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ÇëÎÊLexA::GAD£¬ repo-LexA::GAD£¬lexAop-rCD2::GFP£¬UAS-mCD8::RFP£¬Õâ¶¼ÊÇɶ¡£ ÒÑÓÐ1È˲ÎÓë
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ÔÎÄÈçÏ£º To address this question, we repeated clonal analysis with dual-expression-controlMARCMthatpermitsdifferentiallabel- ing of distinct cells in a given MARCM clone owing to indepen- dent controls over distinct reporter genes by GAL4 versus LexA::GAD, two GAL80-suppressible binary transcriptional ac- tivators (Lai and Lee, 2006). We used repo-LexA::GAD plus the ensheathing glial GAL4 driver, NP6520, in dual-expression- control MARCM, to label GAL80-minus ensheathing glia with bothlexAop-rCD2::GFPandUAS-mCD8::RFPwhilemarkingthe astrocyte-like glia of the same genotype with lexAop-rCD2::GFP only. If a neuropil glial cluster consisted of only ensheathing or astrocyte-likeglia,onewouldexpectexpressionofmCD8::RFPin all or none of the cells in the cluster. In contrast, if ensheathing and astrocyte-like glia derived from common progenitors, one would detect mCD8::RFP in a subset of rCD2::GFP-marked cells in a single cluster. Fifteen clusters of neuropil glia were obtained following induction of somatic recombination at the newly hatched larval stage. Among them, 12 clusters were found to purely consist of either ensheathing glia or astrocyte-like glia, as evidenced by the expression of mCD8::RFP in all or none of the cellsinagivencluster(Fig.7E¨CH).Intheotherthreeclustersthat containedbothmCD8::RFP-positiveand-negativecells,nomat- ter whether ensheathing or astrocyte-like glia predominated, the minority was overwhelmed and composed of only one or two cells per cluster despite the cluster size being roughly ?8 cells. Sincealowlevelofmitoticrecombinationcouldoccurevenwith- out any heat-shock induction (Fig. 4R), it is possible that the presence of one or two heterotypic cells in the otherwise homo- geneous population may occur as a consequence of multiple events of mitotic recombination. Given that ensheathing and astrocyte-like glia rarely coexist in a single cluster, the two sub- types of neuropil glia differ not only in morphology and marker expression but also with different origins of development. |
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