Cellular manganese content is developmentally regulated in human dopaminergic neurons

Kumar, Kevin K.; Lowe, Edward W.; Aboud, Asad A.; Neely, M. Diana; Redha, Rey; Bauer, Joshua A.; Odak, Mihir; Weaver, C. David; Meiler, Jens; Aschner, Michael; Bowman, Aaron B.

doi:10.1038/srep06801

Cited by 73 publications

(70 citation statements)

References 57 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Control iPSC lines CD12 and CC3 as well as iPSC line SM14, from a preclinical PD patient carrying compound heterozygous loss-of-function mutations in PARK2 associated with familial early-onset PD [34–36], were differentiated to BMECs using E6 medium (Fig. 1).…”

Section: Resultsmentioning

confidence: 99%

Accelerated differentiation of human induced pluripotent stem cells to blood–brain barrier endothelial cells

Hollmann

Bailey

Potharazu

et al. 2017

Fluids Barriers CNS

Self Cite

148

186

View full text Add to dashboard Cite

BackgroundDue to their ability to limitlessly proliferate and specialize into almost any cell type, human induced pluripotent stem cells (iPSCs) offer an unprecedented opportunity to generate human brain microvascular endothelial cells (BMECs), which compose the blood–brain barrier (BBB), for research purposes. Unfortunately, the time, expense, and expertise required to differentiate iPSCs to purified BMECs precludes their widespread use. Here, we report the use of a defined medium that accelerates the differentiation of iPSCs to BMECs while achieving comparable performance to BMECs produced by established methods.MethodsInduced pluripotent stem cells were seeded at defined densities and differentiated to BMECs using defined medium termed E6. Resultant purified BMEC phenotypes were assessed through trans-endothelial electrical resistance (TEER), fluorescein permeability, and P-glycoprotein and MRP family efflux transporter activity. Expression of endothelial markers and their signature tight junction proteins were confirmed using immunocytochemistry. The influence of co-culture with astrocytes and pericytes on purified BMECs was assessed via TEER measurements. The robustness of the differentiation method was confirmed across independent iPSC lines.ResultsThe use of E6 medium, coupled with updated culture methods, reduced the differentiation time of iPSCs to BMECs from thirteen to 8 days. E6-derived BMECs expressed GLUT-1, claudin-5, occludin, PECAM-1, and VE-cadherin and consistently achieved TEER values exceeding 2500 Ω × cm2 across multiple iPSC lines, with a maximum TEER value of 4678 ± 49 Ω × cm2 and fluorescein permeability below 1.95 × 10−7 cm/s. E6-derived BMECs maintained TEER above 1000 Ω × cm2 for a minimum of 8 days and showed no statistical difference in efflux transporter activity compared to BMECs differentiated by conventional means. The method was also found to support long-term stability of BMECs harboring biallelic PARK2 mutations associated with Parkinson’s Disease. Finally, BMECs differentiated using E6 medium responded to inductive cues from astrocytes and pericytes and achieved a maximum TEER value of 6635 ± 315 Ω × cm2, which to our knowledge is the highest reported in vitro TEER value to date.ConclusionsGiven the accelerated differentiation, equivalent performance, and reduced cost to produce BMECs, our updated methods should make iPSC-derived in vitro BBB models more accessible for a wide variety of applications.Electronic supplementary materialThe online version of this article (doi:10.1186/s12987-017-0059-0) contains supplementary material, which is available to authorized users.

show abstract

Section: Resultsmentioning

confidence: 99%

Accelerated differentiation of human induced pluripotent stem cells to blood–brain barrier endothelial cells

Hollmann

Bailey

Potharazu

et al. 2017

Fluids Barriers CNS

Self Cite

148

186

View full text Add to dashboard Cite

show abstract

“…TfR is a membrane protein with high affinity for Mn, which is expressed in neurons, microglia, astrocytes and the endothelial cells of the BBB [60]. When TfR recognizes and binds to Tf, the cell membrane expands inwardly and forms an endocytic vesicle, which brings in the Mn [67, 74]. Mn 3+ is a stronger oxidizing agent than Mn 2+ and it may cause severe oxidative stress.…”

Section: Main Textmentioning

confidence: 99%

“…It’s suggested that intracellular Mn levels are actively controlled by the cell and not exclusively by the BBB or blood-cerebrospinal fluid barrier. Furthermore, mechanisms regulating Mn content might be developmentally regulated in DAergic neurons reflecting the changing physiological demand [74]. …”

