Bifurcation of axons from cranial sensory neurons is disabled in the absence of Npr2-induced cGMP signaling

Axonal branching is a prerequisite for the establishment of complex neuronal circuits and their capacity for parallel information processing. Previously, we have identified a cGMP signaling pathway composed of the ligand C-type natriuretic peptide (CNP), its receptor, the guanylyl cyclase natriuretic peptide receptor 2 (Npr2), and the cGMP-dependent kinase I{alpha} (cGKI{alpha}) that regulates axon bifurcation of dorsal root ganglion (DRG) neurons in the spinal cord. Now we asked whether this cascade also controls axon bifurcation elsewhere in the nervous system. An Npr2-lacZ reporter mouse line was generated to clarify the pattern of the CNP receptor expression. It was found that during the period of axonal outgrowth, Npr2 and cGKI{alpha} were strongly labeled in neurons of all cranial sensory ganglia (gV, gVII, gVIII, gIX, and gX). In addition, strong complementary expression of CNP was detected in the hindbrain at the entry zones of sensory afferents. To analyze axon branching in individual Npr2-positive neurons, we generated a mouse mutant expressing a tamoxifen-inducible variant of Cre recombinase expressed under control of the Npr2-promoter (Npr2-CreER(T2)). After crossing this strain with conditional reporter mouse lines, we revealed that the complete absence of Npr2 activity indeed prohibited the bifurcation of cranial sensory axons in their entrance region. Consequently, axons only turned in either an ascending or descending direction, while collateral formation and growth of the peripheral arm was not affected. These findings indicate that in neurons of the cranial sensory ganglia, as in DRG neurons, cGMP signals are necessary for the execution of an axonal bifurcation program

Molecular analysis of sensory axon branching unraveled a cGMP-dependent signaling cascade

Axonal branching is a key process in the establishment of circuit connectivity within the nervous system. Molecular-genetic studies have shown that a specific form of axonal branching - the bifurcation of sensory neurons at the transition zone between the peripheral and the central nervous System - is regulated by a cyclic guanosine monophosphate (cGMP)-dependent signaling cascade which is composed of C-type natriuretic peptide (CNP), the receptor guanylyl cyclase Npr2, and cGMP-dependent protein kinase Iα (cGKIα). In the absence of any one of these components, neurons in dorsal root ganglia (DRG) and cranial sensory ganglia no longer bifurcate, and instead turn in either an ascending or a descending direction. In contrast, collateral axonal branch formation which represents a second type of axonal branch formation is not affected by inactivation of CNP, Npr2, or cGKI. Whereas axon bifurcation was lost in mouse mutants deficient for components of CNP-induced cGMP formation; the absence of the cGMP-degrading enzyme phosphodiesterase 2A had no effect on axon bifurcation. Adult mice that lack sensory axon bifurcation due to the conditional inactivation of Npr2-mediated cGMP signaling in DRG neurons demonstrated an altered shape of sensory axon terminal fields in the spinal cord, indicating that elaborate compensatory mechanisms reorganize neuronal circuits in the absence of bifurcation. On a functional level, these mice showed impaired heat sensation and nociception induced by chemical irritants, whereas responses to cold sensation, mechanical stimulation, and motor coordination are normal. These data point to a critical role of axon bifurcation for the processing of acute pain perception

Energetic beams of negative and neutral hydrogen from intense laser plasma interaction

One of the most striking demonstrations of intermolecular forces is the tension at the surface of liquid n-alkanes. The prediction of surface tension is important in the design of distillation towers, extraction units and tower internals such as bubble caps and trays, since it has a considerable influence on the transfer of mass and energy across interfaces. Surface tension data are needed wherever foaming emulsification, droplet formation or wetting are involved. They are also required in a number of equations for two-phase flow calculations and for determining the flow regime. Petroleum engineers are especially interested in the surface tension in the extraction of crude oil to add surfactants to modify the interfacial properties between crude oil and the geological reservoir to improve production and increase oil yields. In this work, a simple computer program using Arrhenius-type asymptotic exponential function, Vandermoned matrix and Matlab technical computing language, is developed for the estimation of surface tension of paraffin hydrocarbons as a function of molecular weight and temperature. The surface tension is calculated for temperatures in the range of 250 to 440 K and paraffin hydrocarbons molecular weights between 30 and 250. The proposed numerical technique is superior owing to its accuracy and clear numerical background, wherein the relevant coefficients can be retuned quickly if more data become available in the future. Estimations are found to be in excellent agreement with the reliable data in the literature with average absolute deviation being less than 2%

