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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 (Gp-flp-32) is expressed in the brain and ventral nerve cords of pre-parasitic juveniles (J2s).Light microscopy in situ hybridisation images show Gp-flp-32 gene expression in the circumpharyngeal nerve ring (CNR) posterior to the stylet (S) and the pharyngeal bulb (PHB) of pre-parasitic juvenile (J2) G. pallida (A). Gp-flp-32 is also expressed in repeating groups of three to four cell bodies located in the ventral nerve cord (VNC; B and C) which runs posteriorly to the tail where a cluster of three cell bodies also expressing Gp-flp-32 are located in the region of the lumbar ganglia (LG; C and D). Scale bars: A–C = 12 µm; D = 5 µm.
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ppat-1003169-g002: Globodera pallida flp-32 (Gp-flp-32) is expressed in the brain and ventral nerve cords of pre-parasitic juveniles (J2s).Light microscopy in situ hybridisation images show Gp-flp-32 gene expression in the circumpharyngeal nerve ring (CNR) posterior to the stylet (S) and the pharyngeal bulb (PHB) of pre-parasitic juvenile (J2) G. pallida (A). Gp-flp-32 is also expressed in repeating groups of three to four cell bodies located in the ventral nerve cord (VNC; B and C) which runs posteriorly to the tail where a cluster of three cell bodies also expressing Gp-flp-32 are located in the region of the lumbar ganglia (LG; C and D). Scale bars: A–C = 12 µm; D = 5 µm.

Mentions: Gp-flp-32 expression, visualised by the hybridisation of a 201 base pair (bp) probe, was identified both within and connecting the anterior and posterior regions of the nematode (see Fig. 2A–D). Staining was evident in the circumpharyngeal nerve ring (CNR), and within multiple distinct cell bodies in the ventral nerve cord (VNC) and lumbar ganglia (LG) (see Fig. 2A–D). Staining within the CNR was diffuse with no specific neuronal cell bodies staining strongly (Fig. 2A); this pattern was evident in the majority of specimens (>90%) treated with the antisense probe, and is similar to a diffuse ISH staining pattern previously reported in the G. pallida CNR for flp-6 (KSAYMRFG; [17]). In contrast, staining in the VNC was characterised by groups of three to four distinct and strongly reactive cell bodies spaced at regular intervals along the nerve cord, beginning posterior to the CNR and running into the tail (Fig. 2B). Although slightly variable, most specimens exhibited approximately six to eight groups of cells in the VNC, with as many as 18 cell bodies visible at any one time (see Fig. 2B and C). While unequivocal assignment of neuronal cell bodies is difficult, in this scenario it is likely that they belong to VD and/or DD motor neurons which possess cell bodies in the VNC of C. elegans, where they have been shown to express a VRFamide-like flp gene encoding peptides similar to that encoded by Gp-flp-32 [18]. VD neurons are a set of 13 motor neurons which innervate ventral muscle and have cell bodies in the VNC, while DD are a set of six motor neurons (pre-synaptic to VD), which also possess cell bodies in the VNC, but instead innervate dorsal muscle [19], [20]. Together this amounts to 19 identifiable cell bodies within the VNC, and while this number is known to vary in C. elegans according to developmental stage [19], it is similar to the 18 cell bodies visible within the VNC of G. pallida following Gp-flp-32 antisense probe hybridisation.


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 (Gp-flp-32) is expressed in the brain and ventral nerve cords of pre-parasitic juveniles (J2s).Light microscopy in situ hybridisation images show Gp-flp-32 gene expression in the circumpharyngeal nerve ring (CNR) posterior to the stylet (S) and the pharyngeal bulb (PHB) of pre-parasitic juvenile (J2) G. pallida (A). Gp-flp-32 is also expressed in repeating groups of three to four cell bodies located in the ventral nerve cord (VNC; B and C) which runs posteriorly to the tail where a cluster of three cell bodies also expressing Gp-flp-32 are located in the region of the lumbar ganglia (LG; C and D). Scale bars: A–C = 12 µm; D = 5 µm.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3585147&req=5

ppat-1003169-g002: Globodera pallida flp-32 (Gp-flp-32) is expressed in the brain and ventral nerve cords of pre-parasitic juveniles (J2s).Light microscopy in situ hybridisation images show Gp-flp-32 gene expression in the circumpharyngeal nerve ring (CNR) posterior to the stylet (S) and the pharyngeal bulb (PHB) of pre-parasitic juvenile (J2) G. pallida (A). Gp-flp-32 is also expressed in repeating groups of three to four cell bodies located in the ventral nerve cord (VNC; B and C) which runs posteriorly to the tail where a cluster of three cell bodies also expressing Gp-flp-32 are located in the region of the lumbar ganglia (LG; C and D). Scale bars: A–C = 12 µm; D = 5 µm.
Mentions: Gp-flp-32 expression, visualised by the hybridisation of a 201 base pair (bp) probe, was identified both within and connecting the anterior and posterior regions of the nematode (see Fig. 2A–D). Staining was evident in the circumpharyngeal nerve ring (CNR), and within multiple distinct cell bodies in the ventral nerve cord (VNC) and lumbar ganglia (LG) (see Fig. 2A–D). Staining within the CNR was diffuse with no specific neuronal cell bodies staining strongly (Fig. 2A); this pattern was evident in the majority of specimens (>90%) treated with the antisense probe, and is similar to a diffuse ISH staining pattern previously reported in the G. pallida CNR for flp-6 (KSAYMRFG; [17]). In contrast, staining in the VNC was characterised by groups of three to four distinct and strongly reactive cell bodies spaced at regular intervals along the nerve cord, beginning posterior to the CNR and running into the tail (Fig. 2B). Although slightly variable, most specimens exhibited approximately six to eight groups of cells in the VNC, with as many as 18 cell bodies visible at any one time (see Fig. 2B and C). While unequivocal assignment of neuronal cell bodies is difficult, in this scenario it is likely that they belong to VD and/or DD motor neurons which possess cell bodies in the VNC of C. elegans, where they have been shown to express a VRFamide-like flp gene encoding peptides similar to that encoded by Gp-flp-32 [18]. VD neurons are a set of 13 motor neurons which innervate ventral muscle and have cell bodies in the VNC, while DD are a set of six motor neurons (pre-synaptic to VD), which also possess cell bodies in the VNC, but instead innervate dorsal muscle [19], [20]. Together this amounts to 19 identifiable cell bodies within the VNC, and while this number is known to vary in C. elegans according to developmental stage [19], it is similar to the 18 cell bodies visible within the VNC of G. pallida following Gp-flp-32 antisense probe hybridisation.

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