Samantha Mohammad

Blog #1 The Beginning of My MARC Madness

     Hello there and Happy 4^th of July! So I am finishing up my first two weeks of the UCLA MARC Program and I must admit, I think I love it. I have been working in the Mody lab since November 2011, but I was not too crazy about my project. I have always been really interested in epilepsy research because I have a brother who suffers from severe epilepsy, but they had me working on a completely unrelated project until this summer. I have received a new project where I get to work with mice and study the effects of physical exercise on epilepsy susceptibility. I find my project so interesting, and there are so many different viewpoints to look at. My project seems to be growing with every week, even though last week I was mainly doing background research to make sure I understood my project.

A quick more in depth version of my project:

There is a region known as the dentate gyrus which is one of very few regions within the brain which produces newly born granule cells (GCs). It is a known fact that physical exercise reduces ones susceptibility to epileptic attacks but it is unknown exactly how and why. At the Mody lab we are trying to prove that it is because exercise increases the production of immature GCs. To prove this we will have control mice, and mice that have been running, and compare these to transgenic mice without the ability to produce newly-born GCs. Each mouse will be given a drug to cause epileptogenesis and their seizure levels will be scored and compared. We have added another component to prove that running produces more GCs by the addition of histology to the project to stain for immature GCs. I am genuinely excited about my project and to see how the results pan out.

 

Here is a picture of our PFA hood. I overlooked one of the graduate students as she taught me a fixation perfusion method for histology.

This is the rig I used to work on during my old project which was an electrophysiology project. It was very confusing and very intimidating! (despite the cat above the electrodes.

Blog #2
 
Hey again! So these past few weeks have been really busy. I learned to cut on the cryostat which is not too hard but can get pretty tricky due to temperature changes. For those of you that don’t know, a cryostat is used to cut brain slices for histological analysis on microscopes. I am going to be staining for BrdU in my brain samples. BrdU gets incorporated into the DNA of newly born hippocampal granule cells. So I am staining the brain slices to prove that lower amounts of seizures and lower seizures severity corresponds to the lack of newly born granule cells. DNMT mice are not expected to have any newly born granule cells so this will also be a good way to make sure that this holds true for the DNMT mice we have been using for our study since the stain should not have any success.

Back to my original point. Before I can do any of these stains I had to learn to cut my slices. First I perfused the animals which I got to practice my first 2 weeks.

After that I moved onto the cryostat, which gets tricky due to slight changes in temperature as I said previously. This is because if the cryostat is too cold the slices will fold over onto themselves and will become ruined. If the cryostat is too hot the slices will stick to just about any/everything which makes it increasingly difficult to place them in the antifreeze which is where we keep our slices until we are ready to stain them.

I hope to start staining by the end of the summer but the grad students in my lab have told me that BrdU stains are extremely difficult and since I have no previous experience with staining it may be a while before I get to that point.

Blog #3: Weeks 5-6

                So as the weeks have passed I’ve come to get a much better understanding of what research is really like. It’s a lot of patient waiting and a lot of times things do not go exactly how you wanted them to. My experiment has been put on hold due to a lack of new mice. By that I mean I can’t continue forward with beginning new experiments such as testing the DNMT mice, which will have to wait until Fall quarter starts since the new born DNMT mice will not become adults until sometime late September.  In the meantime, between experiments, I have been doing some old field recordings.

                During the school year when I used to do the field recordings, I used to have my mentor prepare everything, but now that I have my DLAM certification for my animal training completed he wanted me to try my best to complete everything on my own. This entailed making new solutions as well as sacrificing the mice. This is the machine used to prepare the slices, and then they are kept on a water bath until they are ready to use.

 

 I learned to prepare the slices for extracellular field recordings, and I am pretty pleased with how much progress I have made as my slices seem to be much better. Unfortunately there are a lot of other things I needed to learn to do on my own such as making electrodes which is done in the back of our lab where we have a little woodshop-like area. I think it’s a really great experience learning how to actually go about making your own devices, so if ever something breaks or goes wrong I will actually be able to deal with it on my own.

