In this interview, we had a chat with Dr. Peter Baas to learn more about his team, their research on the neuronal cytoskeleton, and their experiences with the Cellaxess ACE system. Peter heads a research group at the Department of Neurobiology and Anatomy, and is also the Director of the Graduate Program in Neuroscience at Drexel University College of Medicine, Philadelphia.
Hi Peter, tell us a little more about your research.
My research is dedicated to studying the mechanisms that organize microtubule arrays in developing neurons, and the pathological consequences of the disruption of normal microtubule organization during disease. We are looking at various aspects of microtubules during neurodegenerative diseases as well as during nerve injury and regeneration. Most of these studies require transfection of cultured neurons with various DNA constructs and also the introduction of RNAi.
What methods of transfection are you currently using in your lab?
Each type of experiment we do present its own unique challenges and so having a wide array of transfection options is important for us. Neurons, however, are notoriously difficult to transfect, and electroporation is by far the best technique, as lipid-based transfection agents can be toxic.
Our previous electroporation equipment only allowed neurons to be transfected in suspension, prior to plating for culture. However, many of the most important experiments that we would like to conduct demand that we transfect neurons after they have been cultured for several days or even weeks, and so this presented many problems for us. This is the reason why we adopted the Cellaxess ACE system as it is especially designed to transfect adherent cells, allowing us to add this important capability to our cadre of tools.
What would you say are the main benefits of using an in-situ transfection system such as the Cellaxess ACE in your research?
The Cellaxess ACE has opened the door to many new opportunities, as we can now transfect adherent neurons of virtually any type and at any age or configuration in the culture dish, avoiding the potential toxicity of lipid-based transfection techniques.
This has completely changed the way we carry out many of our research projects, for example we have improved co-transfection efficiencies. Delivering both DNA constructs directly from the electrode ensures the majority of the transfected cells express both constructs, and such high probability of co-transfection is not achieved by other methods in our experiments.
As for our RNAi studies, we now have the ability to give cells a “boost” by initially transfecting all the cells prior to plating with our old technique, and then introducing a boost of RNAi into select groups of cells at a later time point using the Cellaxess ACE system. This is a fast and efficient way of maintaining RNAi levels throughout longer experiments. For developmental studies, we can also transfect and then observe a few hours later to document the behaviors of small amounts of newly synthesized proteins.
Where do you see your research leading in the future?
Neuroscience research is essential for driving our understanding of many common and debilitating diseases.
In our research, we are especially excited about being able to conduct studies on much older neuronal cultures with synaptic connections, as they are a good model for diseases that afflict the adult nervous system. We are currently studying Hereditary Spastic Paraplegia, and are eager to be able to transfect cultures of neurons to express the mutated human proteins that give rise to this disease.
Biography
Dr Baas earned his PhD in 1987 from Michigan State University, and then trained as a Postdoctoral Fellow at Temple University. From there, he was on the faculty of the University of Wisconsin for ten years before joining the faculty of Drexel University in 2000. Dr. Baas is interested in all aspects of the neuronal cytoskeleton, with a particular emphasis on the regulation of microtubules in developing neurons. He is also interested in various aspects of microtubules during neurodegenerative diseases as well as during nerve injury and regeneration. He is currently the Director of the Graduate Program in Neuroscience at Drexel University.