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An ancient role for a prokineticin domain in invertebrate hematopoiesis
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Physiology and Developmental Biology, Comparative Physiology.
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Physiology and Developmental Biology, Comparative Physiology.
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Physiology and Developmental Biology, Comparative Physiology.
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Physiology and Developmental Biology, Comparative Physiology.
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2005 (English)In: Journal of Immunology, ISSN 0022-1767, E-ISSN 1550-6606, Vol. 174, no 10, 6153-6160 p.Article in journal (Refereed) Published
Abstract [en]

Hemopoietic development requires firm control of cell proliferation and differentiation. Although recent research has revealed conserved function of transcription factors and signaling pathways regulating lineage commitment in hemopoietic development in Drosophila melanogaster and vertebrates, little is known about hemopoietic cytokines among the invertebrate phyla. In the present study, we show that differentiation and growth of hemopoietic stem cells in vitro from an invertebrate, Pacifastacus leniusculus, require an endogenous cytokine-like factor, astakine, containing a prokineticin (PK) domain. Astakine induces a strong hematopoiesis response in live animals. An astakine homologue was also found in the shrimp, Penaeus monodon. So far, PK domains are only identified in vertebrates, in which they, for example, direct angiogenic growth. Our finding of the first PK-like cytokine characterized from any invertebrate provides novel information concerning the evolution of growth factors and blood cell development.

Place, publisher, year, edition, pages
2005. Vol. 174, no 10, 6153-6160 p.
Keyword [en]
Amino Acid Sequence, Animals, Astacoidea, Base Sequence, Cell Differentiation/physiology, Cell Division/physiology, Cell Movement/physiology, Cells; Cultured, Cloning; Molecular, Cytokines/genetics/*isolation & purification/*physiology/secretion, Cytoplasmic Granules/secretion, Evolution; Molecular, Hematopoiesis/genetics/*physiology, Hematopoietic Stem Cells/cytology/physiology, Hemocytes/cytology/physiology/secretion, Molecular Sequence Data, Penaeidae, Protein Structure; Tertiary/genetics, Research Support; Non-U.S. Gov't, Sequence Homology; Amino Acid, Structural Homology; Protein, Vascular Endothelial Growth Factor; Endocrine-Gland-Derived/chemistry/genetics/isolation & purification/*physiology/secretion
National Category
Natural Sciences
Identifiers
URN: urn:nbn:se:uu:diva-92993PubMedID: 15879111OAI: oai:DiVA.org:uu-92993DiVA: diva2:166334
Available from: 2005-04-14 Created: 2005-04-14 Last updated: 2014-08-13
In thesis
1. White Spot Syndrome Virus Interaction with a Freshwater Crayfish
Open this publication in new window or tab >>White Spot Syndrome Virus Interaction with a Freshwater Crayfish
2005 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Viruses are very abundant in water and hence diseases caused by viruses are common in marine organisms. These diseases create great problems for the commercial farming of crustaceans and mussels. One of the most common and most disastrous diseases for shrimp is caused by the white spot syndrome virus (WSSV), which is spread all around the world and also is infecting many different species of crustaceans including freshwater crayfish. Although during recent years knowledge has been gathered on the ways in which invertebrates defend themselves against bacteria and fungi virtually nothing is known about the defence processes elicited by virus. The aim of this work was to develop a model to use for studies of virus-host interactions in vivo and in vitro.

Temperature was found to be important for the virus infectivity and at lower temperature the virus apparently did not replicate, but if animals kept at low temperature for more than 40 days were transferred to higher temperatures they died quickly due to an increased virus replication. In crayfish infected with the virus it was found that hemocytes did not degranulate and the melanization reaction was also inhibited in the hemocyes. Thus it is apparent that this virus interacts with the immune system and hemocytes in particular and to be able to study this in some greater detail it was necessary to develop a cell culture to study virus-host interactions at the molecular level. Hence, we have developed a stem cell culture from the hematopoietic tissue (hpt) that will differentiate and mature into hemocytes and which can be used to replicate the WSSV in the presence of an endogenous cytokine, astakine. Astakine is the first cytokine like-factor described which is directly involved in hematopoiesis in an invertebrate.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2005. 56 p.
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 47
Keyword
Biology, WSSV, crayfish, hematopoietic tissue, cytokine, innate immunity, hemocytes, Biologi
National Category
Biological Sciences
Identifiers
urn:nbn:se:uu:diva-5776 (URN)91-554-6235-9 (ISBN)
Public defence
2005-05-10, Lindahlsalen, Evolution Biology Centre, Norbyvagen 18A, Uppsala, 10:00 (English)
Opponent
Supervisors
Available from: 2005-04-14 Created: 2005-04-14 Last updated: 2009-04-03Bibliographically approved
2. Hematopoiesis, Kazal Inhibitors and Crustins in a Crustacean
Open this publication in new window or tab >>Hematopoiesis, Kazal Inhibitors and Crustins in a Crustacean
2006 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Hemocytes are important as storage and producers of proteins of the innate immune defence, as well as actors of the cellular immune response. Therefore the hematopoietic process is critical for survival of most invertebrates. In order to search for molecules of importance for hemocyte development in crayfish we investigated proteins in crayfish plasma, which were increased after microbial challenge. As a result we were able to identify, purify and characterize a new invertebrate cytokine named astakine, and could clearly show that this protein is important for hematopoietic development in vivo as well as in an in vitro cell culture system. Astakine contains a prokineticin (PK) domain shown for the first time in an invertebrate, however, unlike the vertebrate PKs, astakine binds to a cell surface F1 ATP synthase β subunit located on the hematopoietic tissue (hpt) cell membranes. Extracellular ATP synthases as receptors have earlier been reported in different vertebrate cells and here we show that extracellular ATP synthase β subunit acts as a receptor for an invertebrate cytokine and is involved in hematopoiesis.

We also found two other groups of proteins, which were increased in plasma after microbial challenge and they were further characterized. A great number of different Kazal type proteinase inhibitors were produced by the hemocytes and this type of proteinase inhibitors have variable reactive sites determining the specificity of their inhibition. In crayfish Kazal inhibitors with similar reactive sites were found as a response to specific microorganisms suggesting that the crayfish Kazal proteinase inhibitors may provide enough variability to participate in diverse innate immune reactions against different pathogens.

Antimicrobial peptides were synthesized by the hemocytes and were likewise released in high amount upon microbial infection and we have characterized the main group of cystein-rich crustin-like antimicrobial peptides and investigated their tissue distribution and expression pattern.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2006. 42 p.
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 215
Keyword
Biochemistry, innate immunity, cytokine, hematopoietic tissue, ATP synthase β subunit, Kazal, antibacterial peptides, Biokemi
Identifiers
urn:nbn:se:uu:diva-7123 (URN)91-554-6643-5 (ISBN)
Public defence
2006-10-06, Lindahlsalen, Evolutionary Biology Centre, 10:00 (English)
Opponent
Supervisors
Available from: 2006-09-25 Created: 2006-09-25 Last updated: 2013-09-18Bibliographically approved

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Söderhäll, Irene

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