Data on spermatogenesis in rat males gestationally exposed to bisphenol A and high fat diets

This data article contains supporting information regarding the research article entitled “High butter-fat diet and bisphenol A additively impair male rat spermatogenesis” (P. Tarapore, M. Hennessy, D. Song, J. Ying, B. Ouyang, V. Govindarajah, et al.,) [1]. Sprague–Dawley females were fed AIN, high fat butter, 17α-ethinyl estradiol, or high fat butter plus four bisphenol A doses (2500 µg/kg bw-d, 250 µg/kg bw-d, 25 µg/kg bw-d, and 2.5 µg/kg bw-d) before and during pregnancy. All diets were switched to AIN after the pups were born. Male offspring received testosterone (T)- and estradiol-17β (E2)-filled implants from postnatal day 70–210 for 20 weeks (T+E2 rat model). The testes were weighed, and examined for impairments in spermatogenesis.


a b s t r a c t
This data article contains supporting information regarding the research article entitled "High butter-fat diet and bisphenol A additively impair male rat spermatogenesis" (P. Tarapore, M. Hennessy, D. Song, J. Ying, B. Ouyang, V. Govindarajah, et al.,) [1]. Sprague-Dawley females were fed AIN, high fat butter, 17α-ethinyl estradiol, or high fat butter plus four bisphenol A doses (2500 mg/ kg bw-d, 250 mg/kg bw-d, 25 mg/kg bw-d, and 2.5 mg/kg bw-d) before and during pregnancy. All diets were switched to AIN after the pups were born. Male offspring received testosterone (T)-and estradiol-17β (E2)-filled implants from postnatal day 70-210 for 20 weeks (Tþ E2 rat model Immuno-histological patterns of expressions of aromatase (Cyp19) and ERα in testis may be useful for future work related to the distribution of these two markers in testis.

Data
We conducted a dose-response analyses to determine the minimal BPA dose that impedes spermatogenesis ( Fig. 1) in male offspring exposed in utero to diets with bisphenol A (BPA) and high fat butter (HFB). Details on diets, animal groups and approach are outlined in Fig. 1A. The number of seminiferous tubules (STs) within the testis (per animal) with progression of spermatogenesis upto the round spermatids (Fig. 1B) or upto spermatozoa (Fig. 1C), was scored and plotted (T þE2 model). The body weights and the weights of testis and spleen were scored ( Figs. 2A-C). In a separate work, data is presented for body weight and weight of the testis, epididymis, spleen, and kidney for offspring prenatally exposed to AIN, BPA, HFB, high fat olive oil (HFO), HFB þBPA, or HFO þ BPA diets (Figs. 2D-G) and T þE2.
We examined the STs of the testis for presence of clusters of cells (using BRDT staining, Fig. 3) and for ERα (Fig. 4) and CYP19 (aromatase, Fig. 5) expression between the diet groups.

Diets and animals
Sprague-Dawley females were fed AIN, high fat butter (39 kcal% fat, HFB), 17α-ethinyl estradiol (EE2 (0.5 mg/kg bw-d), or HFB plus four BPA doses (2500 mg/kg bw-d, 250 mg/kg bw-d, 25 mg/kg bw-d, and 2.5 mg/kg bw-d) before and during pregnancy (Fig. 1A). All diets were switched to AIN after the pups were born. At postnatal day (PND 70), prenatally exposed pups from each diet group were treated with T þE2 via Silastic TM implants [2,3] (T þE2 rat model) for 20 weeks. The animals were weighed, the testis, epididymis, spleen, and kidney were weighed, fixed, paraffin embedded, stained with hematoxylin and eosin and tubules examined for spermatogenesis (Figs. 1B and C). More details on the Tþ E2 model, tissue collection and data analyses are outlined in Tarapore et al., 2016 [1]. For Figs. 2D-G, the BPA administered to the dams in diet was 25 mg/kg bw-d. The sham-implanted, gestational exposed groups exhibited normal spermatogenesis on PND210 (100% offspring showed presence of spermatozoa in 414% of STs). The number of STs with spermatogenesis impaired at the round spermatids was tallied for male offspring exposed to maternal diets indicated. A non-monotonic dose response curve was observed. Significance analyzed with 1-way ANOVA (p ¼ 0.0007) and Dunnett's multiple comparison test. (C) The number of STs with spermatozoa was tallied for male offspring exposed to the maternal diets indicated. A non-monotonic dose response curve was observed. Significance analyzed with 1-way ANOVA, and while the means were significantly different (p¼ 0.0239), significance between groups was not reached. HFB, High Fat Butter; BPA, Bisphenol A; EE2, ethinyl estradiol positive control * p o 0.05, ** p o 0.01, by 1-way ANOVA (parametric) compared to AIN diet.

Immunohistochemistry staining
The procedure and antibody sources are as outlined in Tarapore et al. [1].

Statistical analysis
For Figs. 1 and 2, significance was analyzed with one-way ANOVA and Dunnett's multiple comparison test using the GraphPad Prism software. Body and organ weights of male offspring exposed in utero to various diets. No significant difference was observed in the body weight (A, D), or the weight of the testis (B, E), epididymis (F), spleen (C, G) or prostate (H) of male offspring exposed in utero to the diets indicated in the T þ E2 model. No significance was found using 1-way ANOVA.