A day in the life of Euan MacDonald Centre PhD student Rachel Kline
Jun 2019: Our PhD student Rachel Kline describes a typical day in the lab, with the highs and lows of researching MND in fruit flies.
Hi! My name is Rachel and I am in the second year of my PhD in Dr Tom Wishart’s lab at the Roslin Institute, University of Edinburgh. I am working on identifying proteins that might play a common role in different types of motor neurone diseases (MNDs).
This is important because, as far as we know, most MNDs aren’t the result of just one specific protein going wrong, and so this makes it difficult to develop bespoke treatments. So far, we have found several proteins that are common to childhood and adult-onset MNDs. We are now investigating them in fruit flies to see if they are able to control whether a nerve cell (like a motor neuron) lives or dies. The ultimate goal of our work is to be able to pick out which of these proteins are most protective in preventing motor neurons from dying. We hope, eventually, the strongest common proteins will form the basis of potential medications that could be used to treat many different forms MNDs.
Why fruit flies?
My PhD involves performing many experiments in fruit flies - yes, those pesky insects that seem to pop up and multiply like crazy if you let your bananas get a little too ripe! While it may not seem obvious, flies have a complete neuromuscular system that is, on a simpler level, a mirror of our own: motor neurons originate from their brain and extend throughout their body until they reach the muscle at a point called the neuromuscular junction. In a healthy fly, the signal from the brain responsible for movement is transmitted to the muscle at these neuromuscular junctions.
Thanks to some cool genetics, we can produce gene mutations in these flies that are the same mutations causing human MNDs like amyotrophic lateral sclerosis (ALS) and spinal muscular atrophy (SMA). When we do this, our flies will develop similar symptoms to humans, from a loss of the neuromuscular junctions and motor neurons when you look under the microscope, to difficulty in crawling (when they are maggots) and flying and climbing (when they are adults). We can therefore consider the humble fruit fly an accurate ‘model’ of MND.
Best of all, because flies multiply so rapidly and are so easy to feed and house (again, think of how quickly they - and their maggot offspring - seem to appear in the food compost bin!), we can perform genetic experiments on literally thousands of individual flies within a year!
A typical day in the lab
Read on for an example of a typical day for me, featuring a couple hundred of these fascinating flies...
6:30am: My alarm goes off at its typical Monday time. Mondays are not always the start of the working week for PhD students though, as many frequently work (whether in the lab or at home on the computer) through the weekend. We don’t work 24/7 of course, nobody can, but the option to “work on your own schedule” often means that I find it hard to switch off at night or on the weekend and sometimes end up firing up the computer late in the evening to play about with some data or make more progress on my written work. Also I often need to go in to the lab at the weekend to look after the flies or finish an experiment - flies haven’t yet grasped the concept of a Saturday lie-in! But as PhD students, we love what we do, which is why we dedicate 3-4 years of our life to studying some very niche topics!
7:00- 8:20am: Bus out to our lab at The Roslin Institute. I often use the time to read a scientific research paper that I have saved on my laptop. Scientists across the world share their latest findings by publishing them in research journals, so it’s essential to stay on top of your reading to be aware of what is happening in the MND field and make sure your own experiments are ‘cutting-edge’!
a gel-like food at the bottom
9:00-12:00pm: Today, I am continuing the fly experiment that I have been working on for the past two months and will continue for the next six (yikes). I collect virgin female ‘MND model’ flies (very important, so that we know that their eggs are unfertilised and will only have the genes of the mother) and hope that I have the 150 flies I need to perform the experiment. I am testing about 100 genes in total, so in this experiment I’ll use 15000 flies!
I then add these females to a tube containing male flies that have one of the potentially protective genes (‘candidates’). The outcome of these matings will be offspring that have both an MND gene and a candidate gene in their DNA, so we can assess whether the candidate protects the nerve cells of the new fly. All this work is done by anaesthetising the flies with carbon dioxide, and they wake up happily in their new environment with fresh food, hopefully in the mood to mate and produce meaningful results for me!
The whole process from start to finish for one experiment takes approximately 3 weeks, and that is with months of generating MND “parent” flies beforehand. Some days go well when the correct combination of genes is achieved, but sometimes it can take weeks and weeks to get just one correct mating performed. Needless to say, it is an arduous process and sometimes incredibly frustrating. However nothing beats the buzz of when I examine the offspring and see that they have fewer dying nerve cells, healthier-looking neuromuscular junctions, and most importantly, show an improved ability to crawl and climb. It’s really exciting that a good proportion of our 100 or so candidates seem to be very important in the protection of nerve cells in multiple MNDs! I hope to get through all of these candidates by Christmas 2019, when it will be time for me to start thinking about writing up my thesis.
12:00-1:00pm: Catch-up meeting with my supervisor and lab team. At the moment, the Wishart Lab is a small team, but we are all working on very diverse projects. For this reason, it’s important to take some time to sit down and go over what everyone has been doing; often we end up bouncing ideas off each other or getting some interesting insight on puzzling results.
1:00-2:15pm: After a break for lunch and coffee, I answer some emails. The results from my morning experiment will not be ready until after 2pm, so I take this opportunity to read a good research paper from a lab in the USA. This paper provides evidence that an important ALS-linked gene called TDP-43 influences a second gene responsible for maintaining the structure and shape of the motor neuron.
2:15-5:00pm: Experiment time, part 2. Some of my offspring flies from the previous week’s matings are ready to be analysed for results, and so I begin the long process of anaesthetising the flies again and collecting data. At the same time, I begin to prepare for an experiment in which I look at the neuromuscular junctions of flies at the larval stage (I try to call them this instead of what we all know them as, i.e. maggots!). I like to let the afternoon coffee wear off before doing the necessary dissections, as a steady hand is necessary when you are dissecting out the muscles of 1cm-long fruit fly larvae. For this reason, it's almost 4pm before I start these and get them into the staining chemicals required to see their neuromuscular junctions under a microscope.
junctions, with proteins stained in different colours
5:30-6:00pm- I set up the experiments for tomorrow morning and write up today’s results in my lab book. Today was a particularly interesting day as my results seem to suggest that a couple of my candidates have a very powerful influence on the nerve cell of an MND fly, but don’t seem to do anything in a normal, healthy nerve cell. This is particularly exciting from the perspective of developing future therapies, as we ideally want to only treat the affected cells without any side-effects on healthy ones.
for the Euan MacDonald Centre in 2018
6:00-7:00pm- I go for a run in the Pentlands and muse over these new results. I’ve been running since I was 11 years old, and I love it as it not only energises my body, but also my mind! I find that I often come up with my best ideas for experiments while out running and just letting my mind wander.
8:30-10:30pm- I do not work every evening, and strongly believe in something a colleague once said to me regarding longevity (by taking care of yourself and taking breaks) being the most valuable thing you could do to contribute to the field for the rest of your life. However, this evening, I am under a short deadline to submit my own research paper. Just as it is important to keep on top of the field in terms of knowledge, we need to contribute to the literature as well. I am very keen to get this paper off my desk and off to the journal, from where it will be assessed by peers around the world, who will decide if the experiments have been done properly and the findings are ready to publish.
10:30-11:30pm- Relaxation and winding down. I’m from the USA originally and so I sometimes use this time to catch up with family and friends in their time zone, or else, watch a movie or TV before bed.