The responses of 15 l-vLNs from five brains to 10μM ddFSK, a biologically inactive form of FSK. The responses of 14 l-vLNs from five brains to vehicle (0.1% DMSO in H元 Saline). In all graphs the green triangles indicate the start of bath application. The responses of 14 l-vLNs from five brains to 10μM FSK. The look-up table values represent un-normalized YFP/CFP ratios. The black bar beneath the panels indicates the presence of FSK. A pseudo-colored time-course of Epac1-camps FRET loss in a single l-vLN cell body in response to 10μM FSK. Visible neuronal clock classes are labeled, as are neurons of the ellipsoid body (eb). Anterior aspect confocal Z-series of YFP emission in a uas-Epac1-camps(42A) Cry(39)-Gal4 brain (scale bar = 100μM). Schematic representation of nine classes of circadian clock neurons in a single hemisphere of the adult brain.
#TIME CLOCK 2008 PDF#
Specifically, it experimentally confirms the longstanding hypothesis that PDF is a direct modulator of most neurons in the Drosophila clock network.Įpac1-camps is stably expressed in the neuronal circadian clock network and retains its function as a cAMP reporter. This work introduces a sensitive means of measuring cAMP changes in a living brain with subcellular resolution. Thus, the network of approximately 150 clock neurons displays widespread, though not uniform, PDF receptivity. In contrast, an unrelated Drosophila neuropeptide, DH31, stimulates large cAMP increases in all PDF-expressing clock neurons. We find that a subset of the PDF-expressing neurons respond to PDF with long-lasting cAMP increases and confirm that such responses require the PDF receptor. We address this question by adapting Epac1-camps, a genetically encoded cAMP FRET sensor, for use in the living brain. Whether PDF acts directly on elements of this neural network remains unknown.
The neuropeptide PDF is released by sixteen clock neurons in Drosophila and helps maintain circadian activity rhythms by coordinating a network of approximately 150 neuronal clocks.
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