Student Research at Duke

How many times have I been asked this question? It seems like the answer never gets any easier. From circus clown to veterinarian, my job aspirations have varied greatly over the years. Now that I’m in college, my plans have changed yet again. The more I learn about the field, the more I come to realize that biology is by far the most fascinating subject I’ve ever studied.

I’ve already eliminated the possibility of a career in medicine, which seems to be the plan of the vast majority of bio majors at Duke – I’m not terribly interested in the clinical applications and small-scale individual problems so much as the big picture. And after having the unbelievable opportunity this summer to participate in the Research Fellows program, I think biological research may be just the thing for me.

In writing my application for the program, I had initially thought I might want to get involved in behavioral studies, hence my job working with the mice. However, while the line between biology and psychology is certainly interesting, after some exposure in the West lab, which deals strictly on a molecular scale, I think I may enjoy that equally as much, if not more. I am hoping in the course of my three years left at Duke to make the most of the time I have and to take advantage of the wide range of research opportunities in search of that one area of biology I will spend my life absorbed in. And if that doesn’t work out I suppose I can always fall back on one of my many previous dream jobs, though hopefully I won’t end up as a clown.

Eight weeks certainly seemed like a long time when I first arrived in Durham, but now at the end of the fifth week, I’m starting to realize just how slow the research process is. We are finishing up our second week of activity boxes today and the results are encouraging. Initially the activity boxes were meant to be used as a means of determining whether the mice were capable of performing in the later mazes; however, after running the number through Excel and making LOTS of graphs, we think there may be a difference in the knockout mice’s activity levels during the day versus at night.

So the next step is going to be… more testing! We’ll be running the mice through what’s called an “elevated plus maze” in which two of the arms of the maze are blocked off by high walls whereas the other two arms are completely open. The idea of the maze is to test for anxiety by measuring how much time is spent in the dark, closed-off arms as opposed to the open arms (which to the mice are pretty scary). We’re hoping to see if maybe the differences in activity level that we noted are due to either an elevated or depressed level of anxiety. In other news, we’re lucky to have a new cohort of mice coming in this next week to add to our numbers. As of now, we have only been working with 23 mice, so it’s been difficult finding any significant differences. With only three weeks left, we’ve still got a lot to do - plus mazes and water mazes for the older mice, as well as running the younger rats through both the SHIRPA screens and activity boxes! Yikes!

The scientific community has an important job: to further knowledge in a direction that ultimately improves the quality of life. But how can it do its job when a small minority of researchers undermine the public’s trust when they become blinded by the possibility of fame and money? This is the question that we, as a group of 30 future scientists, have had to grapple with this past week through a series of lectures, films, and readings on integrity at the lab bench (or mouse hole as the case may be).

One of the cases we read about was that of Eric Poehlman, an eminent scientist tenured at the University of Vermont… until he was sentenced to a year and a day in federal prison for falsifying ten years worth of scientific data. (The article, “An Unwelcome Discovery” by Jeneen Interlandi, was quite an interesting read if you care to look at it.) The magnitude of his crime is astonishing; he not only changed numbers to produce more concrete results, but he had the nerve to entirely fabricate data - in one case, testing only two women when his published study claimed to have tested thirty-five. So how did he get away with it? Well, besides being pretty sneaky in changing his data, there is the problem of who is willing to call him out on it. In this case, it was Walter DeNino, a 24 year-old technician in Poehlman’s lab, who was brave enough to alert university officials to the fact that something just wasn’t adding up right.

Most people in today’s academic world have promised to live by some sort of honor code (Duke, for example, is very serious about upholding academic integrity through the Duke Community Standard.) However, there is a large difference between catching a student using notes on an exam and catching a famous scientist publishing false data. The implications of the latter are tremendous; the ensuing investigation affects everyone: the entire lab of undergrads, grad students, and post-docs, the reputation of the university, and even the public’s faith in the scientific community, not to mention the scientist in question is in jeopardy of losing his career and life as he knows it.

That being said, it is obvious why to bring charges against such a well-known researcher takes not only solid evidence, but takes guts, as well. According to Dwight Matthews, a co-worker of Poehlman’s, “everyone loses,” including DeNino whose “career will be ruined because no one is going to protect [him].” This same thought is echoed by Philip Michael, DeNino’s attorney, saying, “A lot of whistle-blowers are retired. For Walter, this is something that will follow him for the rest of his life.”

The important thing to remember about this is that, like I said in the beginning, the object of science is not merely to learn for the sake of expanding our knowledge, although that is a noble goal in its own right. Science, paired with technology, is what keeps our society advancing; it is what continues to raise the standard of living. The consequences of seeking personal recognition over truth are great indeed. Poehlman’s actions have set scientists back years in the field of menopausal studies, an area of research that is not exactly life-or-death. So you can only imagine how detrimental his actions could have been in a different field. For example, we watched the movie, “And the Band Played On,” a film (that I would highly recommend) about the discovery of AIDS and the first attempts of researchers to deal with the epidemic. You would certainly hope that faced with such a crisis, all resources would be directed towards the prevention of the spread of the virus as well as possible treatment; however, the movie made it apparent that various political and personal issues can greatly hinder the process. In a race with the French to discover the AIDS virus, Dr. Robert Gallo attempted to take more credit than was due to him. Instead of working together to promote the AIDS cause, the ensuing legal battle for both the patent and official recognition for the discovery was a huge impediment in controlling the outbreak before it became an epidemic.

