Combining retrograde tracing, intracellular injection, anterograde degeneration and electron microscopy to reveal synaptic links.

Journal of Neuroscience Methods
E BuhlW Singer

Abstract

Synaptic circuitry was investigated by combining retrograde tracing, intracellular staining, anterograde degeneration and electron microscopy in the same piece of tissue. This methodological procedure was successfully applied to disentangle a disynaptic neuronal chain, which originated in the olfactory bulb, was synaptically relayed in the entorhinal cortex and terminated in the ipsilateral hippocampus. Presumed entorhinal relay cells were retrogradely labelled from their hippocampal termination site by means of a fluorescent tracer. Subsequently, the marked projection neurones were intracellularly injected with Lucifer yellow in fixed slice preparations. Following a simple photo-conversion procedure, dye filled cells were processed for electron microscopy. The origin of presynaptic afferents to identified relay cells was revealed ultrastructurally after lesion induced anterograde degeneration of olfactory mitral cell axons. Due to its reliability, technical simplicity and a high degree of selectivity the new approach is considered an appropriate tool for unravelling neuronal networks.

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Citations

Oct 1, 1992·Journal of Neuroscience Methods·G BalerciaM Bentivoglio
Oct 1, 1993·Journal of Neuroscience Methods·X JiangA Burkhalter
May 31, 1990·Neuroscience Letters·M Bentivoglio, H S Su
Sep 18, 2002·Journal of Neuroscience Methods·Christopher J PaceSuzannah Bliss Tieman
Oct 21, 1996·Journal of Neuroscience Methods·L P TolbertJ G Hildebrand
Feb 1, 1997·Journal of Neuroscience Methods·J KaczaJ Seeger
Feb 12, 1999·Brain Research. Brain Research Protocols·G Prusky, T Arjannikova
May 4, 2016·Expert Review of Neurotherapeutics·Sahil GuptaBianca Weinstock-Guttman
Feb 8, 1990·The Journal of Comparative Neurology·W K SchwerdtfegerP Germroth
Apr 27, 2018·Journal of Neuroscience Research·Floris G WouterloodJean-Michel Deniau
Jul 26, 2011·Micron : the International Research and Review Journal for Microscopy·Shannon Modla, Kirk J Czymmek

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