PUMA-STEM Poster Session

Yesterday, two Slug lab members had their first chance to present their ongoing research projects, premiering their work at the 2022 PUMA-STEM summer research conference:

  • Jaqueline Gutierrez presented work on very long-lasting sensitization in Aplysia
  • Monica Lopez presented work on developing earthworms to study long-term sensitization

It was a great event, and both Jaqueline and Monica did themselves proud–they had really nicely designed posters, presented with confidence, and did a fantastic job fielding question.

Go slug lab!

Summer of ’22 — The SlugLab Roaring Back to Life

It’s July of 2022 and for the past 2 months the SlugLab has been lurching back into life.

For the first time since 2019, the sluglab welcomed a new cohort of summer research students: a record 10 students!

Start of the summer celebration with Tavi, Theresa, Emma, Jas, Zayra, Monica, and Jaquelin (all seated); Christian, Steven, and Hannah.

We’ve so far been confronting the many problems associated with getting the lab back up and running. All previous SlugLab students had graduated, so training had to start from scratch with everything: tank maintenance, siphon-withdrawal reflex measurement, sensitization training, dissections, RNA isolation, qPCR, and data analysis.

Not only has getting everyone up to speed been a challenge, there have also been many challenges to confront from raising up the lab from dormancy. We had challenges with our RNA isolation protocol, an unhealthy batch of animals, a tank that shut down mysteriously over the weekend (and a tank monitoring system that didn’t sound the alarm!), a new file sharing service imposed by the university (which has been terrible), a simulator set up wrong… it’s really been an uphill fight almost every step of the way.

While the list of challenges has been lengthy, it’s turned out to be a lot of fun overcoming them. Our new and large group of sluglab scientists has brought tremendous enthusiasm and camaraderie, a surprisingly deep level of artistic talent, donuts, and a whole lot of fun to the lab. We’ve been knocked down, but we’ve made funny memes about it, and got back up again.

Nothing but great concentrations in the SlugLab!

Maybe we’re also smiling because as we finally seem to have kick started the engine in the lab. We switched to hand homogenization and RNA yields have been amazing. We fixed the stimulator, got healthy animals, and doubled-down on training how to measure behavior, and viola–behavioral data has been pretty fantastic. With data starting to roll in we were finally able to have a lab meeting to work through how to analyze qPCR data, and students have been adding plate after plate of new data for us all to ponder.

At this point, it’s late July and things are really cooking! We have developed and pre-registered (https://osf.io/wvx6z/) an experiment to examine the transcriptional correlates of a very long lasting memory, and it looks like we might end the summer with all behavioral data and tissue collection complete (or at least close to it!). This is an exciting experiment. It’s very clear that forming new long-term memories changes gene expression. What is less clear, though, is if these transcriptional changes are needed to help create the memory, or if they are needed both to create and maintain the memory. Neuroscientists have generally assumed an important role in maintenance, and some models specifically imagine transcriptional feed-back loops that help perpetuate transcription to help maintain memory expression​1​. But this would be a costly way to store a memory. Maybe instead, memories can become transcriptionally independent–perhaps by re-allocating resources within a neuron rather than permanently increasing them.

Our lab has had some hints that transcription might not persist throughout maintenance, at least not for the form of long-term sensitization we study in Aplysia. First, we’ve found that transcriptional changes after sensitization fade within 5 days, 2 days earlier than the memory lasts​2​. This might mean that transcription isn’t needed for maintenance, but it could also mean that there is a slight lag between gene expression decaying and memory expression decaying (2 days isn’t that much of a gap). A second line of evidence is that we’ve found that re-activating a seemingly forgotten memory requires no new changes in gene expression (at least none we could detect), suggesting an uncoupling between memory expression and transcription​3​. This is all suggesting, but not at all definitive.

Now we are collecting data that might help illuminate what role (if any) transcription plays in maintaining a long-term sensitization memory. To do this, we’ve cranked up our training protocol to 11– we are training each animal for 4 consecutive days rather than 1. Work in the Byrne lab​4​ and other labs has suggested that this extended training protocol produces very long-lasting sensitization, and indeed we’re seeing robust behavioral expression 11 days after training (in our typical 1-day training protocol, behavior was almost always back to normal within 7 days). With this longer-lasting training protocol we can examine if transcription also lasts a long time (more than 5 days) or it it still fades quickly. Specifically, we’ll conduct microarray on samples harvested 1 and 5 days after the end of training, and compare the levels of gene regulation at those two time points. If we see that the widespread transcritptional changes at 1 day are still present at day 5, this would suggest a potential role in memory maintenance. However, if we see a decay in transcription at day 5, it would suggest something else is going on…. perhaps transcriptional changes are offset by compensatory mechanisms? Or perhaps memories can be maintained without an ongoing transcriptional change?

