Retinal sphingolipids and their very-long-chain fatty acid-containing species

Richard S. Brush, Julie Thu A. Tran, Kimberly R. Henry, Mark E. McClellan, Michael H. Elliott, Nawajes Mandal

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

PURPOSE. Recent evidence suggests that ceramide metabolism plays an important role in retinal photoreceptor cell survival and apoptosis. The purpose of this study was to characterize sphingolipids in the retina with special emphasis on the verylong- chain- containing saturated (VLC-FA) and polyunsaturated (VLC-PUFA) fatty acid- containing species. The VLC-FAs and VLC-PUFAs are synthesized by the ELOVL4 protein, which is involved in human Stargardt's macular dystrophy type 3 (STGD3). METHODS. Total lipids were extracted from retina and other tissues, and different sphingolipid classes were isolated and purified using various combinations of liquid- and solid-phase separation. Purified sphingolipids were analyzed by high-performance thin layer chromatography (HPTLC), gas chromatography (GC), and GC-MS (GC-mass spectrometry). RESULTS. Nonsialylated sphingolipids (NSLs) comprised ~3.5% of total retinal lipids of which 70% was sphingomyelin. Ceramide and glycosylceramides (GCs) constituted ≤1% of total retinal lipids. Gangliosides (GGs), on the other hand, comprised ~3.0% of total retinal lipids. Fatty acid analysis of retinal NSLs indicated an abundance of saturated fatty acids, with the presence of VLC-FAs but not of VLC-PUFAs beyond 24 carbons. However, GG had significant levels of unsaturated, polyunsaturated, and VLC-PUFAs. Retinal rod outer segments (ROS) contained ~1% each of NSL and GG, and their fatty acid profile was not very different from whole retinal NSL and GG, respectively. CONCLUSIONS. Retina has a total of 6% to 7% fatty acids that are N-linked to a sphingosine, which would be 11 to 13 mole % in comparison to phospholipids. The presence of VLC-FAs and VLC-PUFAs in retinal sphingolipids indicates that they may play role in ELOVL4-mediated Stargardt 3.

Original languageEnglish (US)
Pages (from-to)4422-4431
Number of pages10
JournalInvestigative Ophthalmology and Visual Science
Volume51
Issue number9
DOIs
StatePublished - Sep 1 2010

Fingerprint

Sphingolipids
Fatty Acids
Gangliosides
Lipids
Retina
Ceramides
Retinal Photoreceptor Cell Outer Segment
Rod Cell Outer Segment
Retinal Rod Photoreceptor Cells
Vertebrate Photoreceptor Cells
Sphingosine
Sphingomyelins
Macular Degeneration
Thin Layer Chromatography
Gas Chromatography
Gas Chromatography-Mass Spectrometry
Phospholipids
Cell Survival
Carbon
Apoptosis

All Science Journal Classification (ASJC) codes

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

Retinal sphingolipids and their very-long-chain fatty acid-containing species. / Brush, Richard S.; Tran, Julie Thu A.; Henry, Kimberly R.; McClellan, Mark E.; Elliott, Michael H.; Mandal, Nawajes.

In: Investigative Ophthalmology and Visual Science, Vol. 51, No. 9, 01.09.2010, p. 4422-4431.

Research output: Contribution to journalArticle

Brush, Richard S. ; Tran, Julie Thu A. ; Henry, Kimberly R. ; McClellan, Mark E. ; Elliott, Michael H. ; Mandal, Nawajes. / Retinal sphingolipids and their very-long-chain fatty acid-containing species. In: Investigative Ophthalmology and Visual Science. 2010 ; Vol. 51, No. 9. pp. 4422-4431.
@article{42d938f28308451384c5622d9d2809dc,
title = "Retinal sphingolipids and their very-long-chain fatty acid-containing species",
abstract = "PURPOSE. Recent evidence suggests that ceramide metabolism plays an important role in retinal photoreceptor cell survival and apoptosis. The purpose of this study was to characterize sphingolipids in the retina with special emphasis on the verylong- chain- containing saturated (VLC-FA) and polyunsaturated (VLC-PUFA) fatty acid- containing species. The VLC-FAs and VLC-PUFAs are synthesized by the ELOVL4 protein, which is involved in human Stargardt's macular dystrophy type 3 (STGD3). METHODS. Total lipids were extracted from retina and other tissues, and different sphingolipid classes were isolated and purified using various combinations of liquid- and solid-phase separation. Purified sphingolipids were analyzed by high-performance thin layer chromatography (HPTLC), gas chromatography (GC), and GC-MS (GC-mass spectrometry). RESULTS. Nonsialylated sphingolipids (NSLs) comprised ~3.5{\%} of total retinal lipids of which 70{\%} was sphingomyelin. Ceramide and glycosylceramides (GCs) constituted ≤1{\%} of total retinal lipids. Gangliosides (GGs), on the other hand, comprised ~3.0{\%} of total retinal lipids. Fatty acid analysis of retinal NSLs indicated an abundance of saturated fatty acids, with the presence of VLC-FAs but not of VLC-PUFAs beyond 24 carbons. However, GG had significant levels of unsaturated, polyunsaturated, and VLC-PUFAs. Retinal rod outer segments (ROS) contained ~1{\%} each of NSL and GG, and their fatty acid profile was not very different from whole retinal NSL and GG, respectively. CONCLUSIONS. Retina has a total of 6{\%} to 7{\%} fatty acids that are N-linked to a sphingosine, which would be 11 to 13 mole {\%} in comparison to phospholipids. The presence of VLC-FAs and VLC-PUFAs in retinal sphingolipids indicates that they may play role in ELOVL4-mediated Stargardt 3.",
author = "Brush, {Richard S.} and Tran, {Julie Thu A.} and Henry, {Kimberly R.} and McClellan, {Mark E.} and Elliott, {Michael H.} and Nawajes Mandal",
year = "2010",
month = "9",
day = "1",
doi = "10.1167/iovs.09-5134",
language = "English (US)",
volume = "51",
pages = "4422--4431",
journal = "Investigative Ophthalmology and Visual Science",
issn = "0146-0404",
publisher = "Association for Research in Vision and Ophthalmology Inc.",
number = "9",

