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. 2017 Mar 10;9(3):98.
doi: 10.3390/toxins9030098.

The Influence of Low Doses of Zearalenone and T-2 Toxin on Calcitonin Gene Related Peptide-Like Immunoreactive (CGRP-LI) Neurons in the ENS of the Porcine Descending Colon

Krystyna Makowska  1 , Kazimierz Obremski  2 , Lukasz Zielonka  3 , Slawomir Gonkowski  4
Affiliations

The Influence of Low Doses of Zearalenone and T-2 Toxin on Calcitonin Gene Related Peptide-Like Immunoreactive (CGRP-LI) Neurons in the ENS of the Porcine Descending Colon

Krystyna Makowska et al. Toxins (Basel). .

Abstract

The enteric nervous system (ENS) can undergo adaptive and reparative changes in response to physiological and pathological stimuli. These manifest primarily as alterations in the levels of active substances expressed by the enteric neuron. While it is known that mycotoxins can affect the function of the central and peripheral nervous systems, knowledge about their influence on the ENS is limited. Therefore, the aim of the present study was to investigate the influence of low doses of zearalenone (ZEN) and T-2 toxin on calcitonin gene related peptide-like immunoreactive (CGRP-LI) neurons in the ENS of the porcine descending colon using a double immunofluorescence technique. Both mycotoxins led to an increase in the percentage of CGRP-LI neurons in all types of enteric plexuses and changed the degree of co-localization of CGRP with other neuronal active substances, such as substance P, galanin, nitric oxide synthase, and cocaine- and amphetamine-regulated transcript peptide. The obtained results demonstrate that even low doses of ZEN and T-2 can affect living organisms and cause changes in the neurochemical profile of enteric neurons.

Keywords: CGRP; ENS; T-2 toxin; descending colon; pig; zearalenone.

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Conflict of interest statement

The authors declare that they have no conflict of interest.

Figures

Figure 1
Figure 1
Distribution pattern of nervous structures immunoreactive to protein gene-product 9.5 (PGP 9.5)—used as a panneronal marker and calcitonin gene-related peptide (CGRP) in the wall of porcine descending colon under physiological conditions (a), and after T2-toxin (b) and zearalenone (c) administration; I—myenteric plexus; II—outer submucous plexus; III—inner submucous plexus; IV—circular muscle layer and V—submucous/mucous layer. CGRP-positive neurons (I,II,III) and nerve fibers (IV,V) are indicated by arrows. Images I, II, and III are composites of merged images taken separately from green (PGP 9.5) and red (CGRP) fluorescent channels. Images IV and V are performed using a single immunofluorescence technique.
Figure 2
Figure 2
Representative images of co-localization of calcitonin gene-related peptide (CGRP) with other neuronal active substances in the neurons of myenteric plexus (I,II,III) and nerve fibers scattered in the circular muscle layer (IV) of the porcine descending colon under physiological conditions (a), and after T2-toxin (b) and zearalenone (c) administration. I—co-localization of CGRP with substance P (SP); II—co-localization of CGRP with galanin (GAL); III—co-localization of CGRP with cocaine- and amphetamine-regulated transcript (CART) peptide; IV—co-localization of CGRP with neuronal isoform of nitric oxide synthase (nNOS). Images are composites of merged images taken separately from green (CGRP) and red (other substances studied) fluorescent channels. Nervous structures, where CGRP co-localizes with other substances, are indicated by arrows.
Figure 3
Figure 3
Representative images of co-localization of calcitonin gene-related peptide (CGRP) with other neuronal active substances in the neurons of the outer submucous plexus in the wall of porcine descending colon under physiological conditions (a), and after T2-toxin (b) and zearalenone (c) administration. I—co-localization of CGRP with substance P (SP); II—co-localization of CGRP with galanin (GAL); III—co-localization of CGRP with cocaine- and amphetamine- regulated transcript (CART) peptide. Images are composites of merged images taken separately from green (CGRP) and red (other substances studied) fluorescent channels. Neurons, where CGRP co-localizes with other substances are indicated by arrows.
Figure 4
Figure 4
Representative images of co-localization of calcitonin gene related peptide (CGRP) with other neuronal active substances in the neurons of inner submucous plexus (I,II,III) and nerve fibers scattered in submucous/mucous layer (IV) of the porcine descending colon under physiological conditions (a), and after T2-toxin (b) and zearalenone (c) administration. I—co-localization of CGRP with substance P (SP); II—co-localization of CGRP with galanin (GAL); III—co-localization of CGRP with cocaine- and amphetamine- regulated transcript (CART) peptide; IV—co-localization of CGRP with neuronal isoform of nitric oxide synthase (nNOS). Images are composites of merged images taken separately from green (CGRP) and red (other substances studied) fluorescent channels. Nervous structures, where CGRP co-localizes with other substances are indicated by arrows.
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