title: Target tissues and innervation regulate the characteristics of K+ currents in chick ciliary ganglion neurons developing in situ.
abstract: The expression of appropriate ensembles of ionic channels is necessary for the differentiation and normal function of vertebrate neurons. Cell-cell interactions may regulate the expression and properties of ionic channels in embryonic neurons. Previous studies have shown that the expression of A-type K+ channels (IA) and Ca2+-activated K+ channels (lK[Ca]) is abnormal in chick ciliary ganglion neurons developing in vitro in the absence of normal cell-cell interactions. Other voltage-activated currents develop normally under these conditions. The present studies were designed to establish the role of the target tissues and the preganglionic innervation in regulating the expression of these currents in embryonic chick ciliary ganglion neurons developing in situ. Surgical manipulations were used to remove the developing optic vesicle, which contains the target tissues, the mid-dorsal region of the midbrain primordium, which contains the preganglionic nucleus, or both, all prior to the formation of the ciliary ganglion. IA and IK[Ca] were then examined in acutely isolated neurons that developed in ovo in the presence (OV+) or absence (OV-) of the normal target tissues, in the presence (MB+) or absence (MB-) of preganglionic innervation, and in the absence of both preganglionic innervation and target tissues (OV-/MB-). The amplitude of IA was unaffected by the operations. However, the activation and inactivation kinetics of IA were two- to threefold faster in OV- or OV-/MB- cells compared to neurons isolated from control OV+ ganglia at embryonic days 11-14 (E11-E14). There were no changes in the voltage dependence of activation or steady-state inactivation, or in the time course of recovery from inactivation. By contrast, neurons isolated from MB- ganglia expressed an IA with amplitude, voltage dependence, and kinetics that were indistinguishable from those of control MB+ and OV+ ganglia. Therefore, interactions with target tissues in the eye play a role in determining the characteristics of IA in developing ciliary ganglion neurons, whereas preganglionic innervation does not. Furthermore, the amplitude of IK[Ca] was reduced by 90-100% in OV-, MB-, and OV-/MB- neurons isolated at E12-E14 as compared to MB+ and OV+ controls. Voltage-activated Ca2+ currents were present at normal amplitudes in all of these neurons. Thus, the expression of IK[Ca] in chick ciliary ganglion neurons is regulated by both target tissue interactions and preganglionic innervation. Therefore, cell-cell interactions are necessary for the expression of a normal ensemble of ionic channels in chick ciliary ganglion neurons developing in situ.
authors.affiliation: Department of Biological Science, Florida State University, Tallahassee, 32306-4075.
authors.forename: M M
authors.initials: MM
authors.lastname: Dourado
authors.forename: C
authors.initials: C
authors.lastname: Brumwell
authors.forename: M E
authors.initials: ME
authors.lastname: Wisgirda
authors.forename: M H
authors.initials: MH
authors.lastname: Jacob
authors.forename: S E
authors.initials: SE
authors.lastname: Dryer
mesh_terms: D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002642:Chick Embryo; D003714:Denervation; D004594:Electrophysiology; D005123:Eye / Q000196:embryology; D015353:Eye Enucleation; D005726:Ganglia, Parasympathetic / Q000166:cytology / Q000196:embryology* / Q000378:metabolism; D009424:Nervous System Physiological Phenomena; D009474:Neurons / Q000502:physiology; D015221:Potassium Channels / Q000502:physiology*
chemical_list: D015221:Potassium Channels; D002118:Calcium