S1. Wild-type animals immobilize on exogenous tyramine.
Dose dependent immobilization of wild-type animals on exogenous tyramine. The percentage of immobilized animals, at varying concentrations of exogenous tyramine, are depicted over a 20 minute period. Each data point represents the average ± SEM for at least five trials totaling at least 50 animals.
S2. Movie of a wild-type animal on plates containing 30 mM tyramine.
Filming began immediately after the animal was placed on the plate and ended shortly after paralysis. Movie was shot at 15 frames per second (fps) and sped up five times.
S3. Movie of a wild-type animal after 5 minutes on plates containing 30 mM tyramine.
Animals paralyze, but can still move in response to mechanical stimulation. Body movements are apparent, but the head and neck remain paralyzed. Movie was filmed at 15 fps.
S4. Movie of a lgc-55 animal after 5 minutes on plates containing 30 mM tyramine.
lgc-55 mutant worms display sustained head movements, while body movements are inhibited. Movie was filmed at 15 fps.
S5. Co-expression of lgc-55::mCherry and glr-1::GFP
Co-expression of lgc-55::
(Maricq et al., 1995
(Hart et al., 1995
). Confocal images show co-expression of LGC-55 and GLR-1 in the RMD, SMDD and SMDV. The AVB cell-body, similar to the cell labeled by lgc-55
::mCherry, lies on the dorsal side of the animal just anterior to the posterior pharyngeal bulb and has a process that runs ventrally through the amphid commissure (indicated by arrow head) before turning anteriorly and entering the nerve ring (White et al., 1986
). It was previously reported that glr-1
is expressed in the AVJ, RIM, AVD and AVB (Hart et al., 1995
; Maricq et al., 1995
). In the adult animal, we observe glr-1::
GFP expression (middle panels) in three cells just anterior to the posterior bulb. The dorsal most cell in this group is slightly ventral compared to the lgc-55
::mCherry neuron and has a process that extends anteriorly into the nerve ring, similar to the cell body position and axon morphology of the AVJ. Based on cell body position, axon morphology, which includes a process that runs through the amphid commisure and co-expression with sra-11
::GFP (See Figures S6-7
), we conclude lgc-55
is expressed in the AVB. Anterior is to the left. Scale bar, 10 μm.
S6. Co-expression of lgc-55::mCherry and sra-11::GFP
Co-expression of lgc-55::
) (Altun-Gultekin et al., 2001
). Confocal images show LGC-55 expression in the AVB. Co-expressing cell is on the dorsal side of the animal just anterior to the posterior pharyngeal bulb and has a process the projects ventrally before turning anteriorly and entering the nerve ring in both sets of images. Anterior is to the left. Scale bar, 10 μm.
S7. sra-11::GFP and glr-1::GFP Label Different Subsets of Neurons.
(A) Fluorescence microscopy of animals expressing both glr-1::GFP(kyIs29) and sra-11::GFP(otIs123). Animals expressing both GFP markers were examined to determine the presence expression in the AVB. If sra-11::GFP and glr-1::GFP are both expressed in the AVB, three cells should be visible in animals expressing both transgenes. Here, four cells are visible suggesting that sra-11::GFP and glr-1::GFP label different subsets of neurons. (B) Confocal image of sra-11::GFP(zfEx46). Promoter is the same as sra-11::LGC-55, and shows expression in AVB and AIY similar to sra-11::GFP(otIs123).
S8. Movie of gentle anterior touch response of wild-type animals.
Wild-type animals suppress head oscillations in response to anterior touch.
S9. Movie of gentle anterior touch response of lgc-55(tm2913) mutant animals.
lgc-55 mutant animals fail to suppress head oscillations in response to anterior touch.
S10. LGC-55 expression in the RMD and SMD neurons or neck muscles restores sensitivity to exogenous tyramine.
Quantification of head movements of tissue specific rescue lines on 30 mM tyramine. Expression of lgc-55 in muscle (myo-3::LGC-55), RMD and SMD (glr-1::LGC-55) restores sensitivity to exogenous tyramine. Animals expressing lgc-55 in AVB (sra-11::LGC-55) and RMD alone (lim-4::LGC-55) are resistant to exogenous tyramine.
S11. lgc-55 Mutants Have Defects in Reversal Behavior.
Distribution of number of backward body bends in response to anterior touch of wild type (n= 317), tdc-1(n3420) (n=139), lgc-55(tm2913) (n=257), lgc-55 rescue [lgc-55(tm2913); zfEx2, (n=204)], sra-11::LGC-55 [lgc-55(tm2913); zfEx37, (n=190)], lim-4::LGC-55 [lgc-55(tm2913); zfEx44, (n=108)], glr-1::LGC-55 [lgc-55(tm2913); zfEx42, (n=133)], and myo-3::LGC-55 [lgc-55(tm2913); zfEx31, (n=157)] animals. See also .