葛氏鲈塘鳢与七彩神仙鱼心肌细胞钠离子通道的温度响应特征
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东北林业大学野生动物与自然保护地学院 哈尔滨 150040

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赵紫寒,女,硕士研究生;研究方向:动物生理学;E-mail:joyzhaozihan@sina.com。

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中央高校基本科研业务费专项资金项目(No. 2572020BE01),黑龙江省自然科学基金项目(No. LH2022C017);


Temperature Response Characteristics of Sodium Channels in Cardiomyocytes of Perccottus glenii and Symphysodon aequifasciatus
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College of Wildlife and Protected Area, Northeast Forestry University, Harbin 150040, China

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    摘要:

    温度适应对于动物的生存至关重要,而目前对于鱼类这样的外温动物的温度适应相关研究仍然缺乏。葛氏鲈塘鳢(Perccottus glenii)可以在结冰的环境中生存数天,而七彩神仙鱼(Symphysodon aequifasciatus)是被广泛饲养的热带观赏鱼类,本研究将这2种鱼类作为冷水鱼与热带鱼的代表,探究其温度耐受性。为此,测定了它们的临界温度以及不同温度下(0 ~ 38 ℃)的心率,并利用原代细胞急性分离,结合膜片钳电生理技术与控温技术,测定了上述2种鱼类心肌电压门控钠离子通道在不同温度下的电生理特征。结果显示,葛氏鲈塘鳢和七彩神仙鱼的温度耐受范围具有较大差异,分别为﹣2.0 ~ 27.4 ℃以及13.1 ~ 39.3 ℃。葛氏鲈塘鳢的心率在0 ~ 19 ℃之间随着温度的升高而稳定升高,在19 ℃时达到最高,其后逐渐降低,这与葛氏鲈塘鳢心肌细胞上的电压门控钠离子通道在20 ℃时电流峰值最大且通道的开放概率最大这一电生理特征具有一致性。而七彩神仙鱼的心率则在14 ~ 31 ℃之间稳定升高,这与其心肌细胞电压门控钠离子通道电流峰值和开放概率在15 ~ 30 ℃的实验范围内随温度的升高而升高的电生理特性也一致。以上结果说明,这2种鱼类的心率、心肌细胞电压门控钠离子通道复合电流对于温度的响应与其自身的温度耐受范围以及原产栖息地温度紧密相关。因此,鱼类心肌细胞上的电压门控钠离子通道可能存在对温度的适应机制,保证鱼类循环系统在不同温度下正常运行。

    Abstract:

    [Objectives]Temperature adaptation is very important for the survival of animals. However, there is still a lack of correlational research on the temperature adaptation of ectothermic animals such as fish. Perccottus glenii can survive for several days in frozen environments, and Symphysodon aequifasciatus is a widely-reared tropical ornamental fish. These two species of fish were regarded as the representative of the cold-water fish and tropical fish in this study. [Methods] To investigate their temperature tolerance, we performed linear regression on the percentage of experimental fish that were out of balance at different temperatures and then interpolated the temperature value corresponding to 50% individual imbalance to obtain the critical temperature of the two fish species. We also measured their heart rate across the temperature gradient. Then the cardiomyocytes of P. glenii and S. aequifasciatus were acutely isolated, and whole-cell patch-clamp and temperature control techniques were also used to explore the electrophysiological characteristics of voltage-gated sodium channels in two kinds of fish cardiomyocytes at different temperatures (15 ℃, 20 ℃, 25 ℃, and 30 ℃). PatchMaster was used to record the original data, and Igor Pro 6.37 was used to fit the data and draw curves, the significance was inspected using one-way ANOVA. [Results] The temperature tolerance ranges of P. glenii and S. aequifasciatus were﹣2.0 to 27.4 ℃ and 13.1 to 39.3 ℃, respectively. The heart rate of P. glenii increased steadily from 0 ℃ to 19 ℃, reached the highest at 19 ℃, and then decreased gradually (Fig. 3), which was consistent with the electrophysiological characteristics of the voltage-gated sodium channel in the myocardial cells:voltage-gated sodium channels in the cardiomyocytes of P. glenii had the highest peak current (normalized current:﹣0.93 ± 0.11) and the highest opening probability (G50:﹣29.48 ± 0.33 mV) at the temperature of 20 ℃ (Fig. 4 and Fig. 6). The heart rate of S. aequifasciatus increased steadily from 14 ℃ to 31 ℃, which was consistent with the electrophysiological characteristics that the peak current and open probability of voltage-gated sodium channel in cardiomyocytes increased with the increase of temperature within the experimental range between 15 ℃ and 30 ℃, and voltage-gated sodium channels of S. aequifasciatus had the highest peak current (normalized current:﹣1 ± 0) and the highest opening probability (G50:﹣31.46 ± 0.33 mV) at 30 ℃ (Fig. 5 and Fig. 6). [Conclusion] These results indicate that the temperature response of the heart rate and voltage-gated sodium channel complex current in the cardiomyocytes of these two fish species is closely related to their own temperature tolerance range and the temperature of their native habitat. Therefore, the voltage-gated sodium channel of fish cardiomyocytes may play an important role in the temperature adaptation process of the two species of fish and maybe a key protein in the temperature adaptation of the two species of fish.

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赵紫寒,尹传玲,王云飞,程鲲,黄璞祎.2023.葛氏鲈塘鳢与七彩神仙鱼心肌细胞钠离子通道的温度响应特征.动物学杂志,58(6):926-936.

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  • 收稿日期:2022-09-20
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  • 在线发布日期: 2023-12-19
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