Fujimoto H, Erickson R P, Toné S
Laboratory of Cell Biology, Mitsubishi-Kasei Institute of Life Science, Tokyo, Japan.
Mol Reprod Dev. 1988;1(1):27-34. doi: 10.1002/mrd.1080010106.
The expression of the mRNA for mouse testicular lactate dehydrogenase (LDH-X) was examined by RNA:cDNA hybridization in situ in the testis and by Northern analyses of meiotic and postmeiotic spermatogenic cell populations. Silver grains accumulated in cells inside the second layer from the periphery of the seminiferous tubule, confirming previous findings that LDH-X mRNA first appears in the spermatocyte and continues to accumulate until the late spermatid stage. Northern analyses showed that meiotic and postmeiotic cells contained 1.2 and 1.3 kb classes of hybridizing mRNA, respectively. RNase H digestion of oligo (dT)-hybridized RNA and poly(U)-Sepharose column chromatography with differential elution by formamide revealed that the difference in size of the two classes of mRNAs was due to the poly(A) tail length of the LDH-X mRNA. When the distribution of the LDH-X mRNA was examined across polysome gradients, both mRNAs were partially associated with polysomes. These results suggest that the changes in the polyadenylation of LDH-X mRNA were associated with the meiotic division during spermatogenesis in the mouse. They raise the possibility that the stable accumulation of the LDH-X mRNAs in the postmeiotic cells is enhanced by poly(A) tails of increased length.
通过RNA:cDNA原位杂交技术在小鼠睾丸中检测小鼠睾丸乳酸脱氢酶(LDH-X)mRNA的表达,并对减数分裂期及减数分裂后精子发生细胞群体进行Northern分析。银颗粒聚集在曲细精管外周第二层以内的细胞中,证实了先前的研究结果,即LDH-X mRNA首先出现在精母细胞中,并持续积累直至精子细胞后期。Northern分析表明,减数分裂期及减数分裂后细胞分别含有1.2 kb和1.3 kb的杂交mRNA类别。用寡聚(dT)杂交RNA进行RNase H消化,并用甲酰胺进行差异洗脱的聚(U)-琼脂糖柱层析显示,两类mRNA大小的差异是由于LDH-X mRNA的多聚腺苷酸尾长度不同。当在多核糖体梯度上检测LDH-X mRNA的分布时,两种mRNA均部分与多核糖体相关联。这些结果表明,LDH-X mRNA多聚腺苷酸化的变化与小鼠精子发生过程中的减数分裂有关。它们增加了这样一种可能性,即减数分裂后细胞中LDH-X mRNA的稳定积累是由长度增加的多聚腺苷酸尾增强的。