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This revealed that ∼70% of dnj-14 mutants exhibited alterations in head neuron staining compared with only 9% of wild-type worms, representing an ∼8-fold increase.
The previously described male-specific head neuron expression for transcript c [ 25], the daf-19 transcript labelled simply as male-specific in WormBase, was observed with the recombineered fusion specific for F33H1.1c (WBID Expr9747).
To quantify these phenotypes, worms from 9 days of adulthood and older were imaged and scored for head neuron abnormalities ('neuron loss') and the presence of dorsal nerve cord GFP aggregates ('punctae') (Supplementary Material, Fig. S2).
Examination of strain CTD1052 (Additional file 1: Table S7), transformed with the single EEED8.6::Mc-mCherry construct (fNH060), demonstrated clear, distinct reporter expression in head neuron cell bodies and anterior dendritic extensions.
We also observed an increase in the proportion of dnj-14 animals with large GFP punctae in the dorsal nerve cord (Fig. 3A and Supplementary Material, Fig. S3); however, this phenotype was more variable and the difference between wild-type and mutant worms less obvious for than for the head neuron abnormalities described above.
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ALG-2 appears to be the primary neuronal Argonaute in C. elegans head neurons (58).
For example, C36B1.12, the worm ortholog of three presenilin-like genes, was almost exclusively expressed in head neurons, suggesting an ancient conserved role important to neuronal function.
The src-1 and csk-1 genes are co-expressed in some head neurons, the anchor cell and the tail region, while kin-22 and csk-1 genes are co-expressed in pharyngeal muscles and tail region.
Prominent expression was found in four head neurons and some pharyngeal cells.
Fourth, we showed that hmt-1 is expressed in coelomocytes, head neurons, and intestinal cells.
In the case of fluorescence quantification, head neurons of 10 20 worms per genotype were imaged with identical exposure times.
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