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First published online 6 October 2004
doi: 10.1242/dev.01409

Development 131, 5417-5427 (2004)
Published by The Company of Biologists 2004
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Phosphatidylserine receptor is required for the engulfment of dead apoptotic cells and for normal embryonic development in zebrafish

Jiann-Ruey Hong1,*, Gen-Hwa Lin2, Cliff Ji-Fan Lin2,3, Wan-ping Wang2, Chien-Chung Lee2, Tai-Lang Lin2 and Jen-Leih Wu2,*,{dagger}

1 Laboratory of Molecular Virology and Biotechnology, Institute of Biotechnology, National Cheng-Kung University, Tainan 701, Taiwan
2 Laboratory of Marine Molecular Biology and Biotechnology, Institute of Zoology, Academia Sinica, Nankang, Taipei 115, Taiwan
3 Graduate Institute of Life Sciences, National Defense Medical Center, Taipei 117, Taiwan

{dagger} Author for correspondence (e-mail: jlwu{at}gate.sinica.edu.tw)

Accepted 19 August 2004

During development, the role of the phosphatidylserine receptor (PSR) in the removal of apoptotic cells that have died is poorly understood. We have investigated this role of PSR in developing zebrafish. Programmed cell death began during the shield stage, with dead cells being engulfed by a neighboring cell that showed a normal-looking nucleus and the nuclear condensation multi-micronuclei of an apoptotic cell. The zebrafish PSR engulfing receptor was cloned (zfpsr), and its nucleotide sequence was compared with corresponding sequences in Drosophila melanogaster (76% identity), human (74%), mouse (72%) and Caenorhabditis elegans (60%). The PSR receptor contained a jmjC domain (residues 143-206) that is a member of the cupin metalloenzyme superfamily, but in this case serves an as yet unknown function(s). psr knockdown by a PSR morpholino oligonucleotide led to accumulation of a large number of dead apoptotic cells in whole early embryo. These cells interfered with embryonic cell migration. In addition, normal development of the somite, brain, heart and notochord was sequentially disrupted up to 24 hours post-fertilization. Development could be rescued in defective embryos by injecting psr mRNA. These results are consistent with a PSR-dependent system in zebrafish embryos that engulfs apoptotic cells mediated by PSR-phagocytes during development, with the system assuming an important role in the normal development of tissues such as the brain, heart, notochord and somite.

Key words: Phosphatidylserine receptor, Apoptotic corpses, Zebrafish, Knockdown, Brain, Organogenesis


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© The Company of Biologists Ltd 2004