PubMed Health⌕ Search

Biomedical subjects

Nicholas C Spitzer

Publications and source records attributed to Nicholas C Spitzer.

14 recordsLinked to original sources
PubMed2006

Activity-dependent neurotransmitter-receptor matching at the neuromuscular junction.

Signaling in the nervous system requires matching of neurotransmitter receptors with cognate neurotransmitters at synapses. The vertebrate neuromuscular junction is the best studied cholinergic synapse, but the mechanisms by which acetylcholine is matched with acetylcholine receptors are not fully understood. Because alterations in neuronal calcium spike activity alter transmitter specification in embryonic spinal neurons, we hypothesized that receptor expression in postsynaptic cells follows changes in transmitter expression to achieve this specific match. We find that embryonic vertebrate striated muscle cells normally express receptors for glutamate, GABA, and glycine as well as for acetylcholine. As maturation progresses, acetylcholine receptor expression prevails. Receptor selection is altered when early neuronal calcium-dependent activity is perturbed, and remaining receptor populations parallel changes in transmitter phenotype. In these cases, glutamatergic, GABAergic, and glycinergic synaptic currents are recorded from muscle cells, demonstrating that activity regulates matching of transmitters and their receptors in the assembly of functional synapses.

Animals↗
PubMed2006

Electrical activity in early neuronal development.

The construction of the brain during embryonic development was thought to be largely independent of its electrical activity. In this view, proliferation, migration and differentiation of neurons are driven entirely by genetic programs and activity is important only at later stages in refinement of connections. However, recent findings demonstrate that activity plays essential roles in early development of the nervous system. Activity has similar roles in the incorporation of newly born neurons in the adult nervous system, suggesting that there are general rules underlying activity-dependent development. The extensive involvement of activity makes it likely that it is required at all developmental stages as a necessary partner with genetic programs.

Animals↗
PubMed2006

Second messenger pas de deux: the coordinated dance between calcium and cAMP.

Dynamic calcium signaling is a well-established precept in biology. Different cell types exhibit spontaneous as well as stimulus-triggered transient changes in the concentration of intracellular calcium. Does this behavior extend to other second messengers? Optical dissection of various signal transduction pathways with fluorescent reporter molecules that enable visualization of changes in concentration of other second messengers is well under way. Recent research using technologically refined probes provides improved temporal and spatial resolution of adenosine 3',5'-monophosphate (cAMP) dynamics to generate insights into the bidirectional interplay between intracellular fluctuations of cAMP and calcium. cAMP oscillations are generated in response to hormones, and cells can recognize and differentially respond to transient versus sustained changes in this second messenger. Second messenger reporters are now available to track multiple players and so provide a dynamic picture of signaling networks.

Adenylyl Cyclases↗
PubMed2005

Local calcium transients contribute to disappearance of pFAK, focal complex removal and deadhesion of neuronal growth cones and fibroblasts.

Cell adhesion is crucial for migration of cells during development, and cell-substrate adhesion of motile cells is accomplished through the formation and removal of focal complexes that are sites of cell-substrate contact. Because Ca2+ signaling regulates the rate of axon outgrowth and growth cone turning, we investigated the potential role of Ca2+ in focal complex dynamics. We describe a novel class of localized, spontaneous transient elevations of cytosolic Ca2+ observed both in Xenopus neuronal growth cones and fibroblasts that are 2-6 mum in spatial extent and 2-4 s in duration. They are distributed throughout growth cone lamellipodia and at the periphery of fibroblast pseudopodia, which are regions of high motility. In both cell types, these Ca2+ transients lead to disappearance of phosphorylated focal adhesion kinase (pFAK) and deadhesion from the substrate as assessed by confocal and internal reflection microscopy, respectively. The loss of pFAK is inhibited by cyclosporin A, suggesting that these Ca2+ transients exert their effects via calcineurin. These results identify an intrinsic mechanism for local cell detachment that may be modulated by agents that regulate motility.

Animals↗
PubMed2005

Homeostatic activity-dependent paradigm for neurotransmitter specification.

Calcium-signaling plays a central role in specification of the chemical transmitters neurons express, adjusting the numbers of cells that express excitatory and inhibitory transmitters as if to achieve homeostatic regulation of excitability. Here we review the extent to which this activity-dependent regulation is observed for a range of different transmitters. Strikingly the homeostatic paradigm is observed both for classical and for peptide transmitters and in mature as well as in embryonic nervous systems. Transmitter homeostasis adds another dimension to homeostatic regulation of function in the nervous system that includes regulation of levels of voltage-gated ion channels, densities of neurotransmitter receptors, and synapse numbers and strength.

Animals↗
PubMed2004

Activity-dependent homeostatic specification of transmitter expression in embryonic neurons.

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.

Action Potentials↗
PubMed2004

Calcium transients regulate patterned actin assembly during myofibrillogenesis.

