A dye mixture (Neurobiotin and Alexa 488) reveals extensive dye-coupling among neurons in leeches; physiology confirms the connections |
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Authors: | Ruey-Jane?Fan Antonia?Marin-Burgin Kathleen?A?French Email author" target="_blank">W?Otto FriesenEmail author |
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Institution: | (1) Department of Biology, University of Virginia, Charlottesville, VA 22904-4328, USA;(2) Division of Biological Sciences, Neurobiology Section, University of California, San Diego La Jolla, CA 92093-0357, USA |
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Abstract: | Although the neuronal circuits that generate leech movements have been studied for over 30 years, the list of interneurons
(INs) in these circuits remains incomplete. Previous studies showed that some motor neurons (MNs) are electrically coupled
to swim-related INs, e.g., rectifying junctions connect IN 28 to MN DI-1 (dorsal inhibitor), so we searched for additional
neurons in these behavioral circuits by co-injecting Neurobiotin and Alexa Fluor 488 into segmental MNs DI–1, VI–2, DE–3 and
VE–4. The high molecular weight Alexa dye is confined to the injected cell, whereas the smaller Neurobiotin molecules diffuse
through gap junctions to reveal electrical coupling. We found that MNs were each dye-coupled to approximately 25 neurons,
about half of which are likely to be INs. We also found that (1) dye-coupling was reliably correlated with physiologically
confirmed electrical connections, (2) dye-coupling is unidirectional between MNs that are linked by rectifying connections,
and (3) there are novel electrical connections between excitatory and inhibitory MNs, e.g. between excitatory MN VE-4 and
inhibitory MN DI-1. The INs found in this study provide a pool of novel candidate neurons for future studies of behavioral
circuits, including those underlying swimming, crawling, shortening, and bending movements. |
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Keywords: | Hirudo Circuits Gap junctions Diode Motoneurons |
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