Picture: Fig. 3A, B, C, D. |
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In the transgenic animals carrying crm-1a::gfp reporter, consistent signals were detected in neurons in the ventral nerve cord. By the positions and clustering of the cell body as well as the axonal outgrowth along the ventral nerve cord, crm-1a is found to be expressed in the DA neurons 2 to 7, the DB neurons 3 to 7 and additional neurons in the VA, VB and AS classes along the nerve cord (VNC). Expression was also observed in neurons around the pharynx. In the male tail, gfp signals in the RnA neuronal cells of sensory rays 2 and 4 and the PVR neuron with the entire axonal process along the VNC were also observed. |
Entire axonal process of PVR along the VNC. |
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Embryonic expression of exc-7 was first observed at the bean stage. By reverse lineaging with use of SIMI-Biocell software, we confirm the identity of one of the expressing cells at this stage as the excretory canal cell. In L1 animals, broad expression in the head, ventral nerve cord (VNC), and tail was observed. In young adults, expression is notably observed in vulva cells. In the nervous system specifically, expression is observed in many neurons throughout the body, but unlike Drosophila Elav, exc-7::gfp it is not panneuronally expressed. We confirmed previously reported expression in cholinergic VNC MNs, but absence of GABAergic VNC MNs, consistent with previous reports (Fujita et al., 1999; Loria et al., 2003) and consistent with exc-7 functioning in cholinergic, but not GABAergic neurons to control alternative splicing (Norris et al., 2014). exc-7::gfp is also expressed in some non-neuronal cell types, including muscle and hypodermis, but not in the gut. A previous report showed that exc-7 is only transiently and weakly expressed in the excretory cell, which, based on exc-7's excretory mutant phenotype, has puzzled researchers (Fujita et al., 2003). We find that the gfp tagged exc-7 locus is strongly and continuously expressed in the excretory canal cell. |
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Staining embryos with this anti-VAB-7 antibody confirmed that VAB-7 is expressed in posterior muscle and epidermal cells. However, a second phase of VAB-7 expression was discovered in embryonic ventral nerve cord (VNC) neurones, beginning at the 1.5-fold stage. At the threefold stage VAB-7 is expressed in nine neuronal nuclei (seven in the VNC and two nuclei in the head), and in the five most posterior epidermal nuclei, which form the hyp8 to hyp11 cells. VAB-7 is expressed in the seven DB class motoneurones; VAB-7 staining is also seen in two additional head neurones that have not been identified. DB neurones express VAB-7 throughout development. In addition, VAB-7 is expressed continuously in the hypodermal syncitium hyp10, and from the L1 stage in four unidentified neurones in the tail. Finally, at the adult stage, VAB-7 is detected in three VC neurones: VC1, VC2 and VC6. |
nuclei |
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Picture: Fig 6. |
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Apart from cells in the neuroblast lineage that generate ASE, nhr-67::mCherry was expressed in multiple other neuroblast lineages in the developing embryo. Expression was usually observed in the grandmother or mother of a neuron, but not earlier. Within the ASEL and ASER-generating lineage branches, nhr-67 was expressed in neuroblasts that generate closely or distantly related cousins of ASEL and ASER. For example, the sister neuroblast of the ASE-generating neuroblast creates the AUA and ASJ neurons and it expressed nhr-67. The cousin of the ASE-mother cell generates the AWB and ADF sensory neurons. nhr-67 was expressed in these cells. In late stage embryos, a few other, postmitotic neurons started to express nhr-67. Embryonic nhr-67 expression was not restricted to the nervous system, but was observed in a small subset of mesodermal and hypodermal cells. No expression was detected in endodermal cells or the germ line. nhr-67 was expressed in the excretory canal cell. Postembryonically, nhr-67 expression persisted only in a few neurons in the head ganglia until the first larval stage and faded shortly thereafter in most, but not all, of these neurons, with expression persisting through adulthood only in the CEPD/V, RMED/V, AVL and RIS neurons. During mid-larval development, nhr-67 was transiently and dynamically expressed in the AC cells of the vulva. Expression was also found in the VU cells and somatic gonad, but not in vulA, vulB or vulC. Within the ASEL/R generating lineages, nhr-67::mCherry was first observed in the grandmother cells of ASEL and ASER. Transgenic animals that co-express a functional nhr-67::mCherry reporter and a functional che-1::yfp reporter revealed that nhr-67 precedes che-1 expression. nhr-67 expression was maintained in the ASEL and ASER neurons until the first larval stage after which it became undetectable, whereas che-1 expression was maintained throughout the life of the animal. In spite of its genetically deduced role in asymmetric gene expression in ASEL and ASER, nhr-67 expression is bilaterally symmetric in ASEL and ASER. |
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At 335 minutes post fertilization. |
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efn-2 expression showed widespread left-right asymmetry inembryos, both in amphid neurons and in other cells. |
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acr-5 ORF |
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