Molecular characterization and expression analysis of ADAM12 during chicken embryonic development

Abstract

ADAM12 is a member of the disintegrin and metalloprotease (ADAM) family of molecules, which consist of multiple domains. ADAM12 is involved in different physiological and pathological processes. In the present study, full-length sequences of two chicken ADAM12 isoforms were cloned and identified by reverse transcription-polymerase chain reaction (RT-PCR), rapid amplification of cDNA ends methods and bioinformatics analysis. The long isoform consists of all domains characteristic for ADAMs and is strongly expressed in different tissues, whereas the short isoform lacks large parts of the metalloprotease and disintegrin domains and is only expressed weakly. Results from semi-quantitative RT-PCR show that the complete ADAM12 is stably expressed throughout chicken embryonic development, while the short isoform is only regionally detectable in the lung and brain. Results from in situ hybridization show that chicken ADAM12 is expressed exclusively in tissues and organs derived from the neural tube, the neural crest or the mesoderm, with a highly regulated spatiotemporal expression pattern. Our data confirm and extend studies of ADAM12 in other species, and suggest that ADAM12 may play a role in the development of several organs, including the formation of feather buds.

Differential regional expression of multiple ADAMs during feather bud formation

“…We have found that the developing brain and spinal cord express each of seven members of the ADAM family (ADAM9, ADAM10, ADAM12, ADAM13, ADAM17, ADAM22, and ADAM23) in a characteristic spatiotemporal pattern (Lin et al, 2008a, 2010b; Yan et al, 2010). Our preliminary data showed that developing avian feathers express ADAM12 and ADAM19 also (Lin et al, 2010a; Yan et al, 2011). Therefore, in the present study, we investigate the expression patterns of the seven ADAMs during feather bud formation in detail by in situ hybridization.…”

Fast preparation of a polyclonal antibody against chicken protocadherin 1

“…Protein levels are measured indirectly. Indirect methods include immunohistochemistry, semiquantitative PCR (Lin et al, 2010), Western blotting, and ELISA (Zhao et al, 2012). Due to the complexity of protein composition in chicken embryo, it is difficult to isolate Pcdh1, so indirect detection is more applicable.…”