Dataset Information

Effects of Interleukin-1? and -6 on the Expression of Ion Transporters Involved in Eggshell Mineralization in Cultured Hen Uterine Mucosal Tissue.

ABSTRACT: This study determined the effects of pro-inflammatory cytokines (interleukin (IL)-1? and IL-6) on the expression of eggshell mineralization-related ion transporters in the hen uterus mucosa. Uterine mucosal tissues collected from White Leghorn laying hens were cultured for 1.5 or 3 h in TCM-199 medium with or without 100 ng/mL recombinant chicken IL-1? or IL-6. Total RNA and protein were extracted from the cultured tissues for real-time polymerase chain reaction (PCR) and western blot analyses and some tissues were processed into paraffin sections for immunostaining with calcium-binding protein D28K (CaBP-D28K) antibody. The gene expression of CaBP-D28K, PMCA1, PMCA2 (plasma membrane calcium-transporting ATPase 1 and 2; calcium pumps), CA2 (carbonic anhydrase 2), and SLC26A9 (solute carrier family 26 member 9; HCO3 - transporter) was analyzed by real-time PCR and protein density of CaBP-D28K by western blotting. Expression of CaBP-D28K, PMCA1, PMCA2, CA2, and SLC26A9 was significantly higher in the tissues treated with IL-1? and IL-6 than in the control group at 1.5 h of incubation. Immunoreactive CaBP-D28K was localized in the uterine tubular gland cells in all groups, but its level was significantly lower in the tissues incubated for 1.5 h with IL-1? and IL-6 than in the control group. No significant differences were observed in the expression of all tested genes and CaBP-D28k content between the cytokine-treated and control groups at 3 h of incubation. These results suggest that IL-1? and IL-6 may not suppress the expression of genes related to Ca2+ and HCO3- transportation for eggshell formation, while CaBP-D28K protein content in uterine glandular cells was reduced by these cytokines during the early exposure phase. Thus, IL-1? and IL-6 induced by infections may disrupt the transportation of Ca2+ for eggshell formation through decreased CaBP-D28K content in the uterus.

SUBMITTER: Nii T

PROVIDER: S-EPMC6756487 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Effects of Interleukin-1β and -6 on the Expression of Ion Transporters Involved in Eggshell Mineralization in Cultured Hen Uterine Mucosal Tissue.

Nii Takahiro T Isobe Naoki N Yoshimura Yukinori Y

The journal of poultry science 20171125 2

This study determined the effects of pro-inflammatory cytokines (interleukin (IL)-1β and IL-6) on the expression of eggshell mineralization-related ion transporters in the hen uterus mucosa. Uterine mucosal tissues collected from White Leghorn laying hens were cultured for 1.5 or 3 h in TCM-199 medium with or without 100 ng/mL recombinant chicken IL-1β or IL-6. Total RNA and protein were extracted from the cultured tissues for real-time polymerase chain reaction (PCR) and western b ...[more]

