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Updated: Jul 27, 2025
A Cell Free Assay to Study Chromatin Decondensation at the End of Mitosis
Published on: December 19, 2015
STABILITY OF ISOLATED MITOTIC APPARATUS DURING STORAGE
1Biology Department, York University, Downsview, Ontario M3J 1P3, Canada.
Abstract:
Mitotic apparatus (MA) were isolated from sea urchin zygotes using various isolation procedures, and various properties of the isolated MA were studied and compared. MA isolated using hexylene glycol had birefringences which depended on the pH of the isolation medium, the lower the pH the higher the MA birefringence. The stability of the MA Birefringence also depended on the pH of the hexylene glycol isolation medium (the lower the pH the slower the rate of decay of birefringence), as did the final birefringence reached after prolonged storage. MA isolated using glycerol-dimethylsulphoxide (MTME) had much more stable birefringence than MA isolated using hexylene glycol; their birefringence decay rates were about 1000 times slower than those of MA isolated using hexylene glycol. Birefringence which remained after extraction of MA with H2 O, or 0.5 M KC1 was also studied; the results depended on the MA isolation medium, on the medium the MA were stored in, and on the amount of time the MA were stored after isolation, as described in detail in the text. These results are discussed, and it is suggested that several components (including, perhaps, oriented ribosomes) contribute to birefringence of isolated MA.
Insights
Researchers isolated sea urchin mitotic apparatus (MA) using different methods. Hexylene glycol isolation affected MA birefringence stability and decay, while glycerol-dimethylsulphoxide offered superior stability.
Area of Science:
- Cell Biology
- Biophysics
- Developmental Biology
Background:
- The mitotic apparatus (MA) is crucial for cell division.
- Understanding the physical properties of the MA, such as birefringence, can provide insights into its structure and function.
Purpose of the Study:
- To compare the properties of mitotic apparatus (MA) isolated using different procedures.
- To investigate the factors influencing the birefringence and its stability in isolated MA.
Main Methods:
- Isolation of mitotic apparatus (MA) from sea urchin zygotes using hexylene glycol and glycerol-dimethylsulphoxide (MTME) methods.
- Measurement and comparison of MA birefringence under varying pH conditions and after extraction with H2O or 0.5 M KCl.
- Analysis of birefringence decay rates and stability over time.
Main Results:
- MA isolated with hexylene glycol exhibited pH-dependent birefringence, with lower pH increasing birefringence and stability.
- MA isolated with glycerol-dimethylsulphoxide (MTME) showed significantly greater birefringence stability, with decay rates approximately 1000 times slower than those isolated with hexylene glycol.
- Residual birefringence after extraction depended on isolation method, storage medium, and storage duration.
Conclusions:
- The isolation method significantly impacts the birefringence properties and stability of the mitotic apparatus (MA).
- Glycerol-dimethylsulphoxide (MTME) is a superior isolation medium for preserving MA birefringence stability.
- Multiple components, potentially including oriented ribosomes, may contribute to the birefringence of isolated MA.
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