Figure 1.
Experimental design, diet composition and heatmap of significantly altered gonadal genes.
(A) The experimental timeline for this study. Four-month-old male and female rats were placed on one of 5 dietary regimes (control (ad libitum), 20% caloric restriction (CR), 40% CR, intermittent fasting (IF), or high fat/high glucose (HFG)). At 10 months of age, rats were euthanized and gonadal tissues were collected. A 17 K mouse gene array was performed and significant gene and pathway expression changes were quantified. (B) The relative proportions of the major nutritional groups in the control and high-fat/glucose (HFG) diets. (C) Regulatory heatmap of the significantly up-regulated (red) and down-regulated (green) genes in gonads collected from male and female rats on the different dietary regimes, compared to gonads collected from ad libitum controls.
Figure 2.
Effects of dietary regimes upon gonadal structure and plasma steroid levels.
Panels A and B depict the deviations in grams from the mean mass of the control ad libitum animals' gonadal size of the wet masses of the testes (A) or ovaries (B) from the multiple animals (n = 8) on the respective dietary regimes. Panels C and D depict the plasma testosterone/estrogen ratios (pg.ml−1/pg.ml−1) values for males (C) or females (D) subjected to the respective dietary regimes. Data is represented as mean±S.E. mean, n = 8 and *p<0.05, **p<0.01.
Figure 3.
Gene changes in the gonads of male and female rats maintained on the 20% CR diet compared to the gonads of male and female rats maintained on a control (ad libitum) diet.
Genes that were significantly up-regulated (red) or down-regulated (green) were clustered into a Venn diagram. There were 9 significantly up-regulated and 4 significantly down-regulated genes in the testes collected from 20% CR male rats compared to the genes from testes collected from control (ad libitum) male rats. Ovaries collected from 20% CR female rats showed 33 significantly up-regulated (red) and 10 significantly down-regulated (green) genes compared to control (ad libitum) female rats. There were no common gene alterations between the 20% CR male and female gonadal tissues. Names of the significantly altered genes can be found in Table S1.
Figure 4.
Gene changes in the gonads of male and female rats maintained on a 40% CR diet compared to the gonads of rats maintained on a control (ad libitum) diet.
Genes that were significantly up-regulated (red) or down-regulated (green) were clustered into a Venn diagram. There were 47 significantly up-regulated and 12 significantly down-regulated genes in the testes collected from 40% CR male rats compared to controls. Ovaries collected from 40% CR female rats showed 24 significantly up-regulated (red) and 32 significantly down-regulated (green) genes compared to the control ad libitum females. There was 1 common gene altered in the gonads of both the male and female rats in this dietary group. This gene, chordc1, was up regulated in female gonadal tissue and down regulated in male gonadal tissue. Names of the significantly altered genes can be found in Table S1.
Figure 5.
Gene changes in the gonads of male and female rats maintained on an IF diet compared to the gonads of rats maintained on a control (ad libitum) diet.
Genes that were significantly up-regulated (red) or down-regulated (green) were clustered into a Venn diagram. There were 82 significantly up-regulated and 53 significantly down-regulated genes in the testes collected from IF male rats compared to the genes from testes collected from control (ad libitum) male rats. Ovaries collected from IF female rats showed 12 significantly up-regulated (red) and 17 significantly down-regulated (green) genes compared to controls. There were 3 genes that was significantly altered in both the males and females in the IF dietary group compared to the control group. The expressed sequence D10Ertd447e was significantly up-regulated in the gonads of IF males and females, compared to control (ad libitum) males and females. The genes Fga and Hspa8 were each down-regulated in IF male and female gonads compared to controls. Names of the significantly altered genes can be found in Table S1.
Figure 6.
Gene changes in the gonads of male and female rats maintained on the HFG diet compared to the gonads of male and female rats maintained on a control (ad libitum) diet.
Genes that were significantly up-regulated (red) or down-regulated (green) were clustered into a Venn diagram. There were 18 significantly up-regulated and 15 significantly down-regulated genes in the testes collected from HFG male rats compared to controls. Ovaries collected from HFG female rats showed 19 significantly up-regulated (red) and 33 significantly down-regulated (green) genes compared to controls. There were no common genes between male and female rats in this dietary group. Names of the significantly altered genes can be found in Table S1.
Figure 7.
Caloric specificity of male and female multi-diet regulated genes.