PMID: 32055167

Similar Datasets

Project description:BACKGROUND: In Gallus gallus, eggshell formation takes place daily in the hen uterus and requires large amounts of the ionic precursors for calcium carbonate (CaCO3). Both elements (Ca2+, HCO3-) are supplied by the blood via trans-epithelial transport. Our aims were to identify genes coding for ion transporters that are upregulated in the uterine portion of the oviduct during eggshell calcification, compared to other tissues and other physiological states, and incorporate these proteins into a general model for mineral transfer across the tubular gland cells during eggshell formation. RESULTS: A total of 37 candidate ion transport genes were selected from our database of overexpressed uterine genes associated with eggshell calcification, and by analogy with mammalian transporters. Their uterine expression was compared by qRTPCR in the presence and absence of eggshell formation, and with relative expression levels in magnum (low Ca2+/HCO3- movement) and duodenum (high rates of Ca2+/HCO3- trans-epithelial transfer). We identified overexpression of eleven genes related to calcium movement: the TRPV6 Ca2+ channel (basolateral uptake of Ca2+), 28 kDa calbindin (intracellular Ca2+ buffering), the endoplasmic reticulum type 2 and 3 Ca2+ pumps (ER uptake), and the inositol trisphosphate receptors type 1, 2 and 3 (ER release). Ca2+ movement across the apical membrane likely involves membrane Ca2+ pumps and Ca2+/Na+ exchangers. Our data suggests that Na+ transport involved the SCNN1 channel and the Na+/Ca2+ exchangers SLC8A1, 3 for cell uptake, the Na+/K+ ATPase for cell output. K+ uptake resulted from the Na+/K+ ATPase, and its output from the K+ channels (KCNJ2, 15, 16 and KCNMA1).We propose that the HCO3- is mainly produced from CO2 by the carbonic anhydrase 2 (CA2) and that HCO3- is secreted through the HCO3-/Cl- exchanger SLC26A9. HCO3- synthesis and precipitation with Ca2+ produce two H+. Protons are absorbed via the membrane's Ca2+ pumps ATP2B1, 2 in the apical membrane and the vacuolar (H+)-atpases at the basolateral level. Our model incorporate Cl- ions which are absorbed by the HCO3-/Cl- exchanger SLC26A9 and by Cl- channels (CLCN2, CFTR) and might be extruded by Cl-/H+ exchanger (CLCN5), but also by Na+ K+ 2 Cl- and K+ Cl- cotransporters. CONCLUSIONS: Our Gallus gallus uterine model proposes a large list of ion transfer proteins supplying Ca2+ and HCO3- and maintaining cellular ionic homeostasis. This avian model should contribute towards understanding the mechanisms and regulation for ionic precursors of CaCO3, and provide insight in other species where epithelia transport large amount of calcium or bicarbonate.

Project description:The importance of phosphate (Pi) as an essential component of hydroxyapatite crystals suggests a key role for membrane proteins controlling Pi uptake during mineralization in the tooth. To clarify the involvement of the currently known Pi transporters (Slc17a1, Slc34a1, Slc34a2, Slc34a3, Slc20a1, Slc20a2, and Xpr1) during tooth development and mineralization, we determined their spatiotemporal expression in murine tooth germs from embryonic day 14.5 to postnatal day 15 and in human dental samples from Nolla stages 6 to 9. Using real-time polymerase chain reaction, in situ hybridization, immunohistochemistry, and X-gal staining, we showed that the expression of Slc17a1, Slc34a1, and Slc34a3 in tooth germs from C57BL/6 mice were very low. In contrast, Slc34a2, Slc20a1, Slc20a2, and Xpr1 were highly expressed, mostly during the postnatal stages. The expression of Slc20a2 was 2- to 10-fold higher than the other transporters. Comparable results were obtained in human tooth germs. In mice, Slc34a2 and Slc20a1 were predominantly expressed in ameloblasts but not odontoblasts, while Slc20a2 was detected neither in ameloblasts nor in odontoblasts. Rather, Slc20a2 was highly expressed in the stratum intermedium and the subodontoblastic cell layer. Although Slc20a2 knockout mice did not show enamel defects, mutant mice showed a disrupted dentin mineralization, displaying unmerged calcospherites at the mineralization front. This latter phenotypical finding raises the possibility that Slc20a2 may play an indirect role in regulating the extracellular Pi availability for mineralizing cells rather than a direct role in mediating Pi transport through mineralizing plasma cell membranes. By documenting the spatiotemporal expression of Pi transporters in the tooth, our data support the possibility that the currently known Pi transporters may be dispensable for the initiation of dental mineralization and may rather be involved later during the tooth mineralization scheme.

Dataset Information

Effects of Interleukin-1? and -6 on the Expression of Ion Transporters Involved in Eggshell Mineralization in Cultured Hen Uterine Mucosal Tissue.

Publications

Effects of Interleukin-1<i>β</i> and -6 on the Expression of Ion Transporters Involved in Eggshell Mineralization in Cultured Hen Uterine Mucosal Tissue.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets