Cytochrome P4501A mRNA and protein induction in striped bass (Morone saxatilis).

The striped bass (Morone saxatilis) supports a valuable recreational fishery and is among the most important piscivorous fish of the San Francisco Estuary. This species has suffered a significant decline in numbers over the past decades, and there is indication that contaminants are important contributors. Polycyclic aromatic hydrocarbons (PAHs) and polyhalogenated aromatic hydrocarbons (PHAHs) including PCBs and dioxins are widespread in the estuary, they typically bioaccumulate through trophic levels, reaching highest levels in top predators and are known to affect the fish health and development. The aim of this study was to investigate the dynamics of cytochrome P4501A (Cyp1a) induction simultaneously at different levels of biological organization (RNA transcription and protein synthesis) as a biomarker of exposure to PAHs and PHAHs. We utilized ß-naphthoflavone (BNF) as a model PAH to induce Cyp1a responses in juvenile striped bass in both dose-response and time-response assessments and determined Cyp1a mRNA and protein levels. Significant responses were measured in both systems at 10 mg ΒΝF kg⁻¹, a concentration used for time-response studies. Messenger RNA levels peaked at 6 h post-injection, while protein levels increased progressively with time, significantly peaking at 96 h post-injection; both remaining elevated throughout the duration of the test (8 days). Our data suggest that rapid induction of gene transcription following exposure and subsequent cumulative protein synthesis could provide a useful means of identifying temporal variants in exposure to Cyp1a inducers in Morone saxatilis. The potential application of this combined Cyp1a gene and protein biomarker in this species for field studies is discussed.

Assessment of three generations of mice derived by ICSI using freeze-dried sperm.

Although the derivation of mice by intracytoplasmic sperm injection (ICSI) using freeze-dried sperm has been demonstrated previously, a comprehensive analysis of their viability, health, and fertility has not. The purpose of the present study was to determine the extent to which ICSI using freeze-dried sperm stored at 4 degrees C for 1-2 months from mice on either an inbred (C57BL/6J) or hybrid (B6D2F1/J) genetic background results in genomic instability and/or phenotypic abnormality in mice and two generations of their progeny. Fertilization rates (number of 2-cells per injected oocytes) using ICSI of fresh and freeze-dried sperm were similar within and between mouse strains, although fewer freeze-dried sperm-derived embryos than fresh sperm-derived embryos developed to blastocysts in vitro (C57BL/6J and B6D2F1/J) and liveborn pups in vivo (B6D2F1/J only). Nevertheless, once born, mice derived by ICSI using freeze-dried sperm in both mouse strains were healthy and reproductively sound. No major differences in litter size, weaning rate, and sex ratio were noted in the two generations of progeny (F2 and F3) of ICSI-derived offspring using freeze-dried sperm compared with that in the natural mating (control) group. Further, there was no evidence that either ICSI or freeze drying induced genomic instability, as determined by microsatellite analysis of the derived mice and subsequent generations when compared with both parental genotypes, nor were there differences in the number or types of pathological changes in any of the three generations of progeny. We conclude that viable, healthy and genomically stable mice can be derived by ICSI using freeze-dried mouse sperm stored in the refrigerator for at least 2 months. Further, because freeze drying is a simpler and more economical technique compared with embryo and sperm cryopreservation, the results of this study justify additional research to continue to develop and enhance the technique for the preservation, storage, and sharing of genetically altered mice.

Tint maps to mouse chromosome 6 and may interact with a notochordal enhancer of Brachyury.

At the proximal part of mouse chromosome 17 there are three well-defined genes affecting the axis of the embryo and consequently tail length: Brachyury, Brachyury the second, and the t-complex tail interaction (T1, T2, and tct). The existence of T1 and tct in fact defines the classical "t-complex" that occupies approximately 40 cM of mouse chromosome 17. Their relationship to each other and various unlinked interacting genes has been enigmatic. The tint gene was the first of the latter to be identified. We report here its genetic mapping using a microsatellite scan together with outcrosses to Mus spretus and M. castaneous followed by a subsequent testcross to T, T1, and T2 mutants. Surprisingly, tint interacts with T2 but not with T1. The implications of our data suggest that T2 may be part of the T1 regulatory region through direct or indirect participation of tint.

Environmental contaminant effects on juvenile striped bass in the San Francisco Estuary, California, USA.

The decline of pelagic organisms in the San Francisco Estuary (SFE) (California, USA) is attributed to several factors, including water diversions, invasive species, and exposure to environmental toxicants. The present study evaluated the effects of environmental contaminants on liver vitellogenin, metallothionein, 7-ethoxyresorufin-O-deethylase (EROD), and benzyloxyresorufin O-deethylase (BROD) activity in juvenile striped bass (Morone saxitilis) in the SFE. Analysis of juvenile striped bass liver extracts revealed site-specific elevations of vitellogenin, metallothionein, and EROD biomarkers across the estuary. Although some striped bass in the estuary showed EROD activity similar to unhandled hatchery controls, several sites in the estuary showed significantly higher EROD activity that was in the range of beta-naphthoflavone (BNF)-injected, positive controls. Overall, EROD activity averaged 283% higher in estuary fish than in hatchery controls. Chemical analyses of extracts from semipermeable membrane devices (SPMDs) deployed in the estuary for one month showed elevated polyaromatic hydrocarbon (PAH) levels. Semipermeable membrane devices extract injections-induced metallothionein and BROD in striped bass livers. These data show that environmental exposures are impacting EROD and other biomarkers in the SFE striped bass population. Previous studies in our laboratory have associated poor larval development with maternal transfer of environmental contaminants. Further studies are needed to monitor contaminant exposures by the use of biomarkers and to integrate them into a more effective pelagic species recovery plan in the SFE.

Genetic susceptibility of mice to Candida albicans vaginitis correlates with host estrogen sensitivity.

We compared susceptibility to Candida vaginitis in derived murine substrains differing