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1.
J Environ Qual ; 2024 Jun 13.
Article in English | MEDLINE | ID: mdl-38872323

ABSTRACT

Corn (Zea mays) crops harvested as grain in autumn do not provide opportunity for cover crop establishment, which may be remedied by interseeding cover crops into growing corn. Grazing cover crops after corn grain harvest could provide added revenues and increase nutrient cycling in the system while providing additional ecosystem services. However, tradeoffs between cash crop productivity and cover crop inclusion, and use as grazed forage, are not fully understood. This 4-year Long-Term Agroecosystem Research Integrated Common Experiment project evaluated the effect of interseeding cereal rye (Secale cereale) into corn for grazing after corn grain harvest on corn grain yield and late-season grazing. Cereal rye was interseeded into corn in early June. After corn grain harvest, six paddocks at each location were randomly allotted to grazed (GRAZ) or not grazed (NG). The GRAZ paddocks were grazed with beef cattle in late autumn and again in early spring if regrowth allowed. Paddocks were flown with an unmanned aerial system (UAS) to characterize spatial forage yield and quality. Cereal rye provided an additional 20-30 grazing days in the autumn for 24 beef cows on 4.8 ha. Early spring growth shows potential to provide even greater forage yields than autumn, but growth is less dependable. Corn grain yields did not decrease except in 2019 (dry year) when yields were 40% lower. There were no significant differences in soil health indicators between GRAZ and NG paddocks. The UAS shows promise as a tool for monitoring forage yield and quality and optimizing grazing management.

2.
Front Plant Sci ; 14: 1029141, 2023.
Article in English | MEDLINE | ID: mdl-37575930

ABSTRACT

There has been considerable interest in use of Miscanthus (Miscanthus x giganteus) as a feedstock for bioenergy production due to its potential to reduce greenhouse gas emissions associated with cellulosic feedstock production and more recently for alternative uses as a biomass crop. To date, data on Miscanthus production in the US has been based on small scale research plots due to the lack of commercial scale production fields. Research plot yields are often much higher than commercial fields for a variety of reasons including reduced spatial variability and location on better quality farmland. The objectives of this study were to quantify the inputs for production of Miscanthus at the commercial farm scale, evaluating methods to characterize fuel use for establishment and management of Miscanthus production and using satellite data to characterize spatial yield variation of production fields. We logged energy use on agricultural machinery from Miscanthus production planted on more than 1000 ha of land and modeled N2O emissions and changes in soil carbon using DayCent. Although fuel use was higher for land preparation in fields with perennial vegetation, fuel to harvest Miscanthus dominated greenhouse gas (GHG) emissions (>90%) from agriculture machinery for crop management. The N2O emissions and changes in soil carbon were the largest source and sink of GHG emissions associated with Miscanthus production, respectively. Although ~ 50% of the established lands had Miscanthus yields < 5 Mg/ha, yields needed to be > 5 Mg/ha for ΔSOC to be positive. Given the large impact of yield on ΔSOC, net GHG for Miscanthus production with yields of 5 to 25 Mg/ha ranged ~130 to -260 kg CO2e/Mg biomass. Use of both energy use for Miscanthus harvest and satellite imagery were good methods to characterize spatial variability of commercial production fields. This demonstrates the potential to use this within field yield data to better understand factors driving subfield yield variability and use of satellite data to quantify early yield predictions.

