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WBPicture0000012234 Description: Figure 5. Temporal and cellular expression of the osm-3 gene during C. elegans development. An osm-3::lacZ fusion gene,that included a nuclear localization signal (see Figure 4 for the construct details) was used to study the cellular expression of the osm-3 gene during larval and adult stages of the nematode. A, lacZ staining of the 8 pairs of amphid neurons ADF,ADL, ASE, ASG, ASH, ASI, ASJ, and ASK, marked as amp. The cell bodies are located bilaterally on either side in the left and right lateral ganglia, with the exception of ADL cell bodies, which are located in the dorsal region of the lateral ganglia in front of the second pharyngeal bulb. Stained cells and other morphological landmarks were observed under Nomarski microscopy. In addition to the 16 amphid neurons, 6 neuronal cell bodies most likely belonging to the set of inner labial neurons IL2 (IL2DL, IL2DR, IL2L, IL2R, IL2VL, and IL2VR), located in front of the nerve ring in a circumpharyngeal manner are brightly stained. IL2 forms a set of 6 ciliated neurons together with the set of 6 IL1 neurons forming the inner labial sensilla. However, the osm-3::lacZ fusion gene expresses only in 1 of the 2 inner labial neurons classes, most probably the IL2s. B, The same animal as in A, staining of the 8 pairs of amphid neurons and 6 IL2 neurons stained with the nuclear staining dye DAPI, viewed under ultraviolet epi-illumination using UV excitation and barrier filters. C, An FITC-stained larval animal about the same age as that in A, showing the cellular disposition of the amphidneurons that accumulate fluorescein dyes. D, lacZ staining of the 6 IL2 neuronal cell bodies (IL2DL, IL2DR, IL2L, IL2R,IL2VL, IL2VR) in the head of an adult hermaphrodite. The cell bodies are located in a 6-fold symmetry in a ring-like disposition in front of the nerve ring, very similar to the disposition of the IL1 neuronal cell bodies. Staining of the amphid neurons in the lateral ganglia, as seen in young larval stages L1-L2, totally disappears from late L3 to L4 and adult stages.E, Same adult animal as in D, stained with the DAPI nuclear dye. F, Left lateral view, lacZ staining of the pair of PHAL and PHBL, phasmid neurons located in the left lumbar ganglion in the tail of the L4 larva. Similar staining of the pair of PHAR and PHBR neurons can be observed on the right side of the animal in the right lumbar ganglion. Staining due to the fusion gene expression can be observed in all larval L1 to L4 and adult stages. G, Same developmental stage larva as shown in F, stained with the FITC dye. The anteriorly and posteriorly directed processes from the PHAL and PHBL cell bodies are projected into the pre-anal ganglion and the phasmid opening into the tail spike, respectively. Counterstaining of the fusion gene expressing animals with the nuclear dye DAPI further aided in cell identification (not shown). H, Schematic cartoon of the osm-3::lacZ expressing neurons in C. elegans. Only the left lateral view is shown. In the head 3 IL2 neurons IL2DL, IL2L, IL2VL, 8 amphid neurons ADFL, ADLL, ASEL, ASGL, ASHL, ASIL, ASJL, ASKL,and in the tail 2 phasmid neurons PHAL, and PHBL are stained. Thus, in a hermaphrodite up to L1-L2 larval stages all26 neurons open to outside environment express the fusion gene. Later in L3 and L4 stages staining of the 16 amphid neurons disappears and only 10 (6 IL2s, and 4 phasmid) neurons express the fusion gene. Scale bar represents 20 microns.Nematode strains were maintained on NGM nutrient agar plates at 20 C as described (Brenner, 1974). Germline transformation of C. elegans was done as previously described (Fire, 1986) by microinjecting osm-3::lacZ fusion gene recombinant DNA into the germ cells of adult gonad syncitium, as described by Mello et al. (1991). Injection solution contained 100 mg/ml of the fusion gene DNA in injection buffer (2% w/v) polyethylene glycol, 20 mM potassium citrate,adjusted to pH 7.5 with KOH). As a cotransformation marker, the DNA from a dominant allele rol-6(su1006) in the plasmidpRF4 (Kramer et al., 1990) at a concentration of 100 mg/ml in injection buffer was used. The transformed animals assume a distinct right-hand roller phenotype. Handling of the injected worms, histochemical staining, and photomicrography was as described (Fire, 1992; Fukushige et al., 1993).
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  Depict: Expr_pattern: Expr1583
    Anatomy (26)
  Acknowledgment: Template: WormBase thanks <Journal_URL> for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Reprinted from <Article_URL>. Copyright (<Publication_year>) with permission from <Publisher_URL>.
    Publication_year: 1995
    Article_URL: DOI id 10.1006/jmbi.1994.0146
    Journal_URL: JournalofMolecularBiology
    Publisher_URL: Elsevier
  Reference: WBPaper00002154