Blog #4

Hey everyone! So as it happens I had forgotten to do some old scoring using the Racine scale which gave me a little more project related things to do. Basically I will watch the mice given Kainate undergo status epilepticus for 4 hours and score them once every 5 minutes scoring them based on their most severe seizure.

MARC also kept me busy analyzing my data making graphs for my results and writing up my abstract. It’s weird to think the school year is right around the corner again, which I am both looking forward to and regretting since I am really afraid of starting my core Neuroscience classes.

On a brighter note, I am half Palestinian, and have observed Ramadan since about 3^rd grade. This whole month of August I have been fasting from sunrise to sunset. I missed a few days but am still proud of how much I actually completed this year, especially on days where I’d work out because no water and working out leads to a very exhausted Samantha. Once the month of fasting is over though, there is a holiday called the Eid. Unfortunately, my parents found a great opportunity to go to Vegas while all of my siblings came home to celebrate so that they could take a much needed vacation while we took care of our autistic/epileptic brother Kimi. A couple of us just decided to go to dinner and we all celebrated at home. 

Alyson Ramirez

Blog #1: 7/2/12

          Hello MARC-ians! (MARC-iotes?  MARC-ers?) Today starts the second week of my summer program at UCLA, and it couldn’t have possibly been better. I was able to enjoy the fantastic weather- a perfect, sunny 75- through the HUGE windows in Terasaki Life Sciences Building, which has become one of my favorite buildings on campus. It’s newly constructed and absolutely gorgeous, and I’m so excited to be able to work in such a state-of-the-art facility this summer.

            And what a summer it’s been so far! I get to continue my research on the development of the central nervous system, and I can tell already that I’ll get so much more accomplished on my project this summer than I would ever be able to during the school year. I’m attempting to induce the lateral motor column motor neuron identity from developing neural progenitor cells by creating a lentivirus that will induce overexpression of Foxp1, a forkhead protein that is important for LMC MN development. Things are going… sort of well at the moment. The beginning stages of developing the myc-tagged Foxp1 virus are going well, and the majority of my cloning is pretty successful. I’m also learning how to differentiate neural progenitor cells from undifferentiated human embryonic stem cells, which is a way more complicated and precise protocol than I ever thought. These cells are just like small children and require so much more care than other cultures I’ve dealt with the past year. I’m a little intimidated, but with the help of graduate students in my lab I’ve been able to do really well so far. I’m having some issues with cloning the HoxC9 insert, as we’re developing a HoxC9 lentivirus as well, but I’ve revisited my negative results and have made some adjustments that will hopefully pan out for the better later this week.

            It’s the fourth of July on Wednesday and I’m really looking forward to experiencing my first fourth in LA- I’ve always been at home or in another country during the fourth, so I’m really excited to spend the day with friends and romp around the city. I’ve recently rented a bike from UCLA’s outdoor adventure center, and it’s been so much fun (and pretty nerve-racking) cycling around campus- LA drivers are a whole ‘nother brand of crazy. It’s so much more efficient than walking though, and you can explore so much more of LA. Summer so far is looking like it’s going to be ridiculous amounts of fun both in and out of lab, and I’m really excited for what the following weeks hold. Until then, my best from the third floor!

Blog #2: 7/19/12

Hola todos!

I cannot believe that it’s already been four weeks! Where did the time go? I’ve accomplished so much in lab but it honestly seems like I’ve been in this program for days, not weeks. Crazy.

            Yesterday was the first time that I honestly understood what it’s going to be like to be in graduate school, and I’m really thankful that I’m decent at time management. Between running around campus for seminars and GRE prep classes, performing experiments in lab, reading scientific journals and editing a bunch of my own papers that needed to be done by the end of the week, I was literally busy from 9 am to 11 pm. Exhausting. 

