Dopaminergic regulation of vestibulo-cerebellar circuits through unipolar brush cells.

While multiple monoamines modulate cerebellar output, the mechanistic details of dopaminergic signaling in the cerebellum remain poorly understood. We show that dopamine type 1 receptors (Drd1) are expressed in unipolar brush cells (UBCs) of the mouse cerebellar vermis. Drd1 activation increases UBC firing rate and post-synaptic NMDAR -mediated currents. Using anatomical tracing and in situ hybridization, we test three hypotheses about the source of cerebellar dopamine. We exclude midbrain dopaminergic nuclei and tyrosine hydroxylase-positive Purkinje (Pkj) cells as potential sources, supporting the possibility of dopaminergic co-release from locus coeruleus (LC) axons. Using an optical dopamine sensor GRABDA2h, electrical stimulation, and optogenetic activation of LC fibers in the acute slice, we find evidence for monoamine release onto Drd1-expressing UBCs. Altogether, we propose that the LC regulates cerebellar cortex activity by co-releasing dopamine onto UBCs to modulate their response to cerebellar inputs. Pkj cells directly inhibit these Drd1-positive UBCs, forming a dopamine-sensitive recurrent vestibulo-cerebellar circuit.

A near-infrared genetically encoded calcium indicator for in vivo imaging.

While calcium imaging has become a mainstay of modern neuroscience, the spectral properties of current fluorescent calcium indicators limit deep-tissue imaging as well as simultaneous use with other probes. Using two monomeric near-infrared (NIR) fluorescent proteins (FPs), we engineered an NIR Förster resonance energy transfer (FRET)-based genetically encoded calcium indicator (iGECI). iGECI exhibits high levels of brightness and photostability and an increase up to 600% in the fluorescence response to calcium. In dissociated neurons, iGECI detects spontaneous neuronal activity and electrically and optogenetically induced firing. We validated the performance of iGECI up to a depth of almost 400 µm in acute brain slices using one-photon light-sheet imaging. Applying hybrid photoacoustic and fluorescence microscopy, we simultaneously monitored neuronal and hemodynamic activities in the mouse brain through an intact skull, with resolutions of ~3 µm (lateral) and ~25-50 µm (axial). Using two-photon imaging, we detected evoked and spontaneous neuronal activity in the mouse visual cortex, with fluorescence changes of up to 25%. iGECI allows biosensors and optogenetic actuators to be multiplexed without spectral crosstalk.

Links Between Single-Trial Changes and Learning Rate in Eyelid Conditioning.

The discovery of single-trial learning effects, where the presence or absence (or the number) of climbing fiber inputs produces measureable changes in Purkinje cell response and in behavior, represents a major breakthrough in cerebellar learning. Among other things, these observations provide strong links between climbing fiber-mediated plasticity and cerebellar learning. They also demonstrate that cerebellar learning is stochastic, with each instantiation of a movement producing a small increment or decrement in gain. The sum of the small changes give rise to the macroscopic properties of cerebellar learning. We used a relatively large data set from another example of cerebellar-dependent learning, classical conditioning of eyelid responses, to attempt a behavioral replication and extension of single-trial learning effects. As a normal part of training, stimulus-alone trials provide instances where the climbing fiber response would be omitted, similar to non-climbing-fiber trials (gain down) during smooth pursuit training. The consequences of the stimulus-alone trial on the amplitude and timing of the conditioned response on the following paired trials were examined. We find that the amplitude of the conditioned response during the trial after a stimulus-alone trial (no climbing fiber input) was measurably smaller than the amplitude on the previous trials, and this single-trial effect on amplitude is larger for longer interstimulus intervals. The magnitude of the single-trial effect parallels the rate of extinction at different interstimulus intervals supporting the previously observed link between single-trial effects and learning.