GSH and Zinc Supplementation Attenuate Cadmium-Induced Cellular Stress and Stimulation of Choline Uptake in Cultured Neonatal Rat Choroid Plexus Epithelia.

Samantha Francis Stuart,Alice Villalobos

doi:10.3390/ijms22168857

Samantha Francis Stuart, Alice Villalobos

Open Access

https://doi.org/10.3390/ijms22168857

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Abstract

Choroid plexus (CP) sequesters cadmium and other metals, protecting the brain from these neurotoxins. These metals can induce cellular stress and modulate homeostatic functions of CP, such as solute transport. We previously showed in primary cultured neonatal rat CP epithelial cells (CPECs) that cadmium induced cellular stress and stimulated choline uptake at the apical membrane, which interfaces with cerebrospinal fluid in situ. Here, in CPECs, we characterized the roles of glutathione (GSH) and Zinc supplementation in the adaptive stress response to cadmium. Cadmium increased GSH and decreased the reduced GSH-to-oxidized GSH (GSSG) ratio. Heat shock protein-70 (Hsp70), heme oxygenase (HO-1), and metallothionein (Mt-1) were induced along with the catalytic and modifier subunits of glutamate cysteine ligase (GCL), the rate-limiting enzyme in GSH synthesis. Inhibition of GCL by l-buthionine sulfoximine (BSO) enhanced stress protein induction and stimulation of choline uptake by cadmium. Zinc alone did not induce Hsp70, HO-1, or GCL subunits, or modulate choline uptake. Zinc supplementation during cadmium exposure attenuated stress protein induction and stimulation of choline uptake; this effect persisted despite inhibition of GSH synthesis. These data indicated up-regulation of GSH synthesis promotes adaptation to cadmium-induced cellular stress in CP, but Zinc may confer cytoprotection independent of GSH.

Highlights

The choroid plexus (CP) epithelia form the blood–cerebrospinal fluid (CSF) barrier
The protocol used in this study to expose cells to 500 nM CdCl2 was similar to that implemented in our prior study of cadmium-induced effects in CP epithelial cells (CPECs) [17]
We previously showed that CPECs exposed for 12 h to 500 nM CdCl2 in serum-free medium (SFM) accumulated cadmium (Cd) [17], which was consistent with accumulation of CdCl2 in SFM (Cd) in the choroid plexus of cadmium-exposed rodents [8,9,10]

Summary

Introduction

The choroid plexus (CP) epithelia form the blood–cerebrospinal fluid (CSF) barrier. These transporting epithelia secrete CSF and selectively exchange inorganic and organic solute between blood and CSF, thereby regulating fluid/electrolyte balance, nutrient availability, and metabolite and xenobiotic clearance in CSF. Modulation of solute transport by physicochemical stressors, such as heavy metals, may compromise regulation of CSF volume and composition and disrupt central neural homeostasis. We previously showed in primary cultured neonatal rat CP epithelial cells (CPECs) that subchronic exposure to 500 nM cadmium induced an oxidative cellular stress response and stimulated apical uptake of choline [17]. Stimulation of choline uptake at the apical membrane of CPECs is analogous to increased removal of choline from the CSF compartment in situ. This could potentially limit central availability of choline and ACh. CP epithelia sequester cadmium and other metals, but mechanisms that might minimize stress modulation of solute transport and other functions critical to CNS homeostasis are not fully characterized

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