Periodic Reporting for period 1 - ALPINE (Ultrastructural analysis of phosphoinositides in nerve terminals: distribution, dynamics and physiological roles in synaptic transmission)
Reporting period: 2018-04-01 to 2020-03-31
As an offshoot of this project, I found that acute brain slices prepared at physiological temperature enhanced the quality of the brain slices compared to the conventional brain slicing method preparing at ice-cold temperature. Using electron microscopic, super-resolution microscopic and electrophysiological approaches, I found that the acute cerebellar slice preparation at ice-cold temperature alters several crucial parameters of synapses including dendritic spine formation, synaptic protein distribution and synaptic vesicle distribution in AZs. In contrast, brain slices prepared at physiological temperature showed no alterations of the synaptic properties shown in “cold-cut” slices, means this “warm-cutting” method improves the slice quality for the following experiments. This result has been published in an open access scientific journal (Frontiers in Cellular Neuroscience) to be known the advantages of the warm-cutting method.
In this project, I found that acute brain slices prepared at physiological temperature enhanced the quality of the preparations for investigation of synaptic functions rather than the conventional brain slice method preparing at ice-cold temperature. The “warm-cutting” method also provides us an additional advantage: skipping the recovery time after slicing. In conventional “cold-cutting” method, brain tissues need a recovery time (~60 min) in buffer at warm temperature before using in following experiments. However, the warm-cutting method does not require this recovery step, and experimenters can use the brain slices after slicing immediately. This is a huge advantage especially for experiments of long-term plasticity induced by in vivo behavior experiments. I have demonstrated that long-term depression (LTD) of electrophysiological synaptic events induced by a motor learning at cerebellum was more detectable in the warm-cut slices than the cold-cut slices, because of the shortening of the duration between the training and the recording. These advantages of the warm-cutting method for acute brain slice preparation will help experimenters in a wide range of neuroscience research, especially for synaptic plasticity induced in vivo.