RT Journal Article
T1 Studying synaptic efficiency by post-hoc immunolabelling
A1 Ramírez-Franco, Jorge
A1 Alonso, Beatris
A1 Bartolomé-Martín, David
A1 Sánchez-Prieto Borja, José
A1 Torres Molina, Magdalena Isabel
AB BackgroundIn terms of vesicular recycling, synaptic efficiency is a key determinant of the fidelity of synaptic transmission. The ability of a presynaptic terminal to reuse its vesicular content is thought to be a signature of synaptic maturity and this process depends on the activity of several proteins that govern exo/endocytosis. Upon stimulation, individual terminals in networks of cultured cerebellar granule neurons exhibit heterogeneous exocytic responses, which reflect the distinct states of maturity and plasticity intrinsic to individual synaptic terminals. This dynamic scenario serves as the substrate for processes such as scaling, plasticity and synaptic weight redistribution. Presynaptic strength has been associated with the activity of several types of proteins, including the scaffolding proteins that form the active zone cytomatrix and the proteins involved in presynaptic exocytosis.MethodsWe have combined fluorescence imaging techniques using the styryl dye FM1-43 in primary cultures of cerebellar granule cells with subsequent post-hoc immunocytochemistry in order to study synaptic efficiency in terms of vesicular release. We describe a protocol to easily quantify these results with minimal user intervention.ResultsIn this study we describe a technique that specifically correlates presynaptic activity with the levels of presynaptic markers. This method involves the use of the styryl dye FM1-43 to estimate the release capacity of a synaptic terminal, and the subsequent post-hoc immunolabelling of thousands of individual nerve terminals. We observed a strong correlation between the release capacity of the nerve terminal and the levels of the RIM1α but not the Munc13-1 protein in the active zone.ConclusionsOur findings support those of previous studies and point out to RIM1α as a crucial factor in determining synaptic efficiency. These results also demonstrate that this technique is a useful tool to analyse the molecular differences underlying the heterogeneous responses exhibited by neuronal networks.
PB Springer Nature
YR 2013
FD 2013
LK https://hdl.handle.net/20.500.14352/129690
UL https://hdl.handle.net/20.500.14352/129690
LA eng
NO Ramírez-Franco, J., Alonso, B., Bartolomé-Martín, D., Sánchez-Prieto, J., & Torres, M. (2013). Studying synaptic efficiency by post-hoc immunolabelling. BMC neuroscience, 14, 127. https://doi.org/10.1186/1471-2202-14-127
NO Contribución de autores:JRF, JSP and MT conceived the study. JRF, BA and DBM designed experiments, and were responsible for collect, analyse and interpret the data; JRF and JSP wrote the manuscript. MT and JSP acquired funding necessary for the completion of the study. All authors read and approved the final manuscript.
NO Ministerio de Economía y Comercio (España)
NO Instituto de Salud Carlos III
NO Comunidad de Madrid
DS Docta Complutense
RD 25 feb 2026