ZFIN ID: ZDB-PUB-140728-25
Detection of nitric oxide by diaminofluorescein visualizes the skeleton in living zebrafish
Renn, J., Pruvot, B., and Muller, M.
Several in vivo stainings, such as Calcein, Alizarin Red and Quercetin are commonly used to visualize ossification in living teleost specimen. These staining techniques represent important tools for bone research in fish, but do not visualize cartilage. In the present study, we show that nitric oxide (NO) labelling by DAF-FM DA visualizes both bone and cartilage in vivo during zebrafish skeletogenesis. NO detection performed in Tg(osterix:mCherry) or in combination with Alizarin Red in wild-type zebrafish reveals that intense staining through NO labelling colocalizes with the appearance of osteoblasts and characterizes ossified structures. Cartilage structures are clearly distinguished in the living larvae, although the labelling is less intensive when compared to ossified structures. This method is the first and easy to handle alternative to cartilage and bone double stainings on fixed samples. In contrast to most live skeletal stainings, which only stain the mineralized bone structures, this protocol in addition allows in vivo visualization of cartilage.