Linear signaling in the Toll-Dorsal pathway of Drosophila: activated Pelle kinase specifies all threshold outputs of gene expression while the bHLH protein Twist specifies a subset

Differential activation of the Toll receptor leads to the formation of a broad Dorsal nuclear gradient that specifies at least three patterning thresholds of gene activity along the dorsoventral axis of precellular embryos. We investigate the activities of the Pelle kinase and Twist basic helix-loop-helix (bHLH) transcription factor in transducing Toll signaling. Pelle functions downstream of Toll to release Dorsal from the Cactus inhibitor. Twist is an immediate-early gene that is activated upon entry of Dorsal into nuclei. Transgenes misexpressing Pelle and Twist were introduced into different mutant backgrounds and the patterning activities were visualized using various target genes that respond to different thresholds of Toll-Dorsal signaling. These studies suggest that an anteroposterior gradient of Pelle kinase activity is sufficient to generate all known Toll-Dorsal patterning thresholds and that Twist can function as a gradient morphogen to establish at least two distinct dorsoventral patterning thresholds. We discuss how the Dorsal gradient system can be modified during metazoan evolution and conclude that Dorsal-Twist interactions are distinct from the interplay between Bicoid and Hunchback, which pattern the anteroposterior axis

Lateral Gene Expression in Drosophila Early Embryos Is Supported by Grainyhead-Mediated Activation and Tiers of Dorsally-Localized Repression

The general consensus in the field is that limiting amounts of the transcription factor Dorsal establish dorsal boundaries of genes expressed along the dorsal-ventral (DV) axis of early Drosophila embryos, while repressors establish ventral boundaries. Yet recent studies have provided evidence that repressors act to specify the dorsal boundary of intermediate neuroblasts defective (ind), a gene expressed in a stripe along the DV axis in lateral regions of the embryo. Here we show that a short 12 base pair sequence (“the A-box”) present twice within the ind CRM is both necessary and sufficient to support transcriptional repression in dorsal regions of embryos. To identify binding factors, we conducted affinity chromatography using the A-box element and found a number of DNA-binding proteins and chromatin-associated factors using mass spectroscopy. Only Grainyhead (Grh), a CP2 transcription factor with a unique DNA-binding domain, was found to bind the A-box sequence. Our results suggest that Grh acts as an activator to support expression of ind, which was surprising as we identified this factor using an element that mediates dorsally-localized repression. Grh and Dorsal both contribute to ind transcriptional activation. However, another recent study found that the repressor Capicua (Cic) also binds to the A-box sequence. While Cic was not identified through our A-box affinity chromatography, utilization of the same site, the A-box, by both factors Grh (activator) and Cic (repressor) may also support a “switch-like” response that helps to sharpen the ind dorsal boundary. Furthermore, our results also demonstrate that TGF-β signaling acts to refine ind CRM expression in an A-box independent manner in dorsal-most regions, suggesting that tiers of repression act in dorsal regions of the embryo

Effects of electrode configuration and geometry on fiber preference in spinal cord stimulation

In contrast to the widespread assumption that dorsal column fibers are the primary targets of spinal cord stimulation by a dorsal epidural electrode, it appears that dorsal root fibers are recruited as well, and even preferentially under various conditions. This will, however, limit the coverage of the painful body areas with paresthesia, a prerequisite for the management of chronic pain. In order to favor the preferential stimulation of dorsal column fibers, advantage was taken of the different positions and orientations of fibers in the dorsal columns and dorsal roots. Using an SCS computer model, electrode configurations have been designed for the selective stimulation of the human dorsal column

Paresthesia thresholds in spinal cord stimulation: a comparison of theoretical results with clinical data

