title: DNA damage precedes apoptosis during the regression of the interdigital tissue in vertebrate embryos.
abstract: DNA damage independent of caspase activation accompanies programmed cell death in different vertebrate embryonic organs. We analyzed the significance of DNA damage during the regression of the interdigital tissue, which sculpts the digits in the embryonic limb. Interdigit remodeling involves oxidative stress, massive apoptosis and cell senescence. Phosphorylation of H2AX mediated by ATM precedes caspase dependent apoptosis and cell senescence during interdigit regression. The association of γH2AX with other downstream DNA repair factors, including MDC1, Rad50 and 53BP1 suggests a defensive response of cells against DNA damage. The relative distribution of cells γH2AX-only positive, TUNEL-only positive, and cells double positive for both markers is consistent with a sequence of degenerative events starting by damage of the DNA. In support of this interpretation, the relative number of γH2AX-only cells increases after caspase inhibition while the relative number of TUNEL-only cells increases after inhibition of ATM. Furthermore, cultured interdigits survived and maintained intense chondrogenic potential, even at advanced stages of degeneration, discarding a previous commitment to die. Our findings support a new biological paradigm considering embryonic cell death secondary to genotoxic stimuli, challenging the idea that considers physiological cell death a cell suicide regulated by an internal death clock that pre-programmes degeneration.
authors.affiliation: Departamento de Anatomía y Biología Celular and IDIVAL, Universidad de Cantabria, Santander 39011, Spain.
authors.forename: Juan A
authors.initials: JA
authors.lastname: Montero
authors.affiliation: Departamento de Anatomía y Biología Celular and IDIVAL, Universidad de Cantabria, Santander 39011, Spain.
authors.forename: Cristina
authors.initials: C
authors.lastname: Sanchez-Fernandez
authors.affiliation: Departamento de Anatomía y Biología Celular and IDIVAL, Universidad de Cantabria, Santander 39011, Spain.
authors.forename: Carlos I
authors.initials: CI
authors.lastname: Lorda-Diez
authors.affiliation: Departamento de Anatomía y Biología Celular and IDIVAL, Universidad de Cantabria, Santander 39011, Spain.
authors.forename: Juan A
authors.initials: JA
authors.lastname: Garcia-Porrero
authors.affiliation: Departamento de Anatomía y Biología Celular and IDIVAL, Universidad de Cantabria, Santander 39011, Spain.
authors.forename: Juan M
authors.initials: JM
authors.lastname: Hurle
mesh_terms: D000818:Animals; D017209:Apoptosis; D020169:Caspases / Q000378:metabolism; D016923:Cell Death; D016922:Cellular Senescence / Q000235:genetics; D002642:Chick Embryo; D020219:Chondrogenesis / Q000235:genetics; D004249:DNA Damage; D004260:DNA Repair; D047108:Embryonic Development / Q000235:genetics*; D005455:Fluorescent Antibody Technique; D006657:Histones / Q000378:metabolism; D007150:Immunohistochemistry; D051379:Mice; D018384:Oxidative Stress; D014714:Vertebrates
chemical_list: D006657:Histones; D020169:Caspases