At this point we have no idea how the new study will work out… will transcription persist as long as behavior? Will it fade early? We don’t know, but we’re excited to find out. At this point, it looks like we might end the summer with all behavioral data collected and tissue harvested… so it won’t be too much longer now before we have an answer (hopefully).

It’s been a grueling but fantastic summer.

  1. 1.
    Zhang Y, Smolen P, Baxter DA, Byrne JH. The sensitivity of memory consolidation and reconsolidation to inhibitors of protein synthesis and kinases: Computational analysis. Learn Mem. Published online August 24, 2010:428-439. doi:10.1101/lm.1844010
  2. 2.
    Patel U, Perez L, Farrell S, et al. Transcriptional changes before and after forgetting of a long-term sensitization memory in Aplysia californica. Neurobiology of Learning and Memory. Published online November 2018:474-485. doi:10.1016/j.nlm.2018.09.007
  3. 3.
    Rosiles T, Nguyen M, Duron M, et al. Registered Report: Transcriptional Analysis of Savings Memory Suggests Forgetting is Due to Retrieval Failure. eNeuro. Published online September 14, 2020:ENEURO.0313-19.2020. doi:10.1523/eneuro.0313-19.2020
  4. 4.
    Wainwright ML, Byrne JH, Cleary LJ. Dissociation of Morphological and Physiological Changes Associated With Long-Term Memory in Aplysia. Journal of Neurophysiology. Published online October 2004:2628-2632. doi:10.1152/jn.00335.2004

The Sluglab is back!

Today we received a shipment of Aplysia–the first shipment we’ve had since February of 2020.

It’s been a long, frustrating, and anxiety-ridden time for the animal colony to be empty. It’s not that the lab has been inactive–in fact, we published what I think is our best paper ever just a few months ago​1​ . But it has been a long stretch without being able to provide the our typical level of involvement and excitement for our student researchers in the slug lab.

It feels really good to know that we are getting back on track. In fact, in addition to welcoming new slugs we’ve welcomed 5 new lab members: Lucas Eggers, Cynthia Espino, Daniel Mason, Delaney Mcriley, & Steven Proutsos. They join continuing member Melissa Nguyen to round out the Fall 2021 edition of the Sluglab. Let’s kick some a**! (scientifically)

First batch in a long time: Dr. Bob, Dr. C-J, and new lab member Cynthia Espino, October 2021

Our first project with this batch of animals will be to explore for epigenetic markers accompanying long-term sensitization.

Over the last summer, C-J has worked like crazy on protocols for measuring methylation. We’ve found that it is surprisingly easy to full yourself, to obtain signals due to non-specific binding. What we’ve settled on is a process to check specificity of primer sets exhaustively by using synthetic DNA that we can manually methylate. Using this approach we’re pretty sure a key CPG island in the CREB1 promoter is *not* methylated in either control or trained animals. And our summer results also identified a CPG island in the egr promoter that seems to be default methylated, but with no change after sensitization. Our goal with these new animals is to now survey other methylation sites in the promoters of highly learning-regulated transcripts. Having lab meetings back in person has been fantastic (masks, of course, and DU has a vaccine mandate which has been very well implemented); very excited to see where research involvement takes our latest batch of slug lab members.

  1. 1.
    Rosiles T, Nguyen M, Duron M, et al. Registered Report: Transcriptional Analysis of Savings Memory Suggests Forgetting is Due to Retrieval Failure. eNeuro. Published online September 14, 2020:ENEURO.0313-19.2020. doi:10.1523/eneuro.0313-19.2020

SFN 2019 for the Slug Lab

We’ve just wrapped up a great Society for Neuroscience conference for the slug lab. This year’s meeting (2019) was held right here in Chicago, which provided lots of opportunities for our talented crop of students.

We presented a poster examining the time course of forgetting and transcriptional changes at the undergraduate session and at the main meeting. Leading the poster presentation were Tania Rosiles and Melissa Nguyen. After warming up in the undergrad session they were bombarded with tough questions at the main meeting–and they handled themselves amazingly well, doing an awesome job presenting the research. Here they, basking in the knowledge that they had completely crushed it:

Tania Rosiles and Melissa Nguyen presenting that the 2019 Society for Neuroscience Meeting.

I guess they made a big splash, because later at the meeting, guess who asked me for a selfie?

Bob, Nobel-Prize Winner Eric Kandel, and Lisa Gabel. Kandel’s the one in the middle.

Ok – maybe it was me asking Kandel for a selfie, but either way it was cool to briefly meet the godfather of sea slug studies at the meeting.

Sluglab 2019 – It will be an unforgettable summer

It’s summer and the slug lab is rocking. We have 8 students working in the lab (!), and a number of really exciting projects.