3.
Semin Ophthalmol ; 37(6): 690-698, 2022 Aug.
Article in English | MEDLINE | ID: mdl-35657058

ABSTRACT

INTRODUCTION: A 43-year-old Caucasian male presented to our ophthalmology clinic with blurry vision and metamorphopsia in his right eye, 24 hours after receiving the first dose of the Pfizer-BioNTech COVID-19 vaccine. METHODS: Clinical examination and imaging tests were consistent with acute unilateral central serous chorioretinopathy (CSCR) that completely resolved after 2 months without any treatment. He had no significant ophthalmic or medical history. He also lacked the classical risk factors for CSCR such as recent psychosocial stressors, Type-A personality traits, history of exogenous steroid use, connective tissue disorders and obstructive sleep apnea. RESULTS: This appears to be only the second reported case of CSCR, temporally associated with a recombinant COVID-19 mRNA vaccine. We also present a summary of published reports demonstrating intraocular complications associated with the novel recombinant COVID-19 mRNA vaccines. CONCLUSION: Findings in this report should not deter COVID-19 vaccinations given the rarity of aforementioned ocular complications and the greater benefit of protection from COVID-19 infection. Medical practitioners, however, should remain mindful of potential ocular complications, given the greater likelihood of occurrence with increasing vaccination booster rates.


Subject(s)
BNT162 Vaccine , COVID-19 , Central Serous Chorioretinopathy , Acute Disease , Adult , BNT162 Vaccine/adverse effects , COVID-19/prevention & control , Central Serous Chorioretinopathy/chemically induced , Central Serous Chorioretinopathy/diagnosis , Humans , Male , Vaccination/adverse effects , Vision Disorders/diagnosis
4.
Optom Vis Sci ; 98(11): 1287-1294, 2021 11 01.
Article in English | MEDLINE | ID: mdl-34510152

ABSTRACT

SIGNIFICANCE: Multiple vision-related quality of life (VRQol) instruments exist, but questionnaires designed specifically for myopic children that are appropriate for assessing the impact of refractive error are rare. PURPOSE: This study aimed to assess the validity and reliability of the Student Refractive Error and Eyeglasses Questionnaire - Revised (SREEQ-R) in school-aged children with myopia in the United Kingdom. METHODS: Community optometrists in the United Kingdom invited children up to the age of 18 years presenting for an eye examination with current or previous reported use of eyeglasses for myopia to complete the SREEQ-R, which consists of 20 specific items divided into two sections each with three response categories. The "without glasses" section relates to perceptions of uncorrected vision/not wearing glasses, and the "with glasses" relates to corrected vision/wearing glasses. Rasch analysis was used to explore the psychometric performance (content, construct validity, and reliability) of the questionnaire items and scale using Winsteps software (Winsteps.com. Portland, OR). RESULTS: A total of 125 eligible children with a mean ± standard deviation age of 12.7 ± 2.9 years completed the SREEQ-R. All items fit the Rasch model and were retained, and the scale was found to be unidimensional. All children and item infit and outfit mean square statistics fell within the recommended fit criteria. As per the Rasch analysis, the person reliability coefficients were 0.84 and 0.91, whereas item reliabilities were 0.99 and 0.80 for the without glasses and with glasses sections, respectively. The internal consistency for the SREEQ-R was good; Cronbach α values were 0.84 for without glasses and 0.91 for with glasses. CONCLUSIONS: The SREEQ-R had satisfactory validity and reliability evidence. Construct validity of the scale was supported to measure the impact of uncorrected and corrected refractive error on vision-related quality of life in myopic school-aged children in the United Kingdom. The SREEQ-R could be used in future studies to evaluate vision-related quality of life in children with myopia.


Subject(s)
Myopia , Refractive Errors , Adolescent , Child , Eyeglasses , Humans , Myopia/diagnosis , Myopia/therapy , Psychometrics/methods , Quality of Life , Reproducibility of Results , Students , Surveys and Questionnaires
5.
Sci Total Environ ; 714: 136697, 2020 Apr 20.
Article in English | MEDLINE | ID: mdl-31982745

