Identification of the alternative spliced form of the alpha 2/delta subunit of voltage sensitive Ca2+ channels expressed in PC12 cells
| Autor: | Sarah Halevi, Batia Gilad, Michael Trus, Nina Shenkar, Daphne Atlas |
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| Rok vydání: | 1995 |
| Předmět: |
Protein subunit
Molecular Sequence Data Gene Expression Biology PC12 Cells Antibodies Complementary DNA medicine Animals Northern blot RNA Messenger Messenger RNA Neurotransmitter Agents General Neuroscience Alternative splicing HEK 293 cells Nucleic acid sequence Skeletal muscle Sequence Analysis DNA Blotting Northern Molecular biology Rats Alternative Splicing medicine.anatomical_structure Autoradiography Calcium Channels |
| Zdroj: | Neuroscience letters. 193(3) |
| ISSN: | 0304-3940 |
| Popis: | The alpha 2/delta subunit of voltage sensitive Ca2+ channels expressed in PC12 has been cloned and partially sequenced. The message observed in Northern blot analysis displays a 7.5 kb transcript, identical in size to mRNA of rabbit skeletal muscle and rat brain. The nucleotide sequence of the cloned alpha 2 subunit of the PC12 specific cDNA is99% identical to rat brain sequence and 85% to skeletal muscle. Reverse-transcriptase-polymerase chain reaction (RT-PCR) of the alternative splicing region identifies two deleted regions of 57 bp and 21 bp in PC12 expressed alpha 2/delta transcript. The alternative variant alpha 2e of alpha 2/delta subunit which is expressed in PC12 cells was previously identified in human embryonic kidney (HEK293) cells. RT-PCR analysis show two different sized alternative PCR fragments in rat lung and none in rat spleen, kidney and intestine. Antibodies prepared against a 19 amino acid peptide within the alternative spliced region effectively inhibits [3H]dopamine release in PC12 cells. This implies that the alternatively spliced region is positioned extracellularly and is involved in regulation of the L-type Ca2+ channel-mediated transmitter release. |
| Databáze: | OpenAIRE |
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