Ward R D, Mendoza L M, Moy G W, Vacquier V D, Nishioka D
Department of Biology, Georgetown University, Washington, DC 20057-1028.
Zygote. 1994 May;2(2):159-65. doi: 10.1017/s096719940000191x.
Specific mRNAs coding for a 63 kDa sperm membrane protein (63-SMP) were localised in Strongylocentrotus purpuratus testis sections using in situ hybridisation techniques. 35S-labelled antisense RNA probes transcribed from a 766 base pair fragment of the gene coding for the 63-SMP hybridised to all spermatogenic cells in the basal germinal epithelia of testicular acini, except the most peripherally located (least differentiated) spermatogonia. No hybridisation to the luminally located mature spermatozoa or somatic cells of the testis was observed. Using monoclonal antibody J17/30 and indirect immunofluorescence techniques, the 63-SMP was localised to the same subset of spermatogenic cells that contain the 63-SMP mRNA, suggesting that expression of this gene is transcriptionally controlled. In combination with previous studies on the expression of sperm histones and sperm binding, these results show that multiple, perhaps sequential, classes of gene activity contribute to the differentiation of sea urchin sperm.
利用原位杂交技术,编码63 kDa精子膜蛋白(63-SMP)的特定mRNA在紫球海胆睾丸切片中定位。从编码63-SMP的基因的766个碱基对片段转录而来的35S标记反义RNA探针,与睾丸腺泡基底生发上皮中的所有生精细胞杂交,但最外围(分化程度最低)的精原细胞除外。未观察到与管腔中成熟精子或睾丸体细胞的杂交。使用单克隆抗体J17/30和间接免疫荧光技术,63-SMP定位于含有63-SMP mRNA的同一生精细胞亚群,这表明该基因的表达受转录控制。结合先前关于精子组蛋白表达和精子结合的研究,这些结果表明,多种可能是连续的基因活性类别有助于海胆精子的分化。