Search bioRxiv⌕ Search

Biology subjects

Henn, K.

Publications and source records attributed to Henn, K..

3 recordsLinked to original sources

Ultrafast Frame-Free Imaging of Neural Activity with Event Cameras

Frame-based fluorescence imaging has long defined how neural activity is optically measured. This approach requires acquiring all pixels within an image, regardless of whether they carry meaningful neural dynamics, thereby intrinsically coupling spatial and temporal resolution while increasing data output. Here, we introduce an entirely different, frame-free approach that leverages the sparse nature of neural activity using event-based cameras, which asynchronously report fluorescence changes as spatiotemporal events. Compared with a frame-based camera, our method preserves signal fidelity while eliminating the fixed trade-off between spatial resolution, temporal resolution and data rate, thereby reducing data output by orders of magnitude. Applied to hippocampal preparations we demonstrate that the frame-free approach can resolve both single action potentials and fast network dynamics over large fields of view at kilohertz rates, enabling scalable, ultrafast optical recordings.

neuroscience↗

Assembly and lipid-gating of LRRC8A:D volume-regulated anion channels

Volume-regulated anion channels (VRACs) are ubiquitously expressed vertebrate ion channels that open in response to hypotonic swelling. VRACs assemble as heteromers of LRRC8A and LRRC8B-E subunits, with different subunit combinations resulting in channels with different properties. Recent studies have described the structures of LRRC8A:C VRACs, but how other VRACs assemble, and which structural features are conserved or variant across channel assemblies remains unknown. Herein, we used cryo-EM to determine structures of a LRRC8A:D VRAC with a 4:2 subunit stoichiometry, which we captured in two conformations. The presence of LRRC8D subunits increases hydrophobicity and widens the selectivity filter, which may explain the unique substrate selectivity of LRRC8D-containing VRACs. The structures reveal lipids bound inside the channel pore, similar to those observed in LRRC8A:C VRACs. Using electrophysiological experiments, we confirmed that pore lipids block conduction in the closed state, demonstrating that lipid-gating is a general property of VRACs. Finally, we observe that LRRC8D subunit incorporation disrupts packing of the cytoplasmic LRR domains, increasing channel dynamics and opening lateral fenestrations, which we speculate are necessary for pore lipid evacuation and channel activation.

biophysics↗

TMEM165 acts as a proton-activated Ca2+ importer in lysosomes

Lysosomal calcium (Ca2+) release is critical to cell signaling and is mediated by well-known lysosomal Ca2+ channels. Yet, how lysosomes refill their Ca2+ remains hitherto undescribed. Here, from an RNAi screen, we identify an evolutionarily conserved gene, lci-1, that facilitates lysosomal Ca2+ entry in C. elegans and in mammalian cells. Its human homolog TMEM165, previously designated as a Golgi-resident Ca2+/H+ exchanger (CAX), has a minor lysosomal population of unknown function. Using genetics, lysosomal Ca2+ imaging and electrophysiology, we show that TMEM165 acts as a proton-activated, lysosomal Ca2+ importer in lysosomes. Defects in lysosomal Ca2+ channels cause several neurodegenerative diseases, and knowledge of lysosomal Ca2+ importers may provide new avenues to explore the physiology of Ca2+ channels.

cell biology↗