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1.
Cell Rep Methods ; 4(2): 100714, 2024 Feb 26.
Artículo en Inglés | MEDLINE | ID: mdl-38412833

RESUMEN

Anopheles gambiae uses its sense of smell to hunt humans. We report a two-step method yielding cell-type-specific driver lines for enhanced neuroanatomical and functional studies of its olfactory system. We first integrated a driver-responder-marker (DRM) system cassette consisting of a linked T2A-QF2 driver, QUAS-GFP responder, and a gut-specific transgenesis marker into four chemoreceptor genes (Ir25a, Ir76b, Gr22, and orco) using CRISPR-Cas9-mediated homology-directed repair. The DRM system facilitated rapid selection of in-frame integrations via screening for GFP+ olfactory sensory neurons (OSNs) in G1 larval progeny, even at genomic loci such as orco where we found the transgenesis marker was not visible. Next, we converted these DRM integrations into T2A-QF2 driver-marker lines by Cre-loxP excision of the GFP responder, making them suitable for binary use in transcuticular calcium imaging. These cell-type-specific driver lines tiling key OSN subsets will support systematic efforts to decode olfaction in this prolific malaria vector.


Asunto(s)
Anopheles , Malaria , Neuronas Receptoras Olfatorias , Animales , Humanos , Olfato/genética , Anopheles/genética , Mosquitos Vectores/genética
2.
Cold Spring Harb Protoc ; 2024(4): pdb.prot108304, 2024 Apr 01.
Artículo en Inglés | MEDLINE | ID: mdl-37612143

RESUMEN

Anopheles gambiae is a highly anthropophilic (human-preferring) malaria vector that prefers to blood feed frequently and selectively on humans. This mosquito species exhibits a strong innate olfactory preference to seek out human scent over other animals, and certain humans over others-key behavioral traits with the potential to drive heterogeneity in biting risk and malaria transmission. Here, we describe the application of a large-scale, semi-field system in Zambia for the quantification of An. gambiae olfactory preferences toward whole body odor sourced from individual humans. We detail steps for modifying one-person canvas tents to duct odor from sleeping humans into a central, semi-field flight cage arena that is securely screened. Using this system, we describe a protocol to perform two-choice olfactory preference assays with two human volunteers using laboratory-reared An. gambiae and odor-guided thermotaxis assays that leverage infrared video tracking to quantify mosquito landings on heated targets baited with each body odor sample. This multichoice olfactory preference assay has the potential to be applied with expanded cohorts of humans to define the chemosensory basis of An. gambiae host preference and interindividual differences in human attractiveness to mosquitoes and to be used to quantify the effects of protective measures such as personal and spatial repellents on mosquito landing behavior.


Asunto(s)
Anopheles , Malaria , Animales , Humanos , Olor Corporal , Mosquitos Vectores , Olfato
3.
Cold Spring Harb Protoc ; 2024(4): pdb.prot108303, 2024 Apr 01.
Artículo en Inglés | MEDLINE | ID: mdl-37612145

RESUMEN

The African malaria mosquito Anopheles gambiae is strongly attracted to human body odor and skin temperature. Quantitative behavioral assays suitable for use in semi-field environments with this nocturnal mosquito species are essential to gain improved insights into An. gambiae sensory biology, the mechanistic basis of mosquito attraction to humans, and host preference. In this protocol, we describe steps for engineering equipment for a novel behavioral assay for An. gambiae, which we have termed the odor-guided thermotaxis assay (OGTA). The OGTA uses infrared videography to quantify landings of female An. gambiae on an aluminum platform heated to human skin temperature that can be baited with volatile odorants such as carbon dioxide or human whole body odor. The OGTA facilitates high-content recordings of An. gambiae landing behavior during odor-guided thermotaxis under naturalistic semi-field conditions without the requirement for domestic power.


