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Botulinum toxin type A induces changes in the chemical coding of substance P-immunoreactive dorsal root ganglia sensory neurons supplying the porcine urinary bladder.

Bossowska A, Lepiarczyk E, Mazur U, Janikiewicz P, Markiewicz W - Toxins (Basel) (2015)

Bottom Line: Nearly half of these neurons co-expressed PACAP or CGRP (45% and 35%, respectively), while co-localization of SP with GAL, nNOS, SOM or CB was found less frequently (3.7%, 1.8%, 1.2%, and 0.7%, respectively).In BTX-treated pigs (n = 6), toxin-injections caused a decrease in the number of SP-IR cells containing CGRP, SOM or CB (16.2%, 0.5%, and 0%, respectively) and a distinct increase in these nerve cells immunopositive to GAL (27.2%).The present study demonstrates that BTX significantly modifies the chemical phenotypes of SP-IR BPSN.

View Article: PubMed Central - PubMed

Affiliation: Department of Human Physiology, Faculty of Medical Sciences, University of Warmia and Mazury in Olsztyn, Warszawska 30, Olsztyn 10-082, Poland. agnieszka.bossowska@uwm.edu.pl.

ABSTRACT
Botulinum toxin (BTX) is a potent neurotoxin which blocks acetylcholine release from nerve terminals, and therefore leads to cessation of somatic motor and/or parasympathetic transmission. Recently it has been found that BTX also interferes with sensory transmission, thus, the present study was aimed at investigating the neurochemical characterization of substance P-immunoreactive (SP-IR) bladder-projecting sensory neurons (BPSN) after the toxin treatment. Investigated neurons were visualized with retrograde tracing method and their chemical profile was disclosed with double-labelling immunohistochemistry using antibodies against SP, calcitonin gene-related peptide (CGRP), pituitary adenylate cyclase activating polypeptide (PACAP), neuronal nitric oxide synthase (nNOS), galanin (GAL), calbindin (CB), and somatostatin (SOM). In the control group (n = 6), 45% of the total population of BPSN were SP-IR. Nearly half of these neurons co-expressed PACAP or CGRP (45% and 35%, respectively), while co-localization of SP with GAL, nNOS, SOM or CB was found less frequently (3.7%, 1.8%, 1.2%, and 0.7%, respectively). In BTX-treated pigs (n = 6), toxin-injections caused a decrease in the number of SP-IR cells containing CGRP, SOM or CB (16.2%, 0.5%, and 0%, respectively) and a distinct increase in these nerve cells immunopositive to GAL (27.2%). The present study demonstrates that BTX significantly modifies the chemical phenotypes of SP-IR BPSN.

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Representative images of substance P-positive (SP+) dorsal root ganglia (DRG)-urinary bladder-projecting neurons (UBPN) in BTX-treated animals. All images were taken separately from blue (a,d,g,j,m,p), green (b,e,h,k,n,r) and red (c,f,i,l,o,s) fluorescent channels; a–c one fast blue-positive (FB+) neuron (a, blue, arrow), which simultaneously contains SP (b, green, arrow) and pituitary adenylate cyclase activating peptide-PACAP (c, red, arrow). d–f One FB+ neuron (d, blue, arrow), which is simultaneously SP+ (e, green, arrow) and calcitonin gene-related peptide-positive (CGRP+, f, red, arrow). g–i One FB+ neuron (g, blue, arrow), which is simultaneously SP+ (h, green, arrow) and galanin-positive (GAL+, i, red, arrow). j–l One FB+ neuron (j, blue, arrow), which simultaneously contains neuronal nitric oxide synthase-nNOS (k, green, arrow) and SP (l, red, arrow). m–o One FB+ neuron (m, blue, arrow), which is simultaneously SP+ (n, green, arrow) and calbindin-negative (CB-, o, red, arrow). p-s One FB+ neuron (p, blue, arrow), which simultaneously contains SP (r, green, arrow) and somatostatin-SOM (s, red, arrow). Bars 50 μm (a–s).
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toxins-07-04797-f002: Representative images of substance P-positive (SP+) dorsal root ganglia (DRG)-urinary bladder-projecting neurons (UBPN) in BTX-treated animals. All images were taken separately from blue (a,d,g,j,m,p), green (b,e,h,k,n,r) and red (c,f,i,l,o,s) fluorescent channels; a–c one fast blue-positive (FB+) neuron (a, blue, arrow), which simultaneously contains SP (b, green, arrow) and pituitary adenylate cyclase activating peptide-PACAP (c, red, arrow). d–f One FB+ neuron (d, blue, arrow), which is simultaneously SP+ (e, green, arrow) and calcitonin gene-related peptide-positive (CGRP+, f, red, arrow). g–i One FB+ neuron (g, blue, arrow), which is simultaneously SP+ (h, green, arrow) and galanin-positive (GAL+, i, red, arrow). j–l One FB+ neuron (j, blue, arrow), which simultaneously contains neuronal nitric oxide synthase-nNOS (k, green, arrow) and SP (l, red, arrow). m–o One FB+ neuron (m, blue, arrow), which is simultaneously SP+ (n, green, arrow) and calbindin-negative (CB-, o, red, arrow). p-s One FB+ neuron (p, blue, arrow), which simultaneously contains SP (r, green, arrow) and somatostatin-SOM (s, red, arrow). Bars 50 μm (a–s).

