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flp-32 Ligand/receptor silencing phenocopy faster plant pathogenic nematodes.

Atkinson LE, Stevenson M, McCoy CJ, Marks NJ, Fleming C, Zamanian M, Day TA, Kimber MJ, Maule AG, Mousley A - PLoS Pathog. (2013)

Bottom Line: This study investigates the role of flp-32 in G. pallida and shows that: (i) Gp-flp-32 encodes the peptide AMRNALVRFamide; (ii) Gp-flp-32 is expressed in the brain and ventral nerve cord of G. pallida; (iii) migration rate increases in Gp-flp-32-silenced worms; (iv) the ability of G. pallida to infect potato plant root systems is enhanced in Gp-flp-32-silenced worms; (v) a novel putative Gp-flp-32 receptor (Gp-flp-32R) is expressed in G. pallida; and, (vi) Gp-flp-32R-silenced worms also display an increase in migration rate.This work demonstrates that Gp-flp-32 plays an intrinsic role in the modulation of locomotory behaviour in G. pallida and putatively interacts with at least one novel G-protein coupled receptor (Gp-flp-32R).This is the first functional characterisation of a parasitic nematode FLP-GPCR.

View Article: PubMed Central - PubMed

Affiliation: Molecular Biosciences-Parasitology, Institute for Global Food Security, School of Biological Sciences, Queen's University Belfast, Belfast, United Kingdom.

ABSTRACT
Restrictions on nematicide usage underscore the need for novel control strategies for plant pathogenic nematodes such as Globodera pallida (potato cyst nematode) that impose a significant economic burden on plant cultivation activities. The nematode neuropeptide signalling system is an attractive resource for novel control targets as it plays a critical role in sensory and motor functions. The FMRFamide-like peptides (FLPs) form the largest and most diverse family of neuropeptides in invertebrates, and are structurally conserved across nematode species, highlighting the utility of the FLPergic system as a broad-spectrum control target. flp-32 is expressed widely across nematode species. This study investigates the role of flp-32 in G. pallida and shows that: (i) Gp-flp-32 encodes the peptide AMRNALVRFamide; (ii) Gp-flp-32 is expressed in the brain and ventral nerve cord of G. pallida; (iii) migration rate increases in Gp-flp-32-silenced worms; (iv) the ability of G. pallida to infect potato plant root systems is enhanced in Gp-flp-32-silenced worms; (v) a novel putative Gp-flp-32 receptor (Gp-flp-32R) is expressed in G. pallida; and, (vi) Gp-flp-32R-silenced worms also display an increase in migration rate. This work demonstrates that Gp-flp-32 plays an intrinsic role in the modulation of locomotory behaviour in G. pallida and putatively interacts with at least one novel G-protein coupled receptor (Gp-flp-32R). This is the first functional characterisation of a parasitic nematode FLP-GPCR.

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Globodera pallida FLP-32 (AMRNALVRFa) is localised in the nervous system of pre-parasitic juveniles.Confocal microscopy images of G. pallida FLP-32 (AMRNALVRFa) immunoreactivity (IR; green; A) demonstrate staining in the circumpharyngeal nerve ring (CNR) posterior to the pharyngeal bulb (PHB), and in nerve processes (*) running anteriorly towards the stylet protractor muscles (SPM). IR is also evident in the ventral nerve cord (VNC; A) which emanates from the CNR and runs parallel to the body wall muscle (BWM; red) into the tail where a group of AMRNALVRFa-positive cell bodies are located in close proximity to the lumbar ganglia (LG; B). Scale bars = 75 µm.
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ppat-1003169-g003: Globodera pallida FLP-32 (AMRNALVRFa) is localised in the nervous system of pre-parasitic juveniles.Confocal microscopy images of G. pallida FLP-32 (AMRNALVRFa) immunoreactivity (IR; green; A) demonstrate staining in the circumpharyngeal nerve ring (CNR) posterior to the pharyngeal bulb (PHB), and in nerve processes (*) running anteriorly towards the stylet protractor muscles (SPM). IR is also evident in the ventral nerve cord (VNC; A) which emanates from the CNR and runs parallel to the body wall muscle (BWM; red) into the tail where a group of AMRNALVRFa-positive cell bodies are located in close proximity to the lumbar ganglia (LG; B). Scale bars = 75 µm.

Mentions: A custom raised antiserum directed against the single peptide encoded by Gp-flp-32, AMRNALVRFamide, was used to localise Gp-FLP-32 using ICC in G. pallida J2s. The overall pattern of Gp-FLP-32 localisation was similar to the expression pattern of Gp-flp-32 exhibited in ISH experiments, comprising extensive AMRNALVRFamide immunostaining within the nervous system of G. pallida (see Fig. 3A and B). Strong immunoreactivity was visualised within the CNR, with AMRNALVRFamide-immunopositive nerve processes running both anteriorly and posteriorly from the CNR towards the stylet protractor muscles and the VNC respectively (Fig. 3A). While some staining within the CNR was diffuse as previously noted in ISH experiments, there were distinct accumulations of immunoreactivity on both the ventral and dorsal sides of the nerve ring (see Fig. 3A). In addition there was an accumulation of immunopositive staining, albeit weaker than the anterior staining, in the posterior of the nematode close to the tip of the tail (see Fig. 3B). Again this was concurrent with the positioning of Gp-flp-32 expression in the ISH experiments, which would suggest Gp-FLP-32 immunoreactivity in cell bodies of the LG.


flp-32 Ligand/receptor silencing phenocopy faster plant pathogenic nematodes.

