Salazar E L, Macías H, Calzada L
Unidad de Investigación Clínica en Enfermedades Endócrinas, Hospital de Gineco-Obstetricia Dr. Luis Castelazo Ayala, IMSS, México, D.F.
Ginecol Obstet Mex. 1991 Oct;59:308-12.
The electric potential across the membrane (psi) is evaluated through the accumulation of the lipophilic, radiolabelled, triphenylmethylphosphonium (3H-TPMP+), cation on membrane human sperm. The washed sperm cells, were incubated in presence of 3H-TPMP+ in low-K+ medium or high-K+ and allowed to take up the cation to steady-state (ie; 20 min at 37 degrees C). By using this differential, the value obtained was inserted in the Nernst equation, this value yielded a psi of -69 +/- 2 mV. The presence of divalent cations Zn++ and Mg++ in the incubation medium both induced a hyperpolarization of 10% and 8.6%, respectively. The addition of specific reagents such as p-chloromercuribenzene-sulfonate and ethylene-diamine-tetraacetic acid sodium salt both decreased the psi 35% and 58% respectively. The agents to act upon the components of the sperm cell membrane such as the dithiothreitol and progesterone induced hyperpolarization and depolarization of the membrane 16% and 40%, respectively. The presence of propranolol and L-alpha-lysophosphatidylcholine, which affect the ionic gradients present across the plasma membrane, both induced a depolarization from 43% and 92% respectively. Finally, the addition of tetraphenylboron (TPB-) on the incubation medium, enhanced the value of the psi 75%. These studies are transcendent because with the utilization of agents depolarizing, or hyperpolarizing we obtain changes in the psi from -80 +/- 0.6 mV, until -6 +/- 0.6 mV changes of 74 +/- 1.5 mV across of the sperm cell membrane.
通过亲脂性、放射性标记的三苯基甲基鏻(3H-TPMP+)阳离子在人精子膜上的积累来评估跨膜电势(ψ)。将洗涤后的精子细胞在低K+培养基或高K+条件下于3H-TPMP+存在的情况下孵育,并使其摄取阳离子至稳态(即37℃下20分钟)。利用这种差异,将获得的值代入能斯特方程,该值得出的ψ为-69±2mV。孵育培养基中存在二价阳离子Zn++和Mg++分别导致超极化10%和8.6%。添加特定试剂如对氯汞苯磺酸盐和乙二胺四乙酸钠盐分别使ψ降低35%和58%。作用于精子细胞膜成分的试剂如二硫苏糖醇和孕酮分别使膜超极化和去极化16%和40%。影响质膜上离子梯度的普萘洛尔和L-α-溶血磷脂酰胆碱的存在分别导致去极化43%和92%。最后,在孵育培养基中添加四苯基硼(TPB-)使ψ值提高了75%。这些研究具有重要意义,因为通过使用去极化或超极化试剂,我们观察到精子细胞膜的ψ从-80±0.6mV变化到-6±0.6mV,变化幅度达74±1.5mV。