Here’s the lab photo to start the summer.

From left to right: Kiara Rana, Dr. C-J, Dr.Bob, Tania Rosiles, George Garcia, Annette Garcia, Hannah Gordon, Lorena Juarez, Monica Duron, and Melissa Nguyen

We knew this spring that we had recruited a special group of students in to the lab. So far the work this summer has confirmed our hunch–we’ve already completed two rounds of behavioral testing, students are making progress learning qPCR, and yesterday we had a great start to learning electrophysiology. I’m sure we’ll have our ups and downs, but it seems like we’re poised for a fun and productive summer.

Projects we’ll be working on include: 1) investigating if savings memories are re-formed or re-covered, 2) investigating the role of the peptide transmitter FMRF-amide in forgetting, 3) exploring the role of methylation in memory maintenance, and 4) some exciting pilot testing with a paradigm for sensitization in fruit fly larva, in collaboration with Scott Kreher in biology.

Our work this summer continues to be supported by the NIH (our current R15 expired at the end of May, but looks like it will be renewed starting July 1). Huzzah.

In addition, Dominican has received a generous donation from Joe Moskal to start the Moskal scholars program. Joe is a professor of biomedical engineering at Northwestern, a biotech entrepreneur, a Dominican trustee, and an all-around amazing guy. He generously helped Irina and me develop pilot data for our first grant and provided a sparkling letter of support… so it is no exaggeration to say he has already helped make the slug lab what it is today.

This year, Joe took the next step in his efforts to develop and broaden the biotech pipeline by funding the Moskal scholars program. Over the next five years this program will fund students interested in careers in the life and health sciences to spend a summer engaged in intensive research. The goal is for students to have the space, mentoring, and encouragement to develop their skills and passions in the science, and to launch them forward to great things.

Our first two Moskal scholars are Annette Garcia and Tania Rosiles. Tania will be spending her second summer in the slug lab–she’s already gained tremendous lab skills and helped co-author our recent paper on the long-term transcriptional response to sensitization ​(Patel et al., 2018)​. Annette is new to the lab, but was a star in Dr. C-J’s neurobiology class and has already been making big strides in the lab.

The inaugural Moskal Scholars: Annette Garcia and Tania Rosiles

Neither Irina nor I would be where we are today if we hadn’t been fortunate enough to have amazing summer experiences. For Irina it was a summer working at Loyola Medical School. For me, it was a summer at Carnegie Mellon. In both cases it was generous funding from sponsors that enabled us to forgo our usual summer jobs and spend 3 months in intense and life-altering contemplation and study. We are so excited and proud to pay that forward each summer with a new batch of slug lab recruits, and we’re extremely grateful to Joe Moskal for his generosity and support.

One of our annual summer traditions is having DU photographer Ryan Pagelow come to the lab for a group photo and some science B-roll. As always, he does an amazing job. Here’s this year’s album:

 
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  1. Patel, U., Perez, L., Farrell, S., Steck, D., Jacob, A., Rosiles, T., … Calin-Jageman, I. E. (2018). Transcriptional changes before and after forgetting of a long-term sensitization memory in Aplysia californica. Neurobiology of Learning and Memory, 474–485. doi: 10.1016/j.nlm.2018.09.007

SlugLab Alum Derek Stek heading to medical school with a full ride scholarship

Graduates of the sluglab have been moving on to amazing careers. So we were excited to get the news that lab alumni and neuroscience major Derek Stek has just been offered a full scholarship to attend medical school at the School of Medicine and Public Health at the University of Wisconsin. Woo hoo!

The slug lab in summer of 2018. Derek is the tall guy on the left-side of the table; this was just before he shipped out to UCLA.

Derek spent the summer of 2017 working in the lab, and also did summer research programs at the University of Colorado (2016) and UCLA (2018). In the sluglab, Derek learned how to do extract RNA (which does *not* start by vortexing the DNA-ase) and conduct qPCR. He helped track the expression of several transcripts regulated after learning, and was a co-author on the lab’s most recent paper (Patel et al., 2018).

Derek was also a star outside of the classroom, playing a big part on the DU varsity basketball team. This year, as he finishes at DU, Derek has been working with children with autism and learning behavioral therapy.

Congrats, Derek!

  1. Patel, U., Perez, L., Farrell, S., Steck, D., Jacob, A., Rosiles, T., … Calin-Jageman, I. E. (2018). Transcriptional changes before and after forgetting of a long-term sensitization memory in Aplysia californica. Neurobiology of Learning and Memory, 474–485. doi:10.1016/j.nlm.2018.09.007

When you forget

Quanta Magazine recently ran a feature article on the nature of forgetting.  The piece covers several new lines of research on forgetting, including the work we’ve been doing with Aplysia.  It’s a great piece, and it’s amazing to see strong public interest in our work.

