Differential Axonal Conduction Patterns of Mechano-Sensitive and Mechano-Insensitive Nociceptors – A Combined Experimental and Modelling Study

Cutaneous pain sensations are mediated largely by C-nociceptors consisting of both mechano-sensitive (CM) and mechano-insensitive (CMi) fibres that can be distinguished from one another according to their characteristic axonal properties. In healthy skin and relative to CMi fibres, CM fibres show a higher initial conduction velocity, less activity-dependent conduction velocity slowing, and less prominent post-spike supernormality. However, after sensitization with nerve growth factor, the electrical signature of CMi fibres changes towards a profile similar to that of CM fibres. Here we take a combined experimental and modelling approach to examine the molecular basis of such alterations to the excitation thresholds. Changes in electrical activation thresholds and activity-dependent slowing were examined in vivo using single-fibre recordings of CM and CMi fibres in domestic pigs following NGF application. Using computational modelling, we investigated which axonal mechanisms contribute most to the electrophysiological differences between the fibre classes. Simulations of axonal conduction suggest that the differences between CMi and CM fibres are strongly influenced by the densities of the delayed rectifier potassium channel (K_dr), the voltage-gated sodium channels Na_V1.7 and Na_V1.8, and the Na⁺/K⁺-ATPase. Specifically, the CM fibre profile required less K_dr and Na_V1.8 in combination with more Na_V1.7 and Na⁺/K⁺-ATPase. The difference between CM and CMi fibres is thus likely to reflect a relative rather than an absolute difference in protein expression. In support of this, it was possible to replicate the experimental reduction of the ADS pattern of CMi nociceptors towards a CM-like pattern following intradermal injection of nerve growth factor by decreasing the contribution of K_dr (by 50%), increasing the Na⁺/K⁺-ATPase (by 10%), and reducing the branch length from 2 cm to 1 cm. The findings highlight key molecules that potentially contribute to the NGF-induced switch in nociceptors phenotype, in particular Na_V1.7 which has already been identified clinically as a principal contributor to chronic pain states such as inherited erythromelalgia.     

Host-specific effect of P1 exchange between two potyviruses

The potyviruses Plum pox virus (PPV) and Tobacco vein mottling virus (TVMV) have distinct host ranges and induce different symptoms in their common herbaceous hosts. To test the relevance of the P1 protein in host compatibility and pathogenicity, hybrid viruses were constructed in which the P1 coding sequence of PPV was completely or partially replaced by the corresponding sequences from TVMV. Infections induced by these chimeric viruses revealed that the TVMV P1 and a PPV/TVMV hybrid P1 proteins are functionally equivalent in herbaceous plants to the P1 protein of a PPV isolate adapted to these hosts, in spite of having high sequence divergence. Moreover, the presence of TVMV P1 sequences enhanced the competence of a low-infectivity PPV-D-derived chimera in Nicotiana clevelandii . Conversely, all PPV/TVMV hybrids were unable to infect Prunus persicae , a specific host for PPV, suggesting that TVMV P1 is not functionally competent in this plant. Together, these data highlight the importance of the P1 protein in defining the virus host range.     

Waisel, Y., Eshel, A., Kafkafi, U., eds. Plant roots - the hidden half

Research on plant roots represents an exciting and intriguingfield of science. Undoubtedly, the vast major ity of researcherswithin the field are familiar with the first (1991) and second(1996) editions of this book. Since these editions were published,understanding of root biology has been advanced by using molecularbiology methods and molecular genetic tools which offer novelperspectives for the exploitation and understanding of rootstructure and root processes. The third edition of Plant roots– the hidden half includes revised and expanded informationon topics covered in the previous editions, as well as in     

Sexual dimorphism, sex ratio and spatial distribution of male and female shrubs in the dioecious speciesPistacia lentiscus L.

The population sex ratio, spatial distribution, relative growth rate, canopy size and morphology, and nitrogen content of leaves of male and female individuals were compared in seven populations of the dioecious shrubPistacia lentiscus L. occurring in different localities of the Serra da Arrábida Natural Park, Portugal. The expected pattern of male preponderance in stressful habitats was tested. Less perturbed areas, with a well developed vegetation cover, had male-biased sex ratios, while in former agricultural areas there were no significant differences between the number of male and female plants. Our results failed to reveal any significant differences in the morphological variables between sexes except in the number of basal stems. Male plants also had a significantly higher chlorophyll content (chl a+b) than female plants. But when plants were sorted into large and small individuals, larger plants showed a significantly higher leaf area index and a significantly higher number of basal stems. There were no significant differences in the leaf nitrogen content between male and female plants from June to August, but such differences appeared (P<0.001) in September and October corresponding to the production of mature fruits. Differences in sex-ratio may be explained by different resource allocation patterns of the two sexes, which favoured male plants in the less perturbed environments. In agricultural areas farmers could also have favoured female plants in former times.     

