Summary of Study ST002047
This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001294. The data can be accessed directly via it's Project DOI: 10.21228/M8FQ39 This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.
| Study ID | ST002047 |
| Study Title | Lyso-lipid induced oligodendrocytes maturation underlie restoration of optic nerve function |
| Study Summary | Protein hyper-deimination and deficiency of lyso-phospholipids (LPC 18:1) has been associated with the pathology of demyelinating disease in both humans and mice. We uncovered interesting biology of LPC 18:1, in which LPC 18:1 induced optic nerve function restoration through oligodendrocyte maturation and remyelination in mouse model systems. Our in vitro studies show LPC 18:1 protection against neuron-ectopic hyper-deimination and stimulation of oligodendrocyte maturation, while in vivo investigations recorded optic nerve function improvement following optic nerve injections of LPC 18:1, in contrast to LPC 18:0. Thus just a change in a single bond renders a dramatic alternation in biological function. The incorporation of isobaric C13-histidine in newly synthesized myelin proteins and quantitative proteome shifts are consistent with remyelination underlying restoration in optic nerve function. These results suggest that exogenous LPC 18:1 may provide a therapeutic avenue for stemming vision loss in demyelinating diseases. |
| Institute | University of Miami |
| Last Name | Bhattacharya |
| First Name | Sanjoy K. |
| Address | 1638 NW 10th Avenue, Room 706-A, Miami, FL 33136 |
| sbhattacharya@med.miami.edu | |
| Phone | 3054824103 |
| Submit Date | 2021-12-16 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2022-01-21 |
| Release Version | 1 |
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Collection:
| Collection ID: | CO002122 |
| Collection Summary: | RGC cell isolation. Primary retinal ganglion cells (RGCs) were isolated according to the two-step immunopanning protocol described in a previous study (Dvoriantchikova, Degterev et al., 2014). Briefly, whole retinas of 10 postnatal day 11-12 pups were incubated in papain solution (16.5 U/mL; Worthington Biochemical, LS003127) for 30 minutes. Macrophaged and endothelial cells were removed from the cell suspension by panning with anti-macrophage antiserum (Accurate Chemical, AIA31240). RGCs were bound to the panning plates containing the antibody against Thy1.2 (VWR, 102646-060) and were then released by incubation with trypsin solution (Sigma, T9935). RGCs were plated in a plate coated with poly-d-lysine (Sigma, P6407) and Laminin (Sigma, L-6274). RGCs were either not treated (addition of RGC growth media), treated with LPC 18:0 at 10 µM in RGC growth media, or treated with LPC 18:1 at 10 µM in RGC growth media. RGCs were incubated for 24 hours and harvested the next day. OPC cell culture. Oligodendrocyte Precursor Cells (OPC) were purchased from ScienceCell Research Laboratories (R1600, Carlsbad, CA 92008) and cultured according to manufacturer’s recommendations. In brief, plates were coated with poly-d-lysine (Sigma, P6407; 2 µg/cm2) and incubated overnight at 37ºC. OPC growth media was prepared using OPC media (ScienceCell Research Laboratories, 1601) supplemented with OPC growth supplement (ScienceCell Research Laboratories, 1652) and penicillin/streptomycin solution (ScienceCell Research Laboratories, 0503). OPC differentiation medium was prepared using OPC media (ScienceCell Research Laboratories, 1601) supplemented with OPC differentiation supplement (ScienceCell Research Laboratories, 1672), fetal bovine serum (ScienceCell Research Laboratories, 0005) and penicillin/streptomycin solution (ScienceCell Research Laboratories, 0503). Poly-d-lysine coated plate was washed twice with sterile water, OPC growth media was added to each well and plate was placed in incubator to equilibrate for 15 minutes (37ºC and 5% CO2). Frozen vial containing OPCs was warmed in 37ºC water bath and gently rotated to equally suspend cells. 15,000 cells/cm2 were seeded and left overnight (~16 to ~18 hours) in incubator. Media was changed daily for 2 days. OPCs were either not treated (addition of OPC growth media), differentiated (addition of OPC differentiation media), treated with LPC 18:0 at 10 µM in OPC growth media, or treated with LPC 18:1 at 10 µM in OPC growth media. OPCs were incubated for 24 hours and harvested the next day. OPCs and oligodendrocytes were further validated using mRNA analysis following published methods (Jäkel, Agirre et al., 2019). |
| Sample Type: | Cultured cells |
