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Domestication Effects on Stress Induced Steroid Secretion and Adrenal Gene Expression in Chickens
AVIAN Behavioural Genomics and Physiology Group, IFMBiology, Linköping University.
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Chemistry - BMC, Analytical Chemistry.
AVIAN Behavioural Genomics and Physiology Group, IFMBiology, Linköping University.
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Chemistry - BMC, Analytical Chemistry.
Show others and affiliations
2015 (English)In: Scientific Reports, ISSN 2045-2322, E-ISSN 2045-2322, Vol. 5, article id 15345Article in journal (Refereed) Published
Abstract [en]

Understanding the genetic basis of phenotypic diversity is a challenge in contemporary biology. Domestication provides a model for unravelling aspects of the genetic basis of stress sensitivity. The ancestral Red Junglefowl (RJF) exhibits greater fear-related behaviour and a more pronounced HPA-axis reactivity than its domesticated counterpart, the White Leghorn (WL). By comparing hormones (plasmatic) and adrenal global gene transcription profiles between WL and RJF in response to an acute stress event, we investigated the molecular basis for the altered physiological stress responsiveness in domesticated chickens. Basal levels of pregnenolone and dehydroepiandrosterone as well as corticosterone response were lower in WL. Microarray analysis of gene expression in adrenal glands showed a significant breed effect in a large number of transcripts with over-representation of genes in the channel activity pathway. The expression of the best-known steroidogenesis genes were similar across the breeds used. Transcription levels of acute stress response genes such as StAR, CH25 and POMC were upregulated in response to acute stress. Dampened HPA reactivity in domesticated chickens was associated with changes in the expression of several genes that presents potentially minor regulatory effects rather than by means of change in expression of critical steroidogenic genes in the adrenal.

Place, publisher, year, edition, pages
2015. Vol. 5, article id 15345
National Category
Chemical Sciences Genetics
Identifiers
URN: urn:nbn:se:uu:diva-265144DOI: 10.1038/srep15345ISI: 000362885300001OAI: oai:DiVA.org:uu-265144DiVA, id: diva2:862621
Funder
Swedish Research Council, 621-2011-4731Swedish Research Council Formas, 221-2011-1088EU, European Research Council, 322206Available from: 2015-10-23 Created: 2015-10-23 Last updated: 2018-08-06Bibliographically approved
In thesis
1. Targeted analysis of bioactive steroids and oxycholesterols: Method development and application
Open this publication in new window or tab >>Targeted analysis of bioactive steroids and oxycholesterols: Method development and application
2016 (English)Licentiate thesis, comprehensive summary (Other academic)
Abstract [en]

Peripheral steroids and oxycholesterols are important lipid compounds controlling various functions in the human body. Steroid analysis of biological samples is routinely employed in the clinical environment as an essential source of information on endocrine and metabolic disorders. It has been reported that stress related neurosteroids have been implicated in the development and prognoses of neurodegenerative disorders such as Alzheimer’s disease (AD). These compounds have been identified as possible biomarkers in the diagnosis of AD and other neurodegenerative disorders. Therefore, methods for the simultaneous analysis of steroids from the four major classes (estrogens, androgens, progestogens and corticosteroids) are vital in providing useful and more comprehensive data.

Homeostasis of cholesterol in the brain is maintained primarily by metabolism to oxysterols, including oxycholesterols. These oxycholesterols act as a transport form of cholesterol as it readily navigates the blood-brain barrier. Oxycholesterols are generally more bioactive than cholesterol and is of interest in pathophysiology. Moreover, if their production in cells and tissues and/or their introduction with dietary animal fat are excessive, oxycholesterols could indeed contribute to the pathogenesis of various disease processes.

The first study in this thesis focuses on a novel supercritical fluid chromatography–tandem mass spectrometry method for targeted analysis of eighteen peripheral steroids. The method is simple and fast. It has sufficient sensitivity for quantification of 18 different steroids in small volume human plasma. Therefore, this novel method can be applied for screening many steroids within 5 minutes providing the possibility to use for routine healthcare practice. The second study involves the quantification of three adrenal steroids in plasma from domesticated White Leghorn (WL) chickens and Red Junglefowl (RJF) birds. The domestication effects on stress induced steroid secretion and adrenal gene expression in chickens are evaluated. The third study focuses on determination of more than ten oxycholesterols in biological samples with a gas chromatography–mass spectrometry method and a supercritical fluid–tandem mass spectrometry method.

