersey dairy cows with 326.28 ± 30 kg of body weight, 17.66 ± 1.8 of milk yield, and 165.5 ± 6.8 of days in milking were assigned in two 12 × 12 Latin square designs. Air temperature, relative humidity, partial vapor pressure, direct and diffuse short-wave solar radiation and black globe temperature under the shade, and direct sunlight were recorded. Physiological responses as respiratory rate (RR, breaths min-1), ventilation (VE, L s-1), proportion (%) of oxygen (O2) and carbon dioxide (CO2), saturation pressure (PS{TEXH}), and air temperature (TEXH, °C) of the exhaled air were assessed protected from solar radiation and rain. Rectal temperature (TR, °C), skin temperature (TEP, °C), and hair coat surface temperature (TS, °C) were also recorded. The thermal equilibrium was determined from biophysical equations according to the principles of the energy conservation law in a control volume. Exploratory and confirmatory analyses were performed from principal components and by the least square method, respectively. The cows were evaluated under range of ambient air temperature from 26 to 35 °C, relative humidity from 27 to 89%, and short-wave radiation from 0 to 729 W m-2. Exploratory and confirmatory analyses demonstrated that a similar level of nocturnal and diurnal air temperatures evoked distinct (P < 0.05) responses for rectal (TR, °C) and skin (TEP, °C) temperatures, ventilation (VE, L s-1), tidal volume (TV, L breaths-1), and oxygen consumption (∆O2, %) and carbon dioxide output (∆CO2, %), clearly revealing an endogenous rhythm dependence. In conclusion, these findings clarify how the circadian rhythmicity of the thermal environment and animal's biological clock dictate dynamics of heat generated by metabolism, dissipated to the environment and physiological parameters of the housed Jersey cows raised in tropical condition; therefore, it is fundamental to help us to understand how the Jersey dairy cows under tropics are affected by the climatic conditions, leading to better ways of the environmental management.


Assuntos
Regulação da Temperatura Corporal , Leite , Animais , Temperatura Corporal , Bovinos , Feminino , Temperatura Alta , Umidade , Lactação , Temperatura , Clima Tropical
14.
Int J Mol Sci ; 20(14)2019 Jul 19.
Artigo em Inglês | MEDLINE | ID: mdl-31330985

RESUMO

Smith-Magenis syndrome (SMS), linked to Retinoic Acid Induced (RAI1) haploinsufficiency, is a unique model of the inversion of circadian melatonin secretion. In this regard, this model is a formidable approach to better understand circadian melatonin secretion cycle disorders and the role of the RAI1 gene in this cycle. Sleep-wake cycle disorders in SMS include sleep maintenance disorders with a phase advance and intense sleepiness around noon. These disorders have been linked to a general disturbance of sleep-wake rhythm and coexist with inverted secretion of melatonin. The exact mechanism underlying the inversion of circadian melatonin secretion in SMS has rarely been discussed. We suggest three hypotheses that could account for the inversion of circadian melatonin secretion and discuss them. First, inversion of the circadian melatonin secretion rhythm could be linked to alterations in light signal transduction. Second, this inversion could imply global misalignment of the circadian system. Third, the inversion is not linked to a global circadian clock shift but rather to a specific impairment in the melatonin secretion pathway between the suprachiasmatic nuclei (SCN) and pinealocytes. The development of diurnal SMS animal models that produce melatonin appears to be an indispensable step to further understand the molecular basis of the circadian melatonin secretion rhythm.


