Molecular Cloning of Neural Genes 1987

[Paul Coleman]

1987 was the first year for the course "Cloning of Neural Genes". The instructors were Jim Eberwine (then Stanford, now Penn) and Marian Evinger (then Cornell, now Stony Brook). Our class project was to clone the intermediate neurofilament gene, which had not been cloned at the time. Early in the course, Jim Eberwine handed me and my lab partner (Mike McKinney) a piece of rat hippocampus, telling us to make a library from this tissue. I told Jim, of course we will, but I want you to know that it violates my esthetic sensibilities to grind up this tissue in which I know there are so many different cell types. Jim and I discussed this at length and decided to apply his recently developed method of in situ transcription to sections in which we would have stripped away unwanted cells using equipment made by Meridian Instruments. That approach failed (the RNA was degraded) but subsequently Jim, Hermes Yeh and I collaborated on a project in which micropipettes were used to isolate single neurons from culture, extract and then amplify the RNA. This is what is now known as the aRNA method for amplifying and subsequently profiling gene expression from small amounts of starting material (Eberwine et al., P.N.A.S., 1992). Jim has since adapted this method to profile messages expressed in single dendrites, and my lab has adapted it for postmortem human tissue (Chow et al., P.N.A.S., 1998). Of course, many others have used it for a variety of purposes.