ABSTRACT

Climate change is exacerbating environmental pollution from crop production. Spatially and temporally explicit estimates of life-cycle environmental impacts are therefore needed for suggesting location and time relevant environmental mitigations strategies. Emission factors and process-based mechanism models are popular approaches used to estimate life-cycle environmental impacts. However, emission factors are often incapable of describing spatial and temporal heterogeneity of agricultural emissions, whereas process-based mechanistic models, capable of capturing the heterogeneity, tend to be very complicated and time-consuming. Efficient prediction of life-cycle environmental impacts from agricultural production is lacking. This study develops a rapid predictive model to quantify life-cycle global warming (GW) and eutrophication (EU) impacts of corn production using a novel machine learning approach. We used the boosted regression tree (BRT) model to estimate future life-cycle environmental impacts of corn production in U.S. Midwest counties under four emissions scenarios for years 2022-2100. Results from BRT models indicate that the cross-validation (R2) for predicting life cycle GW and EU impacts ranged from 0.78 to 0.82, respectively. Furthermore, results show that future life-cycle GW and EU impacts of corn production will increase in magnitude under all four emissions scenarios, with the highest environmental impacts shown under the high-emissions scenario. Moreover, this study found that changes in precipitation and temperature played a significant role in influencing the spatial heterogeneity in all life-cycle impacts across Midwest counties. The BRT model results indicate that machine learning can be a useful tool for predicting spatially and temporally explicit future life-cycle environmental impacts associated with corn production under different climate scenarios.

6.
BMJ Case Rep ; 12(3)2019 Mar 21.
Article in English | MEDLINE | ID: mdl-30902842

ABSTRACT

A 78-year-old man with vascular risk factors and a family history of glaucoma presents with bilateral superior arcuate visual field loss. MRI brain was reported normal. Intraocular pressure (IOP) and optical coherence tomography of the retinal nerve fibre layer (RNFL) were within normal limits. A tentative diagnosis of normal tension glaucoma was made. Over the next 5 years, IOP remained stable without treatment, serial visual fields noted repeatable bilateral superior depressions with normal RNFL. Referral to a glaucoma subspecialist and subsequently neuro-ophthalmologist prompted repeat MRIs, which demonstrated mild small vessel ischaemia. Standard visual evoked potentials (VEPs) were normal. Multifocal VEPs identified poor response across the entire visual field in both eyes. The combination of visual defects, unremarkable RNFL and reduced multifocal VEPs raised suspicion of bilateral inferior calcarine fissure change. Retrospective review of MRI's in a multidisciplinary meeting confirmed extensive microvascular changes with bilateral inferior calcarine fissure ischaemia.


Subject(s)
Brain Diseases/diagnosis , Glaucoma/diagnosis , Occipital Lobe , Vision, Low/diagnosis , Aged , Diagnosis, Differential , Humans , Male , Visual Field Tests , Visual Fields
7.
J Mol Cell Cardiol ; 127: 116-124, 2019 02.
Article in English | MEDLINE | ID: mdl-30571977

ABSTRACT

In yeast, the Atg2-Atg18 complex regulates Atg9 recycling from phagophore assembly site during autophagy; their function in higher eukaryotes remains largely unknown. In a targeted screening in Drosophila melanogaster, we show that Mef2-GAL4-RNAi-mediated knockdown of Atg2, Atg9 or Atg18 in the heart and indirect flight muscles led to shortened healthspan (declined locomotive function) and lifespan. These flies displayed an accelerated age-dependent loss of cardiac function along with cardiac hypertrophy (increased heart tube wall thickness) and structural abnormality (distortion of the lumen surface). Using the Mef2-GAL4-MitoTimer mitochondrial reporter system and transmission electron microscopy, we observed significant elongation of mitochondria and reduced number of lysosome-targeted autophagosomes containing mitochondria in the heart tube but exaggerated mitochondrial fragmentation and reduced mitochondrial density in indirect flight muscles. These findings provide the first direct evidence of the importance of Atg2-Atg18/Atg9 autophagy complex in the maintenance of mitochondrial integrity and, regulation of heart and muscle functions in Drosophila, raising the possibility of augmenting Atg2-Atg18/Atg9 activity in promoting mitochondrial health and, muscle and heart function.