Asunto(s)
Anopheles , Taxia , Animales , Humanos , Femenino , Odorantes , Olor Corporal , Conducta Animal
4.
Cold Spring Harb Protoc ; 2024(4): pdb.top107663, 2024 Apr 01.
Artículo en Inglés | MEDLINE | ID: mdl-37612146

RESUMEN

The most dangerous mosquito species for human health are those that blood feed preferentially and frequently on humans (anthropophilic mosquitoes). These include prolific disease vectors such as the African malaria mosquito Anopheles gambiae and yellow fever mosquito Aedes aegypti The chemosensory basis for anthropophilic behavior exhibited by these disease vectors, as well as the factors that drive interindividual differences in human attractiveness to mosquitoes, remain largely uncharacterized. Here, we concisely review established methods to quantify mosquito interspecific and intraspecific host preference in the laboratory, as well as semi-field and field environments. Experimental variables for investigator consideration during assays of mosquito host preference across these settings are highlighted.


Asunto(s)
Aedes , Anopheles , Animales , Humanos , Mosquitos Vectores
5.
Front Immunol ; 14: 1194253, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-37809107

RESUMEN

Background: In chronic cystic fibrosis (CF) lung disease, neutrophilic inflammation and T-cell inhibition occur concomitantly, partly due to neutrophil-mediated release of the T-cell inhibitory enzyme Arg1. However, the onset of this tonic inhibition of T cells, and the impact of pulmonary exacerbations (PEs) on this process, remain unknown. Methods: Children with CF aged 0-5 years were enrolled in a longitudinal, single-center cohort study. Blood (n = 35) and bronchoalveolar lavage (BAL) fluid (n = 18) were collected at stable outpatient clinic visits or inpatient PE hospitalizations and analyzed by flow cytometry (for immune cell presence and phenotype) and 20-plex chemiluminescence assay (for immune mediators). Patients were categorized by PE history into (i) no prior PE, (ii) past history of PE prior to stable visit, or (iii) current PE. Results: PEs were associated with increased concentration of both pro- and anti-inflammatory mediators in BAL, and increased neutrophil frequency and G-CSF in circulation. PE BAL samples showed a trend toward an increased frequency of hyperexocytic "GRIM" neutrophils, which we previously identified in chronic CF. Interestingly, expression levels of the T-cell receptor associated molecule CD3 and of the inhibitory programmed death-1 (PD-1) receptor were respectively decreased and increased on T cells from BAL compared to blood in all patients. When categorized by PE status, CD3 and PD-1 expression on blood T cells did not differ among patients, while CD3 expression was decreased, and PD-1 expression was increased on BAL T cells from patients with current PE. Conclusions: Our findings suggest that airway T cells are engaged during early-life PEs, prior to the onset of chronic neutrophilic inflammation in CF. In addition, increased blood neutrophil frequency and a trend toward increased BAL frequency of hyperexocytic neutrophils suggest that childhood PEs may progressively shift the balance of CF airway immunity towards neutrophil dominance.


Asunto(s)
Fibrosis Quística , Niño , Humanos , Receptor de Muerte Celular Programada 1 , Estudios de Cohortes , Linfocitos T , Inflamación
7.
Curr Biol ; 33(12): 2367-2382.e7, 2023 06 19.
Artículo en Inglés | MEDLINE | ID: mdl-37209680

RESUMEN

The African malaria mosquito Anopheles gambiae exhibits a strong innate drive to seek out humans in its sensory environment, classically entering homes to land on human skin in the hours flanking midnight. To gain insight into the role that olfactory cues emanating from the human body play in generating this epidemiologically important behavior, we developed a large-scale multi-choice preference assay in Zambia with infrared motion vision under semi-field conditions. We determined that An. gambiae prefers to land on arrayed visual targets warmed to human skin temperature during the nighttime when they are baited with carbon dioxide (CO2) emissions reflective of a large human over background air, body odor from one human over CO2, and the scent of one sleeping human over another. Applying integrative whole body volatilomics to multiple humans tested simultaneously in competition in a six-choice assay, we reveal high attractiveness is associated with whole body odor profiles from humans with increased relative abundances of the volatile carboxylic acids butyric acid, isobutryic acid, and isovaleric acid, and the skin microbe-generated methyl ketone acetoin. Conversely, those least preferred had whole body odor that was depleted of carboxylic acids among other compounds and enriched with the monoterpenoid eucalyptol. Across expansive spatial scales, heated targets without CO2 or whole body odor were minimally or not attractive at all to An. gambiae. These results indicate that human scent acts critically to guide thermotaxis and host selection by this prolific malaria vector as it navigates towards humans, yielding intrinsic heterogeneity in human biting risk.


