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Changes in chemical coding of sympathetic chain ganglia (SChG) neurons supplying porcine urinary bladder after botulinum toxin (BTX) treatment.

Lepiarczyk E, Bossowska A, Majewski M - Cell Tissue Res. (2015)

Bottom Line: Botulinum toxin (BTX) is a neurotoxin used in medicine as an effective drug in experimental therapy of neurogenic urinary bladder disorders.BTX injections caused a decrease in the number of FB+/DβH+ neurons that were immunopositive to NPY (39.5 ± 4.5 % vs 74.5 ± 11.9 %), VIP (8.9 ± 5.3 % vs 22.3 ± 8.8 %), SOM (5.8 ± 2.3 % vs 17.4 ± 3.7 %) or GAL (0.9 ± 1.2 % vs 5.4 ± 4.4 %) and a distinct increase in the number of FB+/DβH+ neurons that were immunoreactive to L-ENK (3.7 ± 2.9 % vs 1.1 % ± 0.8 %) or nNOS (7.7 ± 3.5 % vs 0.8 ± 0.6 %).Our study provides novel evidence that the therapeutic effects of BTX on the mammalian urinary bladder are partly mediated by SChG neurons.

View Article: PubMed Central - PubMed

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

ABSTRACT
Botulinum toxin (BTX) is a neurotoxin used in medicine as an effective drug in experimental therapy of neurogenic urinary bladder disorders. We have investigated the influence of BTX on the chemical coding of sympathetic chain ganglia (SChG) neurons supplying the porcine urinary bladder. The toxin was injected into the wall of the bladder. SChG neurons were visualized by a retrograde tracing method with fluorescent tracer fast blue (FB) and their chemical coding was investigated by double-labelling immunohistochemistry with antibodies against dopamine β-hydroxylase (DβH; a marker of noradrenergic neurons), neuropeptide Y (NPY), vasoactive intestinal polypeptide (VIP), somatostatin (SOM), galanin (GAL), Leu(5)-enkephalin (L-ENK) and neuronal nitric oxide synthase (nNOS). In both the control (n = 5) and BTX-treated pigs (n = 5), the vast majority (91 ± 2.3 % and 89.8 ± 2.5 %, respectively) of FB-positive (FB+) nerve cells were DβH+. BTX injections caused a decrease in the number of FB+/DβH+ neurons that were immunopositive to NPY (39.5 ± 4.5 % vs 74.5 ± 11.9 %), VIP (8.9 ± 5.3 % vs 22.3 ± 8.8 %), SOM (5.8 ± 2.3 % vs 17.4 ± 3.7 %) or GAL (0.9 ± 1.2 % vs 5.4 ± 4.4 %) and a distinct increase in the number of FB+/DβH+ neurons that were immunoreactive to L-ENK (3.7 ± 2.9 % vs 1.1 % ± 0.8 %) or nNOS (7.7 ± 3.5 % vs 0.8 ± 0.6 %). Our study provides novel evidence that the therapeutic effects of BTX on the mammalian urinary bladder are partly mediated by SChG neurons.

No MeSH data available.


Related in: MedlinePlus

Representative images of sympathetic chain ganglia urinary bladder-projecting neurons (SChG-UBPN) in control pigs. Micrographs were taken separately from the blue fluorescent channel (a, c, e, g) and corresponding images taken separately from the red fluorescent channel demonstrating the results of the preabsorption procedure for neuropeptide Y (P NPY, b), vasoactive intestinal polypeptide (P VIP, d), somatostatin (P SOM, f) or Leu5-enkephalin (P L-ENK, h). The preabsorption of the specific antiserum with an appropriate antigen completely eliminated positive staining (b, d, f, h). Bar 100 μm (a–d, g, h), 50 μm (e, f)
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Fig1: Representative images of sympathetic chain ganglia urinary bladder-projecting neurons (SChG-UBPN) in control pigs. Micrographs were taken separately from the blue fluorescent channel (a, c, e, g) and corresponding images taken separately from the red fluorescent channel demonstrating the results of the preabsorption procedure for neuropeptide Y (P NPY, b), vasoactive intestinal polypeptide (P VIP, d), somatostatin (P SOM, f) or Leu5-enkephalin (P L-ENK, h). The preabsorption of the specific antiserum with an appropriate antigen completely eliminated positive staining (b, d, f, h). Bar 100 μm (a–d, g, h), 50 μm (e, f)

Mentions: Standard controls, i.e. preabsorption for the neuropeptide antisera (20 μg appropriate antigen per 1 ml corresponding antibody at working dilution; all antigens purchased from Peninsula, Sigma or Dianova, Fig. 1) or omission and replacement of the respective primary antiserum with the corresponding non-immune sera, completely abolished immunofluorescence and eliminated specific staining.Fig. 1


Changes in chemical coding of sympathetic chain ganglia (SChG) neurons supplying porcine urinary bladder after botulinum toxin (BTX) treatment.

