Journal
JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY
Volume 66, Issue 1, Pages 47-58Publisher
SAGE PUBLICATIONS LTD
DOI: 10.1369/0022155417739283
Keywords
caspases; endochondral ossification; growth plate; immunohistochemistry; micromass cultures; PCR Array analysis
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Funding
- Czech Science Foundation [GACR 16-18430S]
- IGA UVPS Brno, Czech Republic [117/2017/FVL]
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Caspases have functions particularly in apoptosis and inflammation. Increasing evidence indicates novel roles of these proteases in cell differentiation, including those involved in osteogenesis. This investigation provides a complex screening of osteogenic markers affected by pan caspase inhibition in micromass cultures derived from mouse forelimbs. PCR Array analysis showed significant alterations in expression of 49 osteogenic genes after 7 days of inhibition. The largest change was a decrease in CD36 expression, which was confirmed at organ level by caspase inhibition in cultured mouse ulnae followed by CD36 immunohistochemical analysis. So far, available data point to osteogenic potential of pro-apoptotic caspases. Therefore, the expression of pro-apoptotic caspases (-3, -6, -7, -8, -9) within the growth plate of mouse forelimbs at the stage where the individual zones are clearly apparent was studied. Caspase-9 was reported in the growth plate for the first time as well as caspase-6 and -7 in the resting zone, caspase-7 in the proliferation, and caspase-6 and -8 in the ossification zone. For all caspases, there was a gradient increase in activation toward the ossification zone. The distribution of staining varied significantly from that of apoptotic cells, and thus, the results further support non-apoptotic participation of caspases in osteogenesis.
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