2P-NucTag: on-demand phototagging for molecular analysis of functionally identified cortical neurons.
Jingcheng ShiBoaz NutkovichDahlia KushinskyBovey Y RaoStephanie A HerrlingerTiberiu S MihailaKatayun Cohen-Kashi MalinaCliodhna K O'TooleMargaret E Conde ParedesHyun Choong YongErdem VarolAttila LosonczyIvo SpiegelPublished in: bioRxiv : the preprint server for biology (2024)
Neural circuits are characterized by genetically and functionally diverse cell types. A mechanistic understanding of circuit function is predicated on linking the genetic and physiological properties of individual neurons. However, it remains highly challenging to map the functional properties of transcriptionally heterogeneous neuronal subtypes in mammalian cortical circuits in vivo . Here, we introduce a high-throughput two-photon nuclear phototagging (2P-NucTag) approach optimized for on-demand and indelible labeling of single neurons via a photoactivatable red fluorescent protein following in vivo functional characterization in behaving mice. We demonstrate the utility of this function-forward pipeline by selectively labeling and transcriptionally profiling previously inaccessible 'place' and 'silent' cells in the mouse hippocampus. Our results reveal unexpected differences in gene expression between these hippocampal pyramidal neurons with distinct spatial coding properties. Thus, 2P-NucTag opens a new way to uncover the molecular principles that govern the functional organization of neural circuits.
Keyphrases
- single cell
- spinal cord
- gene expression
- high throughput
- cerebral ischemia
- induced apoptosis
- genome wide
- dna methylation
- living cells
- quantum dots
- stem cells
- spinal cord injury
- cell therapy
- oxidative stress
- skeletal muscle
- signaling pathway
- adipose tissue
- blood brain barrier
- endoplasmic reticulum stress
- copy number
- metabolic syndrome
- brain injury