Effects of infection of EGFP-expressing Escherichia coli on haemocytes in Ciona intestinalis

Ultrastructural and immunocytochemical analysis of circulating hemocytes from Cherax quadricarinatus (von Martens, 1868)

“…This implies that CD34 is considered to be a marker of hematopoietic stem cells in C. quadricarinatus, and that C. quadricarinatus, with hematopoietic stem cells similar to vertebrates, can provide a mechanism study to clarify vertebrate congenital and adaptive immune system origins. The positivity of the stem cell-like cells to anti-CD34 antibody suggests a potential role as a pluripotent cell, able to differentiate into other cell types (Liu et al, 2006).…”

“…After centrifugation at 1332 g for 20 min at 20 °C, the supernatant was removed and the hemocytes washed three times in sterile anticoagulant (Liu et al, 2006). Transmission electron microscopy: Hemolymph samples were washed twice in Piperazine-1, 4-bisethanesulfonic acid (PIPES) buffer (pH 7, containing 5 mmol/L MgSO 4 , 0.5 mmol/L CaCl 2 , 0.05% Triton X-100, 1.5% paraformaldehyde, 0.5% glutaraldehyde and 5 mmol/L ethylene glycol bis [sulfosuccinimidylsuccinate]), fixed in 2.5% glutaraldehyde buffered to pH 7.2 for 2 h at 4 °C, and post-fixed in freshly prepared 1% osmium tetroxide for 1 h at 4 °C.…”

“…Immunocytochemical investigations have been carried out in accordance with Liu et al (2006), with little modifications. The hemocytes samples were spread onto slides and allowed to adhere and then fixed for 30 min at 4°C in 4% paraformaldehyde (Sigma) in MPBS (modified PBS: 25.5 g. L -1 NaCl, 0.6 g. L -1 KCl, 0.9 g. L -1 Na 2 HPO 4 , 0.27 g. L -1 KH 2 PO 4 , 1.0 g. L -1 glucose, pH 7.2).…”

“…CD34 is a transmembrane protein, expressed in mammalian hematopoietic cells (Furukawa, 1998), and is widely adopted as a marker of human hematopoietic stem cells (Liu et al, 2006). In the previous study, it has been showed that CD34 is well-conserved in Metazoa, which is proved to be present in setem-like cells of chondrichthyan Torpedo marmorata (Pica et al, 2000), the gastropod Planorbarius corneus (Franceschi et al, 1991), the bivalve Tapes philippinarum (Cima et al, 2000) and in the hematopoietic organ of the grasshopper Euprepocnemis shiraki (Lim et al, 2004).…”

“…In the previous study, it has been showed that CD34 is well-conserved in Metazoa, which is proved to be present in setem-like cells of chondrichthyan Torpedo marmorata (Pica et al, 2000), the gastropod Planorbarius corneus (Franceschi et al, 1991), the bivalve Tapes philippinarum (Cima et al, 2000) and in the hematopoietic organ of the grasshopper Euprepocnemis shiraki (Lim et al, 2004). In recent research of EST sequence result, integral membrane protein (CD34+) was detected in the expression products of the hemocytes of White Spot Syndrome Virus-infected Kuruma prawn, Penaeus japonicus (Rojtinnakorn et al, 2002;Liu et al, 2006). This implies that CD34 is considered to be a marker of hematopoietic stem cells in C. quadricarinatus, and that C. quadricarinatus, with hematopoietic stem cells similar to vertebrates, can provide a mechanism study to clarify vertebrate congenital and adaptive immune system origins.…”