Abstract
Purpose: :
It is well established that the adult teleost fish neural retina is capable of significant regeneration following injury. However, we are still discovering the molecular and genetic determinants of this process. The current study examines the importance of a target gene, fatty acid binding protein 7 (FABP7), during retinal regeneration in the adult zebrafish as well as during embryonic development of the neural retina. Using the techniques stated below, we wanted to determine if FABP7 plays a significant role during development of the fish retina and is necessarily re-expressed during regeneration of the adult fish retina.
Methods: :
Gene array, real-time qPCR and RT-PCR were utlilized to demonstrate an increase in FABP7 gene expression following injury of the adult zebrafish retina and within the developing embryonic retina. In addition, the spatial distribution of FABP7 was determined using non-isotopic in situ hybridization and immunohistochemistry (IHC). Subsequently, morpholinos were injected into 2-8 cell stage embryos and in the eyes of adult zebrafish as a means to knock-down FABP7 expression. Real-time qPCR, RT-PCR, and in situ hybridization along with IHC were used to examine any expression level changes in embryos and injured adult retinas following incorporation of FABP7 morpholinos.
Results: :
Results observed from using the above techniques indicate that FABP7 is highly expressed during neural retina development and following injury of the adult zebrafish retina. Injection of morpholinos alters gene expression profiles in the developing and injured fish retinas, as well as effecting the number of cells passaging through the cell cycle in the injured retina.
Conclusions: :
The FABP7gene is highly expressed during embryonic development of the neural retina and is upregulated in the adult zebrafish retina following injury. Following MO-FABP7 exposure, alterations in the developing zebrafish retina suggest a necessary role for FABP7 during formation of the retina and perhaps during the regeneration process. These results further suggest a recapitulation of development during the regeneration process of the adult fish retina.
Keywords: development • regeneration • gene/expression