Work by a number of previous students, using a variety of methods, generated much correlative data suggesting that NO levels drop once metamorphosis was triggered. To gain direct molecular evidence in support of this idea, Kenneth Fowler – MS degree 2005 cloned a portion of the larval neuronal NOS gene. Through reverse transcription-based polymerase chain reaction (RT-PCR) analysis, as shown Below, Ken discovered that levels of NOS gene expression decrease during the 24 hours following metamorphic induction (Hens et al., 2006). Ken worked with both Dr. Mark Hens and Dr. Leise while conducting his M.S. thesis research. Supported by NSF grant IBN-0130677.
Above: nNOS RNA levels drop 6 hours after metamorphosis is induced with serotonin (5-HT). Expression of genes for actin and 18s RNA were unchanged during the same time period