The potential distributions produced in the spinal cord and surrounding tissues by dorsal epidural stimulation at the midcervical, midthoracic, and low thoracic levels were calculated with the use of a volume conductor model. Stimulus thresholds of myelinated dorsal column fibers and dorsal root fibers were calculated at each level in models in which the thickness of the dorsal cerebrospinal fluid (CSF) layer was varied. Calculated stimulus thresholds were compared with paresthesia thresholds obtained from measurements at the corresponding spinal levels in patients. The influences of the CSF layer thickness, the contact separation in bipolar stimulation and the laterality of the electrodes on the calculated thresholds were in general agreement with the clinical dat

Ultrasonographic identification of the dorsal atlantoaxial ligament in dogs

Objective: To determine the ability to identify the dorsal atlantoaxial ligament during ultrasonographic examination of dogs. Study design: Canine cadaveric study. Sample population: Canine cervical spines (n = 35). Methods: Thirty-five canine cadavers with an estimated body weight of 6-35 kg were retrieved. Five cervical spines were dissected to assess the length and thickness of the dorsal atlantoaxial ligament and correlate ultrasound-guided injection of ink with anatomical dissection. Ultrasonographic evaluation of the dorsal atlantoaxial ligament was performed on 30 cadavers. The visibility of the dorsal atlantoaxial ligament in both the transverse and the sagittal planes was subjectively scored (0-3). Results: The dorsal atlantoaxial ligament was detectable on ultrasonographic examination in all cadavers: 27/30 and 28/30 were graded as moderately visible (grade 2) or clearly visible (grade 3) in the sagittal and transverse view, respectively. Only 1/30 cadaver specimen of a large breed dog was graded as 1 (indistinct) in both the sagittal and transverse planes. None of the cadavers were graded as 0 (not visible) in any view. Conclusion: The dorsal atlantoaxial ligament can be identified in ultrasonographic examination of normal canine cadavers. Future studies on animals with clinical atlantoaxial instability/subluxation are needed to evaluate the role of this imaging technique in providing a safer diagnosis of this condition. Clinical relevance: Ultrasonographic examination of the dorsal atlantoaxial ligament could potentially provide a noninvasive and safe imaging technique that may contribute to the diagnosis of atlantoaxial instability/subluxation in dogs

Notochord grafts do not suppress formation of neural crest cells or commissural neurons

Grafting experiments previously have established that the notochord affects dorsoventral polarity of the neural tube by inducing the formation of ventral structures such as motor neurons and the floor plate. Here, we examine if the notochord inhibits formation of dorsal structures by grafting a notochord within or adjacent to the dorsal neural tube prior to or shortly after tube closure. In all cases, neural crest cells emigrated from the neural tube adjacent to the ectopic notochord. When analyzed at stages after ganglion formation, the dorsal root ganglia appeared reduced in size and shifted in position in embryos receiving grafts. Another dorsal cell type, commissural neurons, identified by CRABP and neurofilament immunoreactivity, differentiated in the vicinity of the ectopic notochord. Numerous neuronal cell bodies and axonal processes were observed within the induced, but not endogenous, floor plate 1 to 2 days after implantation but appeared to be cleared with time. These results suggest that dorsally implanted notochords cannot prevent the formation of neural crest cells or commissural neurons, but can alter the size and position of neural crest-derived dorsal root ganglia

Surgical techniques in substitution urethroplasty using buccal mucosa for the treatment of anterior urethral strictures

OBJECTIVES: Since the resurgence in the use of buccal mucosa (BM) in substitution urethroplasty in the late 1980s and early 1990s, there has been controversy as to which surgical technique is the most appropriate for its application. METHODS: The authors performed an updated literature review. Several centres have published widely on this topic, and the points considered include the use BM in dorsal onlay grafts, ventral onlay grafts, and tubularised grafts and the role of two-stage procedures. RESULTS: In experienced hands, the outcomes of both dorsal onlay grafts and ventral onlay grafts in bulbar urethroplasty are similar. The dorsal onlay technique is, however, possibly less dependent on surgical expertise and therefore more suitable for surgeons new to the practice of urethroplasty. The complications associated with ventral onlay techniques can be minimised by meticulous surgical technique, but in series with longer follow-up, complications still tend to be more prevalent. In penile urethroplasty, two-stage dorsal onlay of BM (after complete excision of the scarred urethra) still provides the best results, although in certain circumstances a one-stage dorsal onlay procedure is possible. In general, ventral onlay of BM and tube graft procedures in the management of penile strictures are associated with much higher rates of recurrence and should therefore be avoided. CONCLUSIONS: In experienced hands the results of the ventral and dorsal onlay of BM for bulbar urethroplasty are equivalent. Two-stage procedures are preferable in the penile urethra, except under certain circumstances when a one-stage dorsal onlay is feasible

