Elevation difference of Ca<Superscript>2+</Superscript> levels in young leaf cells of bromegrass and induced cold-tolerant enhancement under different controlled chilling temperatures

The changes of Ca2＋ levels in young leaf cells of bromegrass under different controlled chilling temperatures were inves- tigated by an antimonite precipitation cytochemical method. The main results were as follows： under 25/20℃ （day/night） tempera- ture and 14 h photoperiod, electron-dense Ca2＋ antimonite precipitates, indicators of Ca2＋ localization, were mainly localized in the vacuoles, cell walls and intercellular spaces; few Ca2＋ deposits were observed in the cytosol and nuclei. After a 3℃ chilling treatment for 3 h, many Ca2＋ precipitates appeared in the cytosol and nuclei, indicating that Ca2＋ influx had occurred in the cytosol and nuclei. When the 3℃ treatment was prolonged to 8 h, more Ca2＋ deposits appeared in the nuclei and cytosol, but the amount of Ca2＋ deposits in both the cytosol and nuclei decreased markedly after a 24 h treatment and most Ca2＋ deposits were returned to the vacuoles and intercellular spaces after an 8 d treatment. When bromegrass was exposed to 7℃ for 3 h, the Ca2＋ distribution in the cells had no visible changes, compared with that of the 25/20℃ grown control plants. However, when the chilling treatment of 7℃ was prolonged to 8 h, a Ca2＋ influx occurred, where many Ca2＋ deposits were observed in the nuclei and cytosol. More Ca2＋ deposits appeared in the nuclei and cytosol after a 24 h treatment, but the amount of Ca2＋ deposits in the cytosol and nuclei was reduced markedly after an 8 d treatment. After a 14 d treatment, the remaining low level of Ca2＋ was recovered in both the cytosol and nuclei and the Ca2＋ deposits were again located in the vacuoles and the intercellular spaces. The dynamics of subcellular Ca2＋ localization in young leaf cells of bromegrass during a 12℃ chilling treatment were similar to those of the 7℃ treatment. Besides, the results showed that the frost tolerance ofbromegrass exposed to 3℃ for 8 d increased by 6℃, for 7℃ and 8 d by 4℃ and for 12℃ and 14 d by 3℃, compared with the controls.     

Influence of cyclovirobuxine D on intracellular [Ca] regulation and the expression of the calcium cycling proteins in rat myocytes

Our previous studies have shown that cyclovirobuxine D (CVB-D) ameliorated the cardiac function in heart failure rats. Considering the relationship between cardiac function and [Ca^2 +]i, and the role of calcium cycling on regulating [Ca^2 +]i, the present study was designed to evaluate the influence of CVB-D on the calcium transient of myocytes from neonatal rats and adult heart failure (HF) rats. The expression of calcium cycling proteins, including L-type calcium channel (LTCC), ryanodine receptor 2 (RYR2), sarcoplasmic reticulum calcium ATPase 2a (SERCA2a) and sodium–calcium exchanger (NCX), were investigated to explore the underlying mechanism. CVB-D increased the intensity of calcium transient, accelerated the process of calcium transient and attenuation in the neonatal and adult myocytes. Furthermore, CVB-D shortened T_peak and T_attenuation in the adult myocytes and slowed down the heart rate of neonatal myocytes. Besides, CVB-D increased the expression of RYR2 and SERCA2a, decreased the expression of NCX, but showed no significant effect on LTCC. Thus, it was concluded that CVB-D increased the release and uptake of Ca^2 + in systolic and diastolic period, respectively. CVB-D might not only facilitate the utilization of intracellular Ca^2 +, but also prevent the loss of Ca^2 +.     

Alterations of intracellular Ca<Superscript>2+</Superscript> concentration and ultrastructure in spruce apical bud cells during seasonal transition

Potassium antimonite was used to localize Ca^2 in the apical bud cells of spruce from July 1999 to May 2000.During the period of active growth (July 14), Calcium precipitates, an indication of Ca^2 localization, were mainly distributed in vacuoles, intercellular spaces and cell walls. Few Ca^2 deposits localized in the cytosol and nucleus, showing a low level of the cytosolic and nuclear Ca^2 concentration in the warm summer. In August, some Ca^2 deposits appeared in the cytosol and nuclei,indicating that Ca^2 influx occurred in the cytosol and nucleus as the day length became shorter. From September to November, high levels of the cytosolic and nuclear Ca^2 remained. During the mid-winter (December and January), the distribution of Ca^2 deposits and the ultrastructures in the cells were altered dramatically. Plasmolysis occurred in many cells due to the protoplasmic dehydration.In addition plasmalemma invagination and nuclear chromatin aggregation also occurred. A large number of Ca^2 deposits appeared in the space between the plasmalemma and the cell wall. And also some Ca^2 deposits were distributed in the plastids. However, few Ca^2 deposits were observed in the cytosol and nuclei. By spring of the next year (May), when plants were de-acclimated and resumed active growth, Ca^2 subcellular localization essentially restored to that observed in July of the last year, i.e., the cells contained low cytosolic and nuclear Ca^2 concentrations; Ca^2 deposits were mainly distributed in the vacuoles, cell walls and intercellular spaces. The relationships between the seasonal changes of intracellular Ca^2 concentration and the development of dormancy/cold acclimation, as well as plasmolysis associated with dormancy and cold hardiness were discussed.