Analysis of thrombocyte development in CD41-GFP transgenic zebrafish

Hui Feng Lin, David Traver, Hao Zhu, Kimberly Dooley, Barry H. Paw, Leonard I. Zon, Robert I. Handin

Research output: Contribution to journalArticle

203 Citations (Scopus)

Abstract

Thrombocytes are the nucleated equivalent of platelets in nonmammalian vertebrates such as the zebrafish, Danio rerio. We have cloned zebrafish CD41 cDNA (αIIb, glycoprotein IIb [GPIIb]) and its promoter and have generated transgenic zebrafish lines with green fluorescent protein (GFP)-tagged thrombocytes. CD41 mRNA transcripts appeared 42 hours after fertilization (hpf) by reverse-transcriptase-polymerase chain reaction (RT-PCR) and at 48 hpf in circulating hematopoietic cells. Flow sorting of thrombocytes from the mesonephros of adult CD41-GFP zebrafish showed a GFPhigh subset, which had the morphologic appearance of mature thrombocytes, and a GFPlow subset with an immature appearance, suggesting that they may be thrombocyte precursors. Confocal laser microscopy of embryos 40 and 48 hpf also showed a nonmobile population of GFP+ cells in a discrete area between the dorsal aorta and caudal vein. Production of circulating thrombocytes was inhibited by the injection of antisense morpholinos for the stem-cell transcription factor scl and c-mpl, the receptor for thrombopoietin. The nonmobile pool of GFP+ cells was abolished by scl knockdown and partially inhibited by c-mpl knockdown. These studies have shown that it is possible to identify thrombocytes, thrombocyte precursors, and, possibly, early hematopoietic stem cells in zebrafish embryos and track their proliferation and maturation.

Original languageEnglish (US)
Pages (from-to)3803-3810
Number of pages8
JournalBlood
Volume106
Issue number12
DOIs
StatePublished - Dec 1 2005

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Zebrafish
Green Fluorescent Proteins
Blood Platelets
Stem cells
Thrombopoietin Receptors
Morpholinos
Polymerase chain reaction
RNA-Directed DNA Polymerase
Platelets
Sorting
Confocal Microscopy
Glycoproteins
Microscopic examination
Transcription Factors
Complementary DNA
Embryonic Structures
Mesonephros
Messenger RNA
Lasers
Stem Cell Factor

ASJC Scopus subject areas

  • Hematology

Cite this

Lin, H. F., Traver, D., Zhu, H., Dooley, K., Paw, B. H., Zon, L. I., & Handin, R. I. (2005). Analysis of thrombocyte development in CD41-GFP transgenic zebrafish. Blood, 106(12), 3803-3810. https://doi.org/10.1182/blood-2005-01-0179

Analysis of thrombocyte development in CD41-GFP transgenic zebrafish. / Lin, Hui Feng; Traver, David; Zhu, Hao; Dooley, Kimberly; Paw, Barry H.; Zon, Leonard I.; Handin, Robert I.

In: Blood, Vol. 106, No. 12, 01.12.2005, p. 3803-3810.

Research output: Contribution to journalArticle

Lin, HF, Traver, D, Zhu, H, Dooley, K, Paw, BH, Zon, LI & Handin, RI 2005, 'Analysis of thrombocyte development in CD41-GFP transgenic zebrafish', Blood, vol. 106, no. 12, pp. 3803-3810. https://doi.org/10.1182/blood-2005-01-0179
Lin HF, Traver D, Zhu H, Dooley K, Paw BH, Zon LI et al. Analysis of thrombocyte development in CD41-GFP transgenic zebrafish. Blood. 2005 Dec 1;106(12):3803-3810. https://doi.org/10.1182/blood-2005-01-0179
Lin, Hui Feng ; Traver, David ; Zhu, Hao ; Dooley, Kimberly ; Paw, Barry H. ; Zon, Leonard I. ; Handin, Robert I. / Analysis of thrombocyte development in CD41-GFP transgenic zebrafish. In: Blood. 2005 ; Vol. 106, No. 12. pp. 3803-3810.
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