Inuma Michiko, Obara Yoshitaka, Kuro-o Masaki
Cytogenetics Laboratory, Department of Biofunctional Science, Hirosaki University, Hirosaki, Aomori, Japan.
Zoolog Sci. 2007 Jun;24(6):588-95. doi: 10.2108/zsj.24.588.
"Delayed QM-fluorescence" refers to the unusual kinetics of fluorescence from most of the C-heterochromatic regions of the chromosomes of the small Japanese field mouse Apodemus argenteus. When stained with quinacrine mustard (QM-stained), these C-heterochromatic regions emit weak fluorescence immediately after exposure to blue light (BL); they emit bright fluorescence within a few minutes; and the intensity of the fluorescence gradually decreases after maximum fluorescence has been recorded. To elucidate the mechanism of this phenomenon, we used acridine orange staining (AO-staining) and a modified version of the in situ nick-translation method. Focusing on the large C-heterochromatic region (C-block) of the X chromosome, we noted that AO-stained C-blocks emitted greenish fluorescence, while QM-stained and BL-exposed (QM-BL-processed) C-blocks emitted reddish fluorescence upon AO-staining after removal of QM. These findings suggested that the C-block DNA of A. argenteus might undergo a structural change, such as strand breaks, during QM-BL processing. Application of the modified in situ nick-translation method revealed the generation of an appreciable number of nicks in the C-block DNA by QM-BL processing. No such nick formation was observed in the C-blocks of three other mammalian species: Apodemus peninsulae, Microtus montebelli, and Urotrichus talpoides. Our findings support the hypothesis that nick formation due to exposure to BL might play a primary role in inducing delayed QM-fluorescence in the C-blocks of A. argenteus. On the basis of the present and earlier findings, we propose a probable mechanism for delayed QM-fluorescence in A. argenteus chromosomes.
“延迟喹吖因荧光”指的是日本田鼠(Apodemus argenteus)染色体大部分C-异染色质区域荧光的异常动力学现象。用喹吖因氮芥染色(QM染色)后,这些C-异染色质区域在暴露于蓝光(BL)后立即发出微弱荧光;几分钟内发出明亮荧光;在记录到最大荧光后,荧光强度逐渐降低。为阐明这一现象的机制,我们采用了吖啶橙染色(AO染色)和改良的原位缺口平移法。聚焦于X染色体的大C-异染色质区域(C-块),我们注意到AO染色的C-块发出绿色荧光,而QM染色并经BL处理(QM-BL处理)的C-块在去除QM后进行AO染色时发出红色荧光。这些发现表明,在QM-BL处理过程中,日本田鼠的C-块DNA可能会发生结构变化,如链断裂。改良的原位缺口平移法的应用揭示了QM-BL处理在C-块DNA中产生了相当数量的缺口。在其他三种哺乳动物物种:半岛田鼠(Apodemus peninsulae)、蒙特贝利田鼠(Microtus montebelli)和日本鼩鼱(Urotrichus talpoides)的C-块中未观察到这种缺口形成。我们的发现支持了这样一种假说,即暴露于BL导致的缺口形成可能在诱导日本田鼠C-块中的延迟QM荧光中起主要作用。基于目前和早期的发现,我们提出了日本田鼠染色体中延迟QM荧光的一种可能机制。