Subject(s)
Drosophila Proteins/metabolism , Drosophila melanogaster/metabolism , Heart/physiology , Longevity/physiology , Mitochondria, Heart/metabolism , Animals , Autophagy-Related Proteins/metabolism , Cardiomegaly/genetics , Cardiomegaly/pathology , Drosophila melanogaster/ultrastructure , Female , Male , Membrane Proteins/metabolism , Mitochondria, Heart/ultrastructure , Muscles/metabolism
8.
J Environ Qual ; 47(4): 617-624, 2018 Jul.
Article in English | MEDLINE | ID: mdl-30025046

ABSTRACT

Managing cropping systems to sequester soil organic C (SOC) improves soil health and resilience to changing climate. Perennial crops, no-till planting, manure, and cover crops can add SOC; however, their impacts have not been well documented in the northeastern United States. Our objectives were (i) to monitor SOC from a bioenergy cropping study in Pennsylvania that included a corn ( L.)-soybean [ (L.) Merr.]-alfalfa ( L.) rotation, switchgrass ( L.), and reed canarygrass ( L.); (ii) to use the CQESTR model to predict SOC sequestration in the bioenergy crops (with and without projected climate change); and (iii) to use CQESTR to simulate influence of tillage, manure, cover cropping, and corn stover removal in typical dairy forage (silage corn-alfalfa) or grain corn-soybean rotations. Over 8 yr, measured SOC increased 0.4, 1.1, and 0.8 Mg C ha yr in the bioenergy rotation, reed canarygrass, and switchgrass, respectively. Simulated and measured data were significantly correlated ( < 0.001) at all depths. Predicted sequestration (8-14 Mg C ha over 40 yr) in dairy forage rotations was much larger than with corn-soybean rotations (-4.0-0.6 Mg C ha over 40 yr), due to multiple years of perennial alfalfa. No-till increased sequestration in the simulated dairy forage rotation and prevented a net loss of C in corn-soybean rotations. Simulations indicated limited impact of cover crops and manure on long-term SOC sequestration. The low solids content of liquid dairy manure is the likely reason for the less-than-expected impact of manure. Overall, simulations suggest that inclusion of alfalfa provides the greatest potential for SOC sequestration with a typical Pennsylvania crop rotation.


Subject(s)
Carbon Sequestration , Crops, Agricultural , Soil/chemistry , Agriculture , Carbon , Pennsylvania , Zea mays
9.
Clin Exp Optom ; 101(6): 764-770, 2018 11.
Article in English | MEDLINE | ID: mdl-29740867

ABSTRACT

BACKGROUND: To determine the test-retest reproducibility of accommodative facility (AF) measures in an unselected sample of UK primary school children. METHODS: Using ±2.00 DS flippers and a viewing distance of 40 cm, AF was measured in 136 children (range 4-12 years, average 8.1 ± 2.1) by five testers on three occasions (average interval between successive tests: eight days, range 1-21 days). On each occasion, AF was measured monocularly and binocularly, for two minutes. Full datasets were obtained in 111 children (81.6 per cent). RESULTS: Intra-individual variation in AF was large (standard deviation [SD] = 3.8 cycles per minute [cpm]) and there was variation due to the identity of the tester (SD = 1.6 cpm). On average, AF was greater: (i) in monocular compared to binocular testing (by 1.4 cpm, p < 0.001); (ii) in the second minute of testing compared to the first (by 1.3 cpm, p < 0.001); (iii) in older compared to younger children (for example, AF for 4/5-year-olds was 3.3 cpm lower than in children ≥ 10 years old, p = 0.009); and (iv) on subsequent testing occasions (for example, visit-2 AF was 2.0 cpm higher than visit-1 AF, p < 0.001). After the first minute of testing at visit-1, only 36.9 per cent of children exceeded published normative values for AF (≥ 11 cpm monocularly and ≥ 8 cpm binocularly), but this rose to 83.8 per cent after the third test. Using less stringent pass criteria (≥ 6 cpm monocularly and ≥ 3 cpm binocularly), the equivalent figures were 82.9 and 96.4 per cent, respectively. Reduced AF did not co-exist with abnormal near point of accommodation or reduced visual acuity. CONCLUSIONS: The results reveal considerable intra-individual variability in raw AF measures in children. When the results are considered as pass/fail, children who initially exhibit normal AF continued to do so on repeat testing. Conversely, the vast majority of children with initially reduced AF exhibit normal performance on repeat testing. Using established pass/fail criteria, the prevalence of persistently reduced AF in this sample is 3.6 per cent.


