Reynard Louise N, Turner James M A, Cocquet Julie, Mahadevaiah Shantha K, Touré Aminata, Höög Christer, Burgoyne Paul S
MRC National Institute for Medical Research, London NW7 1AA, United Kingdom.
Biol Reprod. 2007 Aug;77(2):329-35. doi: 10.1095/biolreprod.107.061101. Epub 2007 May 2.
The mouse multi-copy X-linked gene Xlr-related, meiosis-regulated (Xmr/Slx) has previously been described as encoding a testis-specific nuclear protein expressed during male meiotic prophase, and during which it becomes concentrated in the inactive X and Y chromatin domain. These conclusions were based on Western blot and immunolocalization analysis using an antibody raised against a related lymphocyte protein, XLR; however, our recently published RNA in situ for Xmr revealed that transcripts are predominantly or exclusively postmeiotic, and this is supported by a growing body of microarray data. This led us to reanalyze the expression of Xmr, both at the RNA level by RT-PCR and by RNA fluorescence in situ hybridization, and at the protein level by using antibodies raised against XMR that do not recognize XLR. In agreement with our previous RNA in situ data, our further transcription analysis showed almost exclusive expression in spermatids, and Western blot and immunostaining with the XMR antibodies showed that the protein is cytoplasmic and restricted to spermatids. Furthermore, the previously used XLR antibody was shown not to cross-react with XMR, and it is suggested that the meiotically expressed nuclear protein recognized by this antibody is another member of the complex Xlr superfamily. As a result of these findings, the gene previously known as Xmr is now officially know as Slx, Sycp3-like, X-linked.
小鼠多拷贝X连锁基因Xlr相关、减数分裂调控基因(Xmr/Slx)此前被描述为编码一种睾丸特异性核蛋白,该蛋白在雄性减数分裂前期表达,并在此时集中于失活的X和Y染色质区域。这些结论基于使用针对相关淋巴细胞蛋白XLR产生的抗体进行的蛋白质印迹和免疫定位分析;然而,我们最近发表的Xmr的RNA原位杂交结果显示,转录本主要或仅在减数分裂后出现,并且越来越多的微阵列数据也支持这一点。这促使我们通过RT-PCR和RNA荧光原位杂交在RNA水平上,以及使用不识别XLR的针对XMR产生的抗体在蛋白质水平上重新分析Xmr的表达。与我们之前的RNA原位杂交数据一致,我们进一步的转录分析显示几乎仅在精子细胞中表达,并且蛋白质印迹和用XMR抗体进行的免疫染色表明该蛋白位于细胞质中且仅限于精子细胞。此外,之前使用的XLR抗体被证明与XMR无交叉反应,并且表明该抗体识别的减数分裂期表达的核蛋白是Xlr超家族复合体的另一个成员。基于这些发现,之前称为Xmr的基因现在正式称为Slx,类Sycp3,X连锁。