Isolation of Sensory Neurons of <em>Aplysia californica</em> for Patch Clamp Recordings of Glutamatergic Currents

Abstract

The marine gastropod mollusk Aplysia californica has a venerable history as a model of nervous system function, with particular significance in studies of learning and memory. The typical preparations for such studies are ones in which the sensory and motoneurons are left intact in a minimally dissected animal, or a technically elaborate neuronal co-culture of individual sensory and motoneurons. Less common is the isolated neuronal preparation in which small clusters of nominally homogeneous neurons are dissociated into single cells in short term culture. Such isolated cells are useful for the biophysical characterization of ion currents using patch clamp techniques, and targeted modulation of these conductances. A protocol for preparing such cultures is described. The protocol takes advantage of the easily identifiable glutamatergic sensory neurons of the pleural and buccal ganglia, and describes their dissociation and minimal maintenance in culture for several days without serum. Video LinkThe video component of this article can be found at

Behavioral aging is associated with reduced sensory neuron excitability in Aplysia californica

“…Primary cultures of PVC and BSC neurons were prepared according to previously reported methods (Fieber, 2000 ; Fieber et al, 2013 ). Cells were isolated from ganglia, dissociated onto 35 × 10 mm polystyrene culture plates coated with poly-D-lysine, and stored at 17°C in a humidified atmosphere.…”

Age-related deficits in synaptic plasticity rescued by activating PKA or PKC in sensory neurons of Aplysia californica

“…Pleural ventral caudal (PVC) SN were isolated from pleural ganglia, dissociated onto 35 × 10 mm polystyrene culture plates coated with poly-D-lysine, and the primary cultures stored at 17°C in a humidified atmosphere as described (Fieber, 2000 ; Fieber et al, 2013 ). Whole-cell voltage clamp recordings were made 24–48 h after plating using 1.5 mm outer diameter borosilicate glass microelectrodes and an Axopatch 200B patch clamp amplifier connected to a PC equipped with a Digidata 1440 A/D converter and pClamp10 software (Molecular Devices, Sunnyvale, CA, USA).…”

Changes in Metabolism and Proteostasis Drive Aging Phenotype in Aplysia californica Sensory Neurons

“…To fill this gap and better understand the behavioral and physiological changes known to occur in this species with age, we performed a transcriptional time series experiment in Aplysia sensory neurons (SN). Specifically, we selected the buccal S (BSC) and pleural ventral caudal (PVC) SN, which comprise a small number of mostly homogeneous neurons (roughly 150 and 200, respectively) that can be reliably identified and sampled between individuals ( Walters et al, 2004 ; Fieber et al, 2013 ). Age-associated declines in excitability of BSC are suggested to contribute to impairments of the biting reflex, likely reflecting the role of BSC in slowed feeding and declines in mass in Aplysia of advanced age ( Fieber et al, 2010 ; Kempsell and Fieber, 2014 ).…”