TY - JOUR
T1 - Evidence for the existence of RNA of Ca2+-channel α2/δ subunit in Xenopus oocytes
AU - Singer-Lahat, Dafna
AU - Lotan, Ilana
AU - Itagaki, Kiyoshi
AU - Schwartz, Arnold
AU - Dascal, Nathan
N1 - Funding Information:
We are grateful to Dr. Terry Snotch for supplying the oligonucleotides a2-1 and a2-2. This study was supported in part by the Muscular Dystrophy Assucia-tion (N.D., I.L.) and National Institutes of health grants POI-HL22619-14 (I-C), T32-HL07382-14 and R37-HL43231 (A.S.). We thank Bayer AG for supplying (-) Bay K 8644.
PY - 1992/10/6
Y1 - 1992/10/6
N2 - Ba2+-currents (IBa) through voltage-dependent Ca2+-channels were studied in Xenopus oocytes injected with RNA from several excitable tissues, using the two-electrode voltage-clamp technique. Previous studies have shown that the expression of cardiac Ca2+-channels can be suppressed by an hybrid-arrest procedure that includes co-injection of the tissue-derived RNA with an 'antisense' oligonucleotide complementary to a part of RNA coding for the Ca2+-channel α1 subunit. In this study, this method was used to investigate the role of the α2/δ subunit. Co-injection of RNA extracted from either rabbit heart, rat brain or rat skeletal muscle (SkM) with 'antisense' oligonucleotides complementary to the α2/δ subunit RNA did not substantially affect the expression of IBa in the oocytes. Using the Northern blot hybridization method, it was shown that native oocytes contain large amounts of α2/δ subunit RNA of Ca2+-channel. It is proposed that te oligonucleotide treatment fails to eliminate the α2/δ RNA because of the vast excess of endogenous α2/δ RNA. These results impose a limit on the use of the hybrid-arrest method.
AB - Ba2+-currents (IBa) through voltage-dependent Ca2+-channels were studied in Xenopus oocytes injected with RNA from several excitable tissues, using the two-electrode voltage-clamp technique. Previous studies have shown that the expression of cardiac Ca2+-channels can be suppressed by an hybrid-arrest procedure that includes co-injection of the tissue-derived RNA with an 'antisense' oligonucleotide complementary to a part of RNA coding for the Ca2+-channel α1 subunit. In this study, this method was used to investigate the role of the α2/δ subunit. Co-injection of RNA extracted from either rabbit heart, rat brain or rat skeletal muscle (SkM) with 'antisense' oligonucleotides complementary to the α2/δ subunit RNA did not substantially affect the expression of IBa in the oocytes. Using the Northern blot hybridization method, it was shown that native oocytes contain large amounts of α2/δ subunit RNA of Ca2+-channel. It is proposed that te oligonucleotide treatment fails to eliminate the α2/δ RNA because of the vast excess of endogenous α2/δ RNA. These results impose a limit on the use of the hybrid-arrest method.
KW - (X. laevis)
KW - Calcium channel
KW - Hybrid-arrest method
KW - Oocyte
KW - RNA
UR - http://www.scopus.com/inward/record.url?scp=0026775691&partnerID=8YFLogxK
U2 - 10.1016/0167-4889(92)90097-U
DO - 10.1016/0167-4889(92)90097-U
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AN - SCOPUS:0026775691
SN - 0167-4889
VL - 1137
SP - 39
EP - 44
JO - BBA - Molecular Cell Research
JF - BBA - Molecular Cell Research
IS - 1
ER -