Analysis of Single Neurons by Perforated Patch Clamp Recordings and MALDI-TOF Mass Spectrometry
| Autor: | Susanne Neupert, Reinhard Predel, Debora Fusca, Peter Kloppenburg |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Patch-Clamp Techniques Physiology Cognitive Neuroscience Population Mass spectrometry Biochemistry Acebutolol Membrane Potentials 03 medical and health sciences Image Processing Computer-Assisted medicine Animals Periplaneta Coloring Agents education Neurons education.field_of_study biology Chemistry Lysine Brain Cell Biology General Medicine biology.organism_classification Immunohistochemistry Matrix-assisted laser desorption/ionization Electrophysiology Electroporation 030104 developmental biology medicine.anatomical_structure nervous system Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Biophysics Neuron Single-Cell Analysis American cockroach Acetylcholine medicine.drug |
| Zdroj: | ACS Chemical Neuroscience. 9:2089-2096 |
| ISSN: | 1948-7193 |
| Popis: | Single-cell mass spectrometry has become an established technique to study specific molecular properties such as the neuropeptide complement of identified neurons. Here, we describe a strategy to characterize, by MALDI-TOF mass spectrometry, neurochemical composition of neurons that were identified by their electrophysiological and neuroanatomical characteristics. The workflow for the first time combined perforated patch clamp recordings with dye loading by electroporation for electrophysiological and neuroanatomical characterization as well as chemical profiling of somata by MALDI-TOF mass spectrometry with subsequent immunohistochemistry. To develop our protocol, we used identified central olfactory neurons from the American cockroach Periplaneta americana. First, the combined approach was optimized using a relative homogeneous, well-characterized neuron population of uniglomerular projection neurons, which show acetylcholine esterase immunoreactivity. The general applicability of this approach was verified on local interneurons, which are a diverse neuron population expressing highly differentiated neuropeptidomes. Thus, this study shows that the newly established protocol is suitable to comprehensively analyze electrophysiological, neuroanatomical, and molecular properties of single neurons. We consider this approach an important step to foster single-cell analysis in a wide variety of neuron types. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|