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

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 transcription 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. This result suggests that the presumed dominance of the brain clock in regulating peripheral clocks needs to be re-examined. This result further demonstrates that LABL allows rapid, affordable, and direct real-time monitoring of clocks in vivo.

Preliminary study on estimation of flow velocity vectors using focused transmit beams

High-frame-rate ultrasound imaging with plane wave transmissions is a predominant method for blood flow imaging, and methods for estimation of blood flow velocity vectors have been developed based on high-frame-rate imaging. On the other hand, in imaging of soft tissues, such as arterial walls and atherosclerotic plaques, high-frame-rate imaging sometimes suffers from high-level clutters. Even in observation of the arterial wall with a focused transmit beam, it would be highly beneficial if blood flow velocity vectors could be estimated simultaneously. We conducted a preliminary study on estimation of blood flow velocity vectors based on a multi-angle Doppler method with focused transmit beam and parallel receive beamforming. It was shown that the lowest estimation error was achieved at a steering angle of 25 degrees by simulation. Also, velocity vectors with typical velocity magnitudes and directions could be obtained by the proposed method in in vivo measurement of a carotid artery.