Ertan Ozyamak
Christien Bowman, MSc, studied psychology during his undergraduate training but realized that he found the quantitative aspects of neuroscience more appealing than the social aspects of his original field. To explore this interest, he started working on pharmacology studies focused on neurological disorders at a pharmaceutical company in Denmark. The industry experience motivated him to identify neuronal mechanisms of brain diseases, leading him to a PhD program in biomedicine at the University of Antwerp. In this interview, we discussed how Bowman uses Chelex 100 Resin as part of a fast and cost-effective method to determine the sex of zebra finch songbirds. This work ultimately helps Bowman study a behavioral paradigm that creates autism-like features in songbirds.
Christien Bowman, MSc is a PhD Researcher at the University of Antwerp in Belgium.
Q: What motivated you to pursue a PhD in neuroscience?
A: During my master’s degree, I collaborated with a pharmaceutical company identifying biomarkers for neurological disorders and successful therapies. During this time, I presented my findings at a neuroscience conference where I met a researcher working on zebra finch songbirds. This was an intriguing possibility for me because at that time I had only seen research performed in rodent models. I also realized that the zebra finch is well-suited to model symptoms of autism spectrum disorder (ASD) in order to hunt for effective therapies. From my time in the field, I was deeply aware that despite decades of research, no comprehensive pharmaceutical therapies for ASD are on the market. Zebra finches have a limited critical period for social vocal learning, similar to the language learning window in humans. How complex a zebra finch’s song is at the end of the learning phase serves as a fixed outcome measure of its experience, like the growth rings in a tree. Because these birds can display social-vocal disorder symptoms, comparable to those seen in severe ASD, I accepted a PhD investigating this model at the University of Antwerp, Belgium.
Q: What projects are you currently working on?
A: I work on a behavioral paradigm that creates autism-like features in songbirds. We separate a juvenile bird early in the learning period from its father who normally teaches the song. Birds that do not learn songs in this critical period produce drastically less complex songs compared to normally-reared adults. At the end of the critical period, we introduce a new male song tutor with or without a pharmaceutical compound to see if we can improve vocal learning. We analyze the similarity of song features between the tutor and the isolated progeny. I developed a program to analyze these features and generated a scoring system to correlate the sound data with MRI-derived metrics that indicate how certain brain structures are organized in songbird brains.
Chelex 100 Resin is a heat-stable bead polymer that captures divalent cations, such as Mg2+ and Ca2+ from solutions. This property allows the inhibition of nucleases in biological samples, preserving RNA or DNA molecules for downstream detection.
Q: How do you distinguish between baby songbirds in your research?
A: DNA sample preparation and sequencing are crucial for that distinction. We isolate male birds for behavior assays because only males sing. The isolation from the father must happen within the first 10 days of birth, otherwise babies would form memories of the father’s songs and learn to sing from them. Therefore, we have to determine whether the bird is a male or female in the first 10 days. Unfortunately, the babies are indistinguishable for the first month because their sex organs are internal. To tell them apart, I perform DNA testing on blood samples. Using traditional DNA extraction kits, it took almost three hours to get DNA and it cost approximately 3.5 euros per sample. Also, one of the most distressing aspects was that it generated a lot of plastic waste. While looking for an eco-friendly option, I came across a simple method based on Bio-Rad’s Chelex 100 Resin. I use Chelex 100 for DNA sample preparation routinely, and it takes only 15 minutes to prepare the DNA and costs only 2 cents per sample. More importantly, the waste is drastically reduced. Because I am the only person on my team running these experiments, I have gained many hours for other tasks because of this product. Overall, it helped me to question the status quo and work on making this protocol efficient, which directly benefitted my PhD project.
DNA Sample Preparation with Chelex 100 Resin
Sample preparation kits are commonly used for purification of DNA. However, as noted by Bowman, these kits are typically costly, time-consuming, and generate large quantities of plastic waste. For many applications, researchers can conserve their resources by simply incubating samples with Chelex 100 Resin instead. This resin captures divalent cations, inhibiting nucleases, producing samples that can be used directly in downstream detection methods.
(Click to enlarge)Comparison of the Chelex 100 Resin–based nucleic acid preparation method and conventional nucleic acid extraction kits. The Chelex 100 Resin workflow requires only four steps: addition of the resin suspension, heat treatment, removal of beads, and use of supernatant in detection applications. In comparison, kit-based methods typically require more steps before downstream analysis.
Discover how to rapidly prepare DNA and RNA samples using Chelex 100 Resin.
Chelex 100 Resin is for research use only.