Fraschini A, Fuhrman Conti A M
Dipartimento di Biologia Animale, Università di Pavia, Italy.
Histochem Cell Biol. 1995 Jan;103(1):47-57. doi: 10.1007/BF01464475.
The phospholipid component of interphase nuclei was analysed in EUE cells (an established cell line from embryonic human epithelium) grown in an isotonic culture medium and during the adaptation process to a hypertonic medium, using a highly specific ultracytochemical procedure, viz. labelling with the phospholipase A2 gold-complex. Within the nucleus, the phospholipids were localized in domains involved in different steps of the synthesis and processing of the RNA. These localizations did not vary at the two key steps of the adaptation process to hypertonic medium: short-term treatment (6 h) representing critical shock condition, and long-term growth (5 days) representing the adapted cells under survival conditions. On the contrary, deep changes of the labelling intensity of phospholipids at these sites occurred at the different times of hypertonic treatment and followed the same course as those observed in the ultramorphological patterns of transcription: the chromatin condensation, as evaluated by image analysis, the permanent nucleolar components, the interchromatin and the perichromatin granules. These data endorse the hypothesis that nuclear phospholipids could be involved in different steps of the transcriptional activity. They are indicative of the deep changes occurring in the EUE cells submitted to hypertonic stress.
利用一种高度特异性的超微细胞化学方法,即磷脂酶A2金复合物标记法,分析了在等渗培养基中生长以及在适应高渗培养基过程中的EUE细胞(一种源自人类胚胎上皮的成熟细胞系)间期细胞核的磷脂成分。在细胞核内,磷脂定位于参与RNA合成和加工不同步骤的区域。在适应高渗培养基的两个关键步骤中,这些定位没有变化:短期处理(6小时)代表临界休克状态,长期生长(5天)代表存活条件下的适应细胞。相反,在高渗处理的不同时间,这些位点的磷脂标记强度发生了深刻变化,并且与在转录的超微形态模式中观察到的变化过程相同:通过图像分析评估的染色质凝聚、永久性核仁成分、染色质间和染色质周颗粒。这些数据支持了核磷脂可能参与转录活性不同步骤的假说。它们表明经受高渗应激的EUE细胞发生了深刻变化。