Neurons differentiating from murine neural crest in culture exhibit sensory or sympathetic-like calcium currents.
The trunk neural crest gives rise to peripheral sensory and sympathetic neurons. In culture, neural crest cells can be induced to differentiate into either neuronal phenotype. Few studies have examined the differentiation of physiological properties in cultures of neural crest cells. Using whole-cell recordings, our study examined the effects of growth factors on high-voltage-activated calcium current profiles exhibited by neurons differentiating in culture. We compared these profiles with those exhibited by sensory and sympathetic neurons. Neural crest cells in culture gave rise to neurons with calcium current profiles identical to either sensory or sympathetic neurons, depending on the growth conditions. On average, the calcium current profile for sensory neurons was 23% (L), 51% (N), and 12% (P), while sympathetic neurons had a similar L-type current (20%), higher N-type (76%), and lower P-type (4%). Neural crest cells cultured with human leukemia inhibitory factor plus somite cells produced neurons with a sympathetic-like calcium current profile (L: 17%, N: 75%, and P: 4%). However, murine leukemia inhibitory factor (L: 25%, N: 52%, and P: 13%) and ciliary neurotrophic factor (L: 18%, N: 49%, and P: 9%) plus somite cells produced neurons with sensory-like calcium current profiles. These growth conditions did not modify the calcium current profiles of neurons cultured from embryonic and neonatal ganglia. Similarly, murine leukemia inhibitory factor produced a greater percentage of neurons (57%) with sensitivity to capsaicin (sensory phenotype) than human leukemia inhibitory factor (3%). Physiological traits can be a useful tool for the determination of neuronal phenotype in culture where other traits may be less stable.
Carey
M B
MB
Department of Biological Structure and Function, Oregon Health Sciences University, Portland 97201, USA.
Matsumoto
S G
SG
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002121:Calcium Channel Blockers / Q000494:pharmacology; D015220:Calcium Channels / Q000032:analysis / Q000502:physiology; D020746:Calcium Channels, L-Type; D020864:Calcium Channels, N-Type; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000032:analysis / Q000096:biosynthesis / Q000502:physiology; D005727:Ganglia, Spinal / Q000166:cytology / Q000196:embryology / Q000502:physiology; D006131:Growth Inhibitors / Q000494:pharmacology; D006719:Homovanillic Acid / Q000378:metabolism; D015850:Interleukin-6; D053605:Leukemia Inhibitory Factor; D008222:Lymphokines / Q000494:pharmacology; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D051379:Mice; D009432:Neural Crest / Q000166:cytology*; D009475:Neurons, Afferent / Q000737:chemistry / Q000166:cytology* / Q000378:metabolism*; D050809:POU Domain Factors; D018408:Patch-Clamp Techniques; D010455:Peptides / Q000494:pharmacology; D020794:Receptor Protein-Tyrosine Kinases / Q000032:analysis / Q000502:physiology; D020801:Receptor, Ciliary Neurotrophic Factor; D017475:Receptors, Nerve Growth Factor / Q000032:analysis / Q000502:physiology; D019170:Somites / Q000166:cytology; D013111:Spider Venoms / Q000494:pharmacology; D017783:Superior Cervical Ganglion / Q000166:cytology / Q000196:embryology / Q000502:physiology*; D050818:Transcription Factor Brn-3; D014157:Transcription Factors / Q000032:analysis / Q000096:biosynthesis / Q000502:physiology; D020867:omega-Agatoxin IVA; D020866:omega-Conotoxin GVIA
D002118:Calcium; D002121:Calcium Channel Blockers; D015220:Calcium Channels; D020746:Calcium Channels, L-Type; D020864:Calcium Channels, N-Type; D004268:DNA-Binding Proteins; D006131:Growth Inhibitors; D006719:Homovanillic Acid; D015850:Interleukin-6; D053605:Leukemia Inhibitory Factor; D008222:Lymphokines; D050809:POU Domain Factors; D010455:Peptides; D020794:Receptor Protein-Tyrosine Kinases; D020801:Receptor, Ciliary Neurotrophic Factor; D017475:Receptors, Nerve Growth Factor; D013111:Spider Venoms; D050818:Transcription Factor Brn-3; D014157:Transcription Factors; D020867:omega-Agatoxin IVA; D020866:omega-Conotoxin GVIA; C507837:LIF protein, human; C507838:Lif protein, mouse