Ionic currents of Kenyon cells from the mushroom body of the honeybee

The mushroom bodies have been suggested to be essentially involved in learning and memory in insects. In the honeybee Apis mellifera they are composed of about 340,000 intrinsic elements, called Kenyon cells, which can be easily separated from all other neurons of the brain. Here we describe a preparation in which we studied ionic currents in the isolated Kenyon cell somata, using tight-seal whole-cell recording. Several outward and inward currents were identified and investigated by the use of pharmacological agents and in ion substitution experiments: a rapidly inactivating A-type potassium current that is completely blocked with 5 mM 4-aminopyridine; a calcium-activated potassium current that is blocked by 1–100 nM charybdotoxin; a delayed rectifier- type potassium current that is only weakly sensitive to tetraethylammonium but is blocked by 100 microM quinidine; a rapidly activating and inactivating, TTX-sensitive sodium current; a persistent sodium current that is both TTX and cadmium sensitive; and a calcium current that is completely blocked at 50 microM cadmium and is affected by verapamil and nifedipine only at high concentrations (100 microM). The currents described here are very similar to currents found in other insect neurons or muscle cells. This preparation will not only facilitate studies concerning the action of transmitters and neuromodulators that are contained within neurons converging onto the Kenyon cells, but will also allow a study of the role of the adenylyl cyclase pathway, elements of which are expressed in Kenyon cells, and are known to be essential for learning in invertebrates.

[1]  R. Latorre,et al.  K+ -channel blockers restore synaptic plasticity in the neuromuscular junction of dunce, a Drosophila learning and memory mutant , 1992, Proceedings of the Royal Society of London. Series B: Biological Sciences.

[2]  G Laurent,et al.  Dendritic excitability and a voltage-gated calcium current in locust nonspiking local interneurons. , 1993, Journal of neurophysiology.

[3]  Y. Zhong,et al.  Differential modulation of potassium currents by cAMP and its long-term and short-term effects: dunce and rutabaga mutants of Drosophila. , 1993, Journal of neurogenetics.

[4]  Ronald L. Davis,et al.  Preferential expression of the drosophila rutabaga gene in mushroom bodies, neural centers for learning in insects , 1992, Neuron.

[5]  R. Menzel,et al.  Localization of short‐term memory in the brain of the bee, Apis mellifera , 1980 .

[6]  Y. Larmet,et al.  Ionic currents in neurones cultured from embryonic cockroach (Periplaneta americana) brains. , 1988, The Journal of experimental biology.

[7]  S. Kreissl,et al.  Dissociated neurons of the pupal honeybee brain in cell culture , 1992, Journal of neurocytology.

[8]  Roger C. Hardie,et al.  Novel potassium channels encoded by the Shaker locus in drosophila photoreceptors , 1991, Neuron.

[9]  C. Wu,et al.  Properties of potassium currents and their role in membrane excitability in Drosophila larval muscle fibers. , 1990, The Journal of experimental biology.

[10]  U. Müller,et al.  Cyclic nucleotide-dependent protein kinases in the neural tissue of the honeybee Apis mellifera , 1991 .

[11]  H. Breer,et al.  Characterization of calcium-dependent potassium channels in antennal receptor neurones of Locusta migratoria , 1992 .

[12]  C. Malécot,et al.  Ionic Species Involved in the Electrical Activity of Single Adult Aminergic Neurones Isolated from the Sixth Abdominal Ganglion of the Cockroach Periplaneta Americana , 1989 .

[13]  M. Brainard,et al.  Single-channel analysis of four distinct classes of potassium channels in Drosophila muscle , 1988, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[14]  R. Aldrich,et al.  Voltage-gated potassium channels in larval CNS neurons of Drosophila , 1988, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[15]  L. Byerly,et al.  Ionic currents of Drosophila neurons in embryonic cultures , 1988, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[16]  M. Hammer An identified neuron mediates the unconditioned stimulus in associative olfactory learning in honeybees , 1993, Nature.

[17]  Ronald L. Davis,et al.  The cyclic AMP phosphodiesterase encoded by the drosophila dunce gene is concentrated in the mushroom body neuropil , 1991, Neuron.

