Improved whole-mount immunofluorescence protocol for consistent and robust labeling of adultDrosophila melanogasteradipose tissue DOI Open Access

Rachael K. Ott,

Alissa Richmond Armstrong

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: April 15, 2024

ABSTRACT Energy storage and endocrine functions of the Drosophila fat body make it an excellent model for elucidating mechanisms that underlie physiological pathophysiological organismal metabolism. Combined with Drosophila’s robust genetic immunofluorescence microscopy toolkits, studies function are ripe cell biological analysis. Unlike larval body, which is easily removed as a single, cohesive sheet tissue, isolating intact adult proves to be more challenging, thus hindering consistent labeling even within single piece adipose tissue. Here, we describe improved approach handling abdomens ensures full access solutions generally used in protocols. In addition, assess quality fluorescence reporter expression antibody immunoreactivity response variations fixative type, fixation incubation time, detergent cellular permeabilization. Overall, provide several recommendations steps whole mount staining protocol results body. SUMMARY STATEMENT Optimization via modifications tissue handling, fixation, permeabilization immunolabeling protocol.

Language: Английский

Dynamic encoding of temperature in the central circadian circuit coordinates physiological activities DOI Creative Commons
Hailiang Li, Zhiyi Li,

Xin Yuan

et al.

Nature Communications, Journal Year: 2024, Volume and Issue: 15(1)

Published: April 2, 2024

Abstract The circadian clock regulates animal physiological activities. How temperature reorganizes circadian-dependent activities remains elusive. Here, using in-vivo two-photon imaging with the control device, we investigated response of Drosophila central circuit to variation and identified that DN1as serves as most sensitive temperature-sensing neurons. gate DN1a’s diurnal response. Trans-synaptic tracing, connectome analysis, functional data reveal bidirectionally targets two neuronal subsets: activity-related E cells sleep-promoting DN3s. Specifically, behavioral demonstrate DN1a-E cell modulates evening locomotion peak in cold temperature, while DN1a-DN3 controls warm temperature-induced nocturnal sleep reduction. Our findings systematically comprehensively illustrate how dynamically integrates light signals effectively coordinate wakefulness at different times day, shedding on conserved neural mechanisms underlying temperature-regulated physiology animals.

Language: Английский

Citations

11
Promoter engineering with programmable upstream activating sequences in Aspergillus Niger cell factory DOI Creative Commons
Xiaomei Zheng, Yuting Guo, Meiling Chen

et al.

Microbial Cell Factories, Journal Year: 2025, Volume and Issue: 24(1)

Published: Jan. 15, 2025

Language: Английский

Citations

0
Taking flight, the use of Drosophila melanogaster for neuroscience research in Uruguay DOI
Gonzalo Budelli, M.J. Ferreiro, Carmen Bolatto

et al.

Neuroscience, Journal Year: 2025, Volume and Issue: unknown

Published: March 1, 2025

Language: Английский

Citations

0
Redesigning and rethinking genetic circuits: the potential of transcriptional rewiring in filamentous fungi DOI

Alejandra Goity,

Luis Larrondo

Current Opinion in Biotechnology, Journal Year: 2025, Volume and Issue: 93, P. 103301 - 103301

Published: April 4, 2025

Language: Английский

Citations

0
Biotechnology-enhanced genetic controls of the global pest Drosophila suzukii DOI Creative Commons
Ying Yan, Hassan M. M. Ahmed, Ernst A. Wimmer

et al.

Trends in biotechnology, Journal Year: 2024, Volume and Issue: unknown

Published: Sept. 1, 2024

Language: Английский

Citations

2
Improved whole-mount immunofluorescence protocol for consistent and robust labeling of adult Drosophila melanogaster adipose tissue DOI Creative Commons

Rachael K. Ott,

Isaiah Williams,

Alissa Richmond Armstrong

et al.

Biology Open, Journal Year: 2024, Volume and Issue: 13(8)

Published: July 23, 2024

Energy storage and endocrine functions of the Drosophila fat body make it an excellent model for elucidating mechanisms that underlie physiological pathophysiological organismal metabolism. Combined with Drosophila's robust genetic immunofluorescence microscopy toolkits, studies function are ripe cell biological analysis. Unlike larval body, which is easily removed as a single, cohesive sheet tissue, isolating intact adult proves to be more challenging, thus hindering consistent labeling even within single piece adipose tissue. Here, we describe improved approach handling abdomens ensures full access solutions generally used in protocols. In addition, assess quality fluorescence reporter expression antibody immunoreactivity response variations fixative type, fixation incubation time, detergent cellular permeabilization. Overall, provide several recommendations steps whole-mount staining protocol results body.

Language: Английский

Citations

1
Transposon and Transgene Tribulations in Mosquitoes: A Perspective of piRNA Proportions DOI Creative Commons
Nelson C. Lau, Vanessa M. Macias

DNA, Journal Year: 2024, Volume and Issue: 4(2), P. 104 - 128

Published: March 30, 2024

Mosquitoes, like Drosophila, are dipterans, the order of “true flies” characterized by a single set two wings. Drosophila prime model organisms for biomedical research, while mosquito researchers struggle to establish robust molecular biology in these that arguably most dangerous vectors human pathogens. Both insects utilize RNA interference (RNAi) pathway generate small RNAs silence transposons and viruses, yet details emerging several RNAi features unique each insect family, such as how culicine mosquitoes have evolved extreme genomic feature differences connected their features. A major technical difference genetic studies is generating stable transgenic animals routine but still variable stability mosquitoes, despite DNA-editing advances. By comparing contrasting pathways this review we propose hypothesis transgene DNAs possibly more intensely targeted chromatin regulatory than Drosophila. We latest findings on pathways, which much less well understood speculate deeper study into modulate viruses with Piwi-interacting (piRNAs) will yield clues improving DNA expression mosquitoes.

Language: Английский

Citations

0
Improved whole-mount immunofluorescence protocol for consistent and robust labeling of adultDrosophila melanogasteradipose tissue DOI Open Access

Rachael K. Ott,

Alissa Richmond Armstrong

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: April 15, 2024

ABSTRACT Energy storage and endocrine functions of the Drosophila fat body make it an excellent model for elucidating mechanisms that underlie physiological pathophysiological organismal metabolism. Combined with Drosophila’s robust genetic immunofluorescence microscopy toolkits, studies function are ripe cell biological analysis. Unlike larval body, which is easily removed as a single, cohesive sheet tissue, isolating intact adult proves to be more challenging, thus hindering consistent labeling even within single piece adipose tissue. Here, we describe improved approach handling abdomens ensures full access solutions generally used in protocols. In addition, assess quality fluorescence reporter expression antibody immunoreactivity response variations fixative type, fixation incubation time, detergent cellular permeabilization. Overall, provide several recommendations steps whole mount staining protocol results body. SUMMARY STATEMENT Optimization via modifications tissue handling, fixation, permeabilization immunolabeling protocol.

Language: Английский

Citations

0