Expression of human TRPC genes in the megakaryocytic cell lines MEG01, DAMI and HEL.
Store-regulated Ca2+ entry represents a major mechanism for Ca2+ influx in non-excitable cells although many details remain to be evaluated including the identification of cation entry channels. Recently human homologues of the Drosophila proteins TRP and TRPL, have been described (TRPC1, TRPC1A, HTRP1) and suggested as candidate cation channels. In this study we sought to examine if the producers of blood platelets, megakaryocytic cells (using the cell lines MEG01, DAMI, HEL), expressed these genes. RNA was prepared from the cell lines and platelets and converted to cDNA. The cDNA was then subjected to 30-35 cycles of PCR using gene specific primers for TRPC1-3. PCR products of the expected sizes were observed for all three TRPC genes in the three cell lines. Direct sequencing confirmed their identity. Additionally for TRPC1, a larger species, and for TRPC2, a smaller species was detected in all three cell lines with sequencing revealing the fragments to contain TRPC sequence, suggesting that they were either products of alternative splicing events or from closely related genes. These results suggest that TRPC genes are expressed in megakaryocytic cell lines and that the TRPC proteins may play a role in mediating cation influx in both megakaryocytes and platelets.
Berg
L P
LP
Molecular Genetics, Thrombosis Research Institute, Chelsea, London, UK.
Shamsher
M K
MK
El-Daher
S S
SS
Kakkar
V V
VV
Authi
K S
KS
D017398:Alternative Splicing; D001483:Base Sequence; D015220:Calcium Channels; D002460:Cell Line; D006801:Humans; D007473:Ion Channels / Q000096:biosynthesis* / Q000235:genetics*; D008533:Megakaryocytes / Q000378:metabolism* / Q000502:physiology; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000096:biosynthesis; D017422:Sequence Analysis, DNA; D050052:TRPC Cation Channels; D050053:TRPM Cation Channels; D014158:Transcription, Genetic
D015220:Calcium Channels; D007473:Ion Channels; D008565:Membrane Proteins; D012333:RNA, Messenger; D050052:TRPC Cation Channels; C491859:TRPC2 protein, human; C104603:TRPC3 cation channel; D050053:TRPM Cation Channels; C121934:TRPM2 protein, human; C096351:transient receptor potential cation channel, subfamily C, member 1