TY - JOUR
T1 - Doc2 Supports Spontaneous Synaptic Transmission by a Ca2+-Independent Mechanism
AU - Pang, Zhiping P.
AU - Bacaj, Taulant
AU - Yang, Xiaofei
AU - Zhou, Peng
AU - Xu, Wei
AU - Südhof, Thomas C.
N1 - Funding Information:
We thank Ira Huryeva for excellent technical support and Dr. Axel Brunger for help with circular dichroism spectra measurements. This paper was supported by a Recovery Act grant from the National Institute of Mental Health ([NIMH] 1R01 MH089054), a NIMH Conte Center Award (P50 MH086403), a NARSAD Young Investigator Award (to Z.P.P.), and a National Institute of Neurological Disorders and Stroke National Research Service Award fellowship (1F32NS067896, to T.B.).
PY - 2011/4/28
Y1 - 2011/4/28
N2 - Two families of Ca2+-binding proteins have been proposed as Ca2+ sensors for spontaneous release: synaptotagmins and Doc2s, with the intriguing possibility that Doc2s may represent high-affinity Ca2+ sensors that are activated by deletion of synaptotagmins, thereby accounting for the increased spontaneous release in synaptotagmin-deficient synapses. Here, we use an shRNA-dependent quadruple knockdown of all four Ca2+-binding proteins of the Doc2 family to confirm that Doc2-deficient synapses exhibit a marked decrease in the frequency of spontaneous release events. Knockdown of Doc2s in synaptotagmin-1-deficient synapses, however, failed to reduce either the increased spontaneous release or the decreased evoked release of these synapses, suggesting that Doc2s do not constitute Ca2+ sensors for asynchronous release. Moreover, rescue experiments revealed that the decrease in spontaneous release induced by the Doc2 knockdown in wild-type synapses is fully reversed by mutant Doc2B lacking Ca2+-binding sites. Thus, our data suggest that Doc2s are modulators of spontaneous synaptic transmission that act by a Ca2+-independent mechanism.
AB - Two families of Ca2+-binding proteins have been proposed as Ca2+ sensors for spontaneous release: synaptotagmins and Doc2s, with the intriguing possibility that Doc2s may represent high-affinity Ca2+ sensors that are activated by deletion of synaptotagmins, thereby accounting for the increased spontaneous release in synaptotagmin-deficient synapses. Here, we use an shRNA-dependent quadruple knockdown of all four Ca2+-binding proteins of the Doc2 family to confirm that Doc2-deficient synapses exhibit a marked decrease in the frequency of spontaneous release events. Knockdown of Doc2s in synaptotagmin-1-deficient synapses, however, failed to reduce either the increased spontaneous release or the decreased evoked release of these synapses, suggesting that Doc2s do not constitute Ca2+ sensors for asynchronous release. Moreover, rescue experiments revealed that the decrease in spontaneous release induced by the Doc2 knockdown in wild-type synapses is fully reversed by mutant Doc2B lacking Ca2+-binding sites. Thus, our data suggest that Doc2s are modulators of spontaneous synaptic transmission that act by a Ca2+-independent mechanism.
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U2 - 10.1016/j.neuron.2011.03.011
DO - 10.1016/j.neuron.2011.03.011
M3 - Article
C2 - 21521611
AN - SCOPUS:79955112041
SN - 0896-6273
VL - 70
SP - 244
EP - 251
JO - Neuron
JF - Neuron
IS - 2
ER -