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Analysis of pregnenolone and dehydroepiandrosterone in rodent brain: cholesterol autoxidation is the key

P Liere, A Pianos, B Eychenne, A Cambourg, K Bodin, W Griffiths, M Schumacher, E.-E Baulieu, J Sjovall, William Griffiths Orcid Logo

The Journal of Lipid Research, Volume: 50, Issue: 12, Pages: 2430 - 2444

Swansea University Author: William Griffiths Orcid Logo

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DOI (Published version): 10.1194/jlr.M900162-JLR200

Abstract

Pregnenolone (PREG) and dehydroepiandrosterone (DHEA), and their respective sulfated forms PREGS and DHEAS, were among the first steroids to be identified in rodent brain. However, unreliable steroid isolation and solvolysis procedures resulted in errors, particularly in the case of brain steroid su...

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Published in: The Journal of Lipid Research
ISSN: 0022-2275
Published: 2009
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URI: https://cronfa.swan.ac.uk/Record/cronfa10496
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first_indexed 2013-07-23T12:03:39Z
last_indexed 2018-02-09T04:39:19Z
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spelling 2013-08-05T09:14:45.3105467 v2 10496 2012-04-04 Analysis of pregnenolone and dehydroepiandrosterone in rodent brain: cholesterol autoxidation is the key 3316b1d1b524be1831790933eed1c26e 0000-0002-4129-6616 William Griffiths William Griffiths true false 2012-04-04 BMS Pregnenolone (PREG) and dehydroepiandrosterone (DHEA), and their respective sulfated forms PREGS and DHEAS, were among the first steroids to be identified in rodent brain. However, unreliable steroid isolation and solvolysis procedures resulted in errors, particularly in the case of brain steroid sulfates analyzed by radioimmunology or GC-MS of liberated free steroids. By using a solid-phase extraction recycling/elution procedure, allowing the strict separation of sulfated, free, and fatty acid esters of PREG and DHEA, PREGS and DHEAS, unlike free PREG, were not detected in rat and mouse brain and plasma. Conversely, considerable amounts of PREG and DHEA were released from unknown precursor(s) present in the lipoidal fraction, distinct from fatty acid ester conjugates. Chromatographic and mass spectrometric studies of the nature of the precursor(s) showed that autoxidation of brain cholesterol (CHOL) was responsible for the release of PREG and DHEA from the lipoidal fraction. When inappropriate protocols were used, CHOL was also the precursor of PREG and DHEA obtained from the fraction assumed to contain sulfated steroids. In contrast, free PREG was definitely confirmed as an endogenous steroid in rat brain. Our study shows that an early removal of CHOL from brain extracts coupled to well-validated extraction and fractionation procedures are prerequisites for reliable measurements of free and conjugated PREG and DHEA by GC-MS or other indirect methods. Journal Article The Journal of Lipid Research 50 12 2430 2444 0022-2275 31 12 2009 2009-12-31 10.1194/jlr.M900162-JLR200 COLLEGE NANME Biomedical Sciences COLLEGE CODE BMS Swansea University 2013-08-05T09:14:45.3105467 2012-04-04T12:26:22.2031600 Faculty of Medicine, Health and Life Sciences Swansea University Medical School - Medicine P Liere 1 A Pianos 2 B Eychenne 3 A Cambourg 4 K Bodin 5 W Griffiths 6 M Schumacher 7 E.-E Baulieu 8 J Sjovall 9 William Griffiths 0000-0002-4129-6616 10
title Analysis of pregnenolone and dehydroepiandrosterone in rodent brain: cholesterol autoxidation is the key
spellingShingle Analysis of pregnenolone and dehydroepiandrosterone in rodent brain: cholesterol autoxidation is the key
William Griffiths
title_short Analysis of pregnenolone and dehydroepiandrosterone in rodent brain: cholesterol autoxidation is the key
title_full Analysis of pregnenolone and dehydroepiandrosterone in rodent brain: cholesterol autoxidation is the key
title_fullStr Analysis of pregnenolone and dehydroepiandrosterone in rodent brain: cholesterol autoxidation is the key
title_full_unstemmed Analysis of pregnenolone and dehydroepiandrosterone in rodent brain: cholesterol autoxidation is the key
title_sort Analysis of pregnenolone and dehydroepiandrosterone in rodent brain: cholesterol autoxidation is the key
author_id_str_mv 3316b1d1b524be1831790933eed1c26e
author_id_fullname_str_mv 3316b1d1b524be1831790933eed1c26e_***_William Griffiths
author William Griffiths
author2 P Liere
A Pianos
B Eychenne
A Cambourg
K Bodin
W Griffiths
M Schumacher
E.-E Baulieu
J Sjovall
William Griffiths
format Journal article
container_title The Journal of Lipid Research
container_volume 50
container_issue 12
container_start_page 2430
publishDate 2009
institution Swansea University
issn 0022-2275
doi_str_mv 10.1194/jlr.M900162-JLR200
college_str Faculty of Medicine, Health and Life Sciences
hierarchytype
hierarchy_top_id facultyofmedicinehealthandlifesciences
hierarchy_top_title Faculty of Medicine, Health and Life Sciences
hierarchy_parent_id facultyofmedicinehealthandlifesciences
hierarchy_parent_title Faculty of Medicine, Health and Life Sciences
department_str Swansea University Medical School - Medicine{{{_:::_}}}Faculty of Medicine, Health and Life Sciences{{{_:::_}}}Swansea University Medical School - Medicine
document_store_str 0
active_str 0
description Pregnenolone (PREG) and dehydroepiandrosterone (DHEA), and their respective sulfated forms PREGS and DHEAS, were among the first steroids to be identified in rodent brain. However, unreliable steroid isolation and solvolysis procedures resulted in errors, particularly in the case of brain steroid sulfates analyzed by radioimmunology or GC-MS of liberated free steroids. By using a solid-phase extraction recycling/elution procedure, allowing the strict separation of sulfated, free, and fatty acid esters of PREG and DHEA, PREGS and DHEAS, unlike free PREG, were not detected in rat and mouse brain and plasma. Conversely, considerable amounts of PREG and DHEA were released from unknown precursor(s) present in the lipoidal fraction, distinct from fatty acid ester conjugates. Chromatographic and mass spectrometric studies of the nature of the precursor(s) showed that autoxidation of brain cholesterol (CHOL) was responsible for the release of PREG and DHEA from the lipoidal fraction. When inappropriate protocols were used, CHOL was also the precursor of PREG and DHEA obtained from the fraction assumed to contain sulfated steroids. In contrast, free PREG was definitely confirmed as an endogenous steroid in rat brain. Our study shows that an early removal of CHOL from brain extracts coupled to well-validated extraction and fractionation procedures are prerequisites for reliable measurements of free and conjugated PREG and DHEA by GC-MS or other indirect methods.
published_date 2009-12-31T03:11:53Z
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score 11.036706

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