Thursday, February 12, 2009

The thrill of victory...

On the off chance that any of my miniscule readership does not already follow Ambivalent Academic's blog, you have to go over and check out yesterday's post over there. In it AA discusses the positive aspects of being a basic academic scientist, and I'd hazard a guess that many of us can strongly relate to those feelings. It serves as a welcome counterpoint to all the crappy and ugly aspects of the practice of science, and all the bitching and complaining that goes on in the blogosphere (and in lunchrooms, coffeebreaks, and post seminar chats between scientists in real life). Now, don't get me wrong, discussing all those negative parts are very useful and necessary. But it's also nice to sit back and reflect on the things we love about science. I find it helps stoke my passion for science, which too often nears extinguishment. And I really do love being a scientist.

Physioprof added a great comment as well, about his excitement at an old breakthrough achieved in grad school. You can feel his enthusiasm, even for an event that must be years old by now. And as he says, we're all chasing that feeling. I remember one of my own as well, which I will share. For my first project in grad school I was recording sodium and calcium currents during action potentials in nociceptors. As there's no good blocker for TTX-resistant sodium currents, I settled on using ionic substitution, replacing external sodium with NMDG. That worked well for the subtraction, but I did notice that the resulting sodium current kept increasing even as the voltage approached ENa, which obviously shouldn't happen. I kept that stuck in the craw of my brain, which chewed over it as I proceeded to look at calcium currents in other cells. (Which is how I normally let it happen; given time the craw digests whatever problem is given to it).

I can still remember sitting at my desk, looking over some other experiments, when it hit me. It was obvious that intracellular Na+ and K+ were making outward currents through unblocked TTX-R channels, and these became sizeable at depolarized potentials near the peak of the AP. In retrospect it isn't so surprising. But it wasn't so obvious to me that the outward currents would really be large enough to make a big difference, relative to the large inward currents when external sodium was high. Turns out they were. Very soon after that I figured out a way to correct my previous results, which became a figure on its own in the final paper. All in all a great feeling.

I think we all just hope that our science doles out sufficient number of these moments to keep us from totally giving up in the face of so many difficulties.


DSK Samways said...

That's absolutely it. I had to wait 'till my postdoc to have my moment though.

My original goal was to identify the selectivity filter conferring Ca2+ permeability in P2X receptors, and the first thing I did was pour over sequence alignments looking for conserved residues in the pore domains. Then it occurred to me that the P2X family have widely varying Ca2+ permeabilities, so looking for strongly conserved residues was probably the wrong approach. I quickly identified acidic residues that were only present in the two family members with the highest Ca2+ permeability/fractional Ca2+ currents and, Bob's Your Mother's Brother, a paper.

Of course, since then I've had a long run of being dead wrong about a lot of things, but so it goes.

Ambivalent Academic said...

Great post Nat (and thanks for the love). I really dig when people share their thrill of victory stories...they happen to be so few and far between that it's nice to live through others' vicariously while waiting for your next hit.

Ambivalent Academic said...

Oh go on then, I'll tell you mine:

Deep in the depths of third-year-grad-student-with-not-a-shred-of-data despair, I was trying to make several transgenic mice. In my lab we do the whole process ourselves. Engineering and cloning the transgene construct, purifying the DNA for injection, harvesting embryos, pronuclear injection, embryo transfer, subsequent screening of founders - the whole friggin' thing. It's quite an undertaking and there are a lot of skills to master and I was stuck (turns out later I wasn't, but thought I was due to some bad info from my advisor). I'd been trying and trying and couldn't get any founders. My advisor was breathing down my neck and asking why I couldn't get it done (yeah, thanks due for your *ahem* wrong protocol, which of course I didn't know was wrong at the time) and I was thusly questioning my ability and reasons for being a scientist, and maybe also my reason for being.

I decided to give it one more go and then if it didn't work (again!), admit defeat and reconsider whether I wanted to stick with the PhD. So I did one more round of pronuclear injections with some modification to the protocol, got a few more litters of potential founders, prepped DNA for genotyping, and ran the PCR, fully expecting another big fat nothing.

To my surprise, when I looked at the gel I had five (5!) bands that indicated the mice were carrying the transgene. Oh. My. God.

I have founders! IhavefoundersIhavefoundersIhavefoundersIhavefoundersIhavefounders

Of course I was thrilled and wanted to tell everyone, but of course it was like stupid o'clock (why do these things always happen at night?). So instead I printed out a picture of the gel and scrawled across the top with a Sharpie "oh FUCK yeah!"

Then I made three photocopies: one for the lab notebook, one to hang over my desk, and one to leave on my advisor's desk. They all have the expletive inscription.

Turns out none of the buggers actually expressed the transgene and I took the project in another direction, but that doesn't make that small victory any less sweet.