Benito Juliana, Houl Jerry H, Roman Gregg W, Hardin Paul E
Department of Biology and Biochemistry, University of Houston, Houston, TX 77204-5001, USA.
J Biol Rhythms. 2008 Aug;23(4):296-307. doi: 10.1177/0748730408318588.
In the fruit fly Drosophila melanogaster, CRYPTOCHROME (CRY) functions as a photoreceptor to entrain circadian oscillators to light-dark cycles and as a transcription factor to maintain circadian oscillator function in certain peripheral tissues. Given the importance of CRY to circadian clock function, we expected this protein to be expressed in all oscillator cells, yet CRY cellular distribution and subcellular localization has not been firmly established. Here we investigate CRY spatial expression in the brain using a newly developed CRY antibody and a novel set of cry deletion mutants. We find that CRY is expressed in s-LNvs, l-LNvs, and a subset of LNds and DN1s, but not DN2s and DN3s. CRY is present in both the nucleus and the cytoplasm of these neurons, and its subcellular localization does not change over the circadian cycle. Although CRY is absent in DN2s and DN3s, cry promoter activity and/or cry mRNA accumulation can be detected in these neurons, suggesting that CRY levels are regulated posttranscriptionally. Oscillators in DN2s and DN3s entrain to environmental light-dark cycles, which implies that they are entrained indirectly by retinal photoreceptors, extraretinal photoreceptors, or other CRY-expressing cells.
在果蝇黑腹果蝇中,隐花色素(CRY)作为一种光感受器,将昼夜节律振荡器与明暗周期同步,并且作为一种转录因子,在某些外周组织中维持昼夜节律振荡器的功能。鉴于CRY对昼夜节律时钟功能的重要性,我们预期这种蛋白质会在所有振荡器细胞中表达,然而CRY的细胞分布和亚细胞定位尚未得到确凿证实。在这里,我们使用新开发的CRY抗体和一组新的cry缺失突变体来研究CRY在大脑中的空间表达。我们发现CRY在小侧神经元(s-LNvs)、大侧神经元(l-LNvs)以及一部分侧神经元(LNds)和DN1神经元中表达,但在DN2和DN3神经元中不表达。CRY存在于这些神经元的细胞核和细胞质中,并且其亚细胞定位在昼夜节律周期中不会改变。尽管DN2和DN3神经元中不存在CRY,但在这些神经元中可以检测到cry启动子活性和/或cry mRNA积累,这表明CRY水平在转录后受到调节。DN2和DN3神经元中的振荡器与环境明暗周期同步,这意味着它们是由视网膜光感受器、视网膜外光感受器或其他表达CRY的细胞间接同步的。