Zinc Transporter 3 (Znt3) as an Active Substance in the Enteric Nervous System of the Porcine Esophagus

Abstract

Zinc transporter 3 (ZnT3), a member of the SLC 30 zinc transporter family, is involved in the transport of zinc ions from the cytoplasm into synaptic vesicles or intracellular organelles. The aim of the present study was to investigate for the first time the percentage of ZnT3-like immunoreactive (ZnT3-LI) neurons in the enteric nervous system (ENS) of the porcine esophagus and denotation of their neurochemical coding. Routine double- and triple-immunofluorescence labeling of cervical, thoracic, and abdominal fragments of esophagus for ZnT3 with protein gene product (PGP 9.5; used as pan-neuronal marker), nitric oxide synthase (NOS), somatostatin, vasoactive intestinal peptide (VIP), vesicular acetylcholine transporter (VAChT), neuropeptide Y (NPY), and galanin (GAL) was performed. The percentage of ZnT3-LI neurons in myenteric ganglia amounted to 50.2 ± 4.7, 63.4 ± 8.3, and 77.1 ± 1.1 % of all PGP 9.5-like immunoreactive neuronal cells in cervical, thoracic, and abdominal esophagus, respectively. In submucous ganglia, these values in particular parts of esophagus amounted to 46.3 ± 6.3, 81.0 ± 8.1, and 74.4 ± 4.4 %. Znt3 co-localized mainly with VAChT, NPY, GAL, NOS, and VIP, but the degree of co-localization depended on the “kind” of enteric ganglia and part of esophagus studied. The obtained results suggest that both ZnT3 and zinc ions may play important and various roles in the neuronal regulation of esophageal functions.

Immunohistochemical characteristics of porcine intrahepatic nerves under physiological conditions and after Bisphenol A administration

“…After at least 2 weeks, the fragments of liver were frozen at -23°C and cut into 10 μm-thick sections using a microtome (Microm, HM 525, Walldorf, Germany). The sections were subjected to a routine single-labelling immunofluorescence technique according to the method described previously by Gonkowski and Wojtkiewicz [17,18,35,59]. A condensed description of the method is as follows: 45 min of drying; incubation with a blocking solution, which included 10% normal goat serum, 0.1% bovine serum albumin, 0.01% NaN3, Triton ×-100 and thimerozal in phosphate buffered saline (PBS) for 1 h; overnight incubation with a polyclonal "primary" antibody directed towards SP, GAL, PACAP, CGRP, or CART; incubation (for 1 h) with species-specific antisera conjugated to fluorescein isothiocyanate (FITC) or biotin, which was visualised by a streptavidin-CY3 complex (the specification of primary and secondary antibodies used in the present study is shown in Table 1).…”

Zinc Signals and Immunity

“…In this regard, Zrt-like, Irt-like protein (ZIP)4 is highly important since it is expressed along the entire GI tract acting as a major processor of zinc uptake into enterocytes from the apical membrane [ 26 ]. Moreover, zinc transporter (ZnT)3, is highly expressed in the human large and porcine small intestine and the esophagus [ 27 , 28 ]. Herein, its concrete function in the GI tract is largely unknown.…”

Neurochemical characterization of intramural nerve fibres in the porcine oesophagus

“…Moreover, it is known that besides extrinsic cholinergic innervation, the oesophageal motor endplates also receive information from enteric neurons located in ganglionated plexuses positioned in the wall of the oesophagus (Wu et al., ). This information in pigs is transmitted with neuronal factors typical of the enteric nervous system (Wojtkiewicz et al., ). The wide range of such substances that may be involved in the innervation of the oesophageal motor endplates was confirmed by this experiment.…”

“…The neurochemical characterization of neuronal structures located in the stomach and intestine is relatively well known both under physiological conditions and during various pathological processes (Brown & Gulbransen, ; Ferri et al., ; Gonkowski et al., ). Contrary to the abovementioned fragments of the GI tract, the knowledge of the chemical coding of the neuronal structures in the oesophagus, especially in pigs, is rather scanty (Wojtkiewicz, Makowska, Bejer‐Olenska, & Gonkowski, ; Wu et al., ). For this reason, the aim of this study was to investigate the neurochemical characterization of nerve fibres located in the muscular and mucosal layers of various parts of the porcine oesophagus.…”