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GABA Immunoreactivity and Pharmacological Effects vary Among Stylet-Bearing Nematodes

Hannah M. Reed

Hannah M. Reed

Department of Crop Sciences, University of Illinois at Urbana-Champaign
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Reed, Hannah M.
, 
Ziduan Han

Ziduan Han

  • Department of Crop Sciences, University of Illinois at Urbana-Champaign
  • College of Plant Protection, Northwest A&F UniversityChina
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Han, Ziduan
 und 
Nathan E. Schroeder

Nathan E. Schroeder

  • Department of Crop Sciences, University of Illinois at Urbana-Champaign
  • Neuroscience Program, University of Illinois at Urbana-Champaign
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Schroeder, Nathan E.
  
18. Nov. 2023
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Artikel-Kategorie: Research Paper
Online veröffentlicht: 18. Nov. 2023
Eingereicht: 06. Juni 2023
DOI: https://doi.org/10.2478/jofnem-2023-0049
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© 2023 Hannah M. Reed et al., published by Sciendo
This work is licensed under the Creative Commons Attribution 4.0 International License.

Figure 1:

The GABA nervous system in stylet-bearing nematodes. 1A. The staining patterns of the GABA-immunoreactive cells in Aphelenchus avenae, Pratylenchus penetrans, Meloidogyne incognita and Heterodera glycines. From the left, the first column shows the light-microscopy images of the examined species. The second and third columns show the antibody staining of the neurotransmitter GABA in the head and tail regions. Arrowheads indicate GABA-immunoreactive neurons. Scale bar = 10 μm. B. The illustration of GABA-immunoreactive neurons that were detected using anti-GABA staining at a given stage in examined nematode species. All nematodes are displayed with anterior to the left and the dorsal side up. Neuron symbols with black filling are strongly stained cells, while gray filling indicates more weakly stained neurons. The staining patterns of the adult Aphenlenchus avenae female, adult Pratylenchus penetrans male, and Meloidogyne incognita second stage juveniles (Heterodera glycines second stage juveniles is from our previous study Han et al., 2018). Neuronal homologs are labeled based on the knowledge of C. elegans (Gendrel et al., 2016). GABA-immune positive cells found in C. elegans with either weak staining or considered involved in GABA clearance are not shown in the illustration. Illustration does not reflect true size.
The GABA nervous system in stylet-bearing nematodes. 1A. The staining patterns of the GABA-immunoreactive cells in Aphelenchus avenae, Pratylenchus penetrans, Meloidogyne incognita and Heterodera glycines. From the left, the first column shows the light-microscopy images of the examined species. The second and third columns show the antibody staining of the neurotransmitter GABA in the head and tail regions. Arrowheads indicate GABA-immunoreactive neurons. Scale bar = 10 μm. B. The illustration of GABA-immunoreactive neurons that were detected using anti-GABA staining at a given stage in examined nematode species. All nematodes are displayed with anterior to the left and the dorsal side up. Neuron symbols with black filling are strongly stained cells, while gray filling indicates more weakly stained neurons. The staining patterns of the adult Aphenlenchus avenae female, adult Pratylenchus penetrans male, and Meloidogyne incognita second stage juveniles (Heterodera glycines second stage juveniles is from our previous study Han et al., 2018). Neuronal homologs are labeled based on the knowledge of C. elegans (Gendrel et al., 2016). GABA-immune positive cells found in C. elegans with either weak staining or considered involved in GABA clearance are not shown in the illustration. Illustration does not reflect true size.

Figure 2:

Behavioral assays in response to GABA and GABA antagonists (A). Characteristic “hook shape” Heterodera glycines second-stage juveniles (marked by red arrowheads) induced via disrupting GABA signaling. The half-maximal effective concentration (EC50) of paralysis caused by piperazine (B) and GABA (C). Piperazine or GABA was dissolved in water with 0.01% Triton-X. Water with 0.01% Triton-X was used as control. Active nematodes were incubated in 10 ml of the chemical at each concentration for 10 min on a petri dish before observation. At a given concentration of each chemical, more than 30 individuals from a species were tested and the experiment was performed twice. Data were pooled from both experiments. Nematodes showing a hook shape or complete paralysis were counted as being affected (D). The locomotion assay of picrotoxin. Five nematodes of a given species were treated with 100 mM picrotoxin dissolved in 5% DMSO for 5 min and then were hand-picked on to the center of a 1-cm diameter circle on a water agar plate. After 20 min, nematodes outside the circle were counted, and the ratio of this number to the total number of test nematodes was calculated
Behavioral assays in response to GABA and GABA antagonists (A). Characteristic “hook shape” Heterodera glycines second-stage juveniles (marked by red arrowheads) induced via disrupting GABA signaling. The half-maximal effective concentration (EC50) of paralysis caused by piperazine (B) and GABA (C). Piperazine or GABA was dissolved in water with 0.01% Triton-X. Water with 0.01% Triton-X was used as control. Active nematodes were incubated in 10 ml of the chemical at each concentration for 10 min on a petri dish before observation. At a given concentration of each chemical, more than 30 individuals from a species were tested and the experiment was performed twice. Data were pooled from both experiments. Nematodes showing a hook shape or complete paralysis were counted as being affected (D). The locomotion assay of picrotoxin. Five nematodes of a given species were treated with 100 mM picrotoxin dissolved in 5% DMSO for 5 min and then were hand-picked on to the center of a 1-cm diameter circle on a water agar plate. After 20 min, nematodes outside the circle were counted, and the ratio of this number to the total number of test nematodes was calculated
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