Nerve growth factor induces neurite outgrowth of PC12 cells by promoting Gβγ-microtubule interaction

Background: Assembly and disassembly of microtubules (MTs) is critical for neurite outgrowth and differentiation. Evidence suggests that nerve growth factor (NGF) induces neurite outgrowth from PC12 cells by activating the receptor tyrosine kinase, TrkA. G protein-coupled receptors (GPCRs) as well as heterotrimeric G proteins are also involved in regulating neurite outgrowth. However, the possible connection between these pathways and how they might ultimately converge to regulate the assembly and organization of MTs during neurite outgrowth is not well understood. Results: Here, we report that Gβγ, an important component of the GPCR pathway, is critical for NGF-induced neuronal differentiation of PC12 cells. We have found that NGF promoted the interaction of Gβγ with MTs and stimulated MT assembly. While Gβγ-sequestering peptide GRK2i inhibited neurite formation, disrupted MTs, and induced neurite damage, the Gβγ activator mSIRK stimulated neurite outgrowth, which indicates the involvement of Gβγ in this process. Because we have shown earlier that prenylation and subsequent methylation/demethylation of γ subunits are required for the Gβγ-MTs interaction in vitro, small-molecule inhibitors (L-28 and L-23) targeting prenylated methylated protein methyl esterase (PMPMEase) were tested in the current study. We found that these inhibitors disrupted Gβγ and ΜΤ organization and affected cellular morphology and neurite outgrowth. In further support of a role of Gβγ-MT interaction in neuronal differentiation, it was observed that overexpression of Gβγ in PC12 cells induced neurite outgrowth in the absence of added NGF. Moreover, overexpressed Gβγ exhibited a pattern of association with MTs similar to that observed in NGF-differentiated cells. Conclusions: Altogether, our results demonstrate that βγ subunit of heterotrimeric G proteins play a critical role in neurite outgrowth and differentiation by interacting with MTs and modulating MT rearrangement. Electronic supplementary material The online version of this article (doi:10.1186/s12868-014-0132-4) contains supplementary material, which is available to authorized users

Lung response to Bordetella pertussis infection in mice identified by gene-expression profiling

Host genetics determines the course of Bordetella pertussis infection in mice. Previously, we found four loci, Tlr4 and three novel loci, designated Bps 1–3, that are involved in the control of B. pertussis infection. The purpose of the present study was to identify candidate genes that could explain genetic differences in the course of B. pertussis infection, assuming that such genes are differentially regulated upon infection. We, therefore, studied the course of mRNA expression in the lungs after B. pertussis infection. Of the 22,000 genes investigated, 1,841 were significantly differentially expressed with 1,182 genes upregulated and 659 genes downregulated. Upregulated genes were involved in immune-related processes, such as the acute-phase response, antigen presentation, cytokine production, inflammation, and apoptosis, while downregulated genes were mainly involved in nonimmune processes, such as development and muscle contraction. Pathway analysis revealed the involvement of granulocyte function, toll-like receptor signaling pathway, and apoptosis. Nine of the differentially expressed genes were located in Bps-1, 13 were located in Bps-2, and 62 were located in Bps-3. We conclude that B. pertussis infection induces a wide and complex response, which appears to be partly specific for B. pertussis and partly nonspecific. We envisage that these data will be helpful in identifying polymorphic genes that affect the susceptibility and course of B. pertussis infection in humans

The Cysteine Rich Necrotrophic Effector SnTox1 Produced by Stagonospora nodorum Triggers Susceptibility of Wheat Lines Harboring Snn1

The wheat pathogen Stagonospora nodorum produces multiple necrotrophic effectors (also called host-selective toxins) that promote disease by interacting with corresponding host sensitivity gene products. SnTox1 was the first necrotrophic effector identified in S. nodorum, and was shown to induce necrosis on wheat lines carrying Snn1. Here, we report the molecular cloning and validation of SnTox1 as well as the preliminary characterization of the mechanism underlying the SnTox1-Snn1 interaction which leads to susceptibility. SnTox1 was identified using bioinformatics tools and verified by heterologous expression in Pichia pastoris. SnTox1 encodes a 117 amino acid protein with the first 17 amino acids predicted as a signal peptide, and strikingly, the mature protein contains 16 cysteine residues, a common feature for some avirulence effectors. The transformation of SnTox1 into an avirulent S. nodorum isolate was sufficient to make the strain pathogenic. Additionally, the deletion of SnTox1 in virulent isolates rendered the SnTox1 mutated strains avirulent on the Snn1 differential wheat line. SnTox1 was present in 85% of a global collection of S. nodorum isolates. We identified a total of 11 protein isoforms and found evidence for strong diversifying selection operating on SnTox1. The SnTox1-Snn1 interaction results in an oxidative burst, DNA laddering, and pathogenesis related (PR) gene expression, all hallmarks of a defense response. In the absence of light, the development of SnTox1-induced necrosis and disease symptoms were completely blocked. By comparing the infection processes of a GFP-tagged avirulent isolate and the same isolate transformed with SnTox1, we conclude that SnTox1 may play a critical role during fungal penetration. This research further demonstrates that necrotrophic fungal pathogens utilize small effector proteins to exploit plant resistance pathways for their colonization, which provides important insights into the molecular basis of the wheat-S. nodorum interaction, an emerging model for necrotrophic pathosystems

