title: [Generation and characterization of the adult neuron-specific ADAM10 knock-out mice].
abstract: A disintegrin and metalloproteinase 10 (ADAM10) is a major sheddase for over 30 different membrane proteins and gets involved in such physiological processes and pathogenesis as embryonic development, cell adhesion, signal transduction, immune reaction, cancer, and Alzheimer's disease. Both ADAM10 knock-out mice and the neural progenitor cell-specific ADAM10 knock-out mice having been reported so far died in the embryonic or perinatal stage, respectively, thus resulting in the failure to investigate ADAM10 function in the adult mouse brain. Through a series of tests, we have succeeded in generating and characterizing the CaMKIIα-Cre/ADAM10(loxP/loxP) mice surviving until adulthood by means of crossing ADAM10(loxP/loxP) mice with newly generated CaMKIIα-Cre transgenic mice. PCR analysis of genomic DNAs from different regions of the ADAM10 cKO mouse brain shows that the deleted ADAM10 alleles are mainly found in the cortex and hippocampus. Real-time RT-PCR findings further confirm that ADAM10 mRNAs decrease in the cortex and hippocampus by 55.7% and 60.8%, respectively. Western-blotting analysis demonstrates 63% and 84.8% loss of mature ADAM10 proteins from the cortex and hippocampus. Immunohistochemical tests show that there is significantly less ADAM10- positive staining in the cortical and hippocampal neurons but not gliocytes of ADAM10 cKO mice compared with control mice. In summary, we established the adult neuron-specific ADAM10 knock-out (cKO) mice for the first time, which prevented ADAM10(-/-) mice from the embryonic and perinatal mortality and laid a firm foundation for the further study of ADAM10 function in the brain of adult mice in vivo.
authors.affiliation: Department of Cell Biology and Genetics, School of Basic Medicine / Center of Cell and Developmental Biology/ Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Fujian Medical University, Fuzhou 350004, China. 290178077@qq.com
authors.forename: Jun
authors.initials: J
authors.lastname: Liu
authors.forename: Chang-Wen
authors.initials: CW
authors.lastname: Zhou
authors.forename: Qiu-Lan
authors.initials: QL
authors.lastname: Wei
authors.forename: Jian-Long
authors.initials: JL
authors.lastname: Zhuang
authors.forename: Zhao-Hua
authors.initials: ZH
authors.lastname: Lin
authors.forename: Jie-Hui
authors.initials: JH
authors.lastname: Zheng
mesh_terms: D051722:ADAM Proteins / Q000235:genetics* / Q000378:metabolism; D000072197:ADAM10 Protein; D053829:Amyloid Precursor Protein Secretases / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D005260:Female; D055786:Gene Knockout Techniques; D008297:Male; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout / Q000196:embryology / Q000235:genetics; D009474:Neurons / Q000201:enzymology*; D013045:Species Specificity
chemical_list: D008565:Membrane Proteins; D053829:Amyloid Precursor Protein Secretases; D051722:ADAM Proteins; D000072197:ADAM10 Protein; C491234:Adam10 protein, mouse