Oral benzo[a]pyrene in Cyp1a1/1b1(-/-) double-knockout mice: Microarray analysis during squamous cell carcinoma formation in preputial gland duct.

Benzo[a]pyrene (BaP) is a prototypical polycyclic aromatic hydrocarbon (PAH) found in combustion processes. Cytochrome P450 1A1 and 1B1 enzymes (CYP1A1, CYP1B1) and other enzymes can activate PAHs to reactive oxygenated intermediates involved in mutagenesis and tumor initiation; also, CYP1 enzymes can detoxify PAHs. Cyp1(+/+) wild-type (WT) and Cyp1b1(-/-) knockout mice receiving oral BaP (12.5 mg/kg/day) remain healthy for >12 months. In contrast, we found that global knockout of the Cyp1a1 gene (1a1KO) results in proximal small intestine (PSI) adenocarcinoma within 8-12 weeks on this BaP regimen; striking compensatory increases in PSI CYP1B1 likely participate in initiation of adenocarcinoma in 1a1KO mice. Cyp1a1/1b1(-/-) double-knockout (DKO) mice on this BaP regimen show no PSI adenocarcinoma, but instead preputial gland duct (PGD) squamous cell carcinoma (SCC) occurs by 12 weeks. Herein, we compare microarray expression of PGD genes in WT, 1a1KO and DKO mice at 0, 4, 8, 12 and 16 weeks of oral BaP; about four dozen genes up- or down-regulated during most critical time-points were further verified by qRT-PCR. In DKO mice, CYP3A59 was unequivocally identified as the BaP-inducible and BaP-metabolizing best candidate responsible for initiation of BaP-induced SCC. Striking increases or decreases were found in 26 cancer-related genes plus eight Serpin genes in DKO, but not in 1a1KO or WT, mice on this BaP regimen; of the 26, 8 were RAS-related oncogenes. The mechanism by which cancer-related genes are responsible for SCC tumor progression in the PGD remains to be elucidated.

Oral benzo[a]pyrene-induced cancer: two distinct types in different target organs depend on the mouse Cyp1 genotype.

Benzo[a]pyrene (BaP) is a prototypical polycyclic aromatic hydrocarbon (PAH) found in combustion processes. Cytochrome P450 1A1 and 1B1 enzymes (CYP1A1 and CYP1B1) can both detoxify PAHs and activate them to cancer-causing reactive intermediates. Following high dosage of oral BaP (125 mg/kg/day), ablation of the mouse Cyp1a1 gene causes immunosuppression and death within ∼28 days, whereas Cyp1(+/+) wild-type mice remain healthy for >12 months on this regimen. In this study, male Cyp1(+/+) wild-type, Cyp1a1(-/-) and Cyp1b1(-/-) single-knockout and Cyp1a1/1b1(-/-) double-knockout mice received a lower dose (12.5 mg/kg/day) of oral BaP. Tissues from 16 different organs-including proximal small intestine (PSI), liver and preputial gland duct (PGD)-were evaluated; microarray cDNA expression and >30 mRNA levels were measured. Cyp1a1(-/-) mice revealed markedly increased CYP1B1 mRNA levels in the PSI, and between 8 and 12 weeks developed unique PSI adenomas and adenocarcinomas. Cyp1a1/1b1(-/-) mice showed no PSI tumors but instead developed squamous cell carcinoma of the PGD. Cyp1(+/+) and Cyp1b1(-/-) mice remained healthy with no remarkable abnormalities in any tissue examined. PSI adenocarcinomas exhibited striking upregulation of the Xist gene, suggesting epigenetic silencing of specific genes on the Y-chromosome; the Rab30 oncogene was upregulated; the Nr0b2 tumor suppressor gene was downregulated; paradoxical overexpression of numerous immunoglobulin kappa- and heavy-chain variable genes was found-although the adenocarcinoma showed no immunohistochemical evidence of being lymphatic in origin. This oral BaP mouse paradigm represents an example of "gene-environment interactions" in which the same exposure of carcinogen results in altered target organ and tumor type, as a function of just 1 or 2 globally absent genes.

Short-photoperiod exposure reduces vasopressin (V1a) receptor binding but not arginine-vasopressin-induced flank marking in male Syrian hamsters.

