Differential tissue growth and patterns of cell death in mouse limb autopod morphogenesis

Dev Dyn. 2001 Apr;220(4):295-306. doi: 10.1002/dvdy.1108.


Programmed cell death (PCD) is considered one of the most important cellular processes in the morphogenesis of organs and tissues during animal development. Although the embryonic limb has been established as a classic model for the study of PCD, detailed studies on this process' contribution to morphogenesis are still lacking. In the present work, using modern computer-aided techniques, we estimated the contribution of PCD to mouse limb morphogenesis. For the detection of apoptotic cell death, we stained whole embryonic limbs with acridine orange or, in some instances, used the TUNEL technique, and visualized the tissues by confocal laser scanning microscopy. We found that cell death patterns are dynamic during limb development, and occur in gradients oriented with the main limb axes, anteroposterior, dorsoventral and distoproximal. Interdigital apoptosis in the autopod was initially detected at the most distal region, and then more proximally as development proceeded. Interestingly, we found that digit separation is more pronounced on the dorsal side, contrary to what is expected from the apoptotic cell distribution, which shows more abundant cell death in the ventral region. Using 2-D and 3-D models, we found that most digit individualization occurs rather by digit growth than by interdigital cell death. Therefore, digits do not mainly individualize by degeneration of preformed interdigital tissue, but probably by a dynamic balance between proliferation and cell death, reducing interdigital growth, which results in protrusion of digits. We determined the expression pattern of fgf-8 during the period of digit individualization, as the product of this gene could participate in defining the limb growth pattern. Initially, fgf-8 expression was coincident with the apical ectodermal ridge, but when cell death was first detected in the interdigits, fgf-8 expression became restricted to the tip of the growing digits. Therefore, FGF-8 could be one of the factors responsible for differential digit-interdigit growth, and might also act as a survival factor on interdigital tissue. We also found that the expression patterns of rar-beta, bmp-2, bmp-4, bmp-7, msx-1, and msx-2 genes, proposed to be involved in the activation of interdigital cell death, did not overlap with, or were not highly expressed in the major zones of cell death in the developing limb.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis*
  • Bone Morphogenetic Protein 2
  • Bone Morphogenetic Protein 4
  • Bone Morphogenetic Protein 7
  • Bone Morphogenetic Proteins / biosynthesis
  • Bone and Bones / embryology
  • Bone and Bones / metabolism
  • Cell Division
  • DNA-Binding Proteins / biosynthesis
  • Extremities / embryology*
  • Fibroblast Growth Factor 8
  • Fibroblast Growth Factors / biosynthesis
  • Homeodomain Proteins / biosynthesis
  • Image Processing, Computer-Assisted
  • In Situ Hybridization
  • In Situ Nick-End Labeling
  • MSX1 Transcription Factor
  • Mice
  • Microscopy, Confocal
  • Receptors, Retinoic Acid / biosynthesis
  • Software
  • Time Factors
  • Tissue Distribution
  • Transcription Factors*
  • Transforming Growth Factor beta*


  • Bmp2 protein, mouse
  • Bmp4 protein, mouse
  • Bone Morphogenetic Protein 2
  • Bone Morphogenetic Protein 4
  • Bone Morphogenetic Protein 7
  • Bone Morphogenetic Proteins
  • DNA-Binding Proteins
  • Fgf8 protein, mouse
  • Homeodomain Proteins
  • MSX1 Transcription Factor
  • MSX2 protein
  • Receptors, Retinoic Acid
  • Transcription Factors
  • Transforming Growth Factor beta
  • retinoic acid receptor beta
  • Fibroblast Growth Factor 8
  • Fibroblast Growth Factors