Lobe Specific Ca2+-Calmodulin Nano-Domain in Neuronal Spines: A Single Molecule Level Analysis

摘要

Calmodulin (CaM) is a ubiquitous Ca²⁺ buffer and second messenger that affects cellular function as diverse as cardiac excitability, synaptic plasticity, and gene transcription. In CA1 pyramidal neurons, CaM regulates two opposing Ca²⁺-dependent processes that underlie memory formation: long-term potentiation (LTP) and long-term depression (LTD). Induction of LTP and LTD require activation of Ca²⁺-CaM-dependent enzymes: Ca²⁺/CaM-dependent kinase II (CaMKII) and calcineurin, respectively. Yet, it remains unclear as to how Ca²⁺ and CaM produce these two opposing effects, LTP and LTD. CaM binds 4 Ca²⁺ ions: two in its N-terminal lobe and two in its C-terminal lobe. Experimental studies have shown that the N- and C-terminal lobes of CaM have different binding kinetics toward Ca²⁺ and its downstream targets. This may suggest that each lobe of CaM differentially responds to Ca²⁺ signal patterns. Here, we use a novel event-driven particle-based Monte Carlo simulation and statistical point pattern analysis to explore the spatial and temporal dynamics of lobe-specific Ca²⁺-CaM interaction at the single molecule level. We show that the N-lobe of CaM, but not the C-lobe, exhibits a nano-scale domain of activation that is highly sensitive to the location of Ca²⁺ channels, and to the microscopic injection rate of Ca²⁺ ions. We also demonstrate that Ca²⁺ saturation takes place via two different pathways depending on the Ca²⁺ injection rate, one dominated by the N-terminal lobe, and the other one by the C-terminal lobe. Taken together, these results suggest that the two lobes of CaM function as distinct Ca²⁺ sensors that can differentially transduce Ca²⁺ influx to downstream targets. We discuss a possible role of the N-terminal lobe-specific Ca²⁺-CaM nano-domain in CaMKII activation required for the induction of synaptic plasticity.