spas-1 is the worm homolog of the human spastic paraplegia gene, spastin. Previous work in C. elegans has shown that SPAS-1 disassembles microtubules in an ATP-dependent manner (Matsushita-Ishiodori et al, 2007). Another study shows that SPAS-1 plays a role in synapse remodeling in motor neurons (Kurup et al, 2015). Here we describe the expression of SPAS-1 in L4 hermaphrodite larvae using the transcriptional reporter, axrEx8[Pspas-1:mCh]: A) Low magnification (10X objective) image of an entire L4 larva. mCherry expression is driven by the spas-1 promoter (sequence provided). Expression is observed in the nerve ring, ventral cord, vulva, and tail ganglia. Expression is also detected in the intestine (asterisks). B) High magnification (100X objective) image of SPAS-1 expression in the developing vulva of an L4 hermaphrodite. C) High magnification (100X objective) image of nerve ring expression. D-F) SPAS-1 expression in the tail ganglia (TG), ventral cord (VC), and mechanosensory neurons (PLM and AVM shown here). Mechanosensory neuron expression is confirmed by overlaying GFP and mCherry fluorescence in axrEx8 [Pspas-1:mCh]; zdIs5 [Pmec-4:GFP + lin-15 (+)] animals. Arrows point at the following mechanosensory neurons: PLM=Posterior Lateral Microtubule cell, AVM=Anterior Ventral Microtubule cell.
Our work shows that SPAS-1 is expressed in several neuronal types as well as in the intestine and developing vulva in L4 larvae. These findings complement expression data by Matsushita-Ishiodori who focused on SPAS-1 expression in embryos.
axrEx8 [Pspas-1:mCh] transcriptional fusion construction details: 480bp upstream of the spas-1 ATG cloned into Pdonr4-1 gateway vector. Expression clone built by LR multisite recombination to drive mCherry expression. Strain REB22, injection concentration 20ng/ul.
spas-1 promoter sequence: provided by the authors and included in the transgene details.
Davidson College faculty study and research funds.
Reviewed ByMelissa Kelley
HistoryReceived: July 12, 2017
Accepted: July 27, 2017
Published: July 31, 2017