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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 control pigs. 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 Two FB+ neurons (g, blue, 1 short arrow, 1 long arrow), which are simultaneously SP+ (h, green, 1 short arrow) or SP-negative (h, green, 1 long arrow) and galanin-positive (GAL+, i, red, 1 short arrow) or GAL-negative (GAL-, i, red, 1 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 calbindin-positive (CB+, n, green, arrow) and SP+ (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-f001: Representative images of substance P-positive (SP+) dorsal root ganglia (DRG)-urinary bladder-projecting neurons (UBPN) in control pigs. 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 Two FB+ neurons (g, blue, 1 short arrow, 1 long arrow), which are simultaneously SP+ (h, green, 1 short arrow) or SP-negative (h, green, 1 long arrow) and galanin-positive (GAL+, i, red, 1 short arrow) or GAL-negative (GAL-, i, red, 1 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 calbindin-positive (CB+, n, green, arrow) and SP+ (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: In the control pigs, almost half of the FB+/SP+ neurons additionally contained pituitary adenylate cyclase activating peptide (PACAP, Figure 1a–c; 45.1% ± 1.3%). Many of the SP-positive BPSN revealed also immunoreactivity to CGRP (Figure 1d–f; 35.6% ± 8.2%). A small number of these neurons (3.7% ± 0.3%) stained for galanin (GAL; Figure 1g–i) but only single FB+/SP+ neuronal somata were neuronal nitric oxide synthase (nNOS; Figure 1j–l)-, somatostatin (SOM; Figure 1p–s)- or calbindin (CB; Figure 1 m–o)-immunopositive (1.8% ± 0.6%, 1.2% ± 0.7% and 0.7% ± 0.7%, respectively).


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 control pigs. 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 Two FB+ neurons (g, blue, 1 short arrow, 1 long arrow), which are simultaneously SP+ (h, green, 1 short arrow) or SP-negative (h, green, 1 long arrow) and galanin-positive (GAL+, i, red, 1 short arrow) or GAL-negative (GAL-, i, red, 1 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 calbindin-positive (CB+, n, green, arrow) and SP+ (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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Related In: Results  -  Collection

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toxins-07-04797-f001: Representative images of substance P-positive (SP+) dorsal root ganglia (DRG)-urinary bladder-projecting neurons (UBPN) in control pigs. 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 Two FB+ neurons (g, blue, 1 short arrow, 1 long arrow), which are simultaneously SP+ (h, green, 1 short arrow) or SP-negative (h, green, 1 long arrow) and galanin-positive (GAL+, i, red, 1 short arrow) or GAL-negative (GAL-, i, red, 1 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 calbindin-positive (CB+, n, green, arrow) and SP+ (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: In the control pigs, almost half of the FB+/SP+ neurons additionally contained pituitary adenylate cyclase activating peptide (PACAP, Figure 1a–c; 45.1% ± 1.3%). Many of the SP-positive BPSN revealed also immunoreactivity to CGRP (Figure 1d–f; 35.6% ± 8.2%). A small number of these neurons (3.7% ± 0.3%) stained for galanin (GAL; Figure 1g–i) but only single FB+/SP+ neuronal somata were neuronal nitric oxide synthase (nNOS; Figure 1j–l)-, somatostatin (SOM; Figure 1p–s)- or calbindin (CB; Figure 1 m–o)-immunopositive (1.8% ± 0.6%, 1.2% ± 0.7% and 0.7% ± 0.7%, respectively).

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