期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：1998
卷号：95
期号：13
页码：7760-7765
DOI：10.1073/pnas.95.13.7760
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：Many neurons of the central nervous system display multiple high voltage-activated Ca2+ currents, pharmacologically classified as L-, N-, P-, Q-, and R-type. Of these current types, the R-type is the least understood. The leading candidate for the molecular correlate of R-type currents in cerebellar granule cells is the 1E subunit, which yields Ca2+ currents very similar to the R-type when expressed in heterologous systems. As a complementary approach, we tested whether antisense oligonucleotides against 1E could decrease the expression of R-type current in rat cerebellar granule neurons in culture. Cells were supplemented with either antisense or sense oligonucleotides and whole-cell patch clamp recordings were obtained after 6-8 days in vitro. Incubation with 1E antisense oligonucleotide caused a 52.5% decrease in the peak R-type current density, from -10 {+/-} 0.6 picoamperes/picofarad (pA/pF) (n = 6) in the untreated controls to -4.8 {+/-} 0.8 pA/pF (n = 11) (P < 0.01). In contrast, no significant changes in the current expression were seen in sense oligonucleotide-treated cells (-11.3 {+/-} 3.2 pA/pF). The specificity of the 1E antisense oligonucleotides was supported by the lack of change in estimates of the P/Q current amplitude. Furthermore, antisense and sense oligonucleotides against 1A did not affect R-type current expression (-11.5 {+/-} 1.7 and -11.7 {+/-} 1.7 pA/pF, respectively), whereas the 1A antisense oligonucleotide significantly reduced whole cell currents under conditions in which P/Q current is dominant. Our results support the hypothesis that members of the E class of 1 subunits support the high voltage-activated R-type current in cerebellar granule cells.
