Hemocyte-lineage marker proteins in a crustacean, the freshwater crayfish, Pacifastacus leniusculus.
To identify proteins associated with development of different hemocyte types in the freshwater crayfish Pacifastacus leniusculus, 2-DE followed by MS analysis was carried out with hematopoietic tissue (Hpt) cells, semigranular cells (SGC) and granular cells (GC). Within the hemocyte lineages one two-domain Kazal proteinase inhibitor (KPI) was found to be specific for SGC, while a superoxide dismutase (SOD) was specific for GC at protein as well as at mRNA level. The proliferation cell nuclear antigen (PCNA) was detected at the mRNA level in Hpt cells only. We also provide evidence that SGC and GC most likely differentiate to maturation as separate lineages. We found that after laminarin or lipopolysaccharide (LPS) injection into crayfish, the transcript levels of PCNA and SOD increased in the Hpt cells, whereas the KPI transcript never was present in Hpt regardless of any challenge. RNA interference of PCNA in the Hpt cells led to that most of the cells did not spread or attach to the tissue culture dish. These results suggest that PCNA, KPI and SOD can be used as markers for Hpt cells, SGC and GC, respectively, and in conjunction with these results, a model is proposed how the Hpt responds to a microbial challenge by proliferation and release of Hpt cells.
Wu
Chenglin
C
Department of Comparative of Physiology, Uppsala University, Uppsala, Sweden.
Söderhäll
Irene
I
Kim
Young-A
YA
Liu
Haipeng
H
Söderhäll
Kenneth
K
D000595:Amino Acid Sequence; D000818:Animals; D003400:Astacoidea / Q000737:chemistry* / Q000166:cytology; D015415:Biomarkers / Q000032:analysis*; D001798:Blood Proteins / Q000032:analysis*; D002454:Cell Differentiation; D019070:Cell Lineage; D049109:Cell Proliferation; D015180:Electrophoresis, Gel, Two-Dimensional; D006413:Hematopoietic System / Q000378:metabolism; D006434:Hemocytes / Q000737:chemistry*; D008954:Models, Biological; D008969:Molecular Sequence Data; D018809:Proliferating Cell Nuclear Antigen / Q000032:analysis; D011480:Protease Inhibitors / Q000032:analysis; D034622:RNA Interference; D013482:Superoxide Dismutase / Q000032:analysis
D015415:Biomarkers; D001798:Blood Proteins; D018809:Proliferating Cell Nuclear Antigen; D011480:Protease Inhibitors; D013482:Superoxide Dismutase