| Changes in the inward current and membrane conductance after fertilization in the mouse eggs |
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Seong-geun Hong1, Choon-ok Park1, Jae-hee Han1, Ik-hyun Kim2, Dae-sik Ha3, Jong-kuk Kwun4 |
1Department of Physiology, College of Medicine, Gyeongsang National University
2Department of Physiology, Graduate School, Gyeongsang National University
3Kyongsangnam-Do, Provincial Goverment Institute of Health and Environment
4Department of Physiology, College of Veterinary Medicine, Seoul National University |
| 수정에 의한 Mouse egg의 세포막전류 변화 |
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홍성근1, 박춘옥1, 한재희1, 김익현2, 하대식3, 권종국4 |
1경상대학교 의과대학 생리학교실
2경상대학교 대학원 의학과
3경상남도 보건환경 연구원
4서울대학교 수의과대학 생리학교실 |
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| Abstract |
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Changes in the both inward current and conductance of membrane by the fertilization were observed using the one microelectrode voltage clamp(or switch clamp) technique. Unfertilized eggs and both 1- and 2-cell stage eggs after fertilization were donated from the superovulated mouse (ICR, more than 6 weeks old) treated with PMSG(pregnant mare serum gonadotropin, Sigma) and HCG(human chorionic gonadotropin, Sigma) and naturally mated ones, respectively in this experiment. Membrane potential was held at -90mV and the voltage step was applied from -80mV to 50mV with interval of 10mV or 20mV for 300ms. since both of amplitudes and time courses in the membrane currents were various according to the states of cells and clamping condition, results were presented by their $averages{pm}SEM$(standard mean error)and ratios or percentages. Inward currents began to appear in response to the step depolarization from -60mV and reached its maximum at -50mV. However, since the potential was not clamped evenly during the voltage step, current-voltage(I-V) relationship might be positively shifted 10 or 20mV. From the steady-state currents plotted in the I-V curve, outward rectification was markedly observed. Peak inward currents$(i_{in})$ at -50mV were $-0.62{pm}0.23nA$(n=4),$-0.52{pm}0.25nA$(n=5) and $-0.37{pm}0.25nA$(n=6), in the 1-cell stage, 2-cell stage fertilized eggs and in the unfertilized eggs, respectively. Pure inward current (difference between steady-state and peak, $i_{in. pure}$) were $-1.01{pm}0.23nA$, $-0.69{pm}0.43nA$ and $-0.68{pm}0.29nA$, respectively in the 1-cell stage fertilized eggs, unfertilized eggs and 2-cell stage fertilized eggs. These results suggested that the outward current in fertilized eggs of 2-cell stage was more increased than those in the unfertilized eggs. Pure inward currents in the all stages of eggs showed a similar fashion in the I-V relationship from -50mV to 50mV and reversal potential at 50mV. Time constant of inactivation$({ au})$ in the inward current was decreased as the membrane potential was depolarized in the unfertilized and 2-cell stage eggs but in the 1-cell stage eggs t was not likely to be affected significantly. Slope conductances were 14.2nS, 8.9n5 and 7.7nS in the 1-cell, 2-cell stage fertilized eggs and the unfertilized eggs, respectively. Membranes between two cells within a zona pellucida seem to be electrical-connected in the 2-cell stage eggs from the observation made in the analysis for the electronic spread and decay to the current stimuli. Both of inward current and membrane conductance were increased after fertilization in the mouse eggs. Inward current seems to be carried by the same ion or through the same channels up to the 2-cell stage and ion that carried inward current was thought to play important function after fertilization in the mouse eggs. |
| Key Words:
inward current, fertilization, mouse egg, switch clamp |
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