Differential expression of splice variants of chemokine CCL27mRNA in lens, cornea, and retina of the normal mouse eye

[摘要] Purpose: Constitutive expression of RNA sequences complementary tothe chemokine CCL27 mRNA has been found in the normal mouse eye.This study examines the nature and location of these endogenous RNAs inocular tissues.Methods: Conventional RT-PCR, 5' RACE, and dideoxy DNA sequencingwere used to examine the sequences of CCL27 related RNAs in the eye.Expression levels of specific RNAs were measured by real time PCR.Tissue distribution of RNA transcripts was determined by RT-PCR usingRNA from microdissected tissues and by in situ hybridization withradiolabeled riboprobes.Results: We detect 5 distinct splice variants derived fromtranscription of the CCL27 gene locus. The most abundant form codesfor a non-secreted protein, PESKY, and is expressed in lens, cornea, andretina. Another variant corresponds to the mRNA of the secretedchemokine and is synthesized in the cornea, but not in retina or lens.The remaining splice variants are novel and may be eye specific, buthave only short open reading frames (<50 amino acids). CCL27transcripts are most abundantly expressed in the retina, as judged by insitu hybridization.Conclusions: PESKY and other CCL27 splice variants of unknownfunction are widely expressed in ocular tissues. Analysis of CCL27transcripts from lens, retina, and cornea indicates that mRNA for thesecreted chemokine, CCL27, is endogenously expressed only in the corneaand may play a role in ocular immune responses involving CD4 lymphocytesin this tissue.

[学科分类] 生物化学/生物物理