Characterization of laminin receptor messenger ribonucleic acid and protein expression in mouse spermatogenic cells.
Characterization of laminin receptor messenger ribonucleic acid and protein expression in mouse spermatogenic cells.
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小鼠生精细胞中层粘连蛋白受体信使核糖核酸和蛋白质表达的表征。
DOI：
10.1095/biolreprod48.3.674
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发表时间：
1993
影响因子：
3.6
通讯作者：
Eddy,EM
中科院分区：
文献类型：
--
作者：
Fulcher,KD;Welch,JE;Davis,CM;O'Brien,DA;Eddy,EM
从小鼠圆形精子细胞表达文库中分离到一个编码小鼠层粘连蛋白受体（MLR）的cDNA。该cDNA含有完整的编码区和3′端非翻译区，但5′端非翻译区缺少前42个碱基。利用cDNA的3′端EcoRI片段进行北方印迹分析，在小鼠睾丸和所有测试的体细胞组织中鉴定出1.2-kb的转录本。1.3和0.9 kb的额外成绩单存在于从小鼠睾丸分离的圆形精子细胞。用核糖核酸酶（RNase）H处理的poly（A）+ RNA进行的北方印迹表明，1.3-kb圆形精子细胞转录本和1.2-kb转录本大小的差异是由于poly（A）+尾长不同所致。0.9 kb圆形精子细胞转录本与1.2 kb MLR cDNA的5′端以外的所有末端杂交，表明使用了替代起始位点或转录本加工发生在这些细胞中。免疫印迹分析确定了生精细胞中对应于67-70 kDa MLR及其43 kDa前体的蛋白质。此外，配体结合研究和亲和层析程序表明，这些大小的生精细胞蛋白结合层粘连蛋白。然而，精母细胞和精子细胞在空间上与睾丸中的层粘连蛋白分离，并且MLR在这些细胞中可能具有其他功能。
A cDNA proposed to encode the mouse laminin receptor (MLR) was isolated from a mouse round spermatid expression library. The cDNA contained complete coding and 3′ untranslated regions but was missing the first 42 bases from the 5′ untranslated region. Northern blot analysis using a 3′EcoRI fragment of the cDNA identified a 1.2-kb transcript in mouse testes and all somatic tissues tested. Additional transcripts of 1.3 and 0.9 kb were present in round spermatids isolated from mouse testes. Northern blots using ribonuclease (RNase) H-treated poly(A)+ RNA indicated that the difference in the size of 1.3-kb round spermatid transcripts and 1.2-kb transcripts was due to differing poly(A)+ tail lengths. The 0.9-kb round spermatid transcript hybridized to all but the 5′ end of the 1.2-kb MLR cDNA, suggesting that an alternate start site is used or that transcript processing occurs in these cells. Immunoblot analysis identified proteins in spermatogenic cells corresponding to the 67–70-kDa MLR and its 43-kDa precursor. In addition, ligand binding studies and affinity chromatography procedures indicated that spermatogenic cell proteins of these sizes bind laminin. However, spermatocytes and spermatids are spatially isolated from laminin in the testes, and MLR may have other functions in these cells.