title: Expression of alternatively spliced CD45 isoforms by channel catfish clonal T and B cells is dependent on activation state of the cell and regulated by protein synthesis and degradation.
abstract: In mammals, expression of the three alternatively spliced exons of the tyrosine phosphatase CD45 is regulated by the developmental and activation state of the cell. In comparison, the channel catfish, Ictalurus punctatus, CD45 homolog contains 18 functional alternatively spliced exons. Since very little is known about CD45 regulation in ectothermic vertebrates, this study examines the regulation of catfish CD45 mRNA isoform expression in clonal T and B cells in response to stimulation. Results show that mitogenic stimulation using catfish serum or concanavalin A induced expression of mRNAs for small CD45 isoforms, and isoform message expression was growth curve dependent, i.e. cells in logarithmic phase express message for smaller CD45 isoforms, whereas stationary phase cells express message for longer CD45 isoforms. In addition, cells treated with the protein synthesis inhibitor cycloheximide expressed message for longer CD45 isoforms, and treatment with lactacystin, which blocks protein degradation, rescued smaller isoform message expression. Collectively these data suggested that expression of CD45 isoforms, in catfish, at least at the mRNA level, is "constitutively dynamic" and highly dependent on extracellular stimuli.
authors.affiliation: Department of Immunology Box 3010, Duke University Medical Center, 352 Jones Building, Research Drive, Durham, NC 27710, USA. kount002@mc.duke.edu
authors.forename: Evgueni
authors.initials: E
authors.lastname: Kountikov
authors.forename: Deepak
authors.initials: D
authors.lastname: Nayak
authors.forename: Melanie
authors.initials: M
authors.lastname: Wilson
authors.forename: Norman W
authors.initials: NW
authors.lastname: Miller
authors.forename: Eva
authors.initials: E
authors.lastname: Bengtén
mesh_terms: D000111:Acetylcysteine / Q000031:analogs & derivatives / Q000494:pharmacology; D017398:Alternative Splicing / Q000187:drug effects / Q000276:immunology; D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D002999:Clone Cells; D003208:Concanavalin A / Q000276:immunology / Q000378:metabolism; D003513:Cycloheximide / Q000494:pharmacology; D025461:Feedback, Physiological; D020869:Gene Expression Profiling; D007059:Ictaluridae; D007114:Immunization; D017493:Leukocyte Common Antigens / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D008213:Lymphocyte Activation / Q000187:drug effects; D020033:Protein Isoforms / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D013601:T-Lymphocytes / Q000166:cytology / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*
chemical_list: D020033:Protein Isoforms; D003208:Concanavalin A; C067713:lactacystin; D003513:Cycloheximide; D017493:Leukocyte Common Antigens; D000111:Acetylcysteine