The highly ordered arrangement of sarcomeric myosin during striated muscle development requires spontaneous calcium (Ca(2+)) transients. Here, we show that blocking transients also compromises patterned assembly of actin thin filaments, titin, and capZ. Because a conserved temporal assembly pattern has been described for these proteins, selective inhibitors of either thick or thin filament formation were used to determine their relative temporal interdependencies. For example, inhibition of myosin light chain kinase (MLCK) by application of a specific inhibitory peptide or phorbol myistate acetate (PMA) disrupts myosin assembly without significantly affecting formation of actin bands. The MLCK inhibitor ML-7, however, disrupted actin as well as myosin. Surprisingly, agents that interfere with actin dynamics, such as cytochalasin D, produced only minor organizational disruptions in actin, capZ, and titin staining. However, cytochalasin D and other actin disrupting compounds significantly perturbed myosin organization. The results indicate that (1) Ca(2+) transients regulate one or more of the earliest steps in sarcomere formation, (2) mature actin filaments can assemble independently of myosin band formation, and (3) myosin thick filament assembly is extremely sensitive to disruption of either the actin or titin filament systems.

Actin Cytoskeleton↗
PubMed2004

Orchestrating neuronal differentiation: patterns of Ca2+ spikes specify transmitter choice.

Appropriate specification of neurotransmitters is a key feature of neuronal network assembly. There is much evidence that genetic programs are responsible for this aspect of cell fate and neuronal differentiation. Are there additional ways in which these processes are shaped? Recent findings demonstrate that altering patterned Ca(2+) spike activity that is spontaneously generated by different classes of embryonic spinal neurons in vivo changes expression of neurotransmitters in a homeostatic manner, as if to achieve a constant level of excitation. Activity-dependent changes in presynaptic transmitter expression pose a matching problem: are there corresponding changes in postsynaptic transmitter receptor expression, or are axons rerouted to novel targets with which functional synapses can be formed?

Action Potentials↗
PubMed2002

Dynamic interactions of cyclic AMP transients and spontaneous Ca(2+) spikes.

Transient increases of intracellular Ca(2+) drive many cellular processes, ranging from membrane channel kinetics to transcriptional regulation, and links of Ca(2+) to other second messengers should activate signalling networks. However, real-time kinetic interactions have been difficult to investigate. Here we report observations of spontaneous increases in concentration of cyclic AMP (cAMP) in embryonic spinal neurons, and their dynamic interactions with Ca(2+) oscillations. Blocking the production of these cAMP transients decreases the intrinsic frequency of spontaneous Ca(2+) spikes, whereas inducing cAMP increases causes spike frequency to increase. Transients of cAMP in turn are absent when Ca(2+) spikes are blocked, and are generated only in response to specific patterns of stimulated spikes that mimic endogenous Ca(2+) kinetics. We present a mathematical model of Ca(2+)-cAMP reciprocity that generates the slow cAMP oscillations and reproduces the dynamics of Ca(2+)-cAMP interactions observed experimentally. The model predicts that this module of coupled second messengers is tuned to optimize production of cAMP transients, and that simultaneous stimulation of Ca(2+) and cAMP systems produces distinct temporal patterns of oscillations of both messengers. Our findings may prove useful in the investigation of the regulation of gene expression by second-messenger transients.

Animals↗
PubMed2002

Adaptation in the chemotactic guidance of nerve growth cones.

Pathfinding by growing axons in the developing nervous system may be guided by gradients of extracellular guidance factors. Analogous to the process of chemotaxis in microorganisms, we found that axonal growth cones of cultured Xenopus spinal neurons exhibit adaptation during chemotactic migration, undergoing consecutive phases of desensitization and resensitization in the presence of increasing basal concentrations of the guidance factor netrin-1 or brain-derived neurotrophic factor. The desensitization is specific to the guidance factor and is accompanied by a reduction of Ca2+ signalling, whereas resensitization requires activation of mitogen-associated protein kinase and local protein synthesis. Such adaptive behaviour allows the growth cone to re-adjust its sensitivity over a wide range of concentrations of the guidance factor, an essential feature for long-range chemotaxis.

Adaptation, Biological↗
PubMed2002

Outside and in: development of neuronal excitability.

Investigation of the development of excitability has revealed that cells are often specialized at early stages to generate Ca(2+) transients. Studies of excitability have converged on the central role of Ca(2+) and K(+) channels in the plasmalemma that regulate Ca(2+) influx and have identified critical functions for receptor-activated channels in the endoplasmic reticulum that allow efflux of Ca(2+) from intracellular stores. The parallels between excitability in these two locations motivate future work, because comparison of their properties identifies shared attributes.

Animals↗
PubMedDate not supplied

Activity-dependent neuronal differentiation prior to synapse formation: the functions of calcium transients.

Spinal cord neurons become excitable prior to synapse formation, and generate spontaneous calcium transients that regulate aspects of their differentiation before neuronal networks are established. Calcium spikes, generated by calcium-dependent action potentials and calcium-induced calcium release (CICR), regulate transcription. Growth cone calcium transients, produced by calcium influx through unidentified channels that triggers CICR, control the rate of axon outgrowth in response to environmental cues. Filopodial calcium transients, generated by calcium influx through channels activated by beta1 integrins, signal information about the molecular identity of the substrate and regulate growth cone turning. All three classes of calcium transients appear to use a frequency code to implement their effects. Oscillations of second messengers in embryonic neurons and perhaps more generally in other differentiating cells may behave like a kinetic quilt, demonstrating patchy fluctuations in concentrations that orchestrate the complex processes of development.

Action Potentials↗