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Neuropeptidomic analysis of the embryonic Japanese quail diencephalon
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Pharmaceutical Biosciences.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Pharmaceutical Biosciences.
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Organism Biology, Environmental Toxicology.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Pharmaceutical Biosciences.
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2010 (English)In: BMC Developmental Biology, ISSN 1471-213X, Vol. 10, 30- p.Article in journal (Refereed) Published
Abstract [en]

Background: Endogenous peptides such as neuropeptides are involved in numerous biological processes in the fully developed brain but very little is known about their role in brain development. Japanese quail is a commonly used bird model for studying sexual dimorphic brain development, especially adult male copulatory behavior in relation to manipulations of the embryonic endocrine system. This study uses a label-free liquid chromatography mass spectrometry approach to analyze the influence of age (embryonic days 12 vs 17), sex and embryonic day 3 ethinylestradiol exposure on the expression of multiple endogenous peptides in the developing diencephalon.

Results: We identified a total of 65 peptides whereof 38 were sufficiently present in all groups for statistical analysis. Age was the most defining variable in the data and sex had the least impact. Most identified peptides were more highly expressed in embryonic day 17. The top candidates for EE2 exposure and sex effects were neuropeptide K (downregulated by EE2 in males and females), gastrin-releasing peptide (more highly expressed in control and EE2 exposed males) and gonadotropin-inhibiting hormone related protein 2 (more highly expressed in control males and displaying interaction effects between age and sex). We also report a new potential secretogranin-2 derived neuropeptide and previously unknown phosphorylations in the C-terminal flanking protachykinin 1 neuropeptide.

Conclusions: This study is the first larger study on endogenous peptides in the developing brain and implies a previously unknown role for a number of neuropeptides in middle to late avian embryogenesis. It demonstrates the power of label-free liquid chromatography mass spectrometry to analyze the expression of multiple endogenous peptides and the potential to detect new putative peptide candidates in a developmental model.

Place, publisher, year, edition, pages
2010. Vol. 10, 30- p.
Keyword [en]
Neuropeptides, Japanese quail, Peptidomics, Developmental neurobiology, estrogen, endocrine disruption, developmental toxicology, toxicology
National Category
Developmental Biology Pharmacology and Toxicology Neurosciences
Research subject
Toxicology; Biology with specialization in Animal Development
Identifiers
URN: urn:nbn:se:uu:diva-96838DOI: 10.1186/1471-213X-10-30ISI: 000276503000001PubMedID: 20298575OAI: oai:DiVA.org:uu-96838DiVA: diva2:171546
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ReproSafe
Available from: 2008-03-14 Created: 2008-03-14 Last updated: 2011-03-03Bibliographically approved
In thesis
1. Genomic and Peptidomic Characterization of the Developing Avian Brain
Open this publication in new window or tab >>Genomic and Peptidomic Characterization of the Developing Avian Brain
2008 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Chicken and Japanese quail are commonly used models in developmental and sex specific neuroendocrine research. There is relatively little known about the mechanisms behind their sex specific brain development, especially regarding the impact of the sex chromosomes (male: ZZ, female ZW) in relation to gonadal hormones. This thesis explores several aspects of these processes. Gene expression analysis with cDNA and Affymetrix arrays on brain tissue from both pre-gonadal embryos and embryos with differentiated gonads indicate a strong sex chromosomal presence in sexual dimorphic somatic tissue development in both chicken and Japanese quail. This sex chromosome pattern seems to remain in adult brain tissue. The data demonstrates that chicken males exhibit a significant level of Z-gene dosage compared to females in both somatic and germ line derived embryonic tissues. Several avian sex determination gene candidates (MHM non-coding RNA, DMRT1, HINTW, and HINTZ) were analyzed by real-time PCR. DMRT1 is dosage compensated in male brain tissue, in contrast to its reported gene dosage in male gonads. Early embryonic ethinylestradiol (EE2) exposure did not affect male or female neural gene expression patterns during later development. A peptidomics analysis on quail embryonic day 12 (ed12) and ed17 diencephalon by LC-MS identified over 60 endogenous peptides and analyzed the expression patterns for 38 of them with regard to age, sex and early EE2 exposure. There was a general upregulation between ed12 and ed17, but no clear sex effects were detected. Multivariate analysis indicates that EE2 exposed individuals differ from control individuals in a gender independent manner, and that Gonadotropin-inhibiting hormone related peptide 2 (GnIH-RP2) is a candidate for EE2 induced peptidomic alterations in male embryonic brain.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2008. 67 p.
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine, ISSN 1651-6206 ; 317
Keyword
Toxicology, Sex differentation, Sex determination, Sex chromosomes, Dosage Compensation, Genetic, Chickens, Coturnix, Gene Expression Profiling, Neuropeptides, Diencephalon, Toxikologi
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:uu:diva-8507 (URN)978-91-554-7117-0 (ISBN)
Public defence
2008-04-04, B42, BMC, Uppsala, 13:15
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Supervisors
Available from: 2008-03-14 Created: 2008-03-14 Last updated: 2012-02-06Bibliographically approved

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Dencker, Lennart

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