Asunto(s)
Anopheles , Malaria , Taxia , Animales , Humanos , Odorantes , Olor Corporal , Dióxido de Carbono , Mosquitos Vectores , Feromonas Humanas , Ácidos Carboxílicos
8.
Immunohorizons ; 6(11): 768-778, 2022 11 01.
Artículo en Inglés | MEDLINE | ID: mdl-36445361

RESUMEN

Juvenile idiopathic arthritis (JIA) is an inflammatory rheumatic disorder. Polymorphonuclear neutrophils (PMNs) are present in JIA synovial fluid (SF), but with variable frequency. SF PMNs in JIA were previously shown to display high exocytic but low phagocytic and immunoregulatory activities. To further assess whether the degree of SF neutrophilia associated with altered immune responses in JIA, we collected SF and blood from 16 adolescent JIA patients. SF and blood leukocytes were analyzed by flow cytometry. SF and plasma were used for immune mediator quantification and metabolomics. Healthy donor blood T cells were cultured in SF to evaluate its immunoregulatory activities. PMN and T cell frequencies were bimodal in JIA SF, delineating PMN high/T cell low (PMNHigh) and PMN low/T cell high (PMNLow) samples. Proinflammatory mediators were increased in SF compared with plasma across patients, and pro- and anti-inflammatory mediators were further elevated in PMNHigh SF. Compared to blood, SF PMNs showed increased exocytosis and programmed death-1/programmed death ligand-1 expression, and SF PMNs and monocytes/macrophages had increased surface-bound arginase-1. SPADE analysis revealed SF monocyte/macrophage subpopulations coexpressing programmed death-1 and programmed death ligand-1, with higher expression in PMNHigh SF. Healthy donor T cells showed reduced coreceptor expression when stimulated in PMNHigh versus PMNLow SF. However, amino acid metabolites related to the arginase-1 and IDO-1 pathways did not differ between the two groups. Hence, PMN predominance in the SF of a subset of JIA patients is associated with elevated immune mediator concentration and may alter SF monocyte/macrophage phenotype and T cell activation, without altering immunoregulatory amino acids.


Asunto(s)
Artritis Juvenil , Líquido Sinovial , Humanos , Arginasa , Leucocitos , Neutrófilos
9.
Cell Rep ; 39(11): 110945, 2022 06 14.
Artículo en Inglés | MEDLINE | ID: mdl-35688145

RESUMEN

SARS-CoV-2-infected subjects are generally asymptomatic during initial viral replication but may suffer severe immunopathology after the virus has receded and monocytes have infiltrated the airways. In bronchoalveolar lavage fluid from severe COVID-19 patients, monocytes express mRNA encoding inflammatory mediators and contain SARS-CoV-2 transcripts. We leverage a human small airway model of infection and inflammation, whereby primary blood monocytes transmigrate across SARS-CoV-2-infected lung epithelium to characterize viral burden, gene expression, and inflammatory mediator secretion by epithelial cells and monocytes. In this model, lung-infiltrating monocytes acquire SARS-CoV-2 from the epithelium and upregulate expression and secretion of inflammatory mediators, mirroring in vivo data. Combined use of baricitinib (Janus kinase inhibitor) and remdesivir (nucleoside analog) enhances antiviral signaling and viral clearance by SARS-CoV-2-positive monocytes while decreasing secretion of proneutrophilic mediators associated with acute respiratory distress syndrome. These findings highlight the role of lung-infiltrating monocytes in COVID-19 pathogenesis and their importance as a therapeutic target.


