Distribution of actin gene isoforms in the Arabidopsis leaf measured in microsamples from intact individual cells.
The contents of single plant cells can be sampled using glass microcapillaries. By combining such single-cell sampling with reverse transcription-polymerase chain reaction (RT-PCR), transcripts of individual genes can be identified and, in principle, quantified. This provides a valuable technique for the analysis and quantification of the intercellular distribution of gene expression in complex tissues. In a proof-of-principle study, the cellular locations of the transcripts of the eight isoforms of actin ( ACT) expressed in Arabidopsis thaliana (L.) Heynh. were analyzed. Cell sap was extracted from epidermal and mesophyll cells of leaves of 3- to 4-week-old plants. Single-cell (SC)-RT-PCR was used to amplify the actin transcripts using specific primer pairs for ACT1, 2, 3, 4, 7, 8, 11 and 12. Only ACT2 and ACT8 were found in epidermal and in mesophyll cells. In individual trichomes, in addition to ACT2 and ACT8, ACT7 and ACT11 transcripts were detectable. By employing the already well-characterized actin system we demonstrate the practicality and power of SC-RT-PCR as a technique for analyzing gene expression at the ultimate level of resolution, the single cell.
Laval
Valérie
V
Plant Molecular Science Group, Institute of Biomedical and Life Sciences, University of Glasgow, Glasgow G12 8QQ, UK.
Koroleva
Olga A
OA
Murphy
Elaine
E
Lu
Chungui
C
Milner
Joel J
JJ
Hooks
Mark A
MA
Tomos
A Deri
AD
D000199:Actins / Q000235:genetics*; D017360:Arabidopsis / Q000166:cytology / Q000235:genetics* / Q000254:growth & development; D022081:Cell Surface Extensions / Q000235:genetics; D002478:Cells, Cultured; D018506:Gene Expression Regulation, Plant; D019441:Plant Epidermis / Q000166:cytology / Q000235:genetics / Q000254:growth & development; D018515:Plant Leaves / Q000166:cytology / Q000235:genetics* / Q000254:growth & development; D020033:Protein Isoforms / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000199:Actins; D020033:Protein Isoforms; D012333:RNA, Messenger