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Analysis of mRNA for class I HLA on human gametogenic cells
Author(s) -
Janitz Michal,
Fiszer Dorota,
MichalczakJanitz Karolina,
Lukaszyk Andrzej,
Fernandez Nelson,
Skorupski Witold,
Kurpisz Maciej
Publication year - 1994
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/mrd.1080380215
Subject(s) - biology , complementary dna , microbiology and biotechnology , human leukocyte antigen , untranslated region , messenger rna , gene , transmembrane protein , coding region , gene expression , signal peptide , genetics , antigen , peptide sequence , receptor
We have studied mRNA expression for Class I HLA (human leukocyte antigen) on male germ cells by amplification of gene fragments in PCR techique and by Northern hybridization. RNA was extracted from fractionated gametogenic cells (isolated from testis) and reversely transcribed. Then, cDNA was amplified for specific HLA sequence (1151 bp) representing whole‐length coding sequence (HLA, ‐A, ‐B, ‐C). The specificity of this product was confirmed in “nested” PCR of 400 bp gene fragment coding for alpha 2 domain, alpha 3 domain, and the transmembrane portion of Class I HLA. The results indicate minimal expression of classical Class I HLA on gametogenic cells. Northern hybridization with 669 bp cDNA fragment (spanning for alpha 3 domain, transmembrane, cytoplasmic, and 3′ untraslated region) resulted in a low intensity signal from gametogenic cell fractions and confirmed our findings obtained by PCR. The minimal expression of classical HLA antigens may create a neutral cover for the male reproductive system, thereby preventing an immunological response during germ cell differentiation. © 1994 Wiley‐Liss, Inc.