Expr1317

The expression of lim-6 in all neurons continues throughout adulthood. The expression of lim-6 in the uterus is dynamic. lim-6 reporter gene constructs (lim-6prom::GFP, lim-6up::GFP, lim-6r::GFP) start to be expressed in two uterine cells during the late L3/early L4 stage and the expression widens during the L4 stage to include the uv2 and uv3 cells, several uterine toroid (ut) cells, which form the lumen of the uterus and at least one cell type (sujn) of the spermatheca-uterine junction. Occasionally, weaker and less consistent expression can be observed in some cells of the distal side of the spermatheca, which connect the spermatheca to the rest of the somatic gonad.

Expr12499

lim-4 is expressed in the SMB neurons rather than the SIA neurons, as previously described (Sagasti et al., 1999; Expr1334). lim-4 expression in the SMB neurons gradually decreased from the L1 larval stage until it became undetectable in the adult stage.

Expr8553

The transgenic animals displayed mCherry in all five pairs of gonadal sheath cells. However, fluorescence in Sh5 was only seen in a subset of the adult animals, suggesting lower endogenous lim-7 expression . As expected, the URA neurons also expressed lim-7(+)::mCherry. Moreover, at least 10 additional cells, which appear to be neuronal, expressed lim-7(+)::mCherry near the isthmus and terminal bulb of the pharynx in a similar region to the URA neurons. Late embryos also appear to express lim-7(+)::mCherry in the same region. The exact identity of these cells requires further confirmation as many different neurons reside in this region in addition to other cell types.

Expr3765

To examine lim-4 expression during the embryogenesis, the eggs were stained at various developing stages with anti-myc antibodies. A GFP reporter was used for the GATA gene elt-2, which is specifically expressed in the gut lineage, to stage the embryos. MYC::LIM-4 immunoreactivity is first detected in two cells approximately 160 min post-fertilization at 8 gut nuclei stage. The two MYC::LIM-4-staining cells was identified as the neuroblasts ABalpppppa and ABpraaappa that respectively give rise to left/right ADF and AWB neurons, based on triple labeling of the embryos with antibodies raised against the LIN-26 protein, which is expressed in the nuclei of the hypodermal cells that border the ADF/AWB mother cells. Like lim-4::gfp, the MYC::LIM-4 immunoreactivity is not detected in ADF at any stage of larvae or adults, but it is observed in AWB and several other neurons in the head throughout life (N > 200). It is difficult to determine if LIM-4 is excluded from ADF or it is present there transiently.

Expr1334

In larvae and adults, lim-4::GFP expression was confined to neurons in the worm's anterior ganglia; embryonic expression was not examined in detail. Expression from both transgenes was observed in the AWB neurons but not in other sensory neurons. lim-4::GFP1 was also expressed in one RME motor neuron (RMEV), two RMD motor neurons (RMDL and RMDR), and the RID, RIV, SAA and SIA interneurons. lim-4::GFP2 was expressed in the same neurons, except for RID and RMEV. Expression in the GABAergic RME motor neurons, which control head movement. In a subsequent study, lim-4 was found to be expressed in SMB neurons, other than SIA neurons (Kim et al., 2015; Expr12499).

Expr8312

GFP fluorescence was shown in the pharyngeal and body wall muscles. The lim-8 promoter gfp fusion was also expressed in vulva, spermathecae, anal sphincter and depressor muscles, head neurons, gonadal sheath, and excretory canal. lim-8 expression was higher in larvae than that in adults, and the actual expression in the adult stage was very weak.

Expr8313

GFP fluorescence was shown in the pharyngeal and body wall muscles. lim-9 GFP fluorescence was also detected in some neuronal processes, vulva, spermathecae, anal sphincter and depressor muscles, gonadal sheath, and excretory canal.

Expr12729

A 393-bp odr-1 promoter region retaining both LIM-4/ SOX-2 and CEH-36/SOX-2 sites was sufficient to drive GFP expression, at the same levels as the reporter driven by a 1,027-bp odr-1 promoter, in both AWB and AWC neurons.

Expr13557

We fused different parts of the 1.2-kb intron to the GFP gene and identified a 300-bp region, lim-6int3_short, that drove expression only in PVT. Strong GFP expression was detected in PVT in embryos as early as in the comma stage, well before the PVP neurons extend their axons.

Expr16568

Since glial spectrin expression was not previously described, we examined this at single-cell resolution in vivo using a single-copy sma-1:zf1:gfp transgene. In embryos, this transgene is broadly expressed. However, day 1 adults exhibited pronounced expression in pharynx, spermatheca-uterine and rectal valves, and PHsh glia. In the anterior head, staining was observed where AMsh glia contact epithelial hyp-3-5 syncytia and AFD-NRE. Since epithelial and glial cells in this region are closely packed with fine cytoplasmic processes, we leveraged the ZF1/ZIF-1 degron system to cell-specifically delineate SMA-1:GFP staining. Proteasomal degradation of SMA-1:ZF1:GFP in AMsh glia by cell-specific expression of ZIF-1 indicated that the SMA-1 ''mesh'' staining was within AMsh glia, and this was depleted in unc-23 mutant animals. Thus, SMA-1 is expressed in the appropriate anatomical region and age, suggesting epithelial UNC-23/BAG2 regulates AMsh glial SMA-1/bH-spectrin cell non-autonomously.