Significantly altered genes that were common between one or more of the diet paradigms are drawn to visualize caloric specificity. (A) Commonly altered genes in the testes of male rats on the various diets showed 29% caloric restriction (CR) specificity (commonly regulated between the 20% CR, 40%, CR, and/or IF diets only) and 71% non-CR specificity. (B) Commonly altered genes in the ovaries of female rats on the various diets showed 50% caloric restriction (CR) specificity and 50% non-CR specificity. Names of the significantly altered genes can be found in Table S1.
Figure 8.
Caloric specificity of male-female multi-diet regulated genes.
Significantly altered genes that were common between one or more of the diet paradigms in the ovaries and the testes were drawn to visualize caloric specificity in cross-gender regulated genes. Names of the significantly altered genes can be found in Table S1.
Figure 9.
Pathway changes in the gonads of male and female rats maintained on the 20% CR diet compared to the gonads of male and female rats maintained on the control (ad libitum) diet.
Significantly regulated, functional pathway clusters were generated from the respective male or female gene sets using PAGE gene set analysis. Pathways that were significantly up-regulated (red) or down-regulated (green) were clustered into a Venn diagram. Pathways in blue were common to both males and females but were differentially regulated, e.g. the SIG_CHEMOTAX pathway, was differentially altered in the 20% CR male (M) and female (F) gonads. This pathway was up-regulated in the males and down regulated in the females in comparison to ad libitum controls. Names of the significantly altered pathways can be found in Table S2.
Figure 10.
Pathway changes in the gonads of male and female rats maintained on the 40% CR diet compared to the gonads of male and female rats maintained on the control (ad libitum) diet.
Significantly regulated, functional pathway clusters were generated from the respective male or female gene sets using PAGE gene set analysis. Pathways that were significantly up-regulated (red) or down-regulated (green) were clustered into a Venn diagram. Pathways in blue were common to both males (M) and females (F) but were differentially regulated. Names of the significantly altered pathways can be found in Table S2.
Figure 11.
Pathway changes in the gonads of male and female rats maintained on the IF diet compared to the gonads of male and female rats maintained on the control (ad libitum) diet.
Significantly regulated, functional pathway clusters were generated from the respective male or female gene sets using PAGE gene set analysis. Pathways that were significantly up-regulated (red) or down-regulated (green) were clustered into a Venn diagram. Pathways in blue were common to both males (M) and females (F) but were differentially regulated. Names of the significantly altered pathways can be found in Table S2.
Figure 12.
Pathway changes in the gonads of male and female rats maintained on the HFG diet compared to the gonads of male and female rats maintained on a control (ad libitum) diet.
Significantly regulated, functional pathway clusters were generated from the respective male or female gene sets using PAGE gene set analysis. Pathways that were significantly up-regulated (red) or down-regulated (green) were clustered into a Venn diagram. Pathways in blue were common to both males (M) and females (F) but were differentially regulated. Names of the significantly altered pathways can be found in Table S2.
Figure 13.
Caloric specificity of male and female multi-diet regulated pathways.
Significantly altered functional pathways that were common between diet paradigms were drawn to visualize caloric specificity. (A) Commonly altered functional pathways in the testes of male rats on the various diets showed 36% caloric restriction (CR) specificity (commonly regulated between the 20% CR, 40%, CR, and/or IF diets only) and 64% non-CR specificity. (B) Similarly, the commonly altered functional pathways in the ovaries showed 35% CR-specificity and 65% non-CR specificity. Names of the significantly altered pathways can be found in Table S2.
Figure 14.
Caloric specificity of male-female common multi-diet regulated pathways.
Significantly altered functional pathways that were common between diet paradigms and both genders were drawn to visualize caloric specificity. Names of the significantly altered pathways can be found in Table S2.
Figure 15.
Correlation of genetic output between complementary animal responses to different dietary paradigms.
Panel A depicts the gene∶pathway ratios created by each gender in response to the imposed dietary regime. Panel B demonstrates the functional cross-over between the KEGG functional pathway output of male IF compared to female 40% CR paradigms.
Table 1.
Webgestalt GO term analysis of gonadal transcriptional response to intermittent fasting compared to 20% caloric restriction.
Table 2.
Webgestalt KEGG pathway analysis of gonadal transcriptional response to intermittent fasting compared to 20% caloric restriction.
Table 3.
Alterations in fertility-related genes in the male and female rats on the different dietary regimes.