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Page updated at 12:43:23 PM on Monday, May 21st, 2012

CNS Development Probed by Random Access Non-linear Optical Electrophysiology

Funded with $350,000.00 for 36 months (from September, 2009 to August, 2012) by HFSPO within HFSP RGP 0027/2009-C

Person in charge: Francesco Saverio Pavone
LENS members: Jacopo Lotti, Leonardo Sacconi

This project aims to investigate the functiogenesis of neural circuits in the perinatal central nervous system.

Two-photon excitation florescence images of sagittal rat cerebellar acute slice, depth 90 μm, dye di-4-AN(F)EPPTEA. Morphologic development since post-natal day 8 (P8) to P27. ML: Molecular Layer. PL: Purkinje Layer. GL: Granule Layer. Red lines: line-scanning recording sites. Scale bar: 10 μm.

Key tools and techniques involved in structural and functional analysis of neural circuits will be new voltage sensitive dyes, non-linear optics and consequent 4D (time plus 3 spatial dimensions) mapping of the system with resolutions that can permit the recording of activity down to the cellular and sub-cellular levels.

The project brings together three research teams with highly complementary and unique strengths. The team of Drs. Sato and Momose-Sato (Tokyo Medical University) has worked for many years to map electrical connectivity in the developing central nervous system and has recently studied the widely-spreading depolarization wave in embryonic nervous systems which originates in the developing brainstem/spinal cord. Prof. Pavone (LENS) is an expert in biophotonics and has developed a unique random access non-linear optical microscope that, in combination with voltage-sensitive dyes of new generation, permit to monitor and record cellular electrical activity with sub-μm spatial and sub-ms temporal resolution to finely investigate the mechanism of the depolarization wave. Finally, Dr. Loew (UConn Health Center) has worked for many years to synthesize voltage sensitive dyes for optical recording of electrical activity, characterizing them in vitro and in vivo and pioneering the use of second harmonic generation (SHG) to image changes in membrane potential.

Thus, the U.S. and Italian labs are creating the appropriate dyes and imaging technologies to allow their Japanese colleagues to probe the developing central nervous system with sufficient resolution to begin to understand how functional connections are established. New methods developed in this research will broadly impact the study of neuronal circuitry in both developing and adult brain.

   

Publications

Only publications with LENS-affiliated authors are listed and for now there is no one.