THE NEURONAL MOLECULAR MECHANISMS OF COGNITIVE FUNCTIONS RELATED TO MEMORY
As we grow, we learn from past experiences by forming memory. Those memories are hopefully utilized to move ourselves forward. However, depending on our condition, e.g. mood, fatigue, body condition, or attention, we cannot act as we hope, although later on, we will notice what the appropriate behavior has to be. In this lab, we are trying to understand how those cognitive processes related to memory is regulated, and what the molecular and neuronal mechanisms are. To study long-lasting form of memory, we have been focused on the molecular events related to transcription, which is significant for the long-term memory. We will further work on specific synapses of memory-related neurons to conduct synapse-specific proteomic analysis. Using our original molecular biology techniques, together with the genetic and optogenetic methods in flies, we will understand how the molecular and neuronal mechanisms are orchestrated to adapt animals' behavior to the given environment or animals' state. Furthermore, we try to expand our understanding to the level of cognition through establishing new behavior paradigms.
TARGETING THE NUCLEUS OF THE SPECIFIC NEURONS
We established a method to collect the specific subtypes on neurons, such as the memory center, mushroom body in flies. We can access to the molecular details occurring in the nucleus, using proteomic, epigenetic (ChIP-seq), and transcriptome (nuclear RNA-seq) analyses.
PROTEOMIC ANALYSIS IN THE MEMORY-RELATED NEURONS
We optimized proteomics analysis to determine the molecular composition and the post-translational modification of the proteins of interest, which is specifically expressed in the memory center, mushroom body. (Although it requires 1,000-2,000 flies), we will have a clear picture of the molecular landscape in mushroom body neurons.
PROTEOMIC APPROACH TO THE SPECIFIC SYNAPSES
The brain is hard wired, but connected in a roughly stereotyped manner. The basic rule of the brain functions should be supported by the specific roles of each synapse, although biochemical dissection is currently impossible. We further optimize and develop a technique to investigate the molecular details of each synapses. This will add biochemical aspects onto the neuronal map, which will advance our understanding of the brain functions.
WATCHING FLY BEHAVIOR
To understand cognitive functions, we have to observe flies in more details. The technology development in tracking system and 3D printing brought us an significant step to accomplish the observation of fly behavior. We can now test anything we want!!
TECHNICAL INTRODUCTION
MEMORY ASSAY
Flies can be trained to remember an odor, when it is presented with reward or punishment. It is efficient and reliable evaluation of memory functions, in an automated system. We can also proceed to the biochemical assay to study the molecular mechanisms.
OPTOGENETIC MANIPULATION
Recent advances in neuroscience allow us to manipulate the neural activity in the living organism, by illuminating light (Optogenetics). By combining with the fly genetic tools, we can manipulate the specific neurons, and investigate the significance of the neurons of interest. Furthermore, we can study molecular changes by this artificial neural activation, through proteomic, epigenetic, and transcriptome analyses.
IMAGING OF THE MOLECULAR ACTION IN THE PRIMARY CULTURED NEURONS
How individual molecules behave in memory formation is the last, but an important investigation we have to conduct. We will not be able to understand the phenomena until we see it; the most likely interpretation based on the past studies may be totally different from the real world. Thus, we built up the system to visualize the molecular action using the primary cultured mushroom body neurons. This approach will lead to new findings regarding the real molecular action in neurons.
