Faculty Bibliography

Ligand-gated ion channels (ionotropic receptors) link to the cortical cytoskeleton via specialized scaffold proteins and thereby to appropriate signal transduction pathways in the cell. We studied the role of filamentous actin in the regulation of Ca influx through glutamate receptor-activated channels in third-order neurons of salamander retina. Staining by Alexa-Fluor 488-phalloidin, to visualize polymerized actin, we show localization of filamentous actin in neurites, and the membrane surrounding the cell soma. With Ca(2+) imaging we found that in dissociated neurons, depolymerization of filamentous actin by latrunculin A, or cytochalasin D significantly reduced glutamate-induced intracellular Ca(2+) accumulation to 53+/-7% of control value. Jasplakinolide, a stabilizer of filamentous actin, by itself slightly increased the glutamate-induced Ca(2+) signal and completely attenuated the inhibitory effect when applied in combination with actin depolymerizing agents. These results indicate that in salamander retinal neurons the actin cytoskeleton regulates Ca(2+) influx through ionotropic glutamate receptor-activated channels, suggesting regulatory roles for filamentous actin in a number of Ca(2+)-dependent physiological and pathological processes

In this paper, we present a novel method for automatically extracting the tagging sheets in tagged cardiac MR images, and tracking their displacement during the heart cycle, using a tunable 3D Gabor filter bank. Tagged MRI is a non-invasive technique for the study of myocardial deformation. We design the 3D Gabor filter bank based on the geometric characteristics of the tagging sheets. The tunable parameters of the Gabor filter bank are used to adapt to the myocardium deformation. The whole 3D image dataset is convolved with each Gabor filter in the filter bank, in the Fourier domain. Then we impose a set of deformable meshes onto the extracted tagging sheets and track them over time. Dynamic estimation of the filter parameters and the mesh internal smoothness are used to help the tracking. Some very encouraging results are shown

The wasp Nasonia vitripennis is emerging as a useful model organism in which to address a variety of biological questions, due, in part, to its ease of laboratory use, unique aspects of its biology and the sequencing of its genome. In order to take full advantage of the potential of this organism, methods for manipulating gene function are needed. To this end, a protocol for parental RNA interference (pRNAi) in N. vitripennis is described. pRNAi entails injecting pupae with double-stranded RNA, allowing the injected wasps to eclose and examining the progeny for developmental defects. This basic protocol is described in the context of the life cycle of N. vitripennis. This technique has been useful in elucidating the function of most, although not all, genes tested to date, and has potential applications beyond embryonic patterning. pRNAi experiments in Nasonia can be completed in as little as 2 weeks.

Vertebrate gap junction channels are formed by a family of more than 20 connexin proteins. These gap junction proteins are expressed with overlapping cellular and tissue specificity, and coding region mutations can cause human hereditary diseases. Here we present a summary of what has been learned from voltage clamp studies performed on cell pairs either endogenously expressing gap junctions or in which connexins are exogenously expressed. General protocols presented here are currently used to transfect mammalian cells with connexins and to study the biophysical properties of the heterologously expressed connexin channels. Transient transfection is accomplished overnight with maximal expression occurring at about 36 h; stable transfectants normally can be generated within three or four weeks through colony selection. Electrophysiological protocols are presented for analysis of voltage dependence and single-channel conductance of gap junction channels as well as for studies of chemical gating of these channels

In this paper, we describe a new approach for reconstructing 3D strains in the myocardium using tagged MR images. We first segment the myocardium using a 3D deformable model driven by image gradients and Gabor filter responses. Tags are automatically detected and tracked as deformable thin plates during systole and early diastole. To keep the tracking results more stable and consistent, we use a combination of gradient information, an intensity probabilistic model, the phase information, and a temporal-spatial smoothness constraint. Based on the tag deformation, we compute a dense displacement in the myocardium around both ventricles. The displacements in x-, y-, and z- directions are calculated separately and are combined to form the final displacement maps. We do not use the information outside the segmented surface of the myocardium to avoid displacement errors caused by noises, artifacts, and correlations between different regions in the myocardium. The strain in the myocardium during the heart cycle is derived from the displacement. This method accepts images of either a tag grid or separate horizontal and vertical tag lines as its input. Experimental results on phantom and real data demonstrate good performance of this method in calculating the myocardial strain

In this paper we present a robust method for segmenting and tracking cardiac contours and tags in 4D cardiac MRI tagged images via spatio-temporal propagation. Our method is based on two main techniques: the Metamorphs segmentation for robust boundary estimation, and the tunable Gabor filter bank for tagging lines enhancement, removal and myocardium tracking. We have developed a prototype system based on the integration of these two techniques, and achieved efficient, robust segmentation and tracking with minimal human interaction