Asunto(s)
Tratamiento Farmacológico de COVID-19 , Azetidinas , Humanos , Mediadores de Inflamación , Pulmón/patología , Monocitos , Purinas , Pirazoles , SARS-CoV-2 , Sulfonamidas
10.
Pediatr Pulmonol ; 57(9): 2189-2198, 2022 09.
Artículo en Inglés | MEDLINE | ID: mdl-35637404

RESUMEN

BACKGROUND: In this pilot study, we investigated whether induced sputum (IS) could serve as a viable alternative to bronchoalveolar lavage (BAL) and yield robust inflammatory biomarkers in toddlers with cystic fibrosis (CF) featuring minimal structural lung disease. METHODS: We collected IS, BAL (right middle lobe and lingula), and blood, and performed chest computed tomography (CT) scans from 2-year-olds with CF (N = 11), all within a single visit. Inflammatory biomarkers included 20 soluble immune mediators and neutrophil elastase (NE), as well as frequency and phenotype of T cells, monocytes/macrophages, and neutrophils. RESULTS: At the molecular level, nine mediators showed similar levels in IS and BAL (CXCL1, CXCL8, IL-1α, IL-1RA, IL-6, CCL2, CXCL10, M-CSF, VEGF-A), four were higher in IS than in BAL (CXCL5, IL-1ß, CXCL11, TNFSF10), and two were present in IS, but undetectable in BAL (IL-10, IFN-γ). Meanwhile, soluble NE had lower activity in IS than in BAL. At the cellular level, T-cell frequency was lower in IS than in BAL. Monocytes/macrophages were dominant in IS and BAL with similar frequencies, but differing expression of CD16 (lower in IS), CD115, and surface-associated NE (higher in IS). Neutrophil frequency and phenotype did not differ between IS and BAL. Finally, neutrophil frequency in IS correlated positively with air trapping. CONCLUSIONS: IS collected from 2-year-olds with CF yields biomarkers of early airway inflammation with good agreement with BAL, notably with regard to molecular and cellular outcomes related to neutrophils and monocytes/macrophages.


Asunto(s)
Fibrosis Quística , Esputo , Biomarcadores , Lavado Broncoalveolar , Líquido del Lavado Bronquioalveolar , Humanos , Neutrófilos , Proyectos Piloto
11.
Front Behav Neurosci ; 16: 819146, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-35418841

RESUMEN

The analysis of kinematics, locomotion, and spatial tasks relies on the accurate detection of animal positions and pose. Pose and position can be assessed with video analysis programs, the "trackers." Most available trackers represent animals as single points in space (no pose information available) or use markers to build a skeletal representation of pose. Markers are either physical objects attached to the body (white balls, stickers, or paint) or they are defined in silico using recognizable body structures (e.g., joints, limbs, color patterns). Physical markers often cannot be used if the animals are small, lack prominent body structures on which the markers can be placed, or live in environments such as aquatic ones that might detach the marker. Here, we introduce a marker-free pose-estimator (LACE Limbless Animal traCkEr) that builds the pose of the animal de novo from its contour. LACE detects the contour of the animal and derives the body mid-line, building a pseudo-skeleton by defining vertices and edges. By applying LACE to analyse the pose of larval Drosophila melanogaster and adult zebrafish, we illustrate that LACE allows to quantify, for example, genetic alterations of peristaltic movements and gender-specific locomotion patterns that are associated with different body shapes. As illustrated by these examples, LACE provides a versatile method for assessing position, pose and movement patterns, even in animals without limbs.

12.
Front Psychol ; 12: 773173, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34867683

RESUMEN

The purpose of this work is to reveal how subjective well-being has been generated in a group of professionals in the healthcare field in Colombia, who carried out postgraduate studies at the time of the pandemic caused by the novel SARS-CoV-2 coronavirus in a synchronous and remote learning course facilitated by employing digital technologies. Two methods were assumed, one was qualitative, taking into account some elements of narrative research and discourse analysis, and the other was quantitative, through a rapid reconnaissance survey. The research assumes the constitution of subjectivity from memory and everyday life, as well as the ethics of care concerning caring for oneself and others, as categories that were (re)signified with the narratives-and as notions that make up a theoretical corpus-to understand subjective well-being.

