Effects Of ClC-3 On The Development Of Teeth And Jaws

ClC-3 belongs to a branch of the voltage gated chloride channel(ClC). The previous study showed that ClC-3 was expressed in osteoblasts and osteoclasts. While ClC-3 was overexpressed, the secretion of bone mineralization factor would be promoted. But whether ClC-3 was involved in the development of jaw and tooth was still unknown. In this study, we would explore the effects on teeth and bones when clcn3 gene defected. The experiment contents as follows:1?The tissues containing tooth germs from 13.5 d?15.5 d?18.5 d BALB/c pregnant mice and postnatal 1 d?5 d mice were dissected and performed regular fixation, embedding and paraffin section, then immunohistochemical stain for CLC-3 were performed. Then the distribution of ClC-3 was observed in mouse tooth germ.2?The gene structure of zebrafish clcn3 was analyzed and two zebrafish clcn3 specific Morpholino were designed and synthezied. Then the optimal concentration of clcn3 Morpholinos were determinged by injection. Then the general changes in morphology, jaws and teeth were observed.The results as follows:1?When the mouse tooth germ was at bud stage, CLC-3 was observed in the dental lamina epithelium. During cap stage, CLC-3 was expressed in the dental papilla cells and dental sac cells. In bell stage, CLC-3 was distributed in the dental papilla cells, dental sac cells, stellate reticulum cells and middle layer cells. The CLC-3 was widely expressed in dental papilla cells, dental sac cells, ameloblasts, odontoblasts, stellate reticulum cells and middle layer cells when the mouse born.2?Clcn3 mRNA was expressed in zebrafish at the different stages. We found that the best clcn3 Morpholino concentration is 0.25 nmol/?l, and the M2 showed the serious changes. After injection, we could see morphology changes of zebrafish as follows: heart edema, hematopoietic dysfunction, short physical development, body bending and activity failure. The ventral view of ceratohyal arranged in inverted “ V ” shape, and branchial arch cartilages showed “ ? ” arrangement. In addition, the length of head was decreased and we could see the asymmetric trabeculae from dorsal view. The number of pharyngeal tooth decreased, there was only one tooth which loosely attached to 5th branchial arch cartilage.So we found that the CLC-3 protein expressed in mouse tooth germ as well as zebrafish at the different stage.The deficiency of clcn3 gene could influence the development of zebrafish jaw and tooth. Further research are need to explore the deeper machanisms.