Developmental changes in delayed rectifier K+ currents in the muscular‐ and neural‐type blastomere of ascidian embryos.

Abstract

1. Developmental changes in the amplitude, kinetic properties, tetraethyl-ammonium (TEA) sensitivity, and ion selectivity of the delayed rectifier K+ currents were investigated in differentiating muscular-type (M) and neural-type (N) blastomeres isolated from the early cleavage-arrested ascidian embryos, using conventional two-microelectrode voltage clamp techniques. 2. No voltage-sensitive outward K+ currents were found in either type of blastomere during the first 35 h of development at 9 degrees C. Thereafter the delayed rectifier K+ current became apparent. The peak amplitude of the K+ current in the M-blastomere increased abruptly from 50 to 60 h and tended to plateau after 60 h, while in the N-blastomere it continued to increase after initial emergence at around 35 h. 3. The threshold potential level of the K+ current in the M-blastomere was initially about -10 mV in a standard external solution (1 mM-K+ solution), but shifted towards the hyperpolarized direction until it reached a steady level at 45 h after fertilization. At the fully differentiated stages, the threshold was around -32 mV and -26 mV in the M- and N-blastomeres, respectively. 4. Throughout development, the reversal potential of the tail current changed with the external K+ concentration in both M- and N-blastomeres as expected for a K(+)-electrode. There was no significant difference in the selectivity ratios for the K+ channel between the two types of blastomeres. The relative selectivities were K+ (1.000): Rb+ (0.774): NH4+ (0.122): Na+ (0.074) and K+ (1.000): Rb+ (0.724): NH4+ (0.155): Na+ (0.074) in the M- and N-blastomeres, respectively. 5. Modified Scatchard plots of TEA-sensitivity data indicated a one-to-one reaction between TEA and the K+ channel. These plots revealed the presence of TEA-resistant K+ channels in addition to TEA-sensitive K+ channels in the M-blastomere, but revealed only TEA-sensitive K+ channels in the N-blastomere. The dissociation constant (Ki) values of these three types of K+ channel did not change during development. In the M-blastomere, the Ki of the TEA-sensitive K+ channel was 1.29 +/- 0.05 mM (mean +/- S.E.M., n = 31) and that of the TEA-resistant K+ channel was 1.4 +/- 0.1 M (mean +/- S.E.M., n = 31) at a test potential of 45 mV. The Ki value of the neural-type K+ current was 1.38 +/- 0.03 mM (mean +/- S.E.M., n = 20) at 45 mV.(ABSTRACT TRUNCATED AT 400 WORDS)