13.
Cell Rep Med ; 2(4): 100239, 2021 04 20.
Artículo en Inglés | MEDLINE | ID: mdl-33948572

RESUMEN

Neutrophils are often considered terminally differentiated and poised for bacterial killing. In chronic diseases such as cystic fibrosis (CF), an unexplained paradox pits massive neutrophil presence against prolonged bacterial infections. Here, we show that neutrophils recruited to CF airways in vivo and in an in vitro transmigration model display rapid and broad transcriptional firing, leading to an upregulation of anabolic genes and a downregulation of antimicrobial genes. Newly transcribed RNAs are mirrored by the appearance of corresponding proteins, confirming active translation in these cells. Treatment by the RNA polymerase II and III inhibitor α-amanitin restores the expression of key antimicrobial genes and increases the bactericidal capacity of CF airway neutrophils in vitro and in short-term sputum cultures ex vivo. Broadly, our findings show that neutrophil plasticity is regulated at the site of inflammation via RNA and protein synthesis, leading to adaptations that affect their canonical functions (i.e., bacterial clearance).


Asunto(s)
Regulador de Conductancia de Transmembrana de Fibrosis Quística/metabolismo , Fibrosis Quística/metabolismo , Neutrófilos/metabolismo , Sistema Respiratorio/microbiología , Fibrosis Quística/genética , Fibrosis Quística/microbiología , Trampas Extracelulares/microbiología , Humanos , Neutrófilos/microbiología , Pseudomonas aeruginosa/patogenicidad , Esputo/metabolismo , Esputo/microbiología
14.
Int J Mol Sci ; 22(5)2021 Mar 03.
Artículo en Inglés | MEDLINE | ID: mdl-33802410

RESUMEN

Cystic fibrosis (CF) lung disease is dominated by the recruitment of myeloid cells (neutrophils and monocytes) from the blood which fail to clear the lung of colonizing microbes. In prior in vitro studies, we showed that blood neutrophils migrated through the well-differentiated lung epithelium into the CF airway fluid supernatant (ASN) mimic the dysfunction of CF airway neutrophils in vivo, including decreased bactericidal activity despite an increased metabolism. Here, we hypothesized that, in a similar manner to neutrophils, blood monocytes undergo significant adaptations upon recruitment to CFASN. To test this hypothesis, primary human blood monocytes were transmigrated in our in vitro model into the ASN from healthy control (HC) or CF subjects to mimic in vivo recruitment to normal or CF airways, respectively. Surface phenotype, metabolic and bacterial killing activities, and transcriptomic profile by RNA sequencing were quantified post-transmigration. Unlike neutrophils, monocytes were not metabolically activated, nor did they show broad differences in activation and scavenger receptor expression upon recruitment to the CFASN compared to HCASN. However, monocytes recruited to CFASN showed decreased bactericidal activity. RNASeq analysis showed strong effects of transmigration on monocyte RNA profile, with differences between CFASN and HCASN conditions, notably in immune signaling, including lower expression in the former of the antimicrobial factor ISG15, defensin-like chemokine CXCL11, and nitric oxide-producing enzyme NOS3. While monocytes undergo qualitatively different adaptations from those seen in neutrophils upon recruitment to the CF airway microenvironment, their bactericidal activity is also dysregulated, which could explain why they also fail to protect CF airways from infection.


Asunto(s)
Adaptación Fisiológica/genética , Microambiente Celular/genética , Fibrosis Quística/genética , Pulmón/patología , Monocitos/patología , Transcripción Genética/genética , Adulto , Células Cultivadas , Femenino , Humanos , Masculino , Neutrófilos/patología , Transducción de Señal/fisiología
15.
Blood Press Monit ; 25(6): 318-323, 2020 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-32740294

