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Volume 17, Number 2, Issue of January 15, 1997 pp. 676-696
Copyright ©1997 Society for Neuroscience

Temporal and Spatial Expression Patterns of Transgenes Containing Increasing Amounts of the Drosophila Clock Gene period and a lacZ Reporter: Mapping Elements of the PER Protein Involved in Circadian Cycling

Received July 8, 1996; revised Oct. 31, 1996; accepted Nov. 5, 1996.

Ralf Stanewsky¹, Brigitte Frisch¹, Christian Brandes¹, Melanie J. Hamblen-Coyle¹, Michael Rosbash^{1, 2}, and Jeffrey C. Hall¹

¹ Department of Biology and ² Howard Hughes Medical Institute, Brandeis University, Waltham, Massachusetts 02254 This article is dedicated to Brigitte Frisch. She found delight in her work and life. She believed that everyone had something important to offer and made us all feel special. Her humor and strength will always be remembered.

Rhythmic oscillations of the PER protein, the product of the Drosophila period (per) gene, in brain neurons of the adult fly are strongly involved in the control of circadian rhythms. We analyzed temporal and spatial expression patterns of three per-reporter fusion genes, which share the same 4 kb regulatory upstream region but contain increasing amounts of per's coding region fused in frame to the bacterial lacZ gene. The fusion proteins contained either the N-terminal half (SG), the N-terminal two-thirds (BG), or nearly all (XLG) of the PER protein. All constructs led to reporter signals only in the known per-expressing cell types within the anterior CNS and PNS. Whereas the staining intensity of SG flies was constantly high at different Zeitgeber times, the in situ signals in BG and XLG flies cycled with ~24 hr periodicity in the PER-expressing brain cells in wild-type and per⁰¹ loss of function flies. Despite the rhythmic fusion-gene expression within the relevant neurons of per⁰¹ BG flies, their locomotor activity in light/dark cycling conditions and in constant darkness was identical to that of per⁰¹ controls, uncoupling protein cycling from rhythmic behavior. The XLG construct restored weak behavioral rhythmicity to (otherwise) per⁰¹ flies, indicating that the C-terminal third of PER (missing in BG) is necessary to fulfill the biological function of this clock protein.

Key words: circadian rhythms; period gene; lacZ reporter; Lateral Neurons; fusion proteins; locomotor behavior; immunochemistry

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