HINTW, a W‐chromosome HINT gene in chick, is expressed ubiquitously and is a robust female cell marker applicable in intraspecific chimera studies. Issue 5 (13th March 2014)
- Record Type:
- Journal Article
- Title:
- HINTW, a W‐chromosome HINT gene in chick, is expressed ubiquitously and is a robust female cell marker applicable in intraspecific chimera studies. Issue 5 (13th March 2014)
- Main Title:
- HINTW, a W‐chromosome HINT gene in chick, is expressed ubiquitously and is a robust female cell marker applicable in intraspecific chimera studies
- Authors:
- Nagai, Hiroki
Sezaki, Maiko
Bertocchini, Federica
Fukuda, Kimiko
Sheng, Guojun - Abstract:
- <abstract abstract-type="main"> <title>Abstract</title> <p>Grafting and transplantation experiments in embryology require proper distinction between host and donor tissues. For the avian model this has traditionally been achieved by using two closely related species (e.g., chick and quail) followed by species‐specific antibody staining. Here, we show that an in situ hybridization probe against the <italic>HINTW</italic> gene is a robust and reliable marker for female‐derived chicken cells. At all pre‐circulation stages tested, all cells in female embryos, independently confirmed by PCR analysis, were strongly positive for <italic>HINTW</italic>, whereas all male embryos were negative. This probe is broadly applicable in intra‐specific chick/chick chimera studies, and as a proof of principle, we utilized this probe to detect female cells in three experimental settings: (1) to mark female donor cells in a node transplantation assay; (2) to distinguish female cells in male/female twins generated by the Cornish pasty culture; and (3) to detect female half of the embryo in artificially generated bilateral gynandromorphs. A rapid, PCR based pre‐screening step increases the efficiency of obtaining desired donor/host sex combination from 25% to 100%. For most avian chimera studies, this female‐specific in situ probe is a low cost alternative to the commonly used QCPN antibody and to ubiquitous‐GFP chicken strains which are not widely available to the research community. genesis<abstract abstract-type="main"> <title>Abstract</title> <p>Grafting and transplantation experiments in embryology require proper distinction between host and donor tissues. For the avian model this has traditionally been achieved by using two closely related species (e.g., chick and quail) followed by species‐specific antibody staining. Here, we show that an in situ hybridization probe against the <italic>HINTW</italic> gene is a robust and reliable marker for female‐derived chicken cells. At all pre‐circulation stages tested, all cells in female embryos, independently confirmed by PCR analysis, were strongly positive for <italic>HINTW</italic>, whereas all male embryos were negative. This probe is broadly applicable in intra‐specific chick/chick chimera studies, and as a proof of principle, we utilized this probe to detect female cells in three experimental settings: (1) to mark female donor cells in a node transplantation assay; (2) to distinguish female cells in male/female twins generated by the Cornish pasty culture; and (3) to detect female half of the embryo in artificially generated bilateral gynandromorphs. A rapid, PCR based pre‐screening step increases the efficiency of obtaining desired donor/host sex combination from 25% to 100%. For most avian chimera studies, this female‐specific in situ probe is a low cost alternative to the commonly used QCPN antibody and to ubiquitous‐GFP chicken strains which are not widely available to the research community. genesis 52:424–430, 2014. © 2014 Wiley Periodicals, Inc.</p> </abstract> … (more)
- Is Part Of:
- Genesis. Volume 52:Issue 5(2014:May)
- Journal:
- Genesis
- Issue:
- Volume 52:Issue 5(2014:May)
- Issue Display:
- Volume 52, Issue 5 (2014)
- Year:
- 2014
- Volume:
- 52
- Issue:
- 5
- Issue Sort Value:
- 2014-0052-0005-0000
- Page Start:
- 424
- Page End:
- 430
- Publication Date:
- 2014-03-13
- Subjects:
- Developmental genetics -- Periodicals
Genetics -- Periodicals
Developmental biology -- Periodicals
Embryology -- Periodicals
Genetic regulation -- Periodicals
576.5 - Journal URLs:
- http://onlinelibrary.wiley.com/journal/10.1002/(ISSN)1526-968X ↗
http://onlinelibrary.wiley.com/ ↗ - DOI:
- 10.1002/dvg.22769 ↗
- Languages:
- English
- ISSNs:
- 1526-954X
- Deposit Type:
- Legaldeposit
- View Content:
- Available online (eLD content is only available in our Reading Rooms) ↗
- Physical Locations:
- British Library DSC - 4111.807500
British Library DSC - BLDSS-3PM
British Library STI - ELD Digital store - Ingest File:
- 3101.xml