Mentions: BTX-injections led to a significant decrease in the number of retrogradely labelled SP-IR cells simultaneously containing CGRP (Figure 2d–f; 16.2% ± 3.8%), SOM (Figure 2p–s; 0.5% ± 0.6%) or CB (Figure 2m–o; 0%) and caused a distinct increase in the SP- and GAL-IR BPSN (Figure 2g–i; 27.2% ± 0.4%). Furthermore, there were no significant differences in the number of FB+/SP+ sensory neurons expressing immunoreactivity to PACAP (Figure 2a–c; 45.7% ± 6.0%) or nNOS (Figure 2j–l; 1.8% ± 2.5%).


Botulinum toxin type A induces changes in the chemical coding of substance P-immunoreactive dorsal root ganglia sensory neurons supplying the porcine urinary bladder.

Bossowska A, Lepiarczyk E, Mazur U, Janikiewicz P, Markiewicz W - Toxins (Basel) (2015)

Representative images of substance P-positive (SP+) dorsal root ganglia (DRG)-urinary bladder-projecting neurons (UBPN) in BTX-treated animals. All images were taken separately from blue (a,d,g,j,m,p), green (b,e,h,k,n,r) and red (c,f,i,l,o,s) fluorescent channels; a–c one fast blue-positive (FB+) neuron (a, blue, arrow), which simultaneously contains SP (b, green, arrow) and pituitary adenylate cyclase activating peptide-PACAP (c, red, arrow). d–f One FB+ neuron (d, blue, arrow), which is simultaneously SP+ (e, green, arrow) and calcitonin gene-related peptide-positive (CGRP+, f, red, arrow). g–i One FB+ neuron (g, blue, arrow), which is simultaneously SP+ (h, green, arrow) and galanin-positive (GAL+, i, red, arrow). j–l One FB+ neuron (j, blue, arrow), which simultaneously contains neuronal nitric oxide synthase-nNOS (k, green, arrow) and SP (l, red, arrow). m–o One FB+ neuron (m, blue, arrow), which is simultaneously SP+ (n, green, arrow) and calbindin-negative (CB-, o, red, arrow). p-s One FB+ neuron (p, blue, arrow), which simultaneously contains SP (r, green, arrow) and somatostatin-SOM (s, red, arrow). Bars 50 μm (a–s).
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toxins-07-04797-f002: Representative images of substance P-positive (SP+) dorsal root ganglia (DRG)-urinary bladder-projecting neurons (UBPN) in BTX-treated animals. All images were taken separately from blue (a,d,g,j,m,p), green (b,e,h,k,n,r) and red (c,f,i,l,o,s) fluorescent channels; a–c one fast blue-positive (FB+) neuron (a, blue, arrow), which simultaneously contains SP (b, green, arrow) and pituitary adenylate cyclase activating peptide-PACAP (c, red, arrow). d–f One FB+ neuron (d, blue, arrow), which is simultaneously SP+ (e, green, arrow) and calcitonin gene-related peptide-positive (CGRP+, f, red, arrow). g–i One FB+ neuron (g, blue, arrow), which is simultaneously SP+ (h, green, arrow) and galanin-positive (GAL+, i, red, arrow). j–l One FB+ neuron (j, blue, arrow), which simultaneously contains neuronal nitric oxide synthase-nNOS (k, green, arrow) and SP (l, red, arrow). m–o One FB+ neuron (m, blue, arrow), which is simultaneously SP+ (n, green, arrow) and calbindin-negative (CB-, o, red, arrow). p-s One FB+ neuron (p, blue, arrow), which simultaneously contains SP (r, green, arrow) and somatostatin-SOM (s, red, arrow). Bars 50 μm (a–s).
Mentions: BTX-injections led to a significant decrease in the number of retrogradely labelled SP-IR cells simultaneously containing CGRP (Figure 2d–f; 16.2% ± 3.8%), SOM (Figure 2p–s; 0.5% ± 0.6%) or CB (Figure 2m–o; 0%) and caused a distinct increase in the SP- and GAL-IR BPSN (Figure 2g–i; 27.2% ± 0.4%). Furthermore, there were no significant differences in the number of FB+/SP+ sensory neurons expressing immunoreactivity to PACAP (Figure 2a–c; 45.7% ± 6.0%) or nNOS (Figure 2j–l; 1.8% ± 2.5%).

