To minimize the chance of off-target effects, we have performed experiments such as dose-response, time course and use of multiple controls including a 5-nucleotide mutated nestin MO. The 5-mis MO control and a standard MO control up to 10 ng did not cause morphological abnormality and apoptotic changes. The results indicate that the defects in brain and eyes are unlikely due to off-target effects of MO as there is consistent dose-response and time-dependent effects. Nestin is a selective NPC intermediate filament involved in cytoskeletal functions such as maintaining cell shape and regulating cell motility. Recent studies suggest that it is involved in other cellular functions. Its role in apoptosis is less well characterized. However, several studies have indicated that the actin cytoskeleton regulates apoptosis and its defects are associated with neurodegenerative diseases. Furthermore, filamentous aggregates of neuronal intermediate filament proteins are considered to be neurophathological signatures of neurodegenerative diseases. It has been reported that deletion of the hepatocyte intermediate filament proteins, keratin 8/keratin 18 Mupirocin heterodimers, increases apoptosis. Our results suggest that Nestin plays an important role in NPC survival in zebrafish embryo development. The mechanism by which nestin protects cell survival is unclear. Studies from murine cells have suggested that nestin may act as a scaffold to control cell functions including apoptosis. It has been reported that in a murine NPC line, Nestin interacts with and sequesters cyclin-dependent kinase 5, which is constitutively expressed in NPCs. Reduced Nestin BAR 501 impurity protein levels lead to enhanced Cdk5 activity, promoting NPC apoptosis. Further studies are needed to clarify whether control of NPC survival by Nestin in zebrafish is also mediated by this mechanism. Our results show that the cranial motor nerve development is disrupted by suppression of nestin in Tg fish which express GFP under the control of a motor neuron-specific islet1 enhancer.