Section: Main Textmentioning

confidence: 99%

“Manganese-induced neurotoxicity: a review of its behavioral consequences and neuroprotective strategies”

Peres

Schettinger

Chen

et al. 2016

BMC Pharmacol Toxicol

Self Cite

279

183

View full text Add to dashboard Cite

Manganese (Mn) is an essential heavy metal. However, Mn’s nutritional aspects are paralleled by its role as a neurotoxicant upon excessive exposure. In this review, we covered recent advances in identifying mechanisms of Mn uptake and its molecular actions in the brain as well as promising neuroprotective strategies. The authors focused on reporting findings regarding Mn transport mechanisms, Mn effects on cholinergic system, behavioral alterations induced by Mn exposure and studies of neuroprotective strategies against Mn intoxication. We report that exposure to Mn may arise from environmental sources, occupational settings, food, total parenteral nutrition (TPN), methcathinone drug abuse or even genetic factors, such as mutation in the transporter SLC30A10. Accumulation of Mn occurs mainly in the basal ganglia and leads to a syndrome called manganism, whose symptoms of cognitive dysfunction and motor impairment resemble Parkinson’s disease (PD). Various neurotransmitter systems may be impaired due to Mn, especially dopaminergic, but also cholinergic and GABAergic. Several proteins have been identified to transport Mn, including divalent metal tranporter-1 (DMT-1), SLC30A10, transferrin and ferroportin and allow its accumulation in the central nervous system. Parallel to identification of Mn neurotoxic properties, neuroprotective strategies have been reported, and these include endogenous antioxidants (for instance, vitamin E), plant extracts (complex mixtures containing polyphenols and non-characterized components), iron chelating agents, precursors of glutathione (GSH), and synthetic compounds that can experimentally afford protection against Mn-induced neurotoxicity.

show abstract

“…Similarly, neurotoxic exposures to Mn have been shown to deplete striatal dopamine levels 7 . Mn is regulated at a cell-level across neuronal differentiation 18 and exposures to Mn have been associated with changes in energy metabolism 19, 20 . Furthermore, both in vitro and in vivo mouse models expressing mutant HTT exhibit reduced total Mn accumulation 21 .…”

Section: Introductionmentioning

confidence: 99%

Untargeted metabolic profiling identifies interactions between Huntington's disease and neuronal manganese status

et al. 2015

Self Cite

View full text Add to dashboard Cite

Manganese (Mn) is an essential micronutrient for development and function of the nervous system. Deficiencies in Mn transport have been implicated in the pathogenesis of Huntington’s disease (HD), an autosomal dominant neurodegenerative disorder characterized by loss of medium spiny neurons of the striatum. Brain Mn levels are highest in striatum and other basal ganglia structures, the most sensitive brain regions to Mn neurotoxicity. Mouse models of HD exhibit decreased striatal Mn accumulation and HD striatal neuron models are resistant to Mn cytotoxicity. We hypothesized that the observed modulation of Mn cellular transport is associated with compensatory metabolic responses to HD pathology. Here we use an untargeted metabolomics approach by performing ultraperformance liquid chromatography-ion mobility-mass spectrometry (UPLC-IM-MS) on control and HD immortalized mouse striatal neurons to identify metabolic disruptions under three Mn exposure conditions, low (vehicle), moderate (non-cytotoxic) and high (cytotoxic). Our analysis revealed lower metabolite levels of pantothenic acid, and glutathione (GSH) in HD striatal cells relative to control cells. HD striatal cells also exhibited lower abundance and impaired induction of isobutyryl carnitine in response to increasing Mn exposure. In addition, we observed induction of metabolites in the pentose shunt pathway in HD striatal cells after high Mn exposure. These findings provide metabolic evidence of an interaction between the HD genotype and biologically relevant levels of Mn in a striatal cell model with known HD by Mn exposure interactions. The metabolic phenotypes detected support existing hypotheses that changes in energetic processes underlie the pathobiology of both HD and Mn neurotoxicity.

show abstract

Cellular manganese content is developmentally regulated in human dopaminergic neurons

Cited by 73 publications

References 57 publications

Accelerated differentiation of human induced pluripotent stem cells to blood–brain barrier endothelial cells

Accelerated differentiation of human induced pluripotent stem cells to blood–brain barrier endothelial cells

“Manganese-induced neurotoxicity: a review of its behavioral consequences and neuroprotective strategies”

Untargeted metabolic profiling identifies interactions between Huntington's disease and neuronal manganese status

Contact Info

Product

Resources

About