Atkinson LE, Stevenson M, McCoy CJ, Marks NJ, Fleming C, Zamanian M, Day TA, Kimber MJ, Maule AG, Mousley A - PLoS Pathog. (2013)

Globodera pallida FLP-32 (AMRNALVRFa) is localised in the nervous system of pre-parasitic juveniles.Confocal microscopy images of G. pallida FLP-32 (AMRNALVRFa) immunoreactivity (IR; green; A) demonstrate staining in the circumpharyngeal nerve ring (CNR) posterior to the pharyngeal bulb (PHB), and in nerve processes (*) running anteriorly towards the stylet protractor muscles (SPM). IR is also evident in the ventral nerve cord (VNC; A) which emanates from the CNR and runs parallel to the body wall muscle (BWM; red) into the tail where a group of AMRNALVRFa-positive cell bodies are located in close proximity to the lumbar ganglia (LG; B). Scale bars = 75 µm.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3585147&req=5

ppat-1003169-g003: Globodera pallida FLP-32 (AMRNALVRFa) is localised in the nervous system of pre-parasitic juveniles.Confocal microscopy images of G. pallida FLP-32 (AMRNALVRFa) immunoreactivity (IR; green; A) demonstrate staining in the circumpharyngeal nerve ring (CNR) posterior to the pharyngeal bulb (PHB), and in nerve processes (*) running anteriorly towards the stylet protractor muscles (SPM). IR is also evident in the ventral nerve cord (VNC; A) which emanates from the CNR and runs parallel to the body wall muscle (BWM; red) into the tail where a group of AMRNALVRFa-positive cell bodies are located in close proximity to the lumbar ganglia (LG; B). Scale bars = 75 µm.
Mentions: A custom raised antiserum directed against the single peptide encoded by Gp-flp-32, AMRNALVRFamide, was used to localise Gp-FLP-32 using ICC in G. pallida J2s. The overall pattern of Gp-FLP-32 localisation was similar to the expression pattern of Gp-flp-32 exhibited in ISH experiments, comprising extensive AMRNALVRFamide immunostaining within the nervous system of G. pallida (see Fig. 3A and B). Strong immunoreactivity was visualised within the CNR, with AMRNALVRFamide-immunopositive nerve processes running both anteriorly and posteriorly from the CNR towards the stylet protractor muscles and the VNC respectively (Fig. 3A). While some staining within the CNR was diffuse as previously noted in ISH experiments, there were distinct accumulations of immunoreactivity on both the ventral and dorsal sides of the nerve ring (see Fig. 3A). In addition there was an accumulation of immunopositive staining, albeit weaker than the anterior staining, in the posterior of the nematode close to the tip of the tail (see Fig. 3B). Again this was concurrent with the positioning of Gp-flp-32 expression in the ISH experiments, which would suggest Gp-FLP-32 immunoreactivity in cell bodies of the LG.

Bottom Line: This study investigates the role of flp-32 in G. pallida and shows that: (i) Gp-flp-32 encodes the peptide AMRNALVRFamide; (ii) Gp-flp-32 is expressed in the brain and ventral nerve cord of G. pallida; (iii) migration rate increases in Gp-flp-32-silenced worms; (iv) the ability of G. pallida to infect potato plant root systems is enhanced in Gp-flp-32-silenced worms; (v) a novel putative Gp-flp-32 receptor (Gp-flp-32R) is expressed in G. pallida; and, (vi) Gp-flp-32R-silenced worms also display an increase in migration rate.This work demonstrates that Gp-flp-32 plays an intrinsic role in the modulation of locomotory behaviour in G. pallida and putatively interacts with at least one novel G-protein coupled receptor (Gp-flp-32R).This is the first functional characterisation of a parasitic nematode FLP-GPCR.

View Article: PubMed Central - PubMed

Affiliation: Molecular Biosciences-Parasitology, Institute for Global Food Security, School of Biological Sciences, Queen's University Belfast, Belfast, United Kingdom.

ABSTRACT
Restrictions on nematicide usage underscore the need for novel control strategies for plant pathogenic nematodes such as Globodera pallida (potato cyst nematode) that impose a significant economic burden on plant cultivation activities. The nematode neuropeptide signalling system is an attractive resource for novel control targets as it plays a critical role in sensory and motor functions. The FMRFamide-like peptides (FLPs) form the largest and most diverse family of neuropeptides in invertebrates, and are structurally conserved across nematode species, highlighting the utility of the FLPergic system as a broad-spectrum control target. flp-32 is expressed widely across nematode species. This study investigates the role of flp-32 in G. pallida and shows that: (i) Gp-flp-32 encodes the peptide AMRNALVRFamide; (ii) Gp-flp-32 is expressed in the brain and ventral nerve cord of G. pallida; (iii) migration rate increases in Gp-flp-32-silenced worms; (iv) the ability of G. pallida to infect potato plant root systems is enhanced in Gp-flp-32-silenced worms; (v) a novel putative Gp-flp-32 receptor (Gp-flp-32R) is expressed in G. pallida; and, (vi) Gp-flp-32R-silenced worms also display an increase in migration rate. This work demonstrates that Gp-flp-32 plays an intrinsic role in the modulation of locomotory behaviour in G. pallida and putatively interacts with at least one novel G-protein coupled receptor (Gp-flp-32R). This is the first functional characterisation of a parasitic nematode FLP-GPCR.

Show MeSH
Related in: MedlinePlus