Subject(s)
Accommodation, Ocular/physiology , Vision Tests/standards , Child , Child, Preschool , Female , Humans , Male , Ocular Motility Disorders/diagnosis , Reproducibility of Results , Schools , Vision, Binocular/physiology , Visual Acuity/physiology
10.
Prog Biophys Mol Biol ; 137: 69-75, 2018 09.
Article in English | MEDLINE | ID: mdl-29649492

ABSTRACT

The frizzled pathway regulates the planar polarity of epithelial cells. In insects this is manifested by the polarity of cuticular structures such as hairs (trichomes) and sensory bristles. A variety of evidence has established that this is achieved by regulating the subcellular location for activating the cytoskeleton in the epithelial cells. How this is accomplished is still poorly understood. In the best-studied tissue, the Drosophila pupal wing two important cytoskeletal regulators have been identified. One, shavenoid (sha), appears to be an activator while the second multiple wing hairs (mwh), appears to be an inhibitor. In vitro biochemistry has confirmed that the Multiple Wing Hairs protein inhibits the elongation of F-actin chains and surprisingly that it also bundles F-actin. These two activities can explain the multifaceted mwh mutant phenotype.


Subject(s)
Cell Polarity , Cytoskeleton/metabolism , Drosophila Proteins/metabolism , Drosophila/cytology , Drosophila/metabolism , Epidermis/metabolism , Animals , Models, Biological
12.
Fly (Austin) ; 11(3): 194-199, 2017 07 03.
Article in English | MEDLINE | ID: mdl-28631994

ABSTRACT

The exoskeleton of insects and other arthropods is a very versatile material that is characterized by a complex multilayer structure. In Sobala and Adler (2016) we analyzed the process of wing cuticle deposition by RNAseq and electron microscopy. In this extra view we discuss the unique aspects of the envelope the first and most outermost layer and the gene expression program seen at the end of cuticle deposition. We discussed the role of undulae in the deposition of cuticle and how the hydrophobicity of wing cuticle arises.


Subject(s)
Drosophila Proteins/genetics , Drosophila melanogaster/genetics , Pupa/genetics , Wings, Animal/metabolism , Animals , Drosophila Proteins/metabolism , Drosophila melanogaster/growth & development , Drosophila melanogaster/metabolism , Gene Expression Regulation, Developmental , Insect Proteins/genetics , Insect Proteins/metabolism , Morphogenesis , Pupa/growth & development , Pupa/metabolism , Wings, Animal/growth & development
14.
G3 (Bethesda) ; 7(4): 1323-1337, 2017 04 03.
Article in English | MEDLINE | ID: mdl-28258110

