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. 2022 Mar 8;11:e74581. doi: 10.7554/eLife.74581

Figure 3. 4-Vinylanisole (4-VA) promotes sexual maturation synchrony in young females by enhancing juvenile hormone/vitellogenin (JH/Vg) signaling pathway at post adult eclosion (PAE) 3–4 days.

(A) The 4-VA effects on sexual maturity of gregarious virgin females at different developmental stages. (B) Dosage effects on electroantennography (EAG) responses of females to 4-VA at different developmental stages. EAG responses to 4-VA with different concentrations were recorded in the antennae of female adults aged at PAE 2 days, PAE 4 days, and PAE 6 days, respectively (n = 7–11). (C) Volcano plot of RNA-seq in the corpus cardiacum-corpora allatum (CC-CA) complex of gregarious females after 4-VA stimulation at PAE 3–4 days. Red dots indicate genes related to JH metabolism. (D) Expression changes of JH metabolism-related genes in the CC-CA by 4-VA stimulation. Red and blue indicate upregulated and downregulated, respectively. (E) JH titers in the hemolymph, (F) the mRNA levels, (G) the protein levels of Vg in the fat body, and (H) the protein levels of Vg in the ovary of gregarious females after 4-VA stimulation at different developmental stages. Dark lines in violin plot indicate median value. White dotted lines indicate upper and lower quartiles, respectively; columns show means ± SEM. Consistency analysis of maturity was analyzed using Levene’s test. The mRNA and protein levels were analyzed using Student’s t-test. The number of biological replicates and p values were shown in the figures. n.s., not significant.

Figure 3—source data 1. Raw data for sexual maturity, juvenile hormone (JH) titer, gene expression, and protein level in 4-vinylanisole (4-VA)-treated females.

Figure 3.

Figure 3—figure supplement 1. Effects of gregarious males with different ages on maturation synchrony of female aged at post adult eclosion (PAE) 3–4 days.

Figure 3—figure supplement 1.

Gregarious females aged at PAE 3 days were reared together with the fifth-instar gregarious males, or male adults aged at PAE 1 day, PAE 3 days, PAE 5 days for 2 days, respectively. Data were analyzed by Levene’s test. The number of biological replicates was shown in the figures. Lines in droplet diagram indicate median quartile.
Figure 3—figure supplement 1—source data 1. Raw data for sexual maturity of females reared with gregarious males with different ages.
Figure 3—figure supplement 2. Enrichment of differentially expressed genes (DEGs) and Kyoto encyclopedia of genes and genomes (KEGG) in the fat body of gregarious females after 4-vinylanisole (4-VA) stimulation at different developmental stages.

Figure 3—figure supplement 2.

Gene expression pattern showed the most dynamic changes in the fat body of females exposed to 4-VA at post adult eclosion (PAE) 3–4 days.
Figure 3—figure supplement 3. Peripheral electrophysiological responses of female locusts to 4-vinylanisole (4-VA).

Figure 3—figure supplement 3.

(A) and (B) Dosage effects on single sensilla responses (SSRs) of basiconic sensillum in females to 4-VA. (C) The SSR of basiconic sensillum in females to 4-VA aged at different developmental stages. Different letters represent significant differences between the two groups (one-way ANOVA, p < 0.05).
Figure 3—figure supplement 3—source data 1. Raw data for electrophysiological responses of female locusts to 4-vinylanisole (4-VA).
Figure 3—figure supplement 4. The mRNA levels of LmigOr35 during post adult eclosion (PAE) 1–8 days.

Figure 3—figure supplement 4.

Different letters represent significant differences (one-way ANOVA, p < 0.05). Points labeled with different letters indicate a significant difference between these groups. Data are shown as means ± SEM.
Figure 3—figure supplement 4—source data 1. Raw data for mRNA levels of LmigOr35 during post adult eclosion (PAE) 1–8 days.
Figure 3—figure supplement 5. Volcano plot of RNA-seq data in the brain of females treated by 4-vinylanisole (4-VA) at post adult eclosion (PAE) 3–4 days.

Figure 3—figure supplement 5.

There were 52 downregulated and 37 upregulated genes in the brain of female locusts upon 4-VA treatment, respectively.
Figure 3—figure supplement 6. The mRNA level of JHAMT and JHE upon 4-vinylanisole (4-VA) treatment.

Figure 3—figure supplement 6.

Student’s t-test, n = 4, *p < 0.05, **p < 0.01. Data are shown as means ± SEM.
Figure 3—figure supplement 6—source data 1. Raw data for mRNA level of JHAMT and JHE upon 4-vinylanisole (4-VA) treatment.