Identification of a novel metalloproteinase and its role in juvenile development of the tobacco hornworm, Manduca sexta (Linnaeus).
Bottom Line: To directly test the hypothesis that Ms-MMP plays a critical role in juvenile development of Manduca sexta, we injected a broad-spectrum MMP inhibitor and recorded its effects on growth and development.Inhibition of MMPs caused a delay in juvenile development and decreased growth rates.Furthermore, elucidating functions for MMPs in lower taxa may yield critical information about the evolution of the numerous MMPs found in vertebrates.
Affiliation: Department of Biological Sciences, North Dakota State University, Fargo, North Dakota 58108-6050, USA.Show MeSH
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Mentions: Using gene-specific primers designed from the Ms-MMP sequence (Table 1), we detected Ms-MMP gene expression in tracheae, fat body and hemocytes (Fig. 3A). Sequence analysis of these PCR products confirmed our initial sequencing results. Western blot analysis of tissue samples revealed most Ms-MMP protein expression in fat body and tracheae (Fig. 3B) as a 46 kDa band, with a smaller band at 35 kDa. In hemolymph, Ms-MMP was detected at 100 and 22 kDa, and hemocytes showed a single band at 22 kDa (Fig. 3B). The bands are specific to Ms-MMP, since subsequent probing of the blot with antibody that was pre-incubated with the peptide-antigen complex resulted in the disappearance of the bands (Fig. 3C).
Affiliation: Department of Biological Sciences, North Dakota State University, Fargo, North Dakota 58108-6050, USA.