Lepiarczyk E, Bossowska A, Majewski M - Cell Tissue Res. (2015)

Representative images of sympathetic chain ganglia urinary bladder-projecting neurons (SChG-UBPN) in control pigs. Micrographs were taken separately from the blue fluorescent channel (a, c, e, g) and corresponding images taken separately from the red fluorescent channel demonstrating the results of the preabsorption procedure for neuropeptide Y (P NPY, b), vasoactive intestinal polypeptide (P VIP, d), somatostatin (P SOM, f) or Leu5-enkephalin (P L-ENK, h). The preabsorption of the specific antiserum with an appropriate antigen completely eliminated positive staining (b, d, f, h). Bar 100 μm (a–d, g, h), 50 μm (e, f)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4544485&req=5

Fig1: Representative images of sympathetic chain ganglia urinary bladder-projecting neurons (SChG-UBPN) in control pigs. Micrographs were taken separately from the blue fluorescent channel (a, c, e, g) and corresponding images taken separately from the red fluorescent channel demonstrating the results of the preabsorption procedure for neuropeptide Y (P NPY, b), vasoactive intestinal polypeptide (P VIP, d), somatostatin (P SOM, f) or Leu5-enkephalin (P L-ENK, h). The preabsorption of the specific antiserum with an appropriate antigen completely eliminated positive staining (b, d, f, h). Bar 100 μm (a–d, g, h), 50 μm (e, f)
Mentions: Standard controls, i.e. preabsorption for the neuropeptide antisera (20 μg appropriate antigen per 1 ml corresponding antibody at working dilution; all antigens purchased from Peninsula, Sigma or Dianova, Fig. 1) or omission and replacement of the respective primary antiserum with the corresponding non-immune sera, completely abolished immunofluorescence and eliminated specific staining.Fig. 1

Bottom Line: Botulinum toxin (BTX) is a neurotoxin used in medicine as an effective drug in experimental therapy of neurogenic urinary bladder disorders.BTX injections caused a decrease in the number of FB+/DβH+ neurons that were immunopositive to NPY (39.5 ± 4.5 % vs 74.5 ± 11.9 %), VIP (8.9 ± 5.3 % vs 22.3 ± 8.8 %), SOM (5.8 ± 2.3 % vs 17.4 ± 3.7 %) or GAL (0.9 ± 1.2 % vs 5.4 ± 4.4 %) and a distinct increase in the number of FB+/DβH+ neurons that were immunoreactive to L-ENK (3.7 ± 2.9 % vs 1.1 % ± 0.8 %) or nNOS (7.7 ± 3.5 % vs 0.8 ± 0.6 %).Our study provides novel evidence that the therapeutic effects of BTX on the mammalian urinary bladder are partly mediated by SChG neurons.

View Article: PubMed Central - PubMed

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

ABSTRACT
Botulinum toxin (BTX) is a neurotoxin used in medicine as an effective drug in experimental therapy of neurogenic urinary bladder disorders. We have investigated the influence of BTX on the chemical coding of sympathetic chain ganglia (SChG) neurons supplying the porcine urinary bladder. The toxin was injected into the wall of the bladder. SChG neurons were visualized by a retrograde tracing method with fluorescent tracer fast blue (FB) and their chemical coding was investigated by double-labelling immunohistochemistry with antibodies against dopamine β-hydroxylase (DβH; a marker of noradrenergic neurons), neuropeptide Y (NPY), vasoactive intestinal polypeptide (VIP), somatostatin (SOM), galanin (GAL), Leu(5)-enkephalin (L-ENK) and neuronal nitric oxide synthase (nNOS). In both the control (n = 5) and BTX-treated pigs (n = 5), the vast majority (91 ± 2.3 % and 89.8 ± 2.5 %, respectively) of FB-positive (FB+) nerve cells were DβH+. BTX injections caused a decrease in the number of FB+/DβH+ neurons that were immunopositive to NPY (39.5 ± 4.5 % vs 74.5 ± 11.9 %), VIP (8.9 ± 5.3 % vs 22.3 ± 8.8 %), SOM (5.8 ± 2.3 % vs 17.4 ± 3.7 %) or GAL (0.9 ± 1.2 % vs 5.4 ± 4.4 %) and a distinct increase in the number of FB+/DβH+ neurons that were immunoreactive to L-ENK (3.7 ± 2.9 % vs 1.1 % ± 0.8 %) or nNOS (7.7 ± 3.5 % vs 0.8 ± 0.6 %). Our study provides novel evidence that the therapeutic effects of BTX on the mammalian urinary bladder are partly mediated by SChG neurons.

No MeSH data available.


Related in: MedlinePlus