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Lines expressing GFP::CED-10 were generated by injection of ced-10::gfp::ced-10 with an unc-76-rescuing construct, P76-16B, both at 75 ng/l, into unc-76(e911) animals. |
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Expr1734
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Cell type identifications were confirmed by the examination of a nuclear-localized GFP expressed from the ced-10 promoter. GFP::CED-10 was expressed broadly, perhaps in all cells, including neurons (CANs, VDs and DDs), engulfing cell types (including the hypodermis, intestine and pharynx) and the distal tip cells. Expression was first seen in early embryogenesis and continued throughout adulthood in all cell types. |
Strong fluorescence was observed within the axons of the nerve ring. The GFP::CED-10 fusion protein accumulated at the plasma membrane. |
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nmgp-1 expresses mostly in sensory neurons and in the egg-laying apparatus of adult hermaphrodites. GFP expression driven by the putative nmgp-1 promoter was detected in several cells, primarily neurons. Within the head, we saw GFP expression in pharyngeal neurons, as many somas and processes within the metacorpus and the posterior bulb ventral ganglia were observed. In addition, some of the processes next to the bulb might correspond to CEP sheath glial cells. In the midbody, we saw labeling of cells within the egg-laying apparatus. Posteriorly, we observed cells in the tail ganglia and possibly phasmid neurons. Neuronal processes along dorsal and ventral cords were also labeled. In addition, to identify specific neurons, we used the NeuroPAL strain (OH15500) developed by Hobert's Lab (Yemini et al., 2021). This strain has a stereotyped fluorescent color map to identify all neurons. We injected it with the same plasmid for GFP expression under the nmgp-1 promoter. The following neurons were identified as expressing GFP: ALA, CEPD, IL1 (head neurons from the nerve ring), the sensory amphid neurons ASK, neurons from the anterior ventral nerve cord (VA6, VB7, DB5, AS5, VD6, DD3, DA4) and posterior ventral cord (VA11, VD11, AS10, DA7, DB7, CB11, VA11), neurons from the preanal ganglion (PVP, PVT, DD6, AS11, VA12, DA8, DA9) dorso-rectal ganglion (DVB, DVA, DVC) and lumbar ganglion (PVQ, PHC). The neurons identified include sensory neurons (amphid and mechanosensory), motor neurons and interneurons. |
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For lin-39 and mab-5, previous work reported their partially overlapping expression in the ventral cord motor neurons (MNs). In this study, we used two fosmid reporters wgIs18[lin39::EGFP] and wgIs27[mab-5::EGFP] to map their expression to the single-cell resolution among the 75 motor neurons (MNs), which include 54 cholinergic MNs, 19 GABAergic MNs, and 2 serotonergic MNs with stereotypical positions. These MNs can be further classified into eight neuron classes (DA, DB, VA, VB, AS, VC, DD, and VD). By crossing the GFP reporters with mCherry strains labelling cholinergic or GABAergic neurons in the ventral cord, we identified lin-39 expression in DA2-5, DB2-7, VA3-8, VB4-9, AS2-8, VC1-6, DD2-6, and VD3-12 and mab-5 expression in DA4-8, DB5-7, VA6-11, VB8-11, AS5-11, VC3-6, DD2-6, and VD2-12. Both lin-39 and mab-5 expression were generally weaker in more anterior MNs. Interestingly, for cholinergic MNs, mab-5 and lin-39 expression only overlapped in a set of mid-body MNs; MNs more anterior to this region expressed only lin-39 and MNs more posterior expressed only mab-5. For GABAergic MNs (DD and VD), however, lin-39and mab-5 expression overlapped entirely. Outside of the ventral nerve cord, lin-39 was expressed in AQR and AIYL/R in the head and AVM, SDQL/R, PDEL/R, and PVDL/R neurons along the body; mab-5 was expressed in the AVL in the head, SDQL in the mid-body region, and PQR neuron in the tail. |
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Expr15628
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Suncellular localization was revealed by another construct with GFP inserted at the carboxy terminus of the UNC-47 protein. This construct rescued the unc-47 mutant phenotype, demonstrating that the construct functions normally. |
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Expr1459
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Animals containing the unc-47::GFP reporter construct showed expression of GFP in all GABAergic neurons, and only in GABAergic neurons. |
Done by another gfp construct: GFP-tagged UNC-47 was localized to synaptic varicosities along the ventral and dorsal cords but not in axons. show that unc-47 expression is associated with synaptic vesicle. |