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. Author manuscript; available in PMC: 2022 Feb 3.
Published in final edited form as: Cell. 2021 Aug 6;184(18):4697–4712.e18. doi: 10.1016/j.cell.2021.07.022

Figure 3. Cer1 is required for P11-acquired learning and vertically inherited memory.

Figure 3.

(A) Schematic of C. elegans Cer1, Cer4, and S. cerevisiae Ty3–1. (LTR, long terminal repeat; PBS, primer binding site; MA, matrix; CA, capsid; NC, nucleocapsid; PR, protease; RT, reverse transcriptase; RH = RNaseH; INT, tntegrase; PPT, PolyPurine. Tract, ORF, open reading frame).

(B) Immunofluorescence of wild-type or Cer1 mutant worm germlines stained for Histone H3 (control) or Cer1 GAG.

(C) Cer1 GAG is detected prominently in fraction 6 by western blot, while other cellular markers are enriched in lighter fractions. Each number represents a fraction from the gradient. The 3* and 6* fractions were further concentrated by ultracentrifugation before western blot.

(D) Western blot of Cer1 GAG in wild type and Cer1 mutant animals. GAG is absent from all fractions in the Cer1(gk870313) mutants (G6369A substitution).

(E and F) Cer1(gk870313) mutants (E) and Cer1 RNAi-treated worms (F) are defective for transgenerational inheritance of avoidance behavior when P0 mothers are trained using PA14 bacteria.

(G and H) Cer1(gk870313) mutants (G) and Cer1 RNAi-treated worms (H) are defective for transgenerational inheritance of avoidance behavior when P0 mothers are trained on the P11 small RNA.

(I) Control or P11-trained worms express Cer1 in the germline, but it is not detectable in neurons.

(J) daf-7p::gfp expression in ASI neurons (white arrows) increases upon P11 small RNA exposure compared to controls. daf-7p::gfp expression does not increase upon P11 exposure in Cer1 RNAi-treated worms. Scale bar, 25 μm.

For choice assays, each dot represents an individual choice assay plate (average of 115 worms per plate) from all replicates. For choice assays n = 29–30 plates per condition. For imaging experiments n = 48–52 neurons imaged. At least three biological replicates for all experiments. Boxplots: center line, median; box range, 25th–75th percentiles; whiskers denote minimum-maximum values. Two-way (E–H and L) analysis of variance (ANOVA), Tukey’s multiple comparison test. ****p < 0.0001, NS, not significant. See also Figure S3 and Table S1 for exact sample sizes (n) and p values.