Differential effects of conductances on the phase resetting curve of a bursting neuronal oscillator.

The intrinsically oscillating neurons in the crustacean pyloric circuit have membrane conductances that influence their spontaneous activity patterns and responses to synaptic activity. The relationship between the magnitudes of these membrane conductances and the response of the oscillating neurons to synaptic input has not yet been fully or systematically explored. We examined this relationship using the phase resetting curve (PRC), which summarizes the change in the cycle period of a neuronal oscillator as a function of the input's timing within the oscillation. We first utilized a large database of single-compartment model neurons to determine the effect of individual membrane conductances on PRC shape; we found that the effects vary across conductance space, but on average, the hyperpolarization-activated and leak conductances advance the PRC. We next investigated how membrane conductances affect PRCs of the isolated pacemaker kernel in the pyloric circuit of Cancer borealis by: (1) tabulating PRCs while using dynamic clamp to artificially add varying levels of specific conductances, and (2) tabulating PRCs before and after blocking the endogenous hyperpolarization-activated current. We additionally used a previously described four-compartment model to determine how the location of the hyperpolarization-activated conductance influences that current's effect on the PRC. We report that while dynamic-clamp-injected leak current has much smaller effects on the PRC than suggested by the single-compartment model, an increase in the hyperpolarization-activated conductance both advances and reduces the noisiness of the PRC in the pacemaker kernel of the pyloric circuit in both modeling and experimental studies.

Phase maintenance in a rhythmic motor pattern during temperature changes in vivo.

Central-pattern-generating neural circuits function reliably throughout an animal's life, despite constant molecular turnover and environmental perturbations. Fluctuations in temperature pose a problem to the nervous systems of poikilotherms because their body temperature follows the ambient temperature, thus affecting the temperature-dependent dynamics of various subcellular components that constitute neuronal circuits. In the crustacean stomatogastric nervous system, the pyloric circuit produces a triphasic rhythm comprising the output of the pyloric dilator, lateral pyloric, and pyloric constrictor neurons. In vitro, the phase relationships of these neurons are maintained over a fourfold change in pyloric frequency as temperature increases from 7°C to 23°C. To determine whether these temperature effects are also found in intact crabs, in the presence of sensory feedback and neuromodulator-rich environments, we measured the temperature dependence of the pyloric frequency and phases in vivo by implanting extracellular electrodes into Cancer borealis and Cancer pagurus and shifting tank water temperature from 11°C to 26°C. Pyloric frequency in the intact crab increased significantly with temperature (Q10 = 2-2.5), while pyloric phases were generally conserved. For a subset of the C. borealis experiments, animals were subsequently dissected and the stomatogastric ganglion subjected to a similar temperature ramp in vitro. We found that the maximal frequency attained at high temperatures in vivo is lower than it is under in vitro conditions. Our results demonstrate that, over a wide temperature range, the phases of the pyloric rhythm in vivo are generally preserved, but that the frequency range is more restricted than it is in vitro.

Co-variation of ionic conductances supports phase maintenance in stomatogastric neurons.

Neuronal networks produce reliable functional output throughout the lifespan of an animal despite ceaseless molecular turnover and a constantly changing environment. Central pattern generators, such as those of the crustacean stomatogastric ganglion (STG), are able to robustly maintain their functionality over a wide range of burst periods. Previous experimental work involving extracellular recordings of the pyloric pattern of the STG has demonstrated that as the burst period varies, the inter-neuronal delays are altered proportionally, resulting in burst phases that are roughly invariant. The question whether spike delays within bursts are also proportional to pyloric period has not been explored in detail. The mechanism by which the pyloric neurons accomplish phase maintenance is currently not obvious. Previous studies suggest that the co-regulation of certain ion channel properties may play a role in governing neuronal activity. Here, we observed in long-term recordings of the pyloric rhythm that spike delays can vary proportionally with burst period, so that spike phase is maintained. We then used a conductance-based model neuron to determine whether co-varying ionic membrane conductances results in neural output that emulates the experimentally observed phenomenon of spike phase maintenance. Next, we utilized a model neuron database to determine whether conductance correlations exist in model neuron populations with highly maintained spike phases. We found that co-varying certain conductances, including the sodium and transient calcium conductance pair, causes the model neuron to maintain a specific spike phase pattern. Results indicate a possible relationship between conductance co-regulation and phase maintenance in STG neurons.

Determination of an optimal organ set to implement deformations to support four-dimensional dose calculations in radiation therapy planning.

Surface-based deformable image registration to generate a four-dimensional (4D) dose calculation in radiation treatment planning requires the selection of a set of organ contours to represent a basis set to generate anatomic deformation. The purpose of the present work was to determine the optimal set of organs needed to generate a basis set for deformation in treatment planning for thoracic tumors such that the required computations are minimized but dose accuracy is high. Using retrospectively reviewed records, we calculated 4D dose distributions based on treatment plans for 10 patients with thoracic tumors using a deformable model algorithm in a research version of a commercial radiation treatment planning system. Various combinations of organs (total lungs, heart, spinal cord, external body surface) were used to generate the basis set for deformations used in the calculations. The external surface contour did not have a noticeable effect on the dose calculation. Total lung, heart, and spinal cord together provided an adequate set of deformation organs to generate accurate dose deformations. The magnitude of calculated dose differences had no obvious relationship to tumor parameters, including site, histologic type, disease stage, extent of motion, or degree of centralization.

Biodegradable fumarate-based polyHIPEs as tissue engineering scaffolds.

PolyHIPEs show great promise as tissue engineering scaffolds due to the tremendous control of pore size and interconnectivity afforded by this technique. Highly porous, fully biodegradable scaffolds were prepared by polymerization of the continuous phase of high internal phase emulsions (HIPEs) containing the macromer poly(propylene fumarate) (PPF) and the cross-linker propylene fumarate diacrylate (PFDA). Toluene was used as a diluent to reduce the viscosity of the organic phase to enable HIPE formation. A range of polyHIPE scaffolds of different pore sizes and morphologies were generated by varying the diluent concentration (40-60 wt %), cross-linker concentration (25-75 wt %), and macromer molecular weight ( M n = 800-1000 g/mol). Although some formulations resulted in macroporous monoliths (pore diameter >500 microm), the majority of the polyHIPEs studied were rigid, microporous monoliths with average pore diameters in the range 10-300 microm. Gravimetric analysis confirmed the porosity of the microporous monoliths as 80-89% with most scaffolds above 84%. These studies demonstrate that emulsion templating can be used to generate rigid, biodegradable scaffolds with highly interconnected pores suitable for tissue engineering scaffolds.