Single-cell resolution long-term luciferase imaging in cultivated Drosophila brains

Schubert, Frank K.; Helfrich‐Förster, Charlotte; Rieger, Dirk

doi:10.17912/micropub.biology.000280

Cited by 1 publication

(1 citation statement)

References 13 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Preparation of the brains was similar to that described previously ( Schubert et al, 2020 ; Versteven et al, 2020 ). Briefly, brains were dissected under red light in ice cold Ca 2+ free Ringer’s solution and placed on a CellView glass bottom imaging dish (35x10 mm; Greiner Bio One) treated with Heptane glue, where the culture medium was added.…”

Section: Methodsmentioning

confidence: 99%

Real time, in vivo measurement of neuronal and peripheral clocks in Drosophila melanogaster

Johnstone

Ogueta

Akay

et al. 2022

eLife

View full text Add to dashboard Cite

Circadian clocks are highly conserved transcriptional regulators that control ~24-hour oscillations in gene expression, physiological function, and behavior. Circadian clocks exist in almost every tissue and are thought to control tissue-specific gene expression and function, synchronized by the brain clock. Many disease states are associated with loss of circadian regulation. How and when circadian clocks fail during pathogenesis remains largely unknown because it is currently difficult to monitor tissue-specific clock function in intact organisms. Here, we developed a method to directly measure the transcriptional oscillation of distinct neuronal and peripheral clocks in live, intact Drosophila, which we term Locally Activatable BioLuminescence, or LABL. Using this method, we observed that specific neuronal and peripheral clocks exhibit distinct transcriptional properties. Loss of the receptor for PDF, a circadian neurotransmitter critical for the function of the brain clock, disrupts circadian locomotor activity but not all tissue-specific circadian clocks. We found that, while peripheral clocks in non-neuronal tissues were less stable after the loss of PDF signaling, they continued to oscillate. We also demonstrate that distinct clocks exhibit differences in their loss of oscillatory amplitude or their change in period, depending on their anatomical location, mutation, or fly age. Our results demonstrate that LABL is an effective tool that allows rapid, affordable, and direct real-time monitoring of individual clocks in vivo.

show abstract

Section: Methodsmentioning

confidence: 99%