For experiments examining syt-lum uptake, immediately after local

For experiments examining syt-lum uptake, immediately after local perfusion, 2 μM TTX was bath-applied for 10 min to isolate spontaneous neurotransmitter release. Neurons were then live labeled with anti-syt-lum for 5 min at RT and processed for immunocytochemistry as described above. The density and intensity of vglut

particles were calculated for each dendritic segment, and the average value was then used for normalizing vlgut density and intensity in all segments (including the treated area). The proportion of vglut particles with syt-lum particles was also determined in each segment. Statistical differences were assessed by ANOVA and Fisher’s LSD post-hoc tests. We thank Richard Tsien, Mia Lindskog, and Rachel Groth for their helpful comments on the manuscript, as well as Hisashi Umemori and members of the Sutton lab for useful discussions. This work was supported JQ1 nmr by RO1MH085798 from The National Institute of Mental Health (M.A.S.) and a grant Small Molecule Compound Library from the Pew Biomedical Scholars Program (M.A.S.). “
“Since the introduction of Dale’s principle of “one neuron releases one fast neurotransmitter” (Dale, 1935), an increasing number of exceptions to this rule have been found in many parts of the nervous system (Burnstock, 2004, Jo and Schlichter, 1999, Jonas et al., 1998, Li et al., 2004, Nishimaru

et al., 2005, Seal and Edwards, 2006, Tsen et al., 2000 and Wojcik et al., 2006), suggesting that corelease of multiple fast neurotransmitters by a single neuron may represent a Thymidine kinase significant mode of neurotransmission. However, the mechanism, circuitry, and function of coneurotransmission in the CNS are poorly understood in general. In the vertebrate retina, starburst amacrine cells (SACs) synthesize and release two classic fast neurotransmitters of opposite excitability, namely acetylcholine (ACh) and gamma-aminobutyric acid (GABA) (Brecha et al., 1988, Kosaka et al., 1988, O’Malley and Masland, 1989 and Vaney and Young, 1988). These cells exist as two mirror-symmetric populations across the inner plexiform layer (IPL), with the somas of one population (conventional or Off SACs) located in the inner nuclear

layer (INL) and those of the other population (displaced or On SACs) in the ganglion cell layer (GCL). The processes (dendrites) of SACs have a radially symmetric (“starburst”) dendritic morphology and ramify in two narrow substrata of the IPL, where the dendrites of neighboring SACs and direction-selective ganglion cells (DSGCs) cofasiculate to form a dense, honeycomb-shaped meshwork (Famiglietti, 1985, Famiglietti, 1992, Famiglietti, 1983, Tauchi and Masland, 1984 and Vaney, 1984). This meshwork is well organized and experimentally approachable, offering a unique opportunity for understanding the mechanism, circuitry, and function of neurotransmitter corelease. SACs are a key component in the direction-selective circuit (Amthor et al.

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