Human Dorsal Striatal Activity during Choice Discriminates Reinforcement Learning Behavior from the Gambler’s Fallacy

Reinforcement learning theory has generated substantial interest in neurobiology, particularly because of the resemblance between phasic dopamine and reward prediction errors. Actor–critic theories have been adapted to account for the functions of the striatum, with parts of the dorsal striatum equated to the actor. Here, we specifically test whether the human dorsal striatum—as predicted by an actor–critic instantiation—is used on a trial-to-trial basis at the time of choice to choose in accordance with reinforcement learning theory, as opposed to a competing strategy: the gambler's fallacy. Using a partial-brain functional magnetic resonance imaging scanning protocol focused on the striatum and other ventral brain areas, we found that the dorsal striatum is more active when choosing consistent with reinforcement learning compared with the competing strategy. Moreover, an overlapping area of dorsal striatum along with the ventral striatum was found to be correlated with reward prediction errors at the time of outcome, as predicted by the actor–critic framework. These findings suggest that the same region of dorsal striatum involved in learning stimulus–response associations may contribute to the control of behavior during choice, thereby using those learned associations. Intriguingly, neither reinforcement learning nor the gambler's fallacy conformed to the optimal choice strategy on the specific decision-making task we used. Thus, the dorsal striatum may contribute to the control of behavior according to reinforcement learning even when the prescriptions of such an algorithm are suboptimal in terms of maximizing future rewards

Developmental potential of trunk neural crest cells in the mouse

The availability of naturally occurring and engineered mutations in mice which affect the neural crest makes the mouse embryo an important experimental system for studying neural crest cell differentiation. Here, we determine the normal developmental potential of neural crest cells by performing in situ cell lineage analysis in the mouse by microinjecting lysinated rhodamine dextran (LRD) into individual dorsal neural tube cells in the trunk. Labeled progeny derived from single cells were found in the neural tube, dorsal root ganglia, sympathoadrenal derivatives, presumptive Schwann cells and/or pigment cells. Most embryos contained labeled cells both in the neural tube and at least one neural crest derivative, and numerous clones contributed to multiple neural crest derivatives. The time of injection influenced the derivatives populated by the labeled cells. Injections at early stages of migration yielded labeled progeny in both dorsal and ventral neural crest derivatives, whereas those performed at later stages had labeled cells only in more dorsal neural crest derivatives, such as dorsal root ganglion and presumptive pigment cells. The results suggest that in the mouse embryo: (1) there is a common precursor for neural crest and neural tube cells; (2) some neural crest cells are multipotent; and (3) the timing of emigration influences the range of possible neural crest derivatives

The double dipole model of theta rhythm generation: Simulation of laminar field potential profiles in dorsal hippocampus of the rat

A set of compartmental models of CA1 pyramidal, granular and polymorph cells of the dorsal hippocampus have been used to simulate membrane potentials generated by synaptic activation at various levels along these cells. From the membrane potential distributions the field potentials in dorsal CA1 and the dorsal blade of the dentate area have been simulated using a model based on volume conduction theory. Field potential profiles similar to laminar profiles, found experimentally in the dorsal hippocampus during theta rhythm, could only be simulated by assuming (almost) simultaneous synaptic excitation of the 3 cell types at given sites. The results lead to 2 alternative models for the simultaneous excitation of CA1 pyramidal cells and dentate granular cells during theta rhythm. Other electrophysiological evidence favours the model in which the two neuronal populations are activated distally near the fissure