ABSTRACT

The Planar cell Polarity Effector (PPE) genes inturned, fuzzy, and fritz are downstream components in the frizzled/starry night signaling pathway, and their function is instructed by upstream Planar Cell Polarity (PCP) core genes such as frizzled and dishevelled PPE proteins accumulate asymmetrically in wing cells and function in a protein complex mediated by direct interactions between In and Frtz and In and Fy. How the PCP proteins instruct the accumulation of PPE protein is unknown. We found a likely direct interaction between Dishevelled and Fritz and Dishevelled and Fuzzy that could play a role in this. We previously found that mild overexpression of frtz rescued a weak in allele. To determine if this was due to extra Frtz stabilizing mutant In or due to Frtz being able to bypass the need for In we generate a precise deletion of the inturned gene (inPD ). We found that mild overexpression of Fritz partially rescued inPD , indicating that fritz has In independent activity in PCP. Previous studies of PPE proteins used fixed tissues, and did not provide any insights into the dynamic properties of PPE proteins. We used CRISPR/Cas9 genome editing technology to edit the fritz gene to add a green fluorescent protein tag. fritzmNeonGreen provides complete rescue activity and works well for in vivo imaging. Our data showed that Fritz is very dynamic in epidermal cells and preferentially distributed to discrete membrane subdomains ("puncta"). Surprisingly, we found it in stripes in developing bristles.


Subject(s)
Cell Polarity , Dishevelled Proteins/metabolism , Drosophila Proteins/metabolism , Drosophila melanogaster/cytology , Drosophila melanogaster/metabolism , Glycoproteins/metabolism , Abdomen , Animals , Animals, Genetically Modified , CRISPR-Cas Systems/genetics , Gene Deletion , Genome, Insect , Green Fluorescent Proteins/metabolism , Hair/metabolism , Imaging, Three-Dimensional , Intracellular Signaling Peptides and Proteins , Phenotype , Protein Binding , Protein Interaction Mapping , Protein Stability , RNA Editing/genetics , Transgenes
15.
Trends Cell Biol ; 27(5): 379-390, 2017 05.
Article in English | MEDLINE | ID: mdl-28153580

ABSTRACT

Why some genes are more popular than others remains an open question, but one example of this phenomenon involves the genes controlling planar cell polarity (PCP), the polarization of cells within a plane of a tissue. Indeed, the so-called 'core' PCP genes such as dishevelled, frizzled, and prickle have been extensively studied both in animal models and by human genetics. By contrast, other genes that influence PCP signaling have received far less attention. Among the latter are inturned, fuzzy, and fritz, but recent work should bring these once obscure regulators into the limelight. We provide here a brief history of planar polarity effector (PPE) and CPLANE (ciliogenesis and planar polarity effector) proteins, discuss recent advances in understanding their molecular mechanisms of action, and describe their roles in human disease.


Subject(s)
Cell Polarity , Cilia/metabolism , Membrane Proteins/metabolism , Organogenesis , Actin Cytoskeleton/metabolism , Animals , Disease , Humans
17.
PLoS Genet ; 12(5): e1006100, 2016 05.
Article in English | MEDLINE | ID: mdl-27232182

ABSTRACT

The cuticular exoskeleton of insects and other arthropods is a remarkably versatile material with a complex multilayer structure. We made use of the ability to isolate cuticle synthesizing cells in relatively pure form by dissecting pupal wings and we used RNAseq to identify genes expressed during the formation of the adult wing cuticle. We observed dramatic changes in gene expression during cuticle deposition, and combined with transmission electron microscopy, we were able to identify candidate genes for the deposition of the different cuticular layers. Among genes of interest that dramatically change their expression during the cuticle deposition program are ones that encode cuticle proteins, ZP domain proteins, cuticle modifying proteins and transcription factors, as well as genes of unknown function. A striking finding is that mutations in a number of genes that are expressed almost exclusively during the deposition of the envelope (the thin outermost layer that is deposited first) result in gross defects in the procuticle (the thick chitinous layer that is deposited last). An attractive hypothesis to explain this is that the deposition of the different cuticle layers is not independent with the envelope instructing the formation of later layers. Alternatively, some of the genes expressed during the deposition of the envelope could form a platform that is essential for the deposition of all cuticle layers.


