Expression of heat-shock protein 70 during limb development and regeneration in the axolotl



Urodele amphibians (e.g., axolotls) have the unique ability, among vertebrates, to regenerate perfectly many parts of their body after amputation. The limb has been the most widely studied regenerating structure in these organisms and provides an ideal model in which to understand how vertebrates can regenerate complex tissues. The present study focuses on Hsp-70, a member of the stress-related heat-shock protein family. This protein is normally induced after a stress or trauma such as heat-shock, ultraviolet irradiation, or wounding. Thus, studying its expression during axolotl limb regeneration, a response to an important traumatic event (amputation), is of great interest to further understand how the regenerative process is mediated. Using molecular biology and biochemical techniques, we have characterized both the spatiotemporal and quantitative expression patterns of Hsp-70 in axolotl development and regeneration. Our results show that Hsp-70 is expressed and regulated during axolotl development as in other vertebrates. Our data also demonstrate an up-regulation of the RNA transcript for Hsp-70 during limb regeneration as early as 24 hr after amputation that is maintained up to early differentiation. We also demonstrate a similar pattern of expression for the protein during regeneration. Finally, we show that axolotl Hsp-70 is induced threefold after heat-shock as observed in other vertebrates. Developmental Dynamics 233:1525–1534, 2005. © 2005 Wiley-Liss, Inc.