Cordiachromes from Auxemma oncocalyx

Further cordiachromes, rel-10,11β-epoxy-11-ethoxy-8-hydroxy-2-methoxy-8aβ-methyl-5,6,7,8,8a,9,10aβ-octahydro-1,4-anthracendione, 6-formyl-2-methoxy-9-methyl-7,8-dihydro-1,4-phenanthrendione, rel-8,11;9,11-diepoxy-1,4-dihydroxy-2-methoxy-8aβ-methyl-5,6,7,8,8a,9,10,10aβ-octahydro-10-anthracenone, rel-9,11-epoxy-1,4,8-trihydroxy-2-methoxy-8aβ-methyl-5,6,7,8,8a,9,10,10aβ-octahydro-10-anthracenone, rel-2″-methoxy-7″-methyl-1″,4″-naphtalendione-(6″→5)-tetrahydropyran-(2-eq→O→2ax)-tetrahydropyran-(5′→6)- 2-methoxy-7-methyl-1,4-naphthalendione, together with the known, allantoin, sitosterol and 3β-O-d-glucopyranosylsitosterol, have been isolated from Auxemma oncocalyx. Their structures were determined from spectral data, including 2D NMR experiments.     

FAPP2 is involved in the transport of apical cargo in polarized MDCK cells

Phosphatidylinositol-4-phosphate (PI(4)P) is the main phosphoinositide in the Golgi complex and has been reported to play a pleiotropic role in transport of cargo from the trans-Golgi network to the plasma membrane (PM) in polarized Madin-Darby canine kidney (MDCK) cells. Overexpression of the chimeric fluorescent protein encoding the pleckstrin homology domain, which is specific for PI(4)P, inhibited both apical and basolateral transport pathways. The transport of apical cargo from the Golgi was shown to be specifically decreased by adenovirus-mediated RNA interference directed against PI(4)P adaptor protein (FAPP) 2. FAPP1 depletion had no effect on transport. On the other hand, FAPP2 was not involved in the Golgi-to-PM transport of cargo that was targeted to the basolateral membrane domain. Thus, we conclude that FAPP2 plays a specific role in apical transport in MDCK cells.     

Virus‐induced gene silencing in transgenic plants: transgene silencing and reactivation associate with two patterns of transgene body methylation

We used bisulfite sequencing to study the methylation of a viral transgene whose expression was silenced upon plum pox virus infection of the transgenic plant and its subsequent recovery as a consequence of so‐called virus‐induced gene silencing (VIGS). VIGS was associated with a general increase in the accumulation of small RNAs corresponding to the coding region of the viral transgene. After VIGS, the transgene promoter was not methylated and the coding region showed uneven methylation, with the 5′ end being mostly unmethylated in the recovered tissue or mainly methylated at CG sites in regenerated silenced plants. The methylation increased towards the 3′ end, which showed dense methylation in all three contexts (CG, CHG and CHH). This methylation pattern and the corresponding silenced status were maintained after plant regeneration from recovered silenced tissue and did not spread into the promoter region, but were not inherited in the sexual offspring. Instead, a new pattern of methylation was observed in the progeny plants consisting of disappearance of the CHH methylation, similar CHG methylation at the 3′ end, and an overall increase in CG methylation in the 5′ end. The latter epigenetic state was inherited over several generations and did not correlate with transgene silencing and hence virus resistance. These results suggest that the widespread CG methylation pattern found in body gene bodies located in euchromatic regions of plant genomes may reflect an older silencing event, and most likely these genes are no longer silenced.     

Resprouting Ability of Six Atlantic Shrub Species

Resprouting from subterranean structures is a principal method of vegetative regeneration that many shrub species show after a disturbance. This study, therefore, aims to determine the resprouting capacity and intensity of six dominant species in an Atlantic shrubland area located in the NW of the Iberian Peninsula and compare their resprouting and germinating strategies. Resprouting intensity is measured using three variables: individual probability of resprouting, number of resprouts and length of these sprouts in three plant-age classes. The intensity for each species was calculated using a simple index (IRI) that included the three measured variables. All studied species, excepting Erica umbellata, could resprout. According to the IRI values, there are three groups of species: strong resprouters (Ulex europaeus, Ulex minor and Pterospartum tridentatum), weak resprouters (Ulex micranthus and Genista triacanthos) and non-resprouters (E. umbellata). The germination of strong resprouters is highly stimulated by fire. Frequent disturbances remove the non- and weak resprouter populations and promote the strong resprouter ones.     

Immunomodulatory effects of alcbholic and hydroalcoholic extracts of Moringa olifera Lam leaves

Effects of 50, 100 and 200 mg/kg body weight of the alcoholic and hydro-alcoholic extract of leaves of M. olifera were studied on various immune paradigms like delayed type hypersensitivity reaction using SRBC as an antigen, determination of antibody titer, neutrophil adhesion test as an indicator for neutrophil index, total leucocyte count in cyclophosphamide induced immunosuppressed animals and carbon clearance assay as a measure of phagocytic activity. Hydro-alcoholic extract of M. olifera substantially enhanced cellular immune response, humoral immune response, neutrophil index and phagoctic activity in doses of 100 and 200 mg/kg body weight. The ethanolic extract (200 mg/kg body weight) was efficient in improving immune response. The results suggest that M. olifera has a significant role to play as an immune stimulator.