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Salk researchers discover that stem cell marker regulates synapse formation
Date:1/30/2011

the current study's first author, took on the task of finding its accomplice.

He began by eliminating prime suspects in the plethora of proteins found on the muscle side of the synapse. "At the beginning it was like shooting in the dark because cdk5 has so many potential targets at the neuromuscular junction," says Yang. After eliminating the obvious candidates, the team finally considered nestin, based on evidence that cdk5 can phosphorylate nestin in some tissues.

To analyze nestin, the group employed mice in which the positive, synapse-stabilizing factor--known as agrin--had been genetically eliminated. As predicted, microscopic examination of diaphragm muscle tissue in agrin mutant mice showed a complete loss of dense receptor clusters that would mark a mature synapse, meaning that without the agrin "cement," synapse-dispersing activity had swept away the clusters.

However, when agrin mutant mice were administered an RNA reagent that literally knocks out nestin expression, the group made a dramatic finding: the pattern of receptor clusters on diaphragm muscle reappeared, reminiscent of synapses of a normal mouse--meaning that getting rid of nestin allows synapses to proceed even in the absence of the stabilizing glue.

"This in vivo experiment represents a critical genetic finding," explains Lee. "Later, we determined that nestin's basic function is to recruit cdk5 and its co-activators to the muscle membrane, leading to cdk5 activation and initiating the dispersion process." Additional experiments confirmed that nestin is expressed on the muscle side of the neuromuscular junction, in other words, in the "right" place, and that nestin phosphorylation is required for its newfound function.

Lee believes that information revealed by the study could enhance development of tissue replacement therapies. "Currently, in efforts to devise therapies for motor neuron disease or spinal cord injury there is a lot of focus is on how
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Contact: Gina Kirchweger
kirchweger@salk.edu
858-453-410-01340
Salk Institute
Source:Eurekalert  

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