Optogenetics in Sinorhizobium meliloti Enables Spatial Control of Exopolysaccharide Production and Biofilm Structure.

Microorganisms play a vital role in shaping the soil environment and enhancing plant growth by interacting with plant root systems. Because of the vast diversity of cell types involved, combined with dynamic and spatial heterogeneity, identifying the causal contribution of a defined factor, such as a microbial exopolysaccharide (EPS), remains elusive. Synthetic approaches that enable orthogonal control of microbial pathways are a promising means to dissect such complexity. Here we report the implementation of a synthetic, light-activated, transcriptional control platform using the blue-light responsive DNA binding protein EL222 in the nitrogen fixing soil bacterium Sinorhizobium meliloti. By fine-tuning the system, we successfully achieved optical control of an EPS production pathway without significant basal expression under noninducing (dark) conditions. Optical control of EPS recapitulated important behaviors such as a mucoid plate phenotype and formation of structured biofilms, enabling spatial control of biofilm structures in S. meliloti. The successful implementation of optically controlled gene expression in S. meliloti enables systematic investigation of how genotype and microenvironmental factors together shape phenotype in situ.

Extracellular single-unit recordings from peripheral nerve axons in vitro by a novel multichannel microelectrode array.

The peripheral nervous system (PNS) is an attractive target for modulation of afferent input (e.g., nociceptive input signaling tissue damage) to the central nervous system. To advance mechanistic understanding of PNS neural encoding and modulation requires single-unit recordings from individual peripheral neurons or axons. This is challenged by multiple connective tissue layers surrounding peripheral nerve fibers that prevent electrical recordings by existing electrodes or electrode arrays. In this study, we developed a novel microelectrode array (MEA) via silicon-based microfabrication that consists of 5 parallel hydrophilic gold electrodes surrounded by silanized hydrophobic surfaces. This novel hydrophilic/hydrophobic surface pattern guides the peripheral nerve filaments to self-align towards the hydrophilic electrodes, which dramatically reduces the technical challenges in conducting single-unit recordings. We validated our MEA by recording simultaneous single-unit action potentials from individual axons in mouse sciatic nerves, including both myelinated A-fibers and unmyelinated C-fibers. We confirmed that our recordings were single units from individual axons by increasing nerve trunk electrical stimulus intensity, which did not alter the spike shape or amplitude. By reducing the technical challenges, our novel MEA will likely allow peripheral single-unit recordings to be adopted by a larger research community and thus expedite our mechanistic understanding of peripheral neural encoding and modulation.