Summary of Study ST002551

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 PR001643. The data can be accessed directly via it's Project DOI: 10.21228/M8B41R 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	ST002551
Study Title	Metabolomics dataset of CNTF induced axon regeneration in mice post optic nerve crush
Study Summary	Axons are processes or extensions of a neuron that help connect one neuron with the next. In the eye all retinal ganglion cells (RGCs) reside within the retina but their axons travel a very long distance traversing through the optic nerve they connect with other neurons in the lateral geniculate nucleus in the brain. Loss of axons results in blindness in glaucoma and traumatic optic neuropathies. Optic nerve crush (ONC) is mouse is an assay system that enable pharmacological induction of axon regeneration from existing RGCs. Lipids form the outer boundary of axons, their synthesis or alterations are associated with metabolite changes. Our motivation was to understand what metabolite changes occurred when ONC axons regenerated due to ciliary neurotrophic factor (CNTF) treatment. We found metabolite profile changes associated with regeneration after crush induced by CNTF. This metabolite dataset was collected from C57Bl/6 mice expressing either AAV2-CNTF to promote regeneration or AAV2-Green Lantern as a control. Animals were subjected to optic nerve crush injury and allowed to recover for either 7 days or 14 days. At the respective time points, animals were euthanized and optic nerves were collected. Nerves underwent two rounds of extraction using a Precellys 24 Touch Homogenizer and a two solvent system of 1:1 Methanol/Water and 8:1:1 Acetonitrile/Methanol/Acetone. Metabolites were analyzed by untargeted liquid chromatography-mass spectrometry (LC MS-MS) using a Vanquish Horizon Binary HPLC coupled to a Q Exactive Orbitrap mass spectrometer. Metabolites were identified using Compound Discoverer 3.3 and quantified using isotopic internal metabolite standards.
Institute	University of Miami
Department	McKnight - Ophthalmology
Laboratory	Bhattacharya Lab
Last Name	Bhattacharya
First Name	Sanjoy
Address	1638 NW 10th Avenue, Room 706-A, Miami, FL 33136
Email	sbhattacharya@med.miami.edu
Phone	3054824103
Submit Date	2023-04-05
Num Groups	4
Total Subjects	23
Raw Data Available	Yes
Raw Data File Type(s)	mzXML
Analysis Type Detail	LC-MS
Release Date	2023-04-21
Release Version	1

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Combined analysis:

Analysis ID	AN004200	AN004201
Analysis type	MS	MS
Chromatography type	HILIC	HILIC
Chromatography system	Thermo Vanquish	Thermo Vanquish
Column	Thermo Accucore Amide HILIC (150 x 2.1mm, 2.6um)	Thermo Accucore Amide HILIC (150 x 2.1mm, 2.6um)
MS Type	ESI	ESI
MS instrument type	Orbitrap	Orbitrap
MS instrument name	Thermo Q Exactive Orbitrap	Thermo Q Exactive Orbitrap
Ion Mode	POSITIVE	NEGATIVE
Units	µg/ml	µg/ml

MS:

MS ID:	MS003947
Analysis ID:	AN004200
Instrument Name:	Thermo Q Exactive Orbitrap
Instrument Type:	Orbitrap
MS Type:	ESI
MS Comments:	The samples were run using a Q ExactiveTM mass spectrometer coupled to a heated electrospray ionization (HESI) source. The spray voltage was set to 3.50 kV, capillary temperature to 350°C, sheath gas to 55, aux gas to 14, sweep gas to 4, and S-Lens RF Level to 30.0. The mass range was set to 67 – 1000 m/z, resolution 140,000 for full scan and 35,000 for ddMS2. AGC target was set to 1e6 for full scan and 2e5 for ddMS2. The max injection time (IT) was 100 seconds for full scan mode and 50 seconds for ddMS2. The number of microscans was 2, and normalized collision energy (NCE) was set to 20, 35, and 50. Samples were run in both positive and negative ion mode separately. The parameters for negative mode were the same except the spray voltage, which was set to 2.50 kV and capillary temperature to 380°C. Metabolites were identified from their Thermo.RAW scans using Compound DiscovererTM 3.3 software. Extraction blanks were used to determine and correct for reagent effects, allow for the creation of exclusions lists, mark background components, and filters the background components from the results table in Compound DiscovererTM 3.3. Pooled QCs were used for initial compound normalization and identification. All non-identified compounds were removed.
Ion Mode:	POSITIVE

MS ID:	MS003948
Analysis ID:	AN004201
Instrument Name:	Thermo Q Exactive Orbitrap
Instrument Type:	Orbitrap
MS Type:	ESI
MS Comments:	The samples were run using a Q ExactiveTM mass spectrometer coupled to a heated electrospray ionization (HESI) source. The spray voltage was set to 3.50 kV, capillary temperature to 350°C, sheath gas to 55, aux gas to 14, sweep gas to 4, and S-Lens RF Level to 30.0. The mass range was set to 67 – 1000 m/z, resolution 140,000 for full scan and 35,000 for ddMS2. AGC target was set to 1e6 for full scan and 2e5 for ddMS2. The max injection time (IT) was 100 seconds for full scan mode and 50 seconds for ddMS2. The number of microscans was 2, and normalized collision energy (NCE) was set to 20, 35, and 50. Samples were run in both positive and negative ion mode separately. The parameters for negative mode were the same except the spray voltage, which was set to 2.50 kV and capillary temperature to 380°C. Metabolites were identified from their Thermo.RAW scans using Compound DiscovererTM 3.3 software. Extraction blanks were used to determine and correct for reagent effects, allow for the creation of exclusions lists, mark background components, and filters the background components from the results table in Compound DiscovererTM 3.3. Pooled QCs were used for initial compound normalization and identification. All non-identified compounds were removed.
Ion Mode:	NEGATIVE