Figure 1. A) Map of the exons, and intron of spch-3 (T27A3.4) with the location of the xc2 deletion in red. B) Map of the new spch-3 allele after mutation, with frameshifted nucleotides to the first stop codon in purple.
Figure 1. A. Map of exons, introns and the 3'UTR of cls-2 (R107.6). B. The eighth exon and 3'UTR of cls-2 (R107.6) with the position of the xc3, xc4, and xc5 mutations indicated in red. C. Alignment of DNA and amino acid sequences in mutant and wildtype worms with mutations in red.
Fig. 3. Immunofluorescent staining of the nematode nervous system with the monoclonal antibody 3A5. Staining of the adult hermaphrodites whole mount preparations with the monoclonal antibody 3A5. Anterior of the animal is to the left. Scale bar is 20 microns. (A) Left lateral view. Staining of nervous system in the head, showing intense staining of the cirumpharyngeal central neuropil, known as the nerve ring (N.R.). Neuronal cell bodies are stained in all major ganglia albeit much less brightly than the neuronal processes emanating from them, including the neurons in anterior ganglion (a.g.), ventral ganglion (v.g.), and along the ventral nerve cord (V.C.). Anteriorly directed dendritic endings in from neurons located in the head ganglia can be seen in front of the anterior ganglion (a.g.). (B) Counter staining of cellular nuclei of the animal stained with 3A5 in panel (A), which is doubly stained with the nuclear dye DAPI to show relative disposition of neurons in the head. (C) Dorsal view of the nerve ring (N.R.) at a higher magnification, consisting of axonal processes from several sensory neurons, inteneurons and motor neurons. Around the nerve ring, intense staining of nerve ring axonal bundles and relatively much weaker staining of neuronal cell bodies. (D) Staining of the left touch receptor neuron (ALM) in the anterior half of the animal, and the ventral cord (V.C.) consisting of cell bodies of motor neurons and various axonal processes that run along the entire body length of the animal. One of the motor neuron commissures comprising dorsally directed axonal fibers from the motor neurons located in the ventral cord, is marked by small arrowheads. (E) Staining of the dorsal cord (D.C.). in the head region that runs dorsally along the anterior-posterior axis of the animal consisting of processes of motor neurons whose cell bodies are located in the ventral cord (V.C,). Neuronal cell bodies of the dorsal ganglion (d.g.) can also be seen behind the nerve ring. (F) Another focal plane of the same animal as shown in panel (E), revealing staining of the lateral nerve cords consisting of axonal processes from neurons located in the head and tail ganglia. Intense staining of the nerve ring (N.R.) which is out of focal plane is also seen. (G) Ventral view of the posterior ganglia in the tail region. Staining of the pair of PLMR touch receptor neuron located in the right lumbar ganglion. Anteriorly and posteriorly directed axonal processes of PLMR are indicated by arrow heads. Also shown is the phasmid sensillum (ph.s.). the cell bodies of phasmid neurons PHA and PHB are situated in the anterior region of lumbar ganglia. Posteriorly directed dendrites from these neurons extend posteriorly into the tail and open to the outside through a hole in the cuticle. The anteriorly directed process extends along the ventral lateral cord. Axonal processes of other lumbar neurons extend along the lumbar commissures (1.c.) to reach the pre-anal ganglion, where they make synapses with neurons located in the pre-anal ganglion and other axonal processes in the ventral cord. (H) The same animal as shown in panel (G), counter stained with the nuclear dye DAPI to reveal relative position of the cellular nuclei in the tail ganglia. Animals were fixed, and stained with 3A5 antibody and the nuclear dye DAPI, as described [15].