Neurotransmitters are essential for interneuronal signalling, and the specification of appropriate transmitters in differentiating neurons has been related to intrinsic neuronal identity and to extrinsic signalling proteins. Here we show that altering the distinct patterns of Ca2+ spike activity spontaneously generated by different classes of embryonic spinal neurons in vivo changes the transmitter that neurons express without affecting the expression of markers of cell identity. Regulation seems to be homeostatic: suppression of activity leads to an increased number of neurons expressing excitatory transmitters and a decreased number of neurons expressing inhibitory transmitters; the reverse occurs when activity is enhanced. The imposition of specific spike frequencies in vitro does not affect labels of cell identity but again specifies the expression of transmitters that are inappropriate for the markers they express, during an early critical period. The results identify a new role of patterned activity in development of the central nervous system.

Stimulation of transient increases in intracellular calcium (Cai2+) activates protein kinases, regulates transcription and influences motility and morphology. Developing neurons generate spontaneous Cai2+ transients, but their role in directing neuronal differentiation and the way in which they encode information are unknown. Here we image Ca2+ in spinal neurons throughout an extended period of early development, and find that two types of spontaneous events, spikes and waves, are expressed at distinct frequencies. Neuronal differentiation is altered when they are eliminated by preventing Ca2+ influx. Reimposing different frequency patterns of Ca2+ elevation demonstrates that natural spike activity is sufficient to promote normal neurotransmitter expression and channel maturation, whereas wave activity is sufficient to regulate neurite extension. Suppression of spontaneous Ca2+ elevations by BAPTA loaded intracellularly indicates that they are also necessary for differentiation. Ca2+ transients appear to encode information in their frequency, like action potentials, although they are 10(4) times longer in duration and less frequent, and implement an intrinsic development programme.

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Calcium ions play critical roles in neuronal development, but the factors that govern spontaneous fluctuations in intracellular calcium are not well understood.

Approximating detailed models with coarse, texture-mapped meshes results in polygonal silhouettes. To eliminate this artifact, we introduce silhouette clipping, a framework for efficiently clipping the rendering of coarse geometry to the exact silhouette of the original model. The coarse mesh is obtained using progressive hulls, a novel representation with the nesting property required for proper clipping. We describe an improved technique for constructing texture and normal maps over this coarse mesh. Given a perspective view, silhouettes are efficiently extracted from the original mesh using a precomputed search tree. Within the tree, hierarchical culling is achieved using pairs of anchored cones. The extracted silhouette edges are used to set the hardware stencil buffer and alpha buffer, which in turn clip and antialias the rendered coarse geometry. Results demonstrate that silhouette clipping can produce renderings of similar quality to high-resolution meshes in less rendering time.