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Not AvailableSamba Mahsuri (BPT5204) is a medium slender grain indica rice variety that is very popular with farmers and consumers across India because of its high yield and excellent cooking quality. However, the variety is susceptible to several diseases and pests, including bacterial blight (BB). We have used PCR based molecular markers in a backcross-breeding program to introgress three major BB resistance genes (Xa21, xa13 and xa5) into Samba Mahsuri from a donor line (SS1113) in which all the three genes are present in a homozygous condition. At each backcross generation, markers closely linked to the three genes were used to select plants possessing these resistance genes (foreground selection) and microsatellite markers polymorphic between donor and recurrent parent were used to select plants that have maximum contribution from the recurrent parent genome (background selection). A selected BC4F1 plant was selfed to generate homozygous BC4F2 plants with different combinations of BB resistance genes. The three-gene pyramid and two-gene pyramid lines exhibited high levels of resistance against the BB pathogen. Under conditions of BB infection, the three-gene pyramid lines exhibited a significant yield advantage over Samba Mahsuri. Most importantly, these lines retain the excellent grain and cooking qualities of Samba Mahsuri without compromising the yield as determined in multi-location trials. This work demonstrates the successful application of marker-assisted selection for targeted introgression of multiple resistance genes into a premium quality rice variety.Not Availabl

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Not AvailableLength–weight relationships for six small indigenous fish species, namely: Trichogaster chuna (Hamilton, 1822), Trichogaster lalius (Hamilton, 1822), Trichogaster fasciata Bloch & Schneider, 1801, Chanda nama Hamilton, 1822, Parambassis lala (Hamilton, 1822), and Macrognathus aral (Bloch & Schneider, 1801) were studied for the first time from Deepor beel, a Ramsar site (589 ha water spread area) located in Assam, India. A total of 617 fish specimens were collected for the present study on a monthly basis from February to August in 2016 from landing centres adjoining the beel. In the present study, b value ranges from 2.778 to 3.215, which is within the normal range. The LWRs for these six fish species from Deepor beel had not yet been reported for FishBase.Not Availabl

Assessment of model-simulated upper ocean biogeochemical dynamics of the Bay of Bengal

The capability of a physical-biogeochemical model configured using Regional Ocean Modeling System (ROMS) in simulating upper ocean biogeochemical dynamics of the Bay of Bengal (BoB) is evaluated with available remote sensing and in situ observations. The accuracy of model-simulated and satellite (MODIS-Aqua) retrieved surface chlorophyll-a (chl-a) are individually assessed against in situ data, measured in surface waters of the western BoB, from March 2008 to November 2015. Model-simulated chl-a showed better correlation (R2 = 0.80) with the in situ observations as compared to that retrieved from satellite (R2 = 0.72). Although, the model underestimates chl-a (slope = 0.84), significant correlation proves its capability to reproduce the in situ trend. The root mean square error (RMSE) between model simulated and satellite retrieved chl-a against measured chl-a are 0.33 and 0.36, respectively. Additionally, a comparison with remote sensing time series data indicates that the model realistically simulates the seasonal variability of chl-a. Further, temperature, salinity, nitrate, chl-a and dissolved oxygen (DO) profiles obtained from two biogeochemical Argo floats deployed in the central BoB, are also compared with model-simulated profiles. In comparison, the model adequately simulates the observed subsurface variability of chl-a as well as persistent Deep Chlorophyll Maximum (DCM) at depths between 20 and 90 m having concentration 0.75–1.0 mg-m−3. The undulations in the subsurface spatial variability of chl-a are appreciably well captured by the model and comparable with the observations albeit the magnitude is overestimated in the model. It is noted that the temporal variability of the DCM and oxycline in the BoB is significantly influenced by the vertical movement of the thermocline