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Chan, I. C. W.

Publications and source records attributed to Chan, I. C. W..

4 recordsLinked to original sources

Odour representations supporting ethology-relevant categorisation and discrimination in the Drosophila mushroom body

Neural representations of sensory stimuli serve multiple distinct purposes, from the rapid recognition of familiar environments, to the precise identification of individual salient cues. In the insect mushroom body (MB), odours are encoded by the activity of Kenyon cells (KCs). The random wiring of olfactory projection neurons (PNs) and KCs in the MB calyx is thought to enhance odour discrimination. Here, we examined the impact of deviations from random wiring and demonstrated their significant roles in shaping odour representations. We confirm that different KC types have distinct PN input biases correlated with the contextual relevance of the odour information delivered by the PNs. By recording the functional responses of different KC types to ethologically defined odour categories, we found that the {beta} and {beta} KCs produce segregated representations of relevant odour groups, potentially enhancing the categorisation of odours based on ethological relevance. Simultaneously, these same KC types displayed distinct representations for food-related odours, supporting precise discrimination. In contrast, {gamma} KCs lacked significant segregation of odour representations by ethological category. Computational simulations refined with our functional data indicated that the specific PN input connection pattern of individual KC types largely accounts for the observed representations. Taken together, we propose that individual KC types process odour information with distinct objectives, supporting both ethological categorisation and discrimination.

neuroscience↗

Three-photon in vivo imaging of neurons and glia in the medial prefrontal cortex with sub-cellular resolution

The medial prefrontal cortex (mPFC) is important for higher cognitive functions, including working memory, decision making, and emotional control. In vivo recordings of neuronal activity in the mPFC have been achieved via invasive electrical and optical approaches. Here we apply low invasive three-photon in vivo imaging in the mPFC of the mouse at unprecedented depth. Specifically, we measure neuronal and astrocytic Ca2+-transient parameters in awake head-fixed mice up to a depth of 1700 {micro}m. Furthermore, we longitudinally record dendritic spine density (0.41 {+/-}0.07 {micro}m-1) deeper than 1 mm for a week. Using 1650 nm wavelength to excite red fluorescent microglia, we quantify their processes motility (58.9 {+/-}2% turnover rate) at previously unreachable depths (1100 {micro}m). We establish three-photon imaging of the mPFC enabling neuronal and glial recordings with subcellular resolution that will pave the way for novel discoveries in this brain region.

neuroscience↗

Neuronal ribosomes dynamically exchange ribosomal proteins in a context-dependent manner.

Owing to their morphological complexity and dense network connections, neurons modify their proteomes locally, using mRNAs and ribosomes present in the neuropil (tissue enriched for dendrites and axons). Although ribosome biogenesis largely takes place in the nucleus and perinuclear region, neuronal ribosomal protein (RP) mRNAs have been frequently detected remotely, in dendrites and axons. Here, using imaging and ribosome profiling, we directly detected the RP mRNAs and their translation in the neuropil. Combining brief metabolic labeling with mass spectrometry, we found that a group of RPs quickly associated with translating ribosomes in the cytoplasm and that this incorporation is independent of canonical ribosome biogenesis. Moreover, the incorporation probability of some RPs was regulated by location (neurites vs. cell bodies) and changes in the cellular environment (in response to oxidative stress). Our results suggest new mechanisms for the local activation, repair and/or specialization of the translational machinery within neuronal processes, potentially allowing remote neuronal synapses a rapid solution to the relatively slow and energy-demanding requirement of nuclear ribosome biogenesis.

molecular biology↗

Subcellular sequencing of single neurons reveals the dendritic transcriptome of GABAergic interneurons

The localization and translation of mRNAs to dendrites and axons maintains and modifies the local proteome of neurons, and is essential for synaptic plasticity. Although significant efforts have allowed the identification of localized mRNAs in excitatory neurons, it is still unclear whether interneurons also localize a large population of mRNAs. In addition, the variability in the population of localized mRNAs within and between cell-types is unknown. Here we developed a method for the transcriptomic characterization of a single neurons subcellular compartments, which combines laser capture microdissection with scRNA-seq. This allowed us to separately profile the dendritic and somatic transcriptomes of individual rat hippocampal neurons and investigate the relation in mRNA abundances between the soma and dendrites of single glutamatergic and GABAergic neurons. We identified two types of glutamatergic and three types of GABAergic interneurons and we found that, like their excitatory counterparts, interneurons contain a rich repertoire of ~4000 mRNAs. The individual somatic transcriptomes exhibited more cell type-specific features than their associated dendritic transcriptomes. The detection and abundance of dendritic mRNAs was not always simply predicted by their somatic counterparts. Finally, using cell-type specific metabolic labelling of isolated neurites, we demonstrated that the processes not only of Glutamatergic but also of GABAergic neurons are capable of local translation, suggesting mRNA localization and local translation is a general property of neurons.

neuroscience↗