Place, publisher, year, edition, pages
Uppsala University, 2016. p. 41
Keywords
Steroids, Oxycholesterols, Supercritical Fluid Chromatography, Mass Spectrometry
National Category
Analytical Chemistry
Research subject
Chemistry with specialization in Analytical Chemistry
Identifiers
urn:nbn:se:uu:diva-305264 (URN)
Presentation
2016-03-04, B7:113a (BMC), Husargatan 3, Uppsala, 13:15 (English)
Opponent
Supervisors
Funder
Swedish Research Council, P29797-1
Available from: 2016-12-21 Created: 2016-10-13 Last updated: 2016-12-21Bibliographically approved
2. Mass spectrometry based analysis of endogenous sterols and hormones
Open this publication in new window or tab >>Mass spectrometry based analysis of endogenous sterols and hormones
2018 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Bioanalytical applications using supercritical fluid chromatography (SFC) as analytical technique are of increasing interest. In essence, bioanalysis involves measurement of bioactive or endogenous compounds in biological matrices. SFC has emerged as an excellent choice for bioanalytical analysis, attributable to its speed, selectivity and efficiency compared with high performance liquid chromatography. Moreover, coupling of SFC with mass spectrometry (MS) provides the additional benefits of specificity and sensitivity.

The aim of this thesis was to exploit these features by developing methods for the analysis of endogenous steroids, cholesterol oxidation products (COPs) and thyroid hormones (THs) by using ultra-performance supercritical fluid chromatography–tandem mass spectrometry (UPSFC–MS/MS) as analytical technique.

Endogenous steroids control many physiological processes, including reproduction, maturation, gene expression and neurological functions in humans and animals. In the first study, three steroids were measured in domesticated White Leghorn (WL) chickens and ancestral Red Junglefowl (RJF) birds. Restraining stress caused a significantly larger increase in corticosterone levels in RJF than in WL, indicating a blunted hypothalamic–pituitary–adrenal (HPA) axis activity in domesticated chickens. The second study was a continuation of the first study and corticosterone levels from the F12 generation of an intercross between WL and RJF birds were measured before and after physical restraint stress. The expression levels of the glucocorticoid receptor (GR) in the hypothalamus and several genes in the adrenal glands were correlated with the post-stress levels of corticosterone in plasma. In the third study, the measurement of steroids was extended to assess more endogenous steroids from the four major classes, i.e. estrogens, androgens, progestogens and corticosterone.

Endogenous COPs are of interest in pathophysiology. COPs are more readily disposed by cells than cholesterol. Therefore, cholesterol is oxidised to the more polar COPs and are generally more bioactive than cholesterol. Moreover, if their production in cells and tissues and/or their introduction with dietary animal fat are excessive, COPs could indeed contribute to the pathogenesis of various disease processes. Fourteen COPs were included in the fourth study and a novel method for their separation was developed.

The last study in this thesis, involved the analysis of five THs. These hormones are vital for growth, developmental and metabolic processes of vertebrate life and play an important role in energy homeostasis. Measurements of circulating thyroid hormone levels are used in thyroid disorder diagnoses or treatment status monitoring. Two rapid methods for the separation of five THs were developed.

In summary, the work in this thesis demonstrates the applicability of UPSFC–MS/MS as an analytical technique in bioanalysis of endogenous compounds.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2018. p. 62
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 1698
Keywords
mass spectrometry, supercritical fluid chromatography, UPSFC–MS/MS, bioanalysis, steroids, oxysterols, thyroid hormones
National Category
Analytical Chemistry
Research subject
Chemistry with specialization in Analytical Chemistry
Identifiers
urn:nbn:se:uu:diva-356767 (URN)978-91-513-0396-3 (ISBN)
Public defence
2018-09-19, B7:101a, BMC, Husargatan 3, Uppsala, 09:15 (English)
Opponent
Supervisors
Available from: 2018-08-30 Created: 2018-08-06 Last updated: 2018-09-10

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