            But on the bright side, things are going really well! My cell cultures are developing and beginning to differentiate, and this week I was able to make virus. Making virus is an involved protocol that requires exact timing (change of media every 8 to 17 hours), so I’ve been pretty busy this week. I’ve also sectioned and stained chick electroporations and by the end of the week should be able to image them using a confocal microscope. We’re hoping to see if overexpression of Foxp1 in developing chick embryos can lead to a change in motor neuron identity. Cool! Being in lab for such an extended period of time each day allows me to accomplish so much more than during the regular school year, and it’s really exciting how quickly I’ve been able to get results and develop my project.

            We had an awesome seminar last week that I keep thinking about- it was a graduate student panel, in which we got to ask five graduate students pretty much any questions we could think of. It was really cool to see things from a graduate student’s perspective, and to hear more about the actual application process and get tips on what and what not to do. Plus, all of the students (previous MARC students, like me!) were so pumped about everything they were doing that I got so incredibly excited about applying to graduate school. Ph.D. here I come!

Blog #3 8/9/12
 

Sup dudes!

The amount of things I have crammed into the next two weeks is absolutely ridiculous. But it’s also incredibly exciting… because I have data! Woo! Learning what it means and being able to convey this to other people is another challenge, but at least I’ve gotten over the first hurdle.

            The viral constructs that I’ve made are working effectively, and when we infect developing cells (both mouse and human) with the lentivirus (a virus that can infect even non-dividing cells) I’ve constructed through cloning, we see an increase in the expression of our target protein, Foxp1! I still need to fine-tune the details to make sure that the increase of Foxp1 is due to the lentivirus infecting these cells, but so far it looks great. This week will be filled with a lot of antibody staining and imaging to get pictures of all of my infections this summer so I can gather information for both my research paper and poster.

            The only downer is that it will be another three weeks before I can infect my human motor neurons (I’ve done mouse already) with the Foxp1 lentivirus, so I won’t have that data for our end-of-the-summer poster session. I’ve set up some other experiments, including more mouse motor neuron infections and a human cell transfection (in which you simply introduce the DNA of interest, and are not actually infecting cells with virus) to again ensure that introduction of Foxp1 will cause a change in cell phenotype. Hopefully those results will be included on my poster!

            I’ve been wobbling around lab this week because I got the incredible opportunity to hike Half Dome in Yosemite on Monday and my calves haven’t quite recovered. It was one of the most terrifying, thrilling, and exciting things I’ve ever done. 12 hours, 17 miles, countless vicious squirrels, 4800 ft. of elevation gain, and one set of really sketchy cables later we were at the top! It was absolutely gorgeous, and was an experience that I might do again. I think I’ll have to wait a while before I recover physically and mentally. I was floored by the marmots that live up at the summit- they look like beavers with no tails (kinda like a prairie dog) and are so much larger than anything you expect to be living on the top of Half Dome. I have no idea how they survive. I have no idea how I survived. The pictures are just great though. Totally worth it.

            Anyway, back to the lab! See you all in a few weeks for the poster session!

Blog #4
8/31/12

Wow. What. A. Summer. I can’t believe 10 weeks have already gone by! Yesterday was our poster session, and today is the last day of the summer program here at UCLA. I’ll still be in lab for about the next five days finishing stuff up, but oh man. I have gotten so much accomplished in just ten weeks! I can’t believe it!

The poster session yesterday was one of the most simultaneously exciting and terrifying things ever. I was absolutely petrified about it before, because for some reason I have really huge problems with public presentations. I had forced my roommates to sit and let me present to them the night before the poster session (they REALLY loved that), so I was pretty prepared but still had no idea what to expect. It ended up being so much fun! My roommates and a lot of my friends came by, as well as almost every person from my lab- I was so excited to see everyone, and it was really cool to be able to finally explain what I do when I disappear to “lab” everyday. I was pretty overwhelmed with the response I got from my friends, and how proud and excited they were for me. It just sort of put things back into perspective, and make me feel fortunate to have had this experience this summer.

