title: [Differentially expressed genes identified in the main olfactory epithelium of mice with deficiency of adenylate cyclase 3 by using suppression subtractive hybridization approach].
abstract: Adenylate cyclase 3 (AC3) is one of the major players in the olfactory signaling within the main olfactory epithelium (MOE) of mice. However, we are not ascertained whether deficiency of AC3 will lead to the differential expression of related genes in the MOE. Forward and reverse subtractive libraries were constructed by suppression subtractive hybridization (SSH) approach, with MOEs from AC3(-/-) and AC3(+/+) mice. These two libraries were primarily screened by Dot blot, differential expressed clones were sequenced and analyzed by bioinformatics, and differential expressed genes were verified by qRT-PCR. A total of 386 differentially expressed clones were picked out after Dot blot. The DNA sequences of 80 clones randomly selected were determined, and 62 clones were identified by blasting in GenBank. We found that 24 up-regulated clones were corresponded to genes of kcnk3, mapk7, megf11, and 38 down-regulated clones were corresponded to tmem88b, c-mip, skp1a, mlycd, etc. Their functions were annotated with Gene Ontology (GO) and found to be mainly focused on molecular binding, cell cycle, processes of biology and cells. Five genes (kcnk3, c-mip, mlycd, tmem88b and trappc5) were verified by qRT-PCR with individuals of AC3(+/+) and AC3(-/-) mice. The data indicate that kcnk3 gene is up-regulated significantly, increasing 1.27 folds compared to control mice, whereas c-mip, mlycd, tmem88b and trappc5 are down-regulated significantly, decreasing 20%, 7%, 32% and 29% compared to the AC3(+/+)mice. The functions of these genes are closely related with K(+) channels, cell differentiation, metabolism of fats, membrane transportation, and so on. It is tempting to speculate that these genes might work together with AC3 to orchestrate the olfactory transduction signaling in the MOE.
authors.affiliation: College of Life Science, Hebei University, Baoding 071002, China.
authors.forename: Cao
authors.initials: C
authors.lastname: Zhenlong
authors.affiliation: College of Life Science, Hebei University, Baoding 071002, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Jiangye
authors.affiliation: College of Life Science, Hebei University, Baoding 071002, China; Research Center of Bioengineering of Hebei Province, Baoding 071002, China.
authors.forename: Zhou
authors.initials: Z
authors.lastname: Yanfen
authors.affiliation: College of Basic Medicine, Hebei University, Baoding 071002, China.
authors.forename: Zhang
authors.initials: Z
authors.lastname: Zhe
authors.affiliation: College of Life Science, Hebei University, Baoding 071002, China.
authors.forename: Ni
authors.initials: N
authors.lastname: Zhihua
authors.affiliation: College of Basic Medicine, Hebei University, Baoding 071002, China.
authors.forename: Hu
authors.initials: H
authors.lastname: Yuanxiang
authors.affiliation: College of Basic Medicine, Hebei University, Baoding 071002, China.
authors.forename: Liu
authors.initials: L
authors.lastname: Weili
authors.affiliation: Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Li
authors.initials: L
authors.lastname: Yongchao
authors.affiliation: Department of Pharmacology, University of Washington, Seattle, WA 98195, USA.
authors.forename: R Storm
authors.initials: RS
authors.lastname: Daniel
authors.affiliation: Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Z Ma
authors.initials: ZM
authors.lastname: Runlin
authors.affiliation: College of Life Science, Hebei University, Baoding 071002, China; Research Center of Bioengineering of Hebei Province, Baoding 071002, China.
authors.forename: Wang
authors.initials: W
authors.lastname: Zhenshan
mesh_terms: D000262:Adenylyl Cyclases / Q000172:deficiency* / Q000235:genetics; D000818:Animals; D020869:Gene Expression Profiling; D051379:Mice / Q000235:genetics* / Q000502:physiology; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D009831:Olfactory Mucosa / Q000378:metabolism*; D011506:Proteins / Q000235:genetics* / Q000378:metabolism; D012903:Smell
chemical_list: D011506:Proteins; D000262:Adenylyl Cyclases; C421784:adenylate cyclase 3