Investigating the role of local axon collaterals of lamina I projection neurons in the spinal dorsal horn
Hungary(HuMSIRC)-University of Debrecen, Debrecen
University of Debrecen, Faculty of Medicine, Department of Anatomy, Histology and Embryology
Peter Szucs, MD, PhD
Peter Szucs, Miklos Sivado, Eva Kokai
4 weeks
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Type of Research Project
- Basic science
What is the background of the project?
Our research group was first to report the existence of local (ipsilateral) axon collaterals from spinal lamina I projection neurons in the superficial dorsal horn of rats. Our recent work is a detailed elaboration of this discovery. The open questions are: what are the targets of the local collaterals? How do they participate in local neuronal networks? What type of modulation do they exert on the target structures? Preliminary data suggest that some of the actions might be via presynaptic inhibition of primary afferent transmitter release.
What is the aim of the project?
To shed light on the postsynaptic contacts and nature of connections from local axon collaterals of projection neurons in the principal nociceptive and sensory relay station, lamina I of the spinal dorsal horn.
What techniques and methods are used?
patch clamp: The patch clamp technique is a laboratory technique in electrophysiology used to study ionic currents in individual isolated living cells, tissue sections, or patches of cell membrane. The technique is especially useful in the study of excitable cells such as neurons retrograde viral labeling : Retrograde tracing is a research method used in neuroscience to trace neural connections from their point of termination, the synapse to their source, the cell body immunocytochemistry: It involves the process of selectively identifying antigens (proteins) in cells of a tissue section by exploiting the principle of antibodies binding specifically to antigens in biological tissues. optogenetics: involves the use of light to control cells in living tissue, typically neurons, that have been genetically modified to express light-sensitive ion channels. Ca-imaging: dynamic imaging of cytosolic Ca2+ concentration in isolated cells or tissue light (visible light passes through the specimen) and electron microscopy (they produce an image of a specimen by using a beam of electrons rather than a beam of light.) 3D neuronal reconstruction: used in neuroscience to determine the pathway of the neurites or neuronal processes, the axons and dendrites, of a neuron neuronal modelling: mathematical description of biophysical models of neuronal cells data analysis: analysis of ionic current s and membrane potential changes by means of specific software (e.g. Axon Calmpfit, Origin, MiniAnalysis, Dempster Electrophysiology package)
What is the role of the student?
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
To learn the basic circuitry of the spinal dorsal horn. Get to know major neuronal populations their markers and basic morphological features. To be able to perform dissection of the spinal cord and brainstem from rodents. To be able to do basic recording solutions, pull recording glass pipettes and if the student shows enthusiasm, to learn to perform simple patch-clamp recordings from intact spinal cord preparations. To be able to follow protocols and carry out immunocytochemistry using multiple labels. To participate in data analysis of ionic current s and membrane potential changes recorded during the lectrophysiology experiments by using specific software (e.g. Axon Calmpfit, Origin, MiniAnalysis, Dempster Electrophysiology package)
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
The tutor will send relevant papers and technical material to the students prior to his/her arrival. These materials can be articles or chapters to read about the project. Lectures, courses and seminars are not provided just small talks or presentation on the topic and daily meetings with the tutor where questions can be asked. Teaching is continuous during the time spent in the laboratory as the student is performing most tasks under supervision when he or she can ask questions from his/her supervisor (laboratory personnel and the tutors).
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- The student will prepare a presentation
- The student’s name will be mentioned in a future publication
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
Knowledge of basic laboratory techniques ( pipetting, preparation of solvents, determining pH, using and adjusting microscopes, basic laboratory calculations etc.) Basic computer skills ( Word, Excel, PowerPoint) Certificate of passed animal handling course in an advantage and preferred but not required. FELASA stands for Federation of European Laboratory Animal Science Associations. The course meets all the mandatory requirements to achieve adequate level of competency on the use of animals in research and education, and to designs and perform animal experiments in the European Union.
Are there any legal limitations in the student’s involvement
Without a valid laboratory animal handling license the student cannot perform certain manipulations of the experimental animals
Type of students accepted
This project accepts:
- Medical students
- Students in biomedical fields
- Szucs P. et al : Axon diversity of lamina I local‐circuit neurons in the lumbar spinal cord. J. Comp. Neurol. 521:2719–2741; 2013.