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Breakthrough in cell-type analysis offers new ways to study development and disease
Date:11/13/2008

netic markers for each kind. But it was an accomplishment that was also a taunt. Ultimately, the researchers wanted to go deeper to understand the precise biochemical characteristics of the cell types they had brought into focus, to learn what makes cells vulnerable to attack - and possibly how to protect them from it - by discovering what's unique to the susceptible cells and the ones that are resistant. Enter TRAP.

Heintz, postdoctoral fellow Myriam Heiman and colleagues attached an EGFP to the surface of the ribosome and used it as a handle to pick out the cell's protein factories and the genetic messages passing through them, called messenger RNAs (mRNAs). Using the GENSAT techniques and findings, they designed new mouse lines that made tagged ribosomes in each of four different cell types. Heiman and colleagues focused on the brain cells that respond to dopamine, an important neurotransmitter involved in muscle movement and emotion regulation, among other things. They used the handle they had made to pluck out the ribosomes and mRNAs from these brain cells and freeze them within minutes of dissection, preserving the "messages" largely as they were inside the living animal and minimizing degradation. Alternative approaches to getting the profiles of cell types in complex tissues have been disappointing because they require the physical isolation of whole cells from the tissues in which they are embedded. TRAP bypasses that logistical nightmare by going straight for the ribosome.

The method proved so sensitive that researchers were able to identify a few hundred genetic messages that differ between two types of dopamine-sensing brain cells that previously had seemed nearly identical. Because these cells are crucial elements of the neural circuit that degenerates in Parkinson's and Huntington's diseases, the newly identified proteins could aid in the design of drugs that would allow these two key cell types to be treated independently of one an
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Contact: Brett Norman
bnorman@rockefeller.edu
212-327-7613
Rockefeller University
Source:Eurekalert

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