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Loss of Thr286 phosphorylation disrupts synaptic CaMKIIα targeting, NMDAR activity and behavior in pre-adolescent
Richard M Gustin1, Brian C Shonesy, Stacey L Robinson
1Department of Pharmacology, Vanderbilt University School of Medicine, Nashville, TN 37232, United States.
Abstract:
In order to provide insight into in vivo roles of CaMKIIα autophosphorylation at Thr286 during postnatal development, behavioral, biochemical, and electrophysiological phenotypes of pre-adolescent Thr286 to Ala CaMKIIα knock-in (T286A-KI) and WT mice were examined. T286A-KI mice displayed cognitive deficits in a novel object recognition test and an anxiolytic phenotype in the elevated plus maze, suggesting disruption of normal developmental processes. At the molecular level, the ratio of total CaMKIIα to CaMKIIβ in hippocampal lysates was significantly decreased≈2-fold in T286A-KI mice, and levels of both isoforms in synaptic subcellular fractions were decreased by≈80%. Total levels of GluA1 AMPA-glutamate receptor subunits and phosphorylation of GluA1 at the CaMKII site (Ser831) in synaptic fractions were unaltered, as were the frequency and amplitude of AMPAR-mediated spontaneous excitatory postsynaptic currents at hippocampal CA3-CA1 synapses. Synaptic levels of NMDA-glutamate receptor GluN1, GluN2A and GluN2B subunits also were unaltered. However, the reduced ratio of CaMKII to NMDAR subunits in synaptic fractions was linked to increased synaptic NMDAR-mediated currents in T286A-KI mice, apparently due to increased functional contributions by GluN2B NMDARs (assessed by Ro 25-6981 sensitivity). Thus, disruption of CaMKII synaptic targeting caused by elimination of Thr286 autophosphorylation leads to synaptic and behavioral deficits during pre-adolescence.
Insights
Autophosphorylation of CaMKIIα at Thr286 is crucial for normal postnatal development. Eliminating this site impairs synaptic targeting, leading to cognitive and anxiety-related behavioral deficits in mice.
Area of Science:
- Neuroscience
- Molecular Biology
- Neurodevelopment
Background:
- Calcium/calmodulin-dependent protein kinase II alpha (CaMKIIα) autophosphorylation at Thr286 is critical for its function.
- The role of CaMKIIα autophosphorylation during postnatal development remains incompletely understood.
Purpose of the Study:
- To investigate the in vivo roles of CaMKIIα autophosphorylation at Thr286 during postnatal development.
- To examine the behavioral, biochemical, and electrophysiological consequences of impaired CaMKIIα autophosphorylation.
Main Methods:
- Utilized CaMKIIα knock-in (T286A-KI) and wild-type (WT) mice.
- Conducted behavioral tests (novel object recognition, elevated plus maze).
- Performed biochemical analyses of hippocampal lysates and synaptic fractions.
- Assessed electrophysiological properties of hippocampal CA3-CA1 synapses.
Main Results:
- T286A-KI mice exhibited cognitive deficits and anxiolytic behavior.
- Reduced CaMKIIα/β ratio and decreased synaptic CaMKII levels were observed in T286A-KI mice.
- Synaptic AMPAR levels and function were unaltered, but NMDAR-mediated currents increased, linked to enhanced GluN2B NMDAR function.
Conclusions:
- Disruption of CaMKIIα autophosphorylation at Thr286 impairs synaptic targeting.
- This impairment leads to significant synaptic and behavioral deficits during pre-adolescence.
- CaMKIIα autophosphorylation is essential for normal neurodevelopmental processes.