A Femtosecond Neutron Source

The possibility to use the ultrashort ion bunches produced by circularly polarized laser pulses to drive a source of fusion neutrons with sub-optical cycle duration is discussed. A two-side irradiation of a thin foil deuterated target produces two countermoving ion bunches, whose collision leads to an ultrashort neutron burst. Using particle-in-cell simulations and analytical modeling, it is evaluated that, for intensities of a few $10^{19} W cm^{-2}$, more than $10^3$ neutrons per Joule may be produced within a time shorter than one femtosecond. Another scheme based on a layered deuterium-tritium target is outlined.Comment: 15 pages, 3 figure

Backbone rigidity and static presentation of guanidinium groups increases cellular uptake of arginine-rich cell-penetrating peptides

In addition to endocytosis-mediated cellular uptake, hydrophilic cell-penetrating peptides are able to traverse biological membranes in a non-endocytic mode termed transduction, resulting in immediate bioavailability. Here we analysed structural requirements for the non-endocytic uptake mode of arginine-rich cell-penetrating peptides, by a combination of live-cell microscopy, molecular dynamics simulations and analytical ultracentrifugation. We demonstrate that the transduction efficiency of arginine-rich peptides increases with higher peptide structural rigidity. Consequently, cyclic arginine-rich cell-penetrating peptides showed enhanced cellular uptake kinetics relative to their linear and more flexible counterpart. We propose that guanidinium groups are forced into maximally distant positions by cyclization. This orientation increases membrane contacts leading to enhanced cell penetration

Search for two Higgs bosons in final states containing two photons and two bottom quarks in proton-proton collisions at 8 TeV

Peer reviewe

A genetic strategy for the analysis of individual axon morphologies in cGMP signalling mutant mice

One of the many physiological functions of cyclic guanosine 3',5' monophosphate (cGMP) signalling is the regulation of a specific mode of axonal branching. The bifurcation of axons from dorsal root ganglion (DRG) neurons at the dorsal root entry zone of the embryonic spinal cord is triggered by a cGMP -signalling pathway comprising the ligand C-type natriuretic peptide (CNP), the cGMP-producing natriuretic peptide receptor 2 (Npr2), and the cGMP-dependent protein kinase Ialpha (cGKIalpha). Absence of any of these components causes a loss of bifurcation and sensory axons instead only turn in either a rostral or a caudal direction. In this chapter we describe a genetic strategy to study the impact of cGMP signalling on the arborization of individual DRG neurons in mice. Expression of an alkaline phosphatase (AP) reporter is selectively induced in Npr2-positive DRG neurons by tamoxifen-dependent activation of a Cre -recombinase under the control of the Npr2 promoter. This approach might also be employed for the analysis of axonal branching in neuronal subsets expressing Npr2 elsewhere in the nervous system

cGMP signaling and branching of sensory axons in the spinal cord

Axonal branching is essential for neurons to establish contacts to different targets. It therefore provides the physical basis for the integration and distribution of information within the nervous system. During embryonic and early postnatal development, several axonal branching modes may be distinguished that might be regulated by activities of the growth cone or by the axon shaft. The various forms of axonal branching are dependent on intrinsic components and are regulated by extrinsic factors that activate specific signaling systems. This article focuses on components implicated in cyclic guanosine monophosphate signaling that regulate axon bifurcation - a specific form of branching - within the spinal cord in animal models. This cascade is composed of the ligand CNP, the guanylyl cyclase Npr2 and the cyclic guanosine monophosphate-dependent kinase I. In the absence of one of these components, axons of dorsal root ganglion neurons do not form T-shaped branches when entering the spinal cord, while collateral (interstitial) branching, another branching mode of the same type of the neuron, is not affected. It will be important to analyze human patients with mutations in the corresponding genes to get insights into the pathophysiological effects of impaired sensory axon branching in the spinal cord