CALCIUM-INDUCED QUIESCENCE IN REACTIVATED SEA-URCHIN SPERM
CALCIUM-INDUCED QUIESCENCE IN REACTIVATED SEA-URCHIN SPERM
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DOI：
10.1083/jcb.84.1.13
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发表时间：
1980-01-01
影响因子：
7.8
通讯作者：
GIBBONS, IR
中科院分区：
文献类型：
--
作者：
GIBBONS, BH;GIBBONS, IR
用Triton X-100对海胆鞭毛进行脱膜后，在EGTA[乙二醇- β]的存在下，鞭毛产生不对称的波浪。（-氨基乙醚）N，N，N‘，N’四乙酸]，在pH 8.1下，在2mm MgSO4存在下，用1mm ATP再活化。在这些重新激活的精子中加入0.1-0.2 mM的游离Ca2+，可诱导70-95%的精子进入静止状态。这种静止可以通过EGTA减少游离Ca2+浓度或通过稀释使MgATP2-浓度降至0.3 mM以下来逆转。静止波形的特征是鞭毛近端区域的急剧主弯曲约为5.6 rad[基数]，中部区域的轻微反向弯曲平均约为0.3 rad，尖端的主弯曲约为1.1 rad。静止的精子在机械上非常脆弱；包括微管滑动在内的破坏，在站立时自发地以缓慢的速度发生，或在轻微搅拌时立即发生。胰蛋白酶的温和消化使鞭毛逐渐出现正常对称的跳动。在静止精子中加入浓度不断增加的钒酸盐，导致近端弯曲角逐渐减小，为50 .mu。在0.1 mM游离Ca2+和10 .mu。在无中间振荡跳动的脱膜精子中，通过添加0.1或1mm ATP诱导出一种具有特征的、有芳香的静止弯曲。在没有钒酸盐的情况下，这两种浓度的ATP分别产生不对称的跳动和静止。活精子的静止是由细胞内Ca2+浓度升高引起的。他们证明，在振荡轴承被抑制的鞭毛中可以发生弯曲，并强调不对称跳动与静止之间的密切关系。
Sperm flagella of the sea urchin T. gratilla beat with asymmetrical being waves after demembranation with Triton X-100 in the presence of EGTA [ethylene glycol-bis(.beta.-aminoethyl ether)N,N,N'',N'' tetraacetic acid] and reactivation at pH 8.1 with 1 mM ATP in the presence of 2 mM MgSO4. Addition of 0.1-0.2 mM free Ca2+ to these reactivated sperm induces 70-95% of them to become quiescent. This quiescence can be reversed by reduction of the free Ca2+ concentration with EGTA or by dilution to reduce the MgATP2- concentration below 0.3 mM. The quiescent waveform is characterized by a sharp principal bend of about 5.6 rad [radix] in the proximal region of the flagellum, a slight reverse bend in the midregion that averages about 0.3 rad and a principal bend of about 1.1 rad in the tip. The quiescent sperm are highly fragile mechanically; disruption, including microtubule sliding, occurs spontaneously at a slow rate upon standing or immediately upon gentle agitation. Mild digestion by trypsin causes a gradual appearance of normal, symmetrical flagellar beating. Addition of increasing concentrations of vanadate to quiescent sperm causes a graded decrease in the proximal bend angle, with 50 .mu.M vanadate reducing it to about 2.6 rad. In the presence of 0.1 mM free Ca2+ and 10 .mu.M vanadate, a characteristic, crescented stationary bend is induced in the demembranated sperm, without intermediate oscillatory beating, by the addition of either 0.1 or 1 mM ATP. In the absence of vanadate, these 2 concentrations of ATP produce asymmetric beating and quiescence, respectively. Quiescence in live sperm is induced by an elevated concentration of intracellular Ca2+. They demonstrate that bending can occur in flagella in which oscillatory bearing is inhibited and emphasize the close relationship between asymmetric beating and quiescence.