Multiple Changes in Peptide and Lipid Expression Associated with Regeneration in the Nervous System of the Medicinal Leech
Figure 4
DD-RP-HPLC of leech axotomized nerve cords acidic extracts at different times after connective nerve trans-section.
After prepurification by solid phase extraction, the 60% AcN eluted material is loaded onto a C18 column. Elution is performed with a linear gradient of AcN (dotted line) and absorbance was monitored at 225 nm. Each individually collected peak is tested for its neurites outgrowth (example in upper left insert) and antimicrobial (example upper right insert) activities. The peaks that showed temporal variation in intensity, whether they produced antimicrobial or neurotrophic activity or not, are further purified and the peptides are identified by MALDI-TOF mass spectrometry and Edman degradation. Sequence results and peptide identification are shown in Table 2.