Abstract

Leech neurons in culture have provided novel insights into the steps in the formation of neurite outgrowth patterns, target recognition and synapse formation. Identified adult neurons from the central nervous system of the leech can be removed individually and plated in culture under well-controlled conditions, where they retain their characteristic physiological properties, grow neurites and form specific chemical or electrical synapses. Different identified neurons develop distinctive outgrowth patterns that depend on their identities and on the molecular composition of the substrate. On native substrates, the patterns displayed by these neurons reproduce characteristics from the adult or the developing neurons. In addition, the substrate may induce selective directed growth between pairs of neurons that normally make contact in the ganglion. Upon contact, pairs of cultured leech neurons form chemical or electrical synapses, or both types depending on the neuronal identities. Anterograde and retrograde signals during membrane contact and synapse formation modify the distribution of synaptic terminals, calcium currents, and responses to 5-hydroxytryptamine.

Highlights

  • During development and regeneration of the nervous system neurons must grow, find their targets and form the correct connections

  • This review focuses on studies with cultured leech neurons, in which many of the steps from neurite outgrowth to synapse formation have been elucidated

  • This similarity between growth of embryonic and adult cultured neurons suggests that an extracellular matrix (ECM) similar to that in the ganglion capsules may provide the initial messages for structuring the outgrowth pattern

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Summary

Introduction

During development and regeneration of the nervous system neurons must grow, find their targets and form the correct connections. Identified neurons from adult leeches can be isolated one by one from the central nervous system (CNS) and kept in culture, where they regenerate neurites and synapses with different targets, allowing the study of these events step by step with single cell resolution [4,5,6,7,8]. After opening the ganglion capsule and incubating with proteolytic enzymes, individual neurons can be removed by applying suction through a glass pipette, sterilized and plated in culture [9]. This procedure permits the experimenter to obtain cells with processes. Cell-to-cell contact and synapse formation induce changes in the distribution of release sites, ion channels and postsynaptic receptors

Sprouting and retracting neurites
Directing growth
Selecting the synaptic attributes
Ganglion capsules
Neurite outgrowth and synapse formation in culture
Conclusions and future directions

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