Mitochondrial dysfunction is implicated in the pathogenesis of multiple neurological diseases, but elucidation of underlying mechanisms is limited experimentally by the inability to damage specific mitochondria in defined neuronal groups. We developed a precision chemoptogenetic approach to target neuronal mitochondria in the intact nervous system in vivo. MG2I, a chemical fluorogen, produces singlet oxygen when bound to the fluorogen-activating protein dL5** and exposed to far-red light. Transgenic zebrafish expressing dL5** within neuronal mitochondria showed dramatic MG2I- and light-dependent neurobehavioral deficits, caused by neuronal bioenergetic crisis and acute neuronal depolarization. These abnormalities resulted from loss of neuronal respiration, associated with mitochondrial fragmentation, swelling and elimination of cristae. Remaining cellular ultrastructure was preserved initially, but cellular pathology downstream of mitochondrial damage eventually culminated in neuronal death. Our work provides powerful new chemoptogenetic tools for investigating mitochondrial homeostasis and pathophysiology and shows a direct relationship between mitochondrial function, neuronal biogenetics and whole-animal behavior.

Larval zebrafish react to changes in ambient illumination with a series of stereotyped motor responses, called the visual motor response (VMR). The VMR has been used widely in zebrafish models to analyze how genetic or environmental manipulations alter neurological function. Prior studies elicited the VMR using white light. In order to elucidate the underlying afferent pathways and to identify light wavelengths that elicit the VMR without also activating optogenetic reagents, we employed calibrated narrow-waveband light sources to analyze the spectral properties of the response. Narrow light wavebands with peaks between 399nm and 632nm triggered the characteristic phases of the VMR, but there were quantitative differences between responses to different light wavelengths at the same irradiant flux density. The O-bend component of the VMR was elicited readily at dark onset following illumination in 399nm or 458nm light, but was less prominent at the transition from 632nm light to dark. Conversely, stable motor activity in light was observed at 458nm, 514nm, and 632nm, but not at 399nm. The differential effect of discrete light wavebands on components of the VMR suggests they are driven by distinct photoreceptor populations. Furthermore, these data enable the selection of light wavebands to drive the VMR in a separate channel to the activation of optogenetic reagents and photosensitizers.

A method is described for analyzing skin impedance data and applied to the determination of a steady‐state electrical model for intact human skin. An algorithm is presented for the analytical procedure, Bode analysis, and a sample impedance function is obtained. Bode plots are employed to synthesize a passive equivalent circuit from sample measurements of “black box” skin impedance magnitude and phase angle, and representative values for the circuit elements are presented.

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