The Expressive Organs And Gene Localization Examination Of The Reindeer β-defensin-1 (reBD-1) In Reindeer Tissues

Defensins are a kind of important component of endogenous antimicrobial producing from organisms and contain five subfamilies ofα-defensins,β-defensins,θ-defensins, insect defensins and plant defensins. Theβ-defensins mostly distributed in the epithelium of skin ang mucosa of vertebrate and formed the first chemistry barrier resisting microorganisms attack, soβ-defensins are proved to be a component of antimicrobial barrier. It has been reported thatβ-defensins express extensively in the epithelial tissues of many organs in mammals such as cow, sheep, goat, pig, mouse, rat, human, monkey.In order to examine the organs that express possibly reBD-1 mRNA in reindeers, we designed a pair of primers, which expected product is 121bp, according to the known reBD-1 cDNA sequence. We extracted the total RNA from the tissues of a reindeer's tongue , esophagus, rumen, reticulum, abomasum, duodenum, ileum, colon, liver, lung, trachea, kidney, bladder, testis, epididymis, heart, spleen and examined the expression of reBD-1 mRNA in above different tissues of the reindeer by RT-PCR. The result indicates that reBD-1 mRNA was expressed in the all organs that was examinated. The most expressed tissues of reBD-1 mRNA was tongue, rumen and testis; the middling grade expressed tissues was oesophagus, duodenum, colon, trachea and spleen and in the reticulum, abomasums, ileum, liver, lung, kidney, bladder, epididymis and heart; the expression of reBD-1 mRNA was relatively low. The extensive expression of reBD-1 in reindeers suggests that reBD-1may play a role in host defense.. In order to make sure the reBD-1 expressive position in tissues of reindeer organs, we choosed reindeer tongue, ileum and kidney as tissue samples and examined the expressive tissues of reBD-1 mRNA in reindeers by RNA in situ hybridization ( RISH ) using the digoxigenin-labeled antisense nucleic acid probe of 300bp which was cloned. The result shows that the reBD-1 mRNA express in the columnar epithelium of intestinal crypts, stratified squamous epithelium of dorsal tongue, epithelium of renal tubule and renal capsule. The hybridization signals of reBD-1 mRNA in stratified squamous epithelium of dorsal tongue are inhomogeneity: hybridization signal mostly lie in the middle-level stratified squamous epithelium and conicae papillae, not in superficial and basal epithelial cells ultimately, which show that the reBD-1 mRNA did not express in stratified squamous epithelium of superficial-level and basal-level, and express in stratified squamous epithelium of middle-level and conicae papillae of tongue. The columnar epithelium of intestinal crypts have hybridize signal, but basal epithelium of intestinal crypts have not hybridization signals. There is hybridization signals in the epithelium of renal tubule and renal capsule, which indicates reBD-1 mRNA also expresses in epithelium in kidney of material organs.