Assuntos
Suscetibilidade a Doenças , Melatonina/biossíntese , Síndrome de Smith-Magenis/etiologia , Síndrome de Smith-Magenis/metabolismo , Animais , Mapeamento Cromossômico , Ritmo Circadiano , Predisposição Genética para Doença , Humanos , Locos de Características Quantitativas , Característica Quantitativa Herdável , Síndrome de Smith-Magenis/diagnóstico
15.
Medicina (B Aires) ; 79(Suppl 1): 44-50, 2019.
Artigo em Espanhol | MEDLINE | ID: mdl-30776279

RESUMO

Adequate sleep is of critical need for a typical synaptic development and brain maturation, a poor quality sleep can have detrimental effects on children's' cognitive attention, memory, mood regulation, and behavior functions. Great concern has been voiced out regarding the high prevalence of poor sleep in children worldwide, the effects of poor sleep may be even more pronounced in children with neurodevelopmental disorders; these children often have difficulties with falling and staying asleep and with night awakenings, this has a strong association with daytime behavior problems. The purpose of this article is to provide an overview of the state of the science of sleep in children with a neurodevelopmental disorder. In this context, it is important to take the circadian cycle into account, a genetically encoded clock that drives cellular rhythms of transcription, translation and metabolism. The circadian clock interacts with the diurnal and nocturnal environment that also drives transcription and metabolism during light/dark, sleep/wake, hot/cold and feast/fast daily and seasonal cycles In conclusion, the sleep problems are a conditioning factor in the evolution and quality of life of children with neurodevelopmental disorders that must be taken into account in all cases and occupy a preferential place in both the diagnostic and the therapeutic stages.


El sueño adecuado es necesario para el desarrollo sináptico y la maduración cerebral, un sueño de mala calidad tiene efectos perjudiciales en las funciones cognitivas, de atención, memoria y conducta de los niños. La preocupación sobre la alta prevalencia de los problemas del sueño es amplia en todo el mundo; las consecuencias de estos problemas son incluso más importantes en los niños portadores de trastornos del neurodesarrollo; estos niños a menudo tienen dificultades de inicio y mantenimiento del sueño y despertares nocturnos frecuentes que afectan a sus problemas de conducta. El propósito de este escrito es revisar el estado del arte de los problemas del sueño en los niños con trastornos del neurodesarrollo. En este punto, es importante tener en cuenta el ritmo circadiano, un reloj que genéticamente dirige los ritmos celulares de transcripción, traslación y metabolismos. Este reloj se combina con el ambiente diurno y nocturno coordinando estos mecanismos durante los ciclos luz/oscuridad, sueño/vigilia, frío/calor, ingesta/ayuno, tanto diariamente como en las diferentes estaciones. En conclusión, los problemas del sueño son un factor condicionante de la evolución y calidad de vida de los niños con trastornos del neurodesarrollo, que debe ser tenido en cuenta en todos los casos y ocupar un lugar preferente tanto en la etapa diagnóstica como en la terapéutica.


Assuntos
Transtornos do Neurodesenvolvimento/fisiopatologia , Transtornos do Sono-Vigília/fisiopatologia , Transtorno do Deficit de Atenção com Hiperatividade/fisiopatologia , Transtorno do Espectro Autista/fisiopatologia , Criança , Relógios Circadianos , Ritmo Circadiano/fisiologia , Humanos , Higiene do Sono/fisiologia , Transtornos do Sono-Vigília/terapia , Síndrome de Smith-Magenis/fisiopatologia
16.
Toxicol Appl Pharmacol ; 332: 149-158, 2017 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-28392392

RESUMO

Dichloromethane (DCM) is a lung and liver carcinogen in mice at inhalation exposures≥2000ppm. The modes of action (MOA) of these responses have been attributed to formation of genotoxic, reactive metabolite(s). Here, we examined gene expression in lung and liver from female B6C3F1 mice exposed to 0, 100, 500, 2000, 3000 and 4000ppm DCM for 90days. We also simulated dose measures - rates of DCM oxidation to carbon monoxide (CO) in lung and liver and expected blood carboxyhemoglobin (HbCO) time courses with a PBPK model inclusive of both conjugation and oxidation pathways. Expression of large numbers of genes was altered at 100ppm with maximal changes in the numbers occurring by 500 or 2000ppm. Most changes in genes common to the two tissues were related to cellular metabolism and circadian clock. At the lower concentrations, the changes in metabolism-related genes were discordant - up in liver and down in lung. These processes included organelle biogenesis, TCA cycle, and respiratory electron transport. Changes in circadian cycle genes - primarily transcription factors - showed strong concentration-related response at higher concentrations (Arntl, Npas2, and Clock were down-regulated; Cry2, Wee1, Bhlhe40, Per3, Nr1d1, Nr1d2 and Dbp) were up-regulated with similar directionality in both tissues. Overall, persistently elevated HbCO from DCM oxidation appears to cause extended periods of hypoxia, leading to altered circadian coupling to cellular metabolism. The dose response for altered circadian processes correlates with the cancer outcome. We found no evidence of changes in genes indicative of responses to cytotoxic, DNA-reactive metabolites.


