Patch clamp electrophysiology in nodose ganglia of adult rat.
The patch clamp technique is widely utilized for studying the electrophysiological properties of enzymatically isolated sensory neurons. Unfortunately, dissociation of the ganglion severs the afferent fibers. As a result, isolated neurons can only be broadly categorized according to somatic action potential waveforms, ion channel subtypes, chemical sensitivities and cell diameter. Such restricted classifications contrast with the continuum of conduction velocities (CVs), discharge patterns, sensory modalities and functional properties of visceral and spinal afferents. Previous reports of patch clamp recordings using intact ganglion have been limited to young animal preparations. This raises concerns regarding postnatal development and impedes the use of chronic models of disease or injury, which often necessitate the use of a more mature animal preparation. Here, we present a methodology for preparing nodose ganglion from adult rat (250-400 g) for study using the patch clamp technique. Successful whole cell recordings were obtained from approximately 50% of the cells selected for study, the majority of which had intact afferent fibers. Measures of somatic discharge and afferent fiber CV at both room and physiological temperatures were consistent with investigations using sharp microelectrodes. Voltage clamp recordings of whole cell Na(+), Ca(2+) and K(+) ion channel currents were comparable to those obtained using isolated neuron preparations. The ability to classify voltage- and ligand-gated ion channel type with afferent fiber CV in an adult preparation adds a valuable new dimension to cellular investigations of the diverse functional and chemical properties of the peripheral afferent nervous system.
Li
B Y
BY
Biomedical Engineering Program, Purdue School of Engineering and Technology, Indiana University, Room SL 174, 723 West Michigan Street, Indianapolis, IN 46202, USA.
Schild
J H
JH
D000818:Animals; D015220:Calcium Channels / Q000502:physiology; D005260:Female; D018698:Glutamic Acid / Q000494:pharmacology; D008297:Male; D009475:Neurons, Afferent / Q000187:drug effects / Q000502:physiology; D009620:Nodose Ganglion / Q000166:cytology / Q000502:physiology*; D018408:Patch-Clamp Techniques; D015221:Potassium Channels / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley; D018094:Receptors, Metabotropic Glutamate / Q000187:drug effects; D012701:Serotonin / Q000494:pharmacology; D015222:Sodium Channels / Q000502:physiology
D015220:Calcium Channels; D015221:Potassium Channels; D018094:Receptors, Metabotropic Glutamate; D015222:Sodium Channels; D012701:Serotonin; D018698:Glutamic Acid