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Representative microscopic images of normal and apoptotic  neutrophils. Aliquots (0.4 ml) of PBN suspensions of A1-a−/− mice (0.5 ×  106/ml) were incubated at 37°C for 12 h in RPMI 1640 medium. Arrows  indicate normal neutrophils and arrowhead indicates an apoptotic neutrophil containing condensed and fragmented nuclei (original magnification,  ×1,000, May-Giemsa stain).
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Figure 1: Representative microscopic images of normal and apoptotic neutrophils. Aliquots (0.4 ml) of PBN suspensions of A1-a−/− mice (0.5 × 106/ml) were incubated at 37°C for 12 h in RPMI 1640 medium. Arrows indicate normal neutrophils and arrowhead indicates an apoptotic neutrophil containing condensed and fragmented nuclei (original magnification, ×1,000, May-Giemsa stain).

Mentions: Neutrophils incubated under specific conditions were stained with May-Giemsa and apoptotic cells were counted under light microscopy (×1,000). Representative microscopic images of normal and apoptotic neutrophils are shown in Fig. 1. Approximately 300–1200 cells were scanned per specimen.

Accelerated Neutrophil Apoptosis in Mice Lacking A1-a, a Subtype of the bcl-2–related A1 Gene

Hamasaki A, Sendo F, Nakayama K, Ishida N, Negishi I, Nakayama K, Hatakeyama S - J. Exp. Med. (1998)

Bottom Line: On the other hand, the extent of tumor necrosis factor alpha-induced acceleration of neutrophil apoptosis did not differ among A1-a-/-, A1-a+/-, and wild-type mice.The descending order of A1 mRNA expression was wild-type, A1-a+/-, and A1-a-/-.Taken together, these results suggest that A1 is involved in inhibition of certain types of neutrophil apoptosis.

Affiliation: Department of Immunology and Parasitology, Yamagata University School of Medicine, Yamagata 990-9585, Japan.

ABSTRACT
To elucidate the role of A1, a new member of the Bcl-2 family of apoptosis regulators active in hematopoietic cell apoptosis, we established mice lacking A1-a, a subtype of the A1 gene in mice (A1-a-/- mice). Spontaneous apoptosis of peripheral blood neutrophils of A1-a-/- mice was enhanced compared with that of either wild-type mice or heterozygous mutants (A1-a+/- mice). Neutrophil apoptosis inhibition induced by lipopolysaccharide treatment in vitro or transendothelial migration in vivo observed in wild-type mice was abolished in both A1-a-/- and A1-a+/- animals. On the other hand, the extent of tumor necrosis factor alpha-induced acceleration of neutrophil apoptosis did not differ among A1-a-/-, A1-a+/-, and wild-type mice. The descending order of A1 mRNA expression was wild-type, A1-a+/-, and A1-a-/-. Taken together, these results suggest that A1 is involved in inhibition of certain types of neutrophil apoptosis.

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