Summary of Study ST000620

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 PR000453. The data can be accessed directly via it's Project DOI: 10.21228/M8H60M This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

Study ID	ST000620
Study Title	Sphingolipid Analysis of hyper and normotensive DBA2J mice aqueous humor and trabecular meshwork
Study Summary	To determine the differential profiles of sphingomyelin, sphingoid base, sphingoid base-1-phosphate, and ceramide and their quantitative differences between trabecular meshwork (TM) and aqueous humor (AH) derived from normotensive and hypertensive intraocular pressure states of DBA/2J mice.
Institute	University of Miami
Last Name	Bhattacharya
First Name	Sanjoy
Address	McKnight Vision Research Building, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, 1638 NW 10th Avenue, Room 706A, Miami, Florida 33136
Email	SBhattacharya@med.miami.edu
Phone	305-482-4103
Submit Date	2017-06-06
Raw Data Available	Yes
Raw Data File Type(s)	raw(Thermo)
Analysis Type Detail	MS(Dir. Inf.)
Release Date	2019-07-17
Release Version	1

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Collection:

Collection ID:	CO000637
Collection Summary:	Breeding pairs of DBA/2J mice originally procured from The Jackson Laboratory (Bar Harbor, ME) were maintained at the animal facility/McKnight vivarium at University of Miami Miller School of Medicine. All animal protocols were reviewed and approved by Institutional Animal Care and Use Committee (IACUC) and in accordance with the ARVO statement for use of animals in ophthalmic and vision research. Mice gender and ages used are as indicated in individual experiments with an n=40 for each time point unless stated otherwise. IOP measurements were obtained using a TonoLab instrument (Colonial Medical Supplies, NH). Average IOP from two weeks of twice daily measurements were calculated. We considered ~3-7.5 month old mice as normotensive (IOP ≤15 mm of Hg) and ~8-12 month old mice as hypertensive when carefully measured IOP showed an IOP ≥18mm of Hg at every single measurement. For the normotensive group no single IOP measurement had been >17 mm of Hg and average IOP over two weeks of measurement had been ≤15 mm of Hg. At 8-9 months of age the IOP elevation in DBA/2J mice is asynchronous and thus any mice that showed an IOP <18 mm of Hg at any single point measurement was excluded from the study. Normotensive and hypertensive AH was sampled from the anterior chamber of DBA/2J mice by paracentesis via syringe (catalog no. 7635-01 Hamilton, Reno, Nevada) with a small 33 gauge removable needle (701RN, catalog no. 7803-05 Hamilton, Reno, Nevada), which is a well established procedure. The syringe and needle were rinsed with LC-MS grade water 5 times preceding and following extraction to flush out impurities. To ensure that samples are contamination free, in between the samples, the wash from a syringe was also evaluated using mass spectrometry. Prior to collection, mice were anesthetized with an injection (0.1ml) of ketamine (100mg/kg) and xylazine (9mg/kg) administered intraperitoneally. A drop of tetracaine hydrochloride ophthalmic solution, 0.5% (Bausch & Lomb, USA) was used to anesthetize the ocular surface just prior to sample collection. A total of ~1-2 μl of AH was collected per animal and used for lipid extraction. Surgical removal of TM samples after euthanasia followed established protocols.

Collection ID:

CO000637

Collection Summary:

Breeding pairs of DBA/2J mice originally procured from The Jackson Laboratory (Bar Harbor, ME) were maintained at the animal facility/McKnight vivarium at University of Miami Miller School of Medicine. All animal protocols were reviewed and approved by Institutional Animal Care and Use Committee (IACUC) and in accordance with the ARVO statement for use of animals in ophthalmic and vision research. Mice gender and ages used are as indicated in individual experiments with an n=40 for each time point unless stated otherwise. IOP measurements were obtained using a TonoLab instrument (Colonial Medical Supplies, NH). Average IOP from two weeks of twice daily measurements were calculated. We considered ~3-7.5 month old mice as normotensive (IOP ≤15 mm of Hg) and ~8-12 month old mice as hypertensive when carefully measured IOP showed an IOP ≥18mm of Hg at every single measurement. For the normotensive group no single IOP measurement had been >17 mm of Hg and average IOP over two weeks of measurement had been ≤15 mm of Hg. At 8-9 months of age the IOP elevation in DBA/2J mice is asynchronous and thus any mice that showed an IOP <18 mm of Hg at any single point measurement was excluded from the study. Normotensive and hypertensive AH was sampled from the anterior chamber of DBA/2J mice by paracentesis via syringe (catalog no. 7635-01 Hamilton, Reno, Nevada) with a small 33 gauge removable needle (701RN, catalog no. 7803-05 Hamilton, Reno, Nevada), which is a well established procedure. The syringe and needle were rinsed with LC-MS grade water 5 times preceding and following extraction to flush out impurities. To ensure that samples are contamination free, in between the samples, the wash from a syringe was also evaluated using mass spectrometry. Prior to collection, mice were anesthetized with an injection (0.1ml) of ketamine (100mg/kg) and xylazine (9mg/kg) administered intraperitoneally. A drop of tetracaine hydrochloride ophthalmic solution, 0.5% (Bausch & Lomb, USA) was used to anesthetize the ocular surface just prior to sample collection. A total of ~1-2 μl of AH was collected per animal and used for lipid extraction. Surgical removal of TM samples after euthanasia followed established protocols.