[18]  R. Wyman,et al.  Ion currents in Drosophila flight muscles , 1983, The Journal of physiology.

[19]  G. Bicker,et al.  Histochemistry of acetylcholinesterase and immunocytochemistry of an acetylcholine receptor‐like antigen in the brain of the honeybee , 1989, The Journal of comparative neurology.

[20]  Ronald L. Davis Mushroom bodies and drosophila learning , 1993, Neuron.

[21]  R C Hardie,et al.  Voltage-sensitive potassium channels in Drosophila photoreceptors , 1991, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[22]  L. Salkoff,et al.  The drosophila shaker gene codes for a distinctive K+ current in a subset of neurons , 1990, Neuron.

[23]  M. Saito,et al.  Expression of ion channels and mutational effects in giant Drosophila neurons differentiated from cell division-arrested embryonic neuroblasts , 1991, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[24]  L. Byerly,et al.  Characterization of single calcium channels in Drosophila embryonic nerve and muscle cells , 1991, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[25]  M. Livingstone,et al.  Loss of calcium/calmodulin responsiveness in adenylate cyclase of rutabaga, a Drosophila learning mutant , 1984, Cell.

[26]  M. Hammer,et al.  Learning and memory in the honeybee , 1995 .

[27]  Y. Jan,et al.  Immunological characterization of K+ channel components from the Shaker locus and differential distribution of splicing variants in drosophila , 1990, Neuron.

[28]  B. Rudy,et al.  Diversity and ubiquity of K channels , 1988, Neuroscience.

[29]  Randolf Menzel,et al.  Chemical codes for the control of behaviour in arthropods , 1989, Nature.

[30]  D. O'Dowd,et al.  Voltage-clamp analysis of sodium channels in wild-type and mutant Drosophila neurons , 1988, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[31]  R. Tsien,et al.  Multiple types of neuronal calcium channels and their selective modulation , 1988, Trends in Neurosciences.

[32]  J. Mauelshagen,et al.  Neural correlates of olfactory learning paradigms in an identified neuron in the honeybee brain. , 1993, Journal of neurophysiology.

[33]  Ronald L. Davis,et al.  Defect in cyclic AMP phosphodiesterase due to the dunce mutation of learning in Drosophila melanogaster , 1981, Nature.

[34]  M. Gho,et al.  Two distinct calcium-activated potassium currents in larval muscle fibres ofDrosophila melanogaster , 1986, Pflügers Archiv.

[35]  P. Mobbs The Brain of the Honeybee Apis Mellifera. I. The Connections and Spatial Organization of the Mushroom Bodies , 1982 .

[36]  S. Schäfer,et al.  Dopamine‐like immunoreactivity in the brain and suboesophageal ganglion of the honeybee , 1989, The Journal of comparative neurology.

[37]  R. Latorre,et al.  A cyclic AMP-activated K+ channel in Drosophila larval muscle is persistently activated in dunce. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[38]  R. Levine,et al.  Calcium and potassium currents in leg motoneurons during postembryonic development in the hawkmoth Manduca sexta. , 1992, The Journal of experimental biology.

[39]  W. Branton,et al.  Spider toxins selectively block calcium currents in drosophila , 1989, Neuron.

[40]  C. Wu,et al.  Voltage clamp analysis of membrane currents in larval muscle fibers of Drosophila: alteration of potassium currents in Shaker mutants , 1985, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[41]  B. Hille Ionic channels of excitable membranes , 2001 .

[42]  A. S. French,et al.  Immunocytochemical localization of sodium channels in an insect central nervous system using a site-directed antibody. , 1993, Journal of neurobiology.

[43]  Satpal Singh,et al.  Complete separation of four potassium currents in drosophila , 1989, Neuron.

[44]  G. Lees,et al.  Calcium channel currents in neurones from locust (Schistocerca gregaria) thoracic ganglia , 1993 .

[45]  Y. Zhong,et al.  Altered synaptic plasticity in Drosophila memory mutants with a defective cyclic AMP cascade. , 1991, Science.