# Presentation: 351040

## Pubertal assays (♀ and ♂)

- Summary of study designs for
- Single-dose
- Multi-dose
- Multi-chemical array

## Pubertalsingle-dosestudy

## Pubertal single-dose:general design

**Gavage**
**6 rats/chemical/block/strain**
**2 blocks per sex**
**6 chemicals per sex, with different means of affecting endocrine system**
**Single (high) dose per chemical**
**2 strains of rat (LE and SD)**

## Slide 4

## Female pubertal single-dose:chemicals and doses (mg/kg/day)

**Ethinyl estradiol**** (potent estrogen)**
**    ****0.005**
**Tamoxifen****(blocks ER; partial antagonist/agonist)**
**  ****10**
**Propylthiouracil****(inhibits T3/T4 formation)**
**240**

**Pimozide****(increases prolactin secretion)**
**  ****30**
**Methoxychlor****(weak estrogen)**
**100**
**Ketoconazole****(inhibits steroidogenesis)**
**100**

## Male pubertal single-dose:chemicals and doses (mg/kg/day)

**Flutamide**** (potent anti-androgen)**
**    ****50**
**Methyl testosterone****(potent androgen)**
**    ****80**
**Propylthiouracil****(inhibits T3/T4 formation)**
**  ****240**

**Pimozide****(increases prolactin secretion)**
**    ****30**
**Dibutylphthalate****(weak anti-androgen)**
**1000**
**Ketoconazole****(inhibits steroidogenesis)**
**  ****100**

## Pubertalmulti-dosestudy

## Pubertal multi-dose:general design

**Gavage**
**15 SD rats per dose**
**Weak estrogen (****♀) ****or anti-androgen (****♂)**** at 3 doses**
**Positive and negative controls****(2 doses of positive control to investigate putative influence of bodyweight changes)**

## Slide 9

## Males
Vehicle control
Vinclozolin
  10
  30
100
Flutamide
  25
  50

**Vehicle control**
**Vinclozolin**
**  ****10**
**  ****30**
**100**
**Flutamide**
**  ****25**
**  ****50**

**Females**
**Vehicle control**
**Methoxychlor**
**  ****12.5**
**  ****25**
**  ****50**
**Ethinyl estradiol**
**    ****0.0025**
**    ****0.0050**

**Pubertal multi-dose:****chemicals and doses**** ****(mg/kg/day)**

## Pubertal multi-dose:endpoints (female)

- Daily body weight, clinical observations
- Age at vaginal opening(pinhole, thread, complete)
- Estrous cyclicity
- Weights: uterus and cervix (wet & dry), ovaries, thyroid/parathyroids; liver, kidney, pituitary, adrenals
- Histopathology of ovaries, uterus, thyroid; gross lesions

## Pubertal multi-dose:endpoints (male)

- Daily body weight, clinical observations
- Age at preputial separation(onset, thread, complete)
- Weights: testes, epididymides, prostate (DL/V separately), seminal vesicles/coagulating glands, levator ani/bulbocavernosus, thyroid/parathyroids; liver, kidney, pituitary, adrenals
- Histopathology of testis, epididymis, thyroid

## Pubertal multi-chemical arraystudy

## Pubertal multi-chemical array:general design

**Gavage**
**15 SD rats per dose**
**10 chemicals with different means of affecting endocrine system**
**2 doses per chemical**
**Vehicle control for each of 2 “components” per sex**

## Pubertalmulti-chemical array study(females)

**Pubertal****multi-chemical array study**(females)

## Female pubertal array:chemicals and doses (mg/kg/day)

**Vehicle control**
**Fenarimol****(inhibits aromatase)**
**  ****50**
**250**
**Octylphenol****(weak estrogen)**
**200**
**400**
**Bisphenol A****(weak estrogen)**
**100**
**400**
**Ketoconazole****(inhibits steroidogenesis)**
**  ****50**
**100**

- Component 1

- Component 2

**Vehicle control**
**Atrazine****(affects hypothalamus)**** **
**  ****75**
**150**
**Propylthiouracil****(inhibits T3/T4 formation)**
**    ****2**
**  ****25**
**Fadrozole****(inhibits aromatase)**
**    ****0.3**
**    ****3.0**

## Female pubertal array:endpoints

- Daily body weight, clinical observations
- T4 and TSH levels
- Age at vaginal opening(pinhole, thread, complete)
- Estrous cyclicity
- Weights: uterus (wet & dry), ovaries, thyroid
- Histopathology of ovaries, uterus, thyroid(vagina if warranted)
- Liver, kidney, pituitary, adrenal weights if warranted

## Pubertalmulti-chemical array study(males)

**Pubertal****multi-chemical array study**(males)

## Male pubertal array:chemicals and doses (mg/kg/day)

**Vehicle control**
**Atrazine****(affects hypothalamus)**** **
**  ****75**
**150**
**Propylthiouracil****(inhibits T3/T4 formation)**
**    ****2**
**  ****25**
**Procymidone****(anti-androgen)**
**  ****50**
**100**

- Component 1

- Component 2

**Vehicle control**
**Linuron**** (anti-androgen +/prenatal, -/adult)**
**  ****50**
**100**
**p,p’-DDE****(anti-androgen)**
**  ****50**
**100**
**Ketoconozole****(inhibits steroidogenesis)**
**  ****50**
**100**

## Daily body weight, clinical observations
T4 and TSH levels
Age at preputial separation(onset, thread, complete)
Weights: testes, epididymides, prostate (whole and DL/V), seminal vesicles/coagulating glands, levator ani/bulbocavernosus, thyroid
Histopathology of testis, epididymis, thyroid
Liver, kidney, pituitary, adrenal weights if warranted

- T4 and TSH levels
- Age at preputial separation(onset, thread, complete)
- Weights: testes, epididymides, prostate (whole and DL/V), seminal vesicles/coagulating glands, levator ani/bulbocavernosus, thyroid
- Histopathology of testis, epididymis, thyroid
- Liver, kidney, pituitary, adrenal weights if warranted

**Male**** pubertal array:****endpoints**