Assuntos
Ritmo Circadiano , Hipóxia/genética , Fígado/efeitos dos fármacos , Pulmão/efeitos dos fármacos , Cloreto de Metileno/toxicidade , Transcriptoma , Animais , Carboxihemoglobina/genética , Carboxihemoglobina/metabolismo , Ritmo Circadiano/efeitos dos fármacos , Ritmo Circadiano/genética , Relação Dose-Resposta a Droga , Feminino , Regulação da Expressão Gênica , Hipóxia/induzido quimicamente , Hipóxia/patologia , Exposição por Inalação/efeitos adversos , Fígado/metabolismo , Pulmão/metabolismo , Camundongos , Camundongos Endogâmicos , Farmacocinética , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
17.
Mol Syst Biol ; 10: 739, 2014 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-25028488

RESUMO

Circadian cycles and cell cycles are two fundamental periodic processes with a period in the range of 1 day. Consequently, coupling between such cycles can lead to synchronization. Here, we estimated the mutual interactions between the two oscillators by time-lapse imaging of single mammalian NIH3T3 fibroblasts during several days. The analysis of thousands of circadian cycles in dividing cells clearly indicated that both oscillators tick in a 1:1 mode-locked state, with cell divisions occurring tightly 5 h before the peak in circadian Rev-Erbα-YFP reporter expression. In principle, such synchrony may be caused by either unidirectional or bidirectional coupling. While gating of cell division by the circadian cycle has been most studied, our data combined with stochastic modeling unambiguously show that the reverse coupling is predominant in NIH3T3 cells. Moreover, temperature, genetic, and pharmacological perturbations showed that the two interacting cellular oscillators adopt a synchronized state that is highly robust over a wide range of parameters. These findings have implications for circadian function in proliferative tissues, including epidermis, immune cells, and cancer.


Assuntos
Ciclo Celular , Ritmo Circadiano , Mamíferos/fisiologia , Adenina/análogos & derivados , Adenina/farmacologia , Animais , Proteínas CLOCK/metabolismo , Ciclo Celular/efeitos dos fármacos , Ritmo Circadiano/efeitos dos fármacos , Criptocromos/metabolismo , Mamíferos/genética , Camundongos , Modelos Biológicos , Células NIH 3T3 , Temperatura , Imagem com Lapso de Tempo
18.
J Heat Transfer ; 137(10): 1030011-10300112, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26424899

RESUMO

Our bodies depend on an exquisitely sensitive and refined temperature control system to maintain a state of health and homeostasis. The exceptionally broad range of physical activities that humans engage in and the diverse array of environmental conditions we face require remarkable strategies and mechanisms for regulating internal and external heat transfer processes. On the occasions for which the body suffers trauma, therapeutic temperature modulation is often the approach of choice for reversing injury and inflammation and launching a cascade of healing. The focus of human thermoregulation is maintenance of the body core temperature within a tight range of values, even as internal rates of energy generation may vary over an order of magnitude, environmental convection, and radiation heat loads may undergo large changes in the absence of any significant personal control, surface insulation may be added or removed, all occurring while the body's internal thermostat follows a diurnal circadian cycle that may be altered by illness and anesthetic agents. An advanced level of understanding of the complex physiological function and control of the human body may be combined with skill in heat transfer analysis and design to develop life-saving and injury-healing medical devices. This paper will describe some of the challenges and conquests the author has experienced related to the practice of heat transfer for maintenance of health and enhancement of healing processes.