RESUMEN

INTRODUCTION: Despite the well documented importance of blood pressure management in patients with spontaneous intracerebral hemorrhage (sICH), little is known about whether emergency departments (EDs) are able to achieve close monitoring and precise management. Our study characterizes ED monitoring and management of blood pressure in sICH patients. METHODS: This is a retrospective study of adults with sICH and elevated intracranial pressure. Patients who were admitted from any referring ED to our CCRU from 1 August 2013 to 30 September 2015 were included. We graphically assessed the association between average minutes between blood pressure measurements and average minutes between administration of antihypertensives. We also performed logistic regression to evaluate factors associated with close blood pressure monitoring and the achievement of goal blood pressure in patients with sICH who presented with hypertension. RESULTS: Of 115 patients, 73 presented to the ED with SBP above 160 mmHg. Length of stay in the ED was significantly associated with a longer period between blood pressure measurements. Longer periods between blood pressure measurements were a significant determinant of failure to achieve blood pressure goal in sICH patients. Longer periods between blood pressure measurements were significantly associated with longer periods between administration of antihypertensives. CONCLUSION: Our study suggests that blood pressure monitoring is related to the frequency of blood pressure interventions and achievement of adequate blood pressure control in patients with sICH. There is significant variability in EDs' achievement of the recommended close blood pressure monitoring and management in patients with sICH.


Asunto(s)
Determinación de la Presión Sanguínea , Hemorragia Cerebral , Adulto , Presión Sanguínea , Hemorragia Cerebral/tratamiento farmacológico , Servicio de Urgencia en Hospital , Humanos , Estudios Retrospectivos
16.
Comput Biol Chem ; 85: 107211, 2020 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-32004971

RESUMEN

As part of the type I IFN signaling, the 2'-5'- oligoadenylate synthetase (OAS) proteins have been involved in the progression of several non-viral diseases. Notably, OAS has been correlated with immune-modulatory functions that promote chronic inflammatory conditions, autoimmune disorders, cancer, and infectious diseases. In spite of this, OAS enzymes have been ignored as drug targets, and to date, there are no reports of compounds that can inhibit their activity. In this study, we have used homology modeling and virtual high-throughput screening to identify potential inhibitors of the human proteins OAS1, OAS2, and OAS3. Altogether, we have found 37 molecules that could exert a competitive inhibition in the ATP binding sites of OAS proteins, independently of the activation state of the enzyme. This latter characteristic, which might be crucial for a versatile inhibitor, was observed in compounds interacting with the residues Asp75, Asp77, Gln229, and Tyr230 in OAS1, and their equivalents in OAS2 and OAS3. Although there was little correlation between specific chemical fragments and their interactions, intermolecular contacts with OAS catalytic triad and other critical amino acids were mainly promoted by heterocycles with π electrons and hydrogen bond acceptors. In conclusion, this study provides a potential set of OAS inhibitors as well as valuable information for their design, development, and optimization.


Asunto(s)
2',5'-Oligoadenilato Sintetasa/antagonistas & inhibidores , Inhibidores Enzimáticos/farmacología , 2',5'-Oligoadenilato Sintetasa/metabolismo , Biología Computacional , Inhibidores Enzimáticos/síntesis química , Inhibidores Enzimáticos/química , Ensayos Analíticos de Alto Rendimiento , Humanos , Modelos Moleculares
17.
Curr Biol ; 29(17): 2961-2969.e4, 2019 09 09.
Artículo en Inglés | MEDLINE | ID: mdl-31447373

RESUMEN

Rhodopsins, the major light-detecting molecules of animal visual systems [1], consist of opsin apoproteins that covalently bind a retinal chromophore with a conserved lysine residue [1, 2]. In addition to capturing photons, this chromophore contributes to rhodopsin maturation [3, 4], trafficking [3, 4], and stabilization [5], and defects in chromophore synthesis and recycling can cause dysfunction of the retina and dystrophy [6-9]. Indications that opsin apoproteins alone might have biological roles have come from archaebacteria and platyhelminths, which present opsin-like proteins that lack the chromophore binding site and are deemed to function independently of light [10, 11]. Light-independent sensory roles have been documented for Drosophila opsins [12-15], yet also these unconventional opsin functions are thought to require chromophore binding [12, 13, 15]. Unconjugated opsin apoproteins act as phospholipid scramblases in mammalian photoreceptor disks [16], yet chromophore-independent roles of opsin apoproteins outside of eyes have, to the best of our knowledge, hitherto not been described. Drosophila chordotonal mechanoreceptors require opsins [13, 15], and we find that their function remains uncompromised by nutrient carotenoid depletion. Disrupting carotenoid uptake and cleavage also left the mechanoreceptors unaffected, and manipulating the chromophore attachment site of the fly's major visual opsin Rh1 impaired photoreceptor, but not mechanoreceptor, function. Notwithstanding this chromophore independence, some proteins that process and recycle the chromophore in the retina are also required in mechanoreceptors, including visual cycle components that recycle the chromophore upon its photoisomerization. Our results thus establish biological function for unconjugated opsin apoproteins outside of eyes and, in addition, document chromophore-independent roles for chromophore pathway components.


