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Complex behaviors arise from neural circuits that assemble from diverse cell types. Sleep is a conserved behavior essential for survival, yet little is known about how the nervous system generates neuron types of a sleep-wake circuit. Here, we focus on the specification of Drosophila 23E10-labeled dorsal fan-shaped body (dFB) long-field tangential input neurons that project to the dorsal layers of the fan-shaped body neuropil in the central complex. We use lineage analysis and genetic birth dating to identify two bilateral type II neural stem cells (NSCs) that generate 23E10 dFB neurons. We show that adult 23E10 dFB neurons express ecdysone-induced protein 93 (E93) and that loss of ecdysone signaling or E93 in type II NSCs results in their misspecification. Finally, we show that E93 knockdown in type II NSCs impairs adult sleep behavior. Our results provide insight into how extrinsic hormonal signaling acts on NSCs to generate the neuronal diversity required for adult sleep behavior. These findings suggest that some adult sleep disorders might derive from defects in stem cell-specific temporal neurodevelopmental programs. 

We report on the construction and characterization of a low-cost Mach–Zehnder optical interferometer in which quadrature signal detection is achieved by means of polarization control. The device incorporates a generic green laser pointer, home-built photodetectors, 3D-printed optical mounts, a circular polarizer extracted from a pair of 3D movie glasses, and a python-enabled microcontroller for analog-to-digital data acquisition. Components fit inside of a [Formula: see text] space and can be assembled on a budget of less than US$500. The device has the potential to make quadrature interferometry accessible and affordable for instructors, students, and enthusiasts alike. 

We report on the construction and characterization of a quadrature-detected optical interferometer that can be assembled on a budget of less than US$500, and in which quadrature detection is achieved by means of polarization control. 

Abstract Improved prodrug‐activating enzymes have the potential to increase the therapeutic efficacy of gene‐directed enzyme prodrug therapy (GDEPT). Yeast cytosine deaminase (yCD) is commonly used to convert the prodrug 5‐fluorocytosine (5‐FC) to the chemotherapeutic 5‐fluorouracil for GDEPT. Mutagenesis studies on yCD aimed at improving its application in GDEPT have been limited to subsets of residues or have sought to improve a single property of the enzyme. We performed comprehensive site‐saturation mutagenesis (CSM) on yCD designed to create all 2,983 possible unique protein mutants with a single amino acid substitution. We identified active variants throughEscherichia coligenetic complementation and screened these mutants, and combinations thereof, for increased ability to sensitizeE. coliand HT1080 fibrosarcoma cells to 5‐FC. Several mutants identified in this study showed increased sensitization ability for bothE. coliand HT1080 cells indicating that CSM is an effective directed evolution tool for identifying unexpectedly beneficial mutations.