To Remember, the Brain Must Actively Forget by Dalmeet Singh, Quanta Magazine

Of course, accolades like this would not be possible if it were not for our amazing students.  Here’s the lab enjoying our end-of-summer party.

 

DU Connections and a New Slug Home

The slug lab is kicking off summer research with a brand new aquarium and mixing station for housing Aplysia.  It’s a great upgrade and an even better story that involves some strong DU connections.

The slug lab has been running for 10 years now (gasp!) and our aquariums were showing signs of wear.  Like the wooden stand on one of the tanks starting to collapse.  Pretty strong sign.  (fortunately a student was in the room as the tank started to tip; she called for help, we drained the tank, and disaster was averted).  By fall of 2017 we were down to just 1 tank and worried about how we’d get back to 2 in time for our blitz of summer research.

DU Connection Number 1: In late fall 2017 we were thrilled to receive a McNichols grant from a Dominican Alumni.  This grant was specifically targeted at funding science education at Dominican. We had some meetings and identified several great projects that could be helped by this generous gift–work to improve our greenhouse, a new measurement instrument for the PChem lab, and… a new tank for the slug lab.  Hooray, and a big thanks to the McNichols family.

DU Connection Number 2: We knew we wanted to upgrade beyond a hobbyist tank.. but there were so many options and it was difficult to determine the best path forward.   Enter DU connection #2, Romney Cirillo and his company Something Fishy.  Romney was a business major at Dominican.  When he graduated he wasn’t entirely sure what to do.  But he remembered Al Rosenbloom’s advice to find something he was passionate about and to find a way to make it a life’s work.  Combining that advice with a great entrepreneurship class he had completed, Romney decided to start a business related to his life-long interest in raising exotic fish.  He recruited a great friend as a business partner.  Something Fishy was quickly born as a full-service aquarium design and maintenance company.   Since its founding, Something Fishy has thrived by providing great service and incredible craftsmanship.  Things were going well, so Romney started giving back to DU–installing a gorgeous salt-water tank in Parmer hall.  That’s how I met Romney and got to know Something Fishy.  So, naturally, when it came time for a new tank, Romney was the first person to call.  Thank goodness for these strong DU connections.

The amazing tank.  Romney and Mike visited the slug lab, walked us through options and designed a customized setup exactly suited for our needs:

  • a bigger tank with a better filter system
  • lower to the ground with a huge lid to make experimental access easier (our students say thank you!)
  • incredibly tough and corrosion resistant tank stand (no more collapses from wooden stands)
  • better insulation to make our chillers work less hard
  • and an incredible mixing station for making salt water, complete with a DI line and an auto-shutoff (no more leaks down into the classroom below).

We just finished our first round of experiments with animals housed in the new tank… wow!  Even on the day of shipment the tank water stayed at crystal clear.  Good water -> healthy animals -> a good platform for good science.

Not only is the tank *way* better than what we had, it looks amazing–the install is clean and sharp; it really looks space age.  Best of all, the cost was not astronomical.

So – thanks to two great DU alum we have a great new home for our animals.

Romney (right) showing me the features of the tank on install day.

Here are some photos, but they don’t really do the tank justice for how cool and clean it all looks.

First fillup. Water has just had salt added.

Smiles even on tank cleaning day! The new tank setup is soooo sharp. (Melissa, Ushma, and Leticia… there are slugs in the tank, but all are hiding in the back).

Slug Lab 2018 – Summer Edition

Summer is here, so the Slug Lab is running full speed.

We’re lucky to be joined once again by a really talented and dedicated team of students.  Here’s most of us at a pizza party to inaugurate a summer of research.  

From far left we have

  • Kayla Hall, joining us from Amherst for the summer
  • Ushma Patel, who has graduated but will stay in the lab for the start of the summer (and then on to study biomedical illustration at UIC)
  • Athira Jacob, lab alum now studying for her MCAT
  • Derek Steck, lab alum who this summer will be in the UCLA summer research program this summer
  • Steven Farrell, who has graduated but is also sticking with the lab for part of the summer while prepping for the MCAT
  • Dr C-J and myself (end of the table)
  • Tanya Rosiles, new lab member
  • Everett Krause, new lab member
  • Leticia Perez, lab alum sticking around for part of the summer before heading to UofI for veterinary medicine
  • Melissa Nguyen, new lab member and post-bac student

And not yet in the photo but planning on joining when she returns from Ireland: Shannon Wilcox

I hope I don’t jinx anything when I say that we’re off to a great start, with a very successful round of behavioral experiments and RNA isolations.  Looking forward to another great slug lab summer.