19.
J Neurogenet ; 28(1-2): 136-45, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24579665

RESUMO

The combination of stable isotope labeling of amino acids in mammals (SILAM) and laser capture microdissection (LCM) for selective proteomic analysis of the targeted tissues holds tremendous potential for refined characterization of proteome changes within complex tissues such as the brain. The authors have applied this approach to measure changes in relative protein abundance in ventral tegmental area (VTA) of the rat brain that correlate to pharmacological perturbations. Enriched (13)C6(15)N2-lysine was introduced in vivo via diet. These animals were sacrificed during the middle of the 12-hour light period to extract isotopically "heavy" proteins, which were then used as a reference for extracts from dosed, unlabeled rats. Animals were administered an orexin peptide (Ox-B), an orexin receptor antagonist (ORA), or a mixture of both (Ox-B + ORA). All samples were obtained at same phase of the sleep cycle. Labeled-pair identification and differential quantitation provided protein identification and expression ratio data. Five proteins were found to exhibit decreased relative abundance after administration of an ORA, including α-synuclein and rat myelin basic protein. Conversely, six proteins showed increased relative abundance upon antagonist treatment, including 2',3'-cyclic nucleotide 3'-phosphodiesterase.


Assuntos
Núcleo Celular/metabolismo , Proteômica , Sono/fisiologia , Área Tegmentar Ventral/citologia , Aminoácidos/metabolismo , Animais , Animais Recém-Nascidos , Peso Corporal/efeitos dos fármacos , Feminino , Peptídeos e Proteínas de Sinalização Intracelular/administração & dosagem , Peptídeos e Proteínas de Sinalização Intracelular/antagonistas & inibidores , Peptídeos e Proteínas de Sinalização Intracelular/química , Tamanho da Ninhada de Vivíparos/efeitos dos fármacos , Lisina/administração & dosagem , Masculino , Neuropeptídeos/administração & dosagem , Neuropeptídeos/antagonistas & inibidores , Neuropeptídeos/química , Orexinas , Peptídeos/administração & dosagem , Gravidez , Mapas de Interação de Proteínas/genética , Proteínas/metabolismo , Desempenho Psicomotor/efeitos dos fármacos , Desempenho Psicomotor/fisiologia , Ratos , Ratos Sprague-Dawley
20.
J Exp Biol ; 217(Pt 15): 2704-9, 2014 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-24803470

RESUMO

Cupiennius salei is a nocturnal spider with eight eyes, which undergo a remarkable circadian cycle: the rhabdomeric membrane of the photoreceptor cells is dismantled during the day and rebuilt at the beginning of the night. Such drastic changes might influence the brightness discrimination ability. We tested this hypothesis by presenting square-shaped flickering stimuli with certain luminances on stationary backgrounds with other luminances to spiders with day- or night-adapted eyes. When the spider, through its three pairs of so-called secondary eyes, perceives a visible contrast between the stimulus and the background, its principal eye muscle activity should increase. We therefore recorded this activity in vivo to assess the brightness discrimination ability of Cupiennius salei. Our results show that this spider has good brightness discrimination ability, which is significantly better with dark-adapted eyes. A Michelson contrast of 0.1 to 0.2 at night, and of 0.2 to 0.3 for day-adapted eyes, is sufficient to elicit a significant response, except below a critical value of luminance (~16 cd m(-2)), where the minimal perceivable contrast needs to be higher. In the Discussion we compare these performances with those of other animals, in particular with jumping spiders.


Assuntos
Adaptação à Escuridão/fisiologia , Aranhas/fisiologia , Visão Ocular/fisiologia , Animais , Músculos Oculomotores/fisiologia , Estimulação Luminosa
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