Subject(s)
Chitin/genetics , Drosophila/genetics , Insect Proteins/biosynthesis , Zona Pellucida Glycoproteins/biosynthesis , Animals , Chitin/metabolism , Drosophila/growth & development , Drosophila/ultrastructure , Gene Expression Regulation, Developmental , Insect Proteins/genetics , Microscopy, Electron, Transmission , Pupa/genetics , Pupa/growth & development , Pupa/ultrastructure , Wings, Animal/growth & development , Wings, Animal/metabolism , Wings, Animal/ultrastructure , Zona Pellucida Glycoproteins/genetics
19.
Ecol Appl ; 25(4): 1142-56, 2015 Jun.
Article in English | MEDLINE | ID: mdl-26465048

ABSTRACT

Crop residues are potentially significant sources of feedstock for biofuel production in the United States. However, there are concerns with maintaining the environmental functions of these residues while also serving as a feedstock for biofuel production. Maintaining soil organic carbon (SOC) along with its functional benefits is considered a greater constraint than maintaining soil erosion losses to an acceptable level. We used the biogeochemical model DayCent to evaluate the effect of residue removal, corn stover, and wheat and barley straw in three diverse locations in the USA. We evaluated residue removal with and without N replacement, along with application of a high-lignin fermentation byproduct (HLFB), the residue by-product comprised of lignin and small quantities of nutrients from cellulosic ethanol production. SOC always decreased with residue harvest, but the decrease was greater in colder climates when expressed on a life cycle basis. The effect of residue harvest on soil N2O emissions varied with N addition and climate. With N addition, N2O emissions always increased, but the increase was greater in colder climates. Without N addition, N2O emissions increased in Iowa, but decreased in Maryland and North Carolina with crop residue harvest. Although SOC was lower with residue harvest when HLFB was used for power production instead of being applied to land, the avoidance of fossil fuel emissions to the atmosphere by utilizing the cellulose and hemicellulose fractions of crop residue to produce ethanol (offsets) reduced the overall greenhouse gas (GHG) emissions because most of this residue carbon would normally be lost during microbial respiration. Losses of SOC and reduced N mineralization could both be mitigated with the application of HLFB to the land. Therefore, by returning the high-lignin fraction of crop residue to the land after production of ethanol at the biorefinery, soil carbon levels could be maintained along with the functional benefit of increased mineralized N, and more GHG emissions could be offset compared to leaving the crop residues on the land.


Subject(s)
Agriculture/methods , Biofuels , Carbon/chemistry , Fossil Fuels , Soil/chemistry , Crops, Agricultural/classification , Ethanol , Nitrous Oxide , Time Factors , United States
20.
Development ; 142(22): 3974-81, 2015 Nov 15.
Article in English | MEDLINE | ID: mdl-26395478

ABSTRACT

Chitin is a polymer of N-acetylglucosamine that is abundant and widely found in the biological world. It is an important constituent of the cuticular exoskeleton that plays a key role in the insect life cycle. To date, the study of chitin deposition during cuticle formation has been limited by the lack of a method to detect it in living organisms. To overcome this limitation, we have developed ChtVis-Tomato, an in vivo reporter for chitin in Drosophila. ChtVis-Tomato encodes a fusion protein that contains an apical secretion signal, a chitin-binding domain (CBD), a fluorescent protein and a cleavage site to release it from the plasma membrane. The chitin reporter allowed us to study chitin deposition in time lapse experiments and by using it we have identified unexpected deposits of chitin fibers in Drosophila pupae. ChtVis-Tomato should facilitate future studies on chitin in Drosophila and other insects.


Subject(s)
Chitin/metabolism , Drosophila/metabolism , Luminescent Proteins/metabolism , Recombinant Fusion Proteins/metabolism , Animals , Genes, Reporter/genetics , Luminescent Proteins/genetics , Microscopy, Confocal , Microscopy, Electron, Transmission , Protein Binding , Pupa/metabolism , Recombinant Fusion Proteins/genetics , Wings, Animal/metabolism , Wings, Animal/ultrastructure
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