Asunto(s)
Apoproteínas/metabolismo , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/fisiología , Mecanorreceptores/metabolismo , Opsinas/metabolismo , Retinaldehído/análogos & derivados , Animales , Retinaldehído/metabolismo
18.
Sci Rep ; 9(1): 3974, 2019 03 08.
Artículo en Inglés | MEDLINE | ID: mdl-30850647

RESUMEN

Sensing environmental temperatures is essential for the survival of ectothermic organisms. In Drosophila, two of the most used methodologies to study temperature preferences (TP) and the genes involved in thermosensation are two-choice assays and temperature gradients. Whereas two-choice assays reveal a relative TP, temperature gradients can identify the absolute Tp. One drawback of gradients is that small ectothermic animals are susceptible to cold-trapping: a physiological inability to move at the cold area of the gradient. Often cold-trapping cannot be avoided, biasing the resulting TP to lower temperatures. Two mathematical models were previously developed to correct for cold-trapping. These models, however, focus on group behaviour which can lead to overestimation of cold-trapping due to group aggregation. Here we present a mathematical model that simulates the behaviour of individual Drosophila in temperature gradients. The model takes the spatial dimension and temperature difference of the gradient into account, as well as the rearing temperature of the flies. Furthermore, it allows the quantification of cold-trapping and reveals unbiased TP. Additionally, our model reveals that flies have a range of tolerable temperatures, and this measure is more informative about the behaviour than commonly used TP. Online simulation is hosted at http://igloo.uni-goettingen.de . The code can be accessed at https://github.com/zerotonin/igloo .


Asunto(s)
Drosophila/fisiología , Locomoción/fisiología , Animales , Sesgo , Frío , Temperatura
20.
Neuron ; 98(1): 67-74.e4, 2018 04 04.
Artículo en Inglés | MEDLINE | ID: mdl-29551493

RESUMEN

Animals rely on mechanosensory feedback from proprioceptors to control locomotory body movements. Unexpectedly, we found that this movement control requires visual opsins. Disrupting the Drosophila opsins NINAE or Rh6 impaired larval locomotion and body contractions, independently of light and vision. Opsins were detected in chordotonal proprioceptors along the larval body, localizing to their ciliated dendrites. Loss of opsins impaired mechanically evoked proprioceptor spiking and cilium ultrastructure. Without NINAE or Rh6, NOMPC mechanotransduction channels leaked from proprioceptor cilia and ciliary Inactive (Iav) channels partly disappeared. Locomotion is shown to require opsins in proprioceptors, and the receptors are found to express the opsin gene Rh7, in addition to ninaE and Rh6. Besides implicating opsins in movement control, this documents roles of non-ciliary, rhabdomeric opsins in cilium organization, providing a model for a key transition in opsin evolution and suggesting that structural roles of rhabdomeric opsins preceded their use for light detection.


Asunto(s)
Proteínas de Drosophila/biosíntesis , Larva/metabolismo , Locomoción/fisiología , Propiocepción/fisiología , Rodopsina/biosíntesis , Animales , Animales Modificados Genéticamente , Proteínas de Drosophila/análisis , Drosophila melanogaster , Femenino , Larva/química , Masculino , Mecanotransducción Celular/fisiología , Rodopsina/análisis
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