Bottom Line: Nearly half of these neurons co-expressed PACAP or CGRP (45% and 35%, respectively), while co-localization of SP with GAL, nNOS, SOM or CB was found less frequently (3.7%, 1.8%, 1.2%, and 0.7%, respectively).In BTX-treated pigs (n = 6), toxin-injections caused a decrease in the number of SP-IR cells containing CGRP, SOM or CB (16.2%, 0.5%, and 0%, respectively) and a distinct increase in these nerve cells immunopositive to GAL (27.2%).The present study demonstrates that BTX significantly modifies the chemical phenotypes of SP-IR BPSN.

View Article: PubMed Central - PubMed

Affiliation: Department of Human Physiology, Faculty of Medical Sciences, University of Warmia and Mazury in Olsztyn, Warszawska 30, Olsztyn 10-082, Poland. agnieszka.bossowska@uwm.edu.pl.

ABSTRACT
Botulinum toxin (BTX) is a potent neurotoxin which blocks acetylcholine release from nerve terminals, and therefore leads to cessation of somatic motor and/or parasympathetic transmission. Recently it has been found that BTX also interferes with sensory transmission, thus, the present study was aimed at investigating the neurochemical characterization of substance P-immunoreactive (SP-IR) bladder-projecting sensory neurons (BPSN) after the toxin treatment. Investigated neurons were visualized with retrograde tracing method and their chemical profile was disclosed with double-labelling immunohistochemistry using antibodies against SP, calcitonin gene-related peptide (CGRP), pituitary adenylate cyclase activating polypeptide (PACAP), neuronal nitric oxide synthase (nNOS), galanin (GAL), calbindin (CB), and somatostatin (SOM). In the control group (n = 6), 45% of the total population of BPSN were SP-IR. Nearly half of these neurons co-expressed PACAP or CGRP (45% and 35%, respectively), while co-localization of SP with GAL, nNOS, SOM or CB was found less frequently (3.7%, 1.8%, 1.2%, and 0.7%, respectively). In BTX-treated pigs (n = 6), toxin-injections caused a decrease in the number of SP-IR cells containing CGRP, SOM or CB (16.2%, 0.5%, and 0%, respectively) and a distinct increase in these nerve cells immunopositive to GAL (27.2%). The present study demonstrates that BTX significantly modifies the chemical phenotypes of SP-IR BPSN.

Show MeSH
Related in: MedlinePlus