. 2017 Jul 5;6:e25751. doi: 10.7554/eLife.25751

Table 1.

Motor neuron subclass markers.

DOI: http://dx.doi.org/10.7554/eLife.25751.005

Gene	Function	Previously reported subclass expression	Confirmed subclass expression	Revised/new subclass expression	Stage for onset of expression (in hermaphrodites)	Sex-specificity of expression	UNC-3 dependency and COE motif number
del-1	DEG/ENaC ion channel	VA1-VA12 VB1-VB11 SABVL/R*		VA1-VA11 VB1-VB11 SABVL/R^†	VA1-VA11: late L2^‡ VB1-VB11: early L2 SABVL/R: 3-fold embryo	Yes, in males VA12 also express del-1	Yes (3 motifs)
unc-129	TGF-beta like molecule	DA1-DA7, DB1-DB7, SABD^§	DA1-DA7, DB1-DB7, SABD^#		DA1-DA7, DB1-DB7, SABD: L1	No	Yes (3 motifs)
hum-2	unconventional myosin	DB1^¶	DB1**		DB1: L1 or earlier	No	N. D (1 motif)
bnc-1	C2H2 Zn finger transcription factor	VA1-VA12, VB2-VB11, SABVL/R^††	VA1-VA12, VB2-VB11, SABVL/R^‡‡		VA1-VA12, VB2-VB11, SABVL/R: late L1	Yes, CP7 and CP8^†† in male VNC	Yes⁷ (4 motifs)
cfi-1	ARID-type transcription factor	DA1-DA8^§§, DB1-DB7	DA1-DA8^##, DB1-DB7	VA2-VA11, VB3-VB11^##	DA1-DA8: not determined VA2-11, VB3-VB11: late L1	Not determined	Yes (>5 motifs)
avr-15	Glutamate-gated cloride channel	DA9, VA12^¶¶		VA11, AS11***	VA11, AS11: late L3	Yes, 4 additional neurons (not ACh) in male PAG	Yes (4 motif)
flp-18	FMRF-like neuropeptide	none		VA11 VA12^†††	VA11: L2/L3 VA12: L2/L3	Yes, 8 additional cells/neurons in male tail	Yes (1 motif)
ser-2	tyramine receptor	DA9^‡‡‡		DA8^§§§	DA8: L1 or earlier	No	No (0 motifs)
mig-13	transmembrane protein	DA9, VA12^###	DA9 VA12^¶¶¶		DA9: L1 or earlier VA12: L2	Yes, 2 additional ACh tail neurons	Yes (1 motif)
glr-4	Glutamate receptor subunit	none		DA9****	DA9: L1 or earlier	No	Yes (2 motifs)
itr-1	Inositol triphosphate receptor	PDA, DA9^††††		DA9^††††	DA9: L2/L3	No	Yes (1 motif)
sra-36	G-protein coupled receptor	none		DA8 DA9 VA11^‡‡‡‡	DA8, DA9: L1 or earlier VA11: L2	No	N. D (2 motifs)

*Reagent: wdIs3 X or wdIs6 II [del-1^prom::gfp]; Winnier et al., 1999.

^†Expression was confirmed in VA1-11, VB1-11, and SABVL/R based on cell body position, axonal trajectory, and expected number of neurons for each class. Also, see Figure 2C.

^‡As described in Winnier at al. (1999), by the end of L2, del-1^prom::gfp is visible in a few anterior VAs. This expression progresses in a wave from anterior to posterior VAs, with all VAs (except VA12) expressing del-1^prom::gfp by the L4/adult stage.

^§Reagent: evIs82B IV [unc-129^prom::gfp]; Colavita et al. (1998), Kratsios et al. (2015).

^#Expression was confirmed in DA1-7, DB1-7, and SABD based on cell body position, axonal trajectory, cell type-specific markers, and expected number of neurons for each class. Also, see Figure 2C.

^¶Reagent: mdIs123 [hum-2^prom::gfp] from James Rand and Stephen Fields; www.wormatlas.org.

**Expression was confirmed in DB1 based on cell body position and axonal trajectory. Also, see Figure 1B.

^††Reagent: bnc-1 (ot845[bnc-1::+mNG+AID]) CRISPR allele; Kerk et al., 2017.

^‡‡Expression was confirmed in VA1-VA12, VB2-VB11, SABVL/R based on cell body position and expected number of neurons for each class.

^§§Reagent: otEx6502 [cfi-1^fosmid::gfp]; Kerk et al., 2017.

^##Expression was confirmed in DA1-8 based on cell body position and the cell-type specific markers juIs14[acr-2::gfp] that labels DA, DB, VA, VB. This analysis also revealed that cfi-1 is not expressed in VA12. No expression in VA1, VB1 and VB2 was inferred based on cell body position and total number (3) of MNs expressing cfi-1 at the retrovesicular ganglion.

^¶¶Reagent: adEx1299[avr-15^prom::gfp]; Dent et al., 1997.

***Expression was confirmed in AS11 based on cell body position, axonal trajectory, and cell type-specific markers. Also, see Figure 9D–E and Figure 1-figure supplement 1 .

^†††Reagent: ynIs59[flp-18^prom::gfp] from Chris Li. Expression in VA11 and VA12 was confirmed with cell type-specific markers. Also, see Figure 1B, Figure 2A–B, and Figure 1—figure supplement 1.

^‡‡‡Reagent: otIs107 [ser-2^prom1::gfp]; Tsalik et al., 2003.

^§§§Expression in DA8 was confirmed with cell type-specific markers. Also, see Figure 1B and Figure 1—figure supplement 1.

^###Reagent: muIs42[mig-13^prom::MIG-13::gfp]; Sym et al., 1999, Klassen and Shen (2007).

^¶¶¶Expression in DA9 and VA12 was confirmed with cell type-specific markers. Also, see Figure 2A–B.

****Reagent: otIs476[glr-4^prom::tagrfp]. Expression in DA9 was confirmed with cell type-specific markers. Also, see Figure 1—figure supplement 1.

^††††Reagent: otIs453 [itr-1^prom::gfp]. Plasmid used for otIs453 transgene is pBT001, which was generated by Gower et al., 2001 and gfp expression in C.elegans in DA9 and PDA is described in Gower et al., 2001 and Klassen and Shen (2007). Expression was confirmed in DA9 using cell type-specific markers. Also, see Figure 2A–B and Figure 1—figure supplement 1.

^‡‡‡‡Reagent: sEx11976[sra-36^prom::gfp]. Expression in VA11, DA8 and DA9 was confirmed with cell type-specific markers. Also, see Figure 1B and Figure 1—figure supplement 1. ^{†, #,} **^{, ‡‡, ##, ¶¶, †††, §§§, ###,} ****^{, ††††}See also ‘Motor Neuron Subclass Identification’ section in Materials and methods.

N. D: Not determined.