TY - JOUR
T1 - m-Calpain is required for preimplantation embryonic development in mice
AU - Dutt, Previn
AU - Croall, Dorothy E.
AU - Arthur, J. Simon C.
AU - De Veyra, Teresa
AU - Williams, Karen
AU - Elce, John S.
AU - Greer, Peter A.
N1 - © 2006 Dutt et al; licensee BioMed Central Ltd.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0),
which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Initial cloning of Capn2 and vector construction was undertaken by DEC as a sabbatical visitor at Queen's University with support from the National Science Foundation (MCB9723636) and the University of Maine.
This work was supported by grants from the Canadian Institutes of Health
Research and The Heart and Stroke Foundation of Canada. P.D. was an
Ontario Graduate Scholar
PY - 2006/1/24
Y1 - 2006/1/24
N2 - Background: μ-calpain and m-calpain are ubiquitously expressed proteases implicated in cellular migration, cell cycle progression, degenerative processes and cell death. These heterodimeric enzymes are composed of distinct catalytic subunits, encoded by Capn1 (μ-calpain) or Capn2 (m-calpain), and a common regulatory subunit encoded by Capn4. Disruption of the mouse Capn4 gene abolished both μ-calpain and m-calpain activity, and resulted in embryonic lethality, thereby suggesting essential roles for one or both of these enzymes during mammalian embryogenesis. Disruption of the Capn1 gene produced viable, fertile mice implying that either m-calpain could compensate for the loss of μ-calpain, or that the loss of m-calpain was responsible for death of Capn4-/- mice. Results: To distinguish between the alternatives described above, we deleted an essential coding region in the mouse Capn2 gene in embryonic stems cells and transmitted this mutant allele through the mouse germline. Breeding of heterozygous animals failed to produce homozygous mutant live offspring or implanted embryos. A nested PCR genotyping protocol was established, and homozygous preimplantation mutant embryos were detected at the morula but not at the blastocysts stage. Conclusion: We conclude that homozygous disruption of the Capn2 gene results in preimplantation embryonic lethality between the morula and blastocyst stage. This establishes that μ-calpain and m-calpain have distinct functions, and that m-calpain is vital for development of the preimplantation murine embryo.
AB - Background: μ-calpain and m-calpain are ubiquitously expressed proteases implicated in cellular migration, cell cycle progression, degenerative processes and cell death. These heterodimeric enzymes are composed of distinct catalytic subunits, encoded by Capn1 (μ-calpain) or Capn2 (m-calpain), and a common regulatory subunit encoded by Capn4. Disruption of the mouse Capn4 gene abolished both μ-calpain and m-calpain activity, and resulted in embryonic lethality, thereby suggesting essential roles for one or both of these enzymes during mammalian embryogenesis. Disruption of the Capn1 gene produced viable, fertile mice implying that either m-calpain could compensate for the loss of μ-calpain, or that the loss of m-calpain was responsible for death of Capn4-/- mice. Results: To distinguish between the alternatives described above, we deleted an essential coding region in the mouse Capn2 gene in embryonic stems cells and transmitted this mutant allele through the mouse germline. Breeding of heterozygous animals failed to produce homozygous mutant live offspring or implanted embryos. A nested PCR genotyping protocol was established, and homozygous preimplantation mutant embryos were detected at the morula but not at the blastocysts stage. Conclusion: We conclude that homozygous disruption of the Capn2 gene results in preimplantation embryonic lethality between the morula and blastocyst stage. This establishes that μ-calpain and m-calpain have distinct functions, and that m-calpain is vital for development of the preimplantation murine embryo.
UR - http://www.scopus.com/inward/record.url?scp=33644558362&partnerID=8YFLogxK
U2 - 10.1186/1471-213X-6-3
DO - 10.1186/1471-213X-6-3
M3 - Article
C2 - 16433929
AN - SCOPUS:33644558362
SN - 1471-213X
VL - 6
JO - BMC Developmental Biology
JF - BMC Developmental Biology
M1 - 3
ER -