In Syrian hamsters, socially relevant information is communicated with a form of scent marking known as flank marking. There is substantial evidence that arginine-vasopressin acting on V1a vasopressin receptors (V1aR) in the medial preoptic-anterior hypothalamic continuum (MPOA-AH) regulates the expression of flank marking. Previous studies have shown that the expression of flank marking is also influenced by the circulating concentrations of gonadal hormones. In hamsters housed in long 'summer-like' photoperiods (i.e. >12.5 h of light/day), castration reduces flank marking and administration of testosterone restores precastration levels of flank marking. When exposed to short 'winter-like' photoperiods (i.e. <12.5 h of light/day), hamsters undergo gonadal regression and the circulating levels of testosterone decline. Surprisingly, flank marking induced during social encounters is not reduced in hamsters exposed to short photoperiods despite the low circulating concentrations of testosterone. In the present study, it was hypothesized that reductions in testosterone, caused by exposure to short photoperiod, would not reduce the ability of vasopressin to stimulate flank marking by its actions in the MPOA-AH. The amount of flank marking induced by vasopressin injected into the MPOA-AH did not significantly differ between hamsters housed in long and short photoperiods; however, short photoperiod-exposed males had significantly less V1aR binding in the MPOA than long photoperiod-exposed males. These results support the hypothesis that the sensitivity of the MPOA-AH to vasopressin is not reduced in short photoperiod-exposed males, despite decreases in serum testosterone. However, by contrast to our predictions, short photoperiod-exposed males have significantly reduced V1aR binding in the MPOA-AH compared to long photoperiod-exposed males.

Perinatal exposure to estradiol masculinizes aspects of sexually dimorphic behavior and morphology in gray short-tailed opossums (Monodelphis domestica).

The effects on adult sexually dimorphic behavior of perinatal exposure to estrogen were examined by treating male and female gray opossums with estradiol (EST), an estrogen receptor antagonist (tamoxifen:TX) or oil control (OIL) during the first week of life, a time period corresponding in this marsupial to late gestation in rodent species. Following gonadectomy and replacement therapy with testosterone in adulthood, males showed more scent-marking behavior than females and EST animals showed more scent marking than TX or OIL animals. Also, phalluses were longer and body weight was higher in males than in females and in EST-treated animals than in TX-treated animals; OIL animals were intermediate in these morphological measures. EST animals of both sexes showed less female-typical screeching threat behavior than OIL or TX animals. Because these hormone manipulations were conducted on the "fetus" directly in this marsupial (rather than via the maternal circulation as in previously studied eutherian species), these findings provide unique confirming evidence for masculinization of aspects of behavior and morphology by early exposure to estradiol in mammals.

Exposure to different spectra of light, continuous light and treatment with melatonin affect reproduction in the Indian desert gerbil Meriones hurrianae (Jerdon).

In two separate experiments, the gonadal response to melatonin treatment and exposure to continuous white light (LL) and different spectra of light (blue, green, yellow and red) was investigated in the Indian desert gerbil Meriones hurrianae. In experiment 1, exposure to green light for 8 weeks increased (p < 0.01) the gonadosomatic index (GSI) and sperm count, whereas exposure to red light decreased (p < 0.01) the GSI and sperm count. The response of accessory organs to the treatment varied. In experiment 2, exposure to LL for 5 weeks decreased the GSI and sperm count, and treatment with melatonin (in the late-light period) not only increased the GSI and sperm count, but also counteracted the inhibitory action of LL and red light. Melatonin treatment stimulated accessory sex organs and the scent gland surface area. The inhibition or stimulation of the reproductive parameters correlated with an increase or decrease in pinealocytes and pinealocyte nuclear diameters. Therefore, the influence of light and its spectra on reproduction may be mediated by the pineal gland, though extrapineal mechanisms cannot be ruled out.

Effects of postnatal exposure to alcohol on reproductive physiology and sexually dimorphic behavior in a marsupial, the gray short-tailed opossum (Monodelphis domestica).

The effects of postnatal exposure to alcohol on reproductive physiology and sexually dimorphic behavior and anatomy in adult male and female gray short-tailed opossums were examined. Female responsiveness to male pheromones and fertility in both sexes were essentially normal in postnatally alcohol-treated animals. However, aspects of sexually dimorphic behavior were masculinized and defeminized in females and demasculinized in males following gonadectomy in adulthood and treatment with male (testosterone) or female (estradiol) hormones. The possible role of alterations in neural aromatase activity by perinatal alcohol exposure in mediating these behavioral effects and the potential use of this marsupial species in perinatal alcohol studies are discussed.

Lack of gonadal control of the dorsal gland and sandbathing in male and female bannertail kangaroo rats, (Dipodomys spectabilis).

Previous studies have demonstrated gonadal control of mammalian scent glands; castration leads to reduced scent-marking rates and smaller gland sizes. I investigated whether gonadal hormones control the size of the dorsal gland (a specialized sebaceous gland) and sandbathing as a scent-marking behavior in adult male and female bannertail kangaroo rats (Dipodomys spectabilis). Gland sizes of males and females were similar in all age classes, except adult males with larger body weights had proportionately larger glands than females. Male gland sizes declined 18% following castration but were not significantly smaller than those of intact males; females showed no change in dorsal gland size either as a result of ovariectomy or after estradiol benzoate implants. Sandbathing rates also did not decline as a result of gonadectomy. Rather, castrated males and overiectomized females sandbathed at higher frequencies than intact males and ovariectomized females with estradiol benzoate implants, respectively. Gonadal hormones apparently have little influence on the function of a specialized scent gland and may inhibit sandbathing as a scent-marking behavior in adult D. spectabilis.