Supplemental Material, Supplemental_Figure_legends - Sex-Dependent Influence of Developmental Toxicant Exposure on Group B <i>Streptococcus</i>-Mediated Preterm Birth in a Murine Model

<p>Supplemental Material, Supplemental_Figure_legends for Sex-Dependent Influence of Developmental Toxicant Exposure on Group B <i>Streptococcus</i>-Mediated Preterm Birth in a Murine Model by Tianbing Ding, Lauren A. Lambert, David M. Aronoff, Kevin G. Osteen, and Kaylon L. Bruner-Tran in Reproductive Sciences</p

Supplemental Material, Suppl_Figure_3 - Sex-Dependent Influence of Developmental Toxicant Exposure on Group B <i>Streptococcus</i>-Mediated Preterm Birth in a Murine Model

<p>Supplemental Material, Suppl_Figure_3 for Sex-Dependent Influence of Developmental Toxicant Exposure on Group B <i>Streptococcus</i>-Mediated Preterm Birth in a Murine Model by Tianbing Ding, Lauren A. Lambert, David M. Aronoff, Kevin G. Osteen, and Kaylon L. Bruner-Tran in Reproductive Sciences</p

Supplemental Material, Ding_Supp_Fig_2_final - Sex-Dependent Influence of Developmental Toxicant Exposure on Group B <i>Streptococcus</i>-Mediated Preterm Birth in a Murine Model

<p>Supplemental Material, Ding_Supp_Fig_2_final for Sex-Dependent Influence of Developmental Toxicant Exposure on Group B <i>Streptococcus</i>-Mediated Preterm Birth in a Murine Model by Tianbing Ding, Lauren A. Lambert, David M. Aronoff, Kevin G. Osteen, and Kaylon L. Bruner-Tran in Reproductive Sciences</p

TUNEL staining in adult testis of control, F1, F2 and F3 males.

<p>TUNEL staining was minimal in control testis (A), but prominent in tissues from F1, F2 and F3 males (B–D). Semi-quantitative analysis of TUNEL staining is shown in E. Compared to control samples, <i>p</i><0.01 for all toxicant exposed mice. Results are representative of at least 6 animals per group and from ≥3 more litters/group. Original magnification, 200x. <b>Inset</b>: Primary antibody omitted using the same tissue shown in Panel D.</p

Analysis of sperm concentration in control and toxicant-exposed mice.

<p>Box plot of sperm number/mg caudal weight. Center lines indicate the median number. Columns and vertical bars indicate the 25–75 percentiles and 10–90 percentiles, respectively. Data from multiple animals per group is presented (Control, N = 7; F1, N = 6; F2, N = 8; F3, N = 13). One additional F1 male was azoospermic and was excluded from the data presented. *<i>p</i> = 0.001.</p

Immunolocalization of AhR protein in caudal sperm smears from adult control, F1, F2 and F3 males.

<p>AhR protein expression was minimal in control animals (A), but this protein was highly expressed in F1 (B) F2 (C) and F3 (D) sperm. Computer assisted AhR intensity (E) was determined using a minimum of 10 individual sperm from each of 6 animals per group. Original magnification, 1000x. <b>Inset</b>: Primary antibody omitted using sperm from a control male. Compared to control, *<i>p</i> = 0.001; **<i>p</i> = 0.0006.</p

Immunolocalization of PGDH in adult testis of control, F1, F2 and F3 males.

<p>PGDH was abundant in testis from control males (A), but markedly reduced in F1 (B) and F2 (C) males. PGDH immunolocalization remained low in the testis of F3 males (D). Semi-quantitative analysis of PGDH staining is shown in Panel E. Compared to control samples, <i>p</i><0.001 for all toxicant exposed mice. Results are representative of at least 6 animals per group and from ≥3 more litters/group. Original magnification, 400x. <b>Inset</b>: Primary antibody omitted using the same tissue shown in Panel D.</p

Analysis of sperm morphology of control C57bl/6 mice, F1 males and two generations of descendants.

<p>Unexposed control mice exhibited the highest number of morphologically normal sperm (53%). There was a significant reduction in the number of morphologically normal sperm in males with a direct toxicant-exposure history (F1: 36%; <i>p</i> = 0.01 and F2: 20%; <i>p</i> = 0.0001, compared to control). Mice with only an indirect exposure also exhibited significant reductions in morphologically normal sperm (F3: 31%; <i>p</i> = 0.0001, compared to control). Tail defects were the most common abnormality in all groups, while spermatocytes from mice with a history of TCDD exposure (direct or indirect) exhibited a slight increase in head, mid-piece and acrosome defects compared to control mice. A minimum of 200 cells were analyzed per animal. N≥6 for all groups.</p

Light microscopy of normal and abnormal murine spermatozoa.

<p>All spermatozoa were classified by standard morphologic assessment. Normal spermatozoan (A); common tail defects (B–E); acrosome defect (F); misshapen head (G); mid-piece defects (H, I) and decapitated sperm (J). Magnification, 1000x.</p

ELISA of PGE₂ in whole testis of adult control, F1, F2 and F3 males.

<p>Only low levels of PGE₂ were detected in the testis of all control animals. In mice with a toxicant exposure history, there was a clear trend toward increased expression of PGE₂; however, production of this prostaglandin was highly variable and did not reach significance (compared to control: F1, <i>p</i> = 0.07; F2, <i>p</i> = 0.10; F3, <i>p</i> = 0.16). N = 6 for all groups.</p