Fig 2. CLR-1/RPTP acts in postsynaptic neurons, and is localized to the synaptic region. (A) Schematics andmicrographs of normal PHB-AVA NLG-1 GRASP fluorescence in wild-type and
clr-1/RPTP
(e1745) mutant animalsexpressing a transgene that drives expression of the
clr-1 cDNA in AVA neurons (pAVA::
clr-1/RPTP), and reducedPHB-AVA NLG-1 GRASP fluorescence in
clr-1/RPTP
(e1745) mutant animals expressing either a construct that drivesexpression of the
clr-1/RPTP cDNA in PHB neurons (pPHB::
clr-1/RPTP), a transgene that drives expression of the
clr-1/RPTP cDNA with the extracellular domain deleted in AVA neurons (pAVA::
clr-1/RPTxcd), or a transgene that drivesexpression of the
clr-1/RPTP cDNA with a mutation that inactivates the phosphatase domain (pAVA::
clr-1/RPTPpd). (B)Quantification of NLG-1 GRASP fluorescence. Expression of
clr-1/RPTP in AVAs, but not PHBs restores NLG-1GRASP fluorescence in
clr-1/RPTP
(e1745) mutants (n>75). Expression of the
clr-1/RPTP cDNA with the extracellulardomain deleted or with a mutation in the active site of the phosphatase domain does not fully restore NLG-1 GRASPfluorescence in
clr-1/RPTP
(e1745) mutants (n>100). Two or more lines were examined with each transgene, andcombined in the graph above. Values for each individual transgenic line are included in S2 Table. NS, not significant, P<0.001, P<0.05, U-test. Comparison to
clr-1/RPTP indicated over individual bars. P-values were adjusted formultiple comparisons using the Hochberg method. 95% confidence intervals for the medians are included in S1 Table.(C) Expression of
clr-1/RPTP in AVAs, but not PHBs, rescues the behavioral defect in
clr-1/RPTP
(e1745) mutants(n>75). Expression of
clr-1/RPTP cDNA with the extracellular domain deleted or with a mutation in the active site ofthe phosphatase domain does not fully rescue the behavioral defect in
clr-1/RPTP
(e1745) mutants (
n60). NS, notsignificant, P<0.001, t-test. Comparison to
clr-1/RPTP indicated over individual bars. P-values were adjusted formultiple comparisons using the Hochberg method. Error bars are SEM. (D) Schematic and micrograph of an animalexpressing the
clr-1/RPTP cDNA linked to YFP in AVA (pAVA::
clr-1/RPTP::YFP). (E) Schematic and micrograph of ananimal expressing the
clr-1/RPTP cDNA linked to mCherry in AVA (pAVA::
clr-1/RPTP::mCherry) and PHB-AVANLG-1 GRASP, and overlay in a
clr-1/RPTP mutant animal. (D-E), CLR-1 localization is brightest in the preanalganglion (yellow box), and the majority of animals show localization in the anterior half of this region, wherePHB-AVA synapses usually form (green fluorescence).