}

TY - JOUR

T1 - Retinal sphingolipids and their very-long-chain fatty acid-containing species

AU - Brush, Richard S.

AU - Tran, Julie Thu A.

AU - Henry, Kimberly R.

AU - McClellan, Mark E.

AU - Elliott, Michael H.

AU - Mandal, Nawajes

PY - 2010/9/1

Y1 - 2010/9/1

N2 - PURPOSE. Recent evidence suggests that ceramide metabolism plays an important role in retinal photoreceptor cell survival and apoptosis. The purpose of this study was to characterize sphingolipids in the retina with special emphasis on the verylong- chain- containing saturated (VLC-FA) and polyunsaturated (VLC-PUFA) fatty acid- containing species. The VLC-FAs and VLC-PUFAs are synthesized by the ELOVL4 protein, which is involved in human Stargardt's macular dystrophy type 3 (STGD3). METHODS. Total lipids were extracted from retina and other tissues, and different sphingolipid classes were isolated and purified using various combinations of liquid- and solid-phase separation. Purified sphingolipids were analyzed by high-performance thin layer chromatography (HPTLC), gas chromatography (GC), and GC-MS (GC-mass spectrometry). RESULTS. Nonsialylated sphingolipids (NSLs) comprised ~3.5% of total retinal lipids of which 70% was sphingomyelin. Ceramide and glycosylceramides (GCs) constituted ≤1% of total retinal lipids. Gangliosides (GGs), on the other hand, comprised ~3.0% of total retinal lipids. Fatty acid analysis of retinal NSLs indicated an abundance of saturated fatty acids, with the presence of VLC-FAs but not of VLC-PUFAs beyond 24 carbons. However, GG had significant levels of unsaturated, polyunsaturated, and VLC-PUFAs. Retinal rod outer segments (ROS) contained ~1% each of NSL and GG, and their fatty acid profile was not very different from whole retinal NSL and GG, respectively. CONCLUSIONS. Retina has a total of 6% to 7% fatty acids that are N-linked to a sphingosine, which would be 11 to 13 mole % in comparison to phospholipids. The presence of VLC-FAs and VLC-PUFAs in retinal sphingolipids indicates that they may play role in ELOVL4-mediated Stargardt 3.

AB - PURPOSE. Recent evidence suggests that ceramide metabolism plays an important role in retinal photoreceptor cell survival and apoptosis. The purpose of this study was to characterize sphingolipids in the retina with special emphasis on the verylong- chain- containing saturated (VLC-FA) and polyunsaturated (VLC-PUFA) fatty acid- containing species. The VLC-FAs and VLC-PUFAs are synthesized by the ELOVL4 protein, which is involved in human Stargardt's macular dystrophy type 3 (STGD3). METHODS. Total lipids were extracted from retina and other tissues, and different sphingolipid classes were isolated and purified using various combinations of liquid- and solid-phase separation. Purified sphingolipids were analyzed by high-performance thin layer chromatography (HPTLC), gas chromatography (GC), and GC-MS (GC-mass spectrometry). RESULTS. Nonsialylated sphingolipids (NSLs) comprised ~3.5% of total retinal lipids of which 70% was sphingomyelin. Ceramide and glycosylceramides (GCs) constituted ≤1% of total retinal lipids. Gangliosides (GGs), on the other hand, comprised ~3.0% of total retinal lipids. Fatty acid analysis of retinal NSLs indicated an abundance of saturated fatty acids, with the presence of VLC-FAs but not of VLC-PUFAs beyond 24 carbons. However, GG had significant levels of unsaturated, polyunsaturated, and VLC-PUFAs. Retinal rod outer segments (ROS) contained ~1% each of NSL and GG, and their fatty acid profile was not very different from whole retinal NSL and GG, respectively. CONCLUSIONS. Retina has a total of 6% to 7% fatty acids that are N-linked to a sphingosine, which would be 11 to 13 mole % in comparison to phospholipids. The presence of VLC-FAs and VLC-PUFAs in retinal sphingolipids indicates that they may play role in ELOVL4-mediated Stargardt 3.

UR - http://www.scopus.com/inward/record.url?scp=77957370497&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77957370497&partnerID=8YFLogxK

U2 - 10.1167/iovs.09-5134

DO - 10.1167/iovs.09-5134

M3 - Article

VL - 51

SP - 4422

EP - 4431

JO - Investigative Ophthalmology and Visual Science

JF - Investigative Ophthalmology and Visual Science

SN - 0146-0404

IS - 9

ER -