So what did I get from this week of ethical inquiry? I already had a firm handle on the concept that cheating is morally wrong and the consequences tend to outweigh the benefits, but it helped illuminate two ideas for me:

1. Such a lack of integrity can be difficult to identify and put a stop to

and

2. The consequences of putting a researcher’s personal success in front of the ultimate goal of advancing scientific knowledge are ridiculously far-reaching.

In other news, we’ve finished testing the mice in the activity boxes, (large cages that track movement when the mice break the beams of light) and we should be moving onto something more exciting this week - maybe even a maze of some sort. The mice are getting pretty adventurous; not only are they comfortable being picked up and poked and prodded (for the most part) but we have had several escapees this week. Apparently they are getting bored with their surroundings and want to take a look around. Luckily they haven’t gotten very far!

So, like I said earlier, I spent the entire first week getting acquainted with the lab setting. We handled the mice, we toured the different labs, we shadowed various researchers and asked questions, but we didn’t really get started on our project until this past week. (I say we because, lucky for me, I am working with another undergrad, Chase, who is at Duke for the summer in the Mechanisms of Behavior program. And I can honestly say that it is really nice to have someone to work with during those long hours spent in Mouse Room 5.) Anyway, one thing I have found out about research is that it doesn’t work around your schedule. For example, we are testing our mice during both the light and dark cycle in case there is a problem with their circadian rhythms. The light cycle has been modified for us so that we aren’t there past midnight, but it isn’t perfect. The lights go off at three o’clock every day and we give them an hour to adjust to the “night,” the result being that we don’t begin dark testing until four each day. Then there is the fact that we need to test them for three consecutive days as they habituate to the activity boxes, which brings us in to work at nine o’clock on a Saturday morning. At the same time, we do get a lot of down time in between trials and it is during these breaks that I’m able to not only catch up on reading Molecular Biology of the Cell (an incredibly heavy 1616 page book lent to me by Dr. West describing in detail everything I could ever want to know about transcription factors and much, much more!) but I also have time to think about what my ultimate goal is for this summer.

So what question am I trying to answer? Specifically, I want to find out if there is any significant difference between the behavior of the CaRF knockout mice and that of normal mice. But what does that really mean? Well, it means that there is a protein called CaRF that has been found to bind specifically to the DNA sequence of Brain-derived neurotrophic factor. (BDNF is an activity-dependent protein that has many important functions, perhaps the most important of which is promoting brain cell survival). By binding to the BDNF gene, CaRF helps to “turn on” the expression of BDNF. However, if that were the end of the story, we would have noticed a much more drastic change in the knockout mice; in fact, there probably would have been a very high mortality rate if CaRF were the only protein able to promote BDNF transcription. What this implies is that not only is CaRF one of multiple transcription factors, but that in the absence of CaRF, another transcription factor completely takes over its role. This functional substitution is an example of redundancy, a concept crucial to the evolution and survival of species. It can be seen all throughout all forms of life where nature has delegated the same job to two separate systems so that in the event of a failure, homeostatic balance is preserved. Perhaps this is why, although we are hoping to find a significant difference in our mice, it would not be so devastating to find concrete evidence of normality after weeks of testing (although I’m still keeping my fingers crossed for something a bit more exciting!)

I would just like to point out that I did, in fact, post a blog about the interview with my PI, Anne West. Unfortunately, however, it would appear as though I am completely technologically inept because the posting seems to have disappeared without a trace. Just for the record, though, it was a very interesting entry because not only is Dr. West’s research absolutely fascinating, but she also gave me an inside look at what it’s like to become a PI, from grad school to post doc to running your own lab and the responsibilities that come with it.

So I’m back at Duke already after a terrific freshman year for an eight week crash couse on laboratory research. I loved the warm NC weather all school year (at least compared to CT) but I didn’t think I’d ever have to experience the summer here. Although to be perfectly honest, so far the weather outside has been more enjoyable than the practically freezing lab rooms. Lucky for me, however, I get to wear a very attractive and conveniently disposable outfit to visit the mice every day. I’ll let you guess whether or not I’m actually smiling…

So what am I actually doing with the mice, you might ask. Well, nothing really yet. This first week has been dedicated to simply handling the mice so that when we pick them up and place them in a maze, they don’t go berzerk from the anxiety of being touched by those darn nitrile gloves. You might imagine that the mice are cute. Well, they are at first, but after the hundredth time that your hands are used as a bathroom, the cuteness starts to wear off.

Despite my flippant attitude, however, the mice really aren’t that bad, and with any luck, it will be a very rewarding summer with all the tests we have in store for them. The mice are a mixture of wild type, heterozygous, and knockouts for the calcium-responsive transcription factor, CaRF, discovered by Anne West and her lab. From casual observation, there doesn’t seem to be anything different with the mice - they aren’t particularly sickly or mutant-looking, but we’re still hoping to find something with a more detailed analysis. Beginning next week we will be performing the SHIRPA Primary Screen on the animals to check for basic behavioral differences between the normal and knockout mice. The tests range anywhere from basic reflexes to measuring the amount of tears and saliva produced. It looks like it’ll be a fun time. Wish me luck!