Presynaptically Silent Synapses Studied with Light Microscopy

Krista L. Moulder; Xiaoping Jiang; Amanda A. Taylor; Ann M. Benz; Steven Mennerick

doi:10.3791/1676

Presynaptically Silent Synapses Studied with Light Microscopy

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Biology

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Presynaptically Silent Synapses Studied with Light Microscopy

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11:02 min

January 4, 2010

DOI: 10.3791/1676-v

Krista L. Moulder¹, Xiaoping Jiang¹, Amanda A. Taylor¹, Ann M. Benz¹, Steven Mennerick^1,2,3

¹Department of Psychiatry,Washington University School of Medicine, ²Department of Anatomy,Washington University School of Medicine, ³Department of Neurobiology,Washington University School of Medicine

Glutamatergic synapses can switch from an active mode to a silent mode. We demonstrate that presynaptic activity status in dissociated culture of rodent neurons is visualized using a fixable form of the FM1-43 dye to visualize active synapses and immunostaining with vGluT-1 antibody to visualize all glutamate synapses.

Some presynaptic terminals in the central nervous system appear non-functional or silent. Here we present a protocol for identifying silent presynaptic terminals. This protocol can be applied to identifying treatments and manipulations that are suspected of silencing or awakening terminals.

The protocol combines vital dye labeling of synaptic vesicles during neuronal depolarization with subsequent labeling of fixed cells with an antibody that labels all presynaptic terminals, both silent and active.Synapsis. Co labeled with dye and antibody are active. Synapsis labeled only by antibody are silent.

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