Enhanced Akt signaling in oocytes of <i>Pten^loxP/loxP</i>; <i>Zp3-Cre+</i> mice.

<p>Oocytes were prepared from ovaries of 3–4 week old mice that were treated with PMSG, as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0006186#s4" target="_blank"><i>Materials and Methods</i></a>. Signaling studies in <i>Pten^loxP/loxP</i>; <i>Zp3-Cre+</i> oocytes showed elevated levels of p-Akt (Ser473), p-Akt (Thr308), and p-Tsc2 (Thr1462) as compared to <i>Pten^loxP/loxP</i> oocytes. Levels of total Akt, Tsc2, and β-actin were used as internal controls. 100–150 oocytes were used for each lane. All experiments were repeated at least three times and representative results are shown.</p

Normal follicular development in <i>Pten^loxP/loxP</i>; <i>Zp3-Cre+</i> mice.

<p>Morphological analysis of ovaries from 13- and 23-day-old, and 16-week-old <i>Pten^loxP/loxP</i>; <i>Zp3-Cre+</i> mice, <i>Pten^loxP/loxP</i>; <i>GCre+</i> mice, and control <i>Pten^loxP/loxP</i> mice. Ovaries were embedded in paraffin and sections of 8-µm thickness were prepared and stained with hematoxylin. Note the overactivation of primordial follicles in <i>Pten^loxP/loxP</i>; <i>GCre+</i> ovaries (C, F, and I, arrows) and the normal follicular development and CL in <i>Pten^loxP/loxP</i>; <i>Zp3-Cre+</i> ovaries (B, E, H and K), which is comparable to the control <i>Pten^loxP/loxP</i> ovaries (A, D, G, and J). CL, corpora lutea.</p

Characterization of <i>Pten</i> deletion by western blot and PCR.

<p>(A) Oocytes were prepared and lysed for western blot as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0006186#s4" target="_blank"><i>Materials and Methods</i></a>. PTEN expression was found to be completely absent in <i>Pten^loxP/loxP</i>; <i>Zp3-Cre+</i> oocytes. For each lane, 150 oocytes were used. β-actin was used as internal control. (B) PCR analysis showing the complete deletion of <i>Pten</i> exon 5 (<i>Pten</i> Δ5) in one allele of the genomic DNA of pups from <i>Pten^loxP/loxP</i>; <i>Zp3-Cre+</i> females.</p

Generation of mutant mice with oocyte-specific deletion of <i>Pten</i>.

<p>A schematic representation of deletion of <i>Pten</i> exon 5 in oocytes of primary and further developed follicles by using the <i>Zp3</i> promoter-mediated Cre transgenic mice. The developmental stages at which the <i>Gdf-9</i> promoter and the <i>Zp3</i> promoter become active are indicated above the illustration of follicles in the figure.</p

Associations between genetic variants in hormone-related genes and breast cancer risk in pre- and post-menopausal women.

a<p>Models were adjusted for ethnicity (Caucasian, African American, Hispanic) and through matching, were also adjusted for age at blood donation, duration of sample storage, and menopausal status.</p>b<p>Models were adjusted for ethnicity (Caucasian, African American, Hispanic), age at first birth/parity (≤20 years, 21–25 years, 26–30 years, >30 years, nulliparous), age at menarche, family history of breast cancer, ever use of HRT, and BMI, and through matching, were also adjusted for age at blood donation, duration of sample storage, and menopausal status.</p>c<p>For each genetic variant (except <i>PGR-12</i> rs1042838), the genotype associated with higher estrogen exposure (see below) was assigned a value of 1 and other genotypes (homozygous and heterozygous for the lower estrogen exposure allele) were assigned 0. A score was created by adding the values. Women with four or five high estrogen variables were grouped because there were too few women with five high estrogen variables to assess separately (3 cases/4 controls).</p><p><b>Notes: For each genotype, variants are listed in order of expected increasing estrogen (or progesterone for </b><b><i>PGR-12</i></b><b>) exposure:</b></p><p><i>SHBG</i> (rs6259) A allele is associated with higher SHBG levels. SHBG binds to estrogens and reduces their bioavailability. G allele = higher estrogen exposure.</p><p><i>PGR-12</i> (rs1042838) T allele has been shown to reduce the <i>PGR</i> transcript stability and the response to progesterone. G allele = higher progesterone exposure.</p><p><i>ESR1</i> (rs2234693) C allele may be associated with increased ERα transcription. C allele = possible higher exposure to estrogen signaling.</p><p><i>CYP19</i> 3′UTR (rs10046) T allele is associated with increased transcriptional activity of aromatase (converts androgens to estrogens). T allele = higher estrogen exposure.</p><p><i>CYP19</i> 5′Flank (rs4775936) A allele is associated with increased transcriptional activity of aromatase (converts androgens to estrogens). A allele = higher estrogen exposure.</p><p><i>UGT1A1</i> (rs8175347) 7 repeat allele is associated with reduced glucuronidation (and clearance) of estrogens. 7 repeat allele = higher estrogen exposure.</p

Distributions of demographic and lifestyle variables by breast cancer status.

a<p>For parous women only.</p>b<p>HRT use during follow-up (for NYUWHS) or use history assessed at enrollment (NSMSC).</p>c<p>p-values were estimated using logistic regression conditional on matching sets.</p

Description of case and control subjects, NSMSC.

<p>a. Missing values: < 5% for height, BMI, age at menarche, age at first full term pregnancy, and OC use; <10% for plasma 25(OH)D level and HRT use; 30% for age at menopause, 38% for alcohol intake, and 13% for ER status</p><p>b. P value for trend was calculated by conditional logistic regression using ordered categories; two-sided</p><p>c. Among ever parous women</p><p>d. Among postmenopausal women</p><p>e. Menopausal status is partially matched. Cases identified/selected more recently have two controls/case matched on menopausal status</p><p>Description of case and control subjects, NSMSC.</p

Season-adjusted 25(OH)D geometric means by genotype (for SNPs associated with 25(OH)D in GWAS) among controls (n = 1292).

<p>a. Number of controls for each genotype with measured 25(OH)D</p><p>b. P value for trend was calculated by ANOVA</p><p>Season-adjusted 25(OH)D geometric means by genotype (for SNPs associated with 25(OH)D in GWAS) among controls (n = 1292).</p

SNPs genotyped in pilot and case-control studies.

<p>SNPs genotyped in pilot and case-control studies.</p

Cinémagazine : hebdomadaire illustré

20 décembre 19291929/12/20 (A9,N51)-1929/12/20