I’ve finished the summer right at the brink of the final part of my research- I’ve spent the past ten weeks making my lentivirus construct, and when I get back from a mini vacation will finally be able to infect my developing human motor neurons to see if we actually get overexpression of our target protein, Foxp1. I know I’ve used the word “excited” about twenty times in this post already but I’m really excited to see what results we get. Excited excited excited.

It’s Labor Day weekend! Woo! It’s finally time to start off my summer, and I’m going hiking, to a baseball game, and of course into lab (just for a little!). Busy weekend ahead and then it’s to the beach to relax before the quarter begins. Hope your summers have been just as great!

 

Jordan Epps

Blog Submission #1
        It's been  26 days since I first landed in NYC. I still can't get over how fast time flies.  It goes without saying that Manhattan is wonderful. I don't think I could ever get tired of that island. I travel to the city often and yet I feel like there's always something new to eplore or see, and even when there isn't there's always a favorite coffee shop, or your favorite spot in central park to go visit. The Bronx on the other hand is a slightly different story. I like to think of it as having all of the negative qualities of Manhattan, but none of the positive ones. The only redeeming qualities are the Bronx Zoo and Botanical Gardens. Luckily, Manhattan is only a 30 minute express bus ride away.
        As for my research, I've been here for three and a half weeks but have only just started actually working on my project. When I first got here I was placed in a computational genetics lab that focuses on RNA-seq data analysis. My P.I. was very friendly, although slightly difficult for me to understand due to a thick asian accent. The work he was having me do was MATLAB programming that could be completed from my computer in my apartment. It was not exactly the lab experience I was looking for. After about a week of trying to force myself to enjoy it, I built up the courage to put in a request to be placed in a new lab. After about a week, Dr. Freedman, the director of SURP, had a new lab in the department of pathology and immunology for me to start working in.
        The lab is led by Dr. Joan Berman and they focus on the effect of HIV on the CNS. More specifically, they are investigating how the virus crosses the blood-brain barrier and the resulting chronic inflammatory response produced by the immune system. My project looks at the effect of buprenorphine, an opioid used clinically as a detoxification and maintenance agent for the treatment of opiate dependence, on human brain microvascular endothelial cells (BMVEC). A portion of HIV infected patients here in the United States are also addicted to heroine and are being treated with buprenorphine. We want to know if the buprenorphine has any effect on the diapedesis of monocytes into the CNS by way of the blood brain barrier and whether or not buprenorhpine interacts with the antiretroviral therapies used to treat the HIV in any way.
        It's unforunate that I won't have more time to spend in this lab than the 4 weeks I have left. In any case I'm looking forward to these last four weeks and I know they are going to be over faster than you can say Albert Einstein College of Medicine Summer Undergraduate Research Program.

 Blog Submission #2
My research here has finally gotten into the full swing now. Last week I learned how to culture human brain microvascular endothelial cells and how to quantify protein levels using a Bradford assay. Additionally, I have learned what it is like to be in a supportive and friendly research environment. Dr. Berman was out last week because of the holiday and other family matters. Instead of having lab meeting on Thursday, which is normally what happens on Thursdays at noon, we all ordered Chinese Food and ate lunch together.  It was great to leave the lab for an hour and spend some time with everyone outside of the research setting. It really is a shame that I've only got three and a half weeks left here. I'm still on my first round of experiments as well; hopefully things will start moving a bit faster once I'm more comfortable with the protocols. Should have my first set of data by the end of this week!
        My life here has also started to feel a little bit more normal. I have finally established my daily and weekly routine. I went to the Union Square Farmer's Market on Saturday which was great. I think I'll be going back there every week for the rest of my stay here. The 4th of July fireworks show in downtown Manhattan was spectacular. Arguably the highlight of last week. This week we're planning to go see Amateur Night at the Apollo Theater in Harlem on Wednesday evening and I just can't wait. Definitely excited for that.