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Microarray and cDNA sequence analysis of transcription during nerve-dependent limb regeneration
James R Monaghan1, Leonard G Epp, Srikrishna Putta
1Department of Biology and Spinal Cord and Brain Injury Research Center, University of Kentucky, Lexington, KY 40506, USA. james.monaghan@uky.edu
BMC Biology
|January 16, 2009
Summary
Limb regeneration in salamanders is not entirely nerve-dependent. Early wound healing processes occur similarly in both innervated and denervated limbs, with nerve influence appearing later in blastema formation.
Area of Science:
- Regenerative Biology
- Molecular Biology
- Developmental Biology
Background:
- Investigating the basis of nerve-dependent limb regeneration in salamanders.
- Utilizing microarray analysis and 454 cDNA sequencing to study Mexican axolotl forelimb regeneration.
Purpose of the Study:
- To determine the role of nerve-derived factors in early limb regeneration.
- To identify transcriptional differences between innervated and denervated limb regeneration.
Main Methods:
- Amputation of innervated (NR) and denervated (DL) axolotl forelimbs.
- Transcriptional profiling at 0, 5, and 14 days post-amputation (dpa) using microarrays and 454 cDNA sequencing.
- Comparison of gene expression patterns between NR and DL limbs.
Main Results:
- Early regeneration (5 and 14 dpa) showed transcriptional similarities between NR and DL limbs, with upregulated extracellular wound-healing genes and downregulated muscle genes.
- Nerve-derived factors influenced later stages, with increased cell proliferation and distinct transcriptional signatures in NR limbs at 14 dpa, absent in DL limbs.
- DL limbs exhibited more diverse and prolonged wound-healing responses.
- 454 sequencing expanded the axolotl EST database, doubling non-redundant orthologous sequences.
Conclusions:
- Many early limb regeneration processes are independent of nerve-derived factors.
- Nerve dependency becomes apparent during blastema formation and cell proliferation.
- Discovery of new gene sequences facilitates future research in axolotl regeneration, wound healing, and related fields.
