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Deficiency of phospholipase C-gamma1 impairs renal development and hematopoiesis
M Shirane1, H Sawa, Y Kobayashi
1Department of Product Research and. Department of Molecular Oncology, Nippon Roche Research Center, Kajiwara 200, Kamakura, Kanagawa 247-8530, Japan.
Abstract:
Phospholipase C-gamma1 (PLC-gamma1) is involved in a variety of intracellular signaling via many growth factor receptors and T-cell receptor. To explore the role of PLC-gamma1 in vivo, we generated the PLC-gamma1-deficient (plc-gamma1(-/-)) mice, which died of growth retardation at embryonic day 8.5-9.5 in utero. Therefore, we examined plc-gamma1(-/-) chimeric mice generated with plc-gamma1(-/-) embryonic stem (ES) cells for further study. Pathologically, plc-gamma1(-/-) chimeras showed multicystic kidney due to severe renal dysplasia and renal tube dilation. Flow cytometric analysis and glucose phosphate isomerase assay revealed very few hematopoietic cells derived from the plc-gamma1(-/-) ES cells in the mutant chimeras. However, differentiation of plc-gamma1(-/-) ES cells into erythrocytes and monocytes/macrophages in vitro was observed to a lesser extent compared with control wild-type ES cells. These data suggest that PLC-gamma1 plays an essential role in the renal development and hematopoiesis in vivo.
Insights
Phospholipase C-gamma1 (PLC-gamma1) is crucial for embryonic development, as PLC-gamma1-deficient mice exhibit severe growth retardation. PLC-gamma1 is essential for kidney development and blood cell formation in vivo.
Area of Science:
- Molecular Biology
- Developmental Biology
- Immunology
Background:
- Phospholipase C-gamma1 (PLC-gamma1) is a key enzyme in intracellular signaling pathways activated by growth factor and T-cell receptors.
- Its precise role in complex biological processes in vivo remains incompletely understood.
Purpose of the Study:
- To investigate the in vivo function of PLC-gamma1 during embryonic development.
- To elucidate the specific roles of PLC-gamma1 in renal development and hematopoiesis.
Main Methods:
- Generation of PLC-gamma1-deficient (plc-gamma1(-/-)) mice and subsequent analysis of plc-gamma1(-/-) chimeric mice derived from embryonic stem (ES) cells.
- Pathological examination of chimeric mice for renal abnormalities.
- Flow cytometry and glucose phosphate isomerase assays to assess hematopoietic cell contribution.
- In vitro differentiation assays of plc-gamma1(-/-) ES cells.
Main Results:
- PLC-gamma1-deficient mice exhibit embryonic lethality due to severe growth retardation.
- PLC-gamma1(-/-) chimeric mice display multicystic kidneys characterized by severe renal dysplasia and tubular dilation.
- A significantly reduced contribution of plc-gamma1(-/-) ES cells to hematopoietic lineages was observed in chimeric mice.
- Impaired in vitro differentiation of plc-gamma1(-/-) ES cells into erythrocytes and monocytes/macrophages compared to wild-type controls.
Conclusions:
- PLC-gamma1 plays an indispensable role in embryonic development, specifically in renal morphogenesis and the development of the hematopoietic system.
- These findings highlight PLC-gamma1 as a critical regulator of kidney formation and blood cell production in vivo.