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Changes in phosphorylation of Ca2+/calmodulin-dependent protein kinase II (CaMKII) in processing of short-term and
1Department of Psychology, Monash University, Clayton, Victoria, Australia. wqzhao@helix.nih.gov
Abstract:
Characteristic autophosphorylation of calcium/ calmodulin-dependent protein kinase II (CaMKII) and its consequences have made this kinase an interesting target in studying the molecular pathway for important neuronal functions including learning and memory formation. In this article, we use immunoprecipitation and immunoblotting methods to detect changes in phosphorylation of CaMKII during memory formation in 1-day-old chicks trained in a single trial passive avoidance task. A 60-kDa protein has been immunoprecipitated from the chick brain with an anti-rabbit CaMKII antibody. This protein shows strong and specific immunoactivities with antibodies against the calmodulin binding site of CaMKII, and the N and C terminals of beta-CaMKII. Commercially available anti-phosphoserine and anti-phosphothreonine antibodies are shown to sensitively detect phosphorylation of purified CaMKII. The basal phosphorylation of CaMKII from the intermediate medial hyperstriatum ventrale (IMHV) and lobus parolfactorius (LPO) regions of the chick brain is shown to be largely right hemisphere-lateralized. When chicks are subjected to a passive avoidance training experience, a specific increase in CaMKII phosphorylation is induced in the IMHV and LPO of the left hemisphere from those chicks whose memory for the training experience is successfully retrieved. While this specific increase in CaMKII phosphorylation is seen in both the left IMHV and left LPO in short-term memory, it is detectable only in the left LPO associated with long-term memory retrieval. The present results provide evidence that in vivo changes in phosphorylation of CaMKII are associated specifically with processing of distinct memory stages, which take place in specific brain regions.
Insights
Calcium/calmodulin-dependent protein kinase II (CaMKII) phosphorylation changes in the chick brain are linked to memory formation. Specific left-hemisphere CaMKII phosphorylation increases in the IMHV and LPO correlate with memory retrieval stages.
Area of Science:
- Neuroscience
- Molecular Biology
- Cognitive Science
Background:
- Calcium/calmodulin-dependent protein kinase II (CaMKII) autophosphorylation is crucial for neuronal functions like learning and memory.
- Understanding CaMKII's role in memory formation requires investigating its phosphorylation state in vivo.
- Previous research highlights CaMKII's importance, but its specific involvement in distinct memory stages needs further elucidation.
Purpose of the Study:
- To investigate changes in CaMKII phosphorylation during memory formation in a chick passive avoidance task.
- To determine the regional and hemispheric specificity of CaMKII phosphorylation changes related to short-term and long-term memory retrieval.
- To provide evidence for in vivo CaMKII phosphorylation changes associated with distinct memory processing stages.
Main Methods:
- Immunoprecipitation and immunoblotting techniques were employed to detect CaMKII phosphorylation.
- Anti-CaMKII, anti-calmodulin binding site, and anti-N/C terminal antibodies were used for protein detection.
- Anti-phosphoserine and anti-phosphothreonine antibodies were utilized to assess CaMKII phosphorylation levels.
Main Results:
- A 60-kDa CaMKII protein was immunoprecipitated from chick brains, showing specific immunoactivities.
- Basal CaMKII phosphorylation in the IMHV and LPO regions was predominantly right hemisphere-lateralized.
- Passive avoidance training induced a specific increase in CaMKII phosphorylation in the left hemisphere's IMHV and LPO, correlating with successful memory retrieval.
Conclusions:
- In vivo CaMKII phosphorylation changes are specifically associated with the processing of distinct memory stages.
- Short-term memory retrieval involves increased CaMKII phosphorylation in both the left IMHV and LPO.
- Long-term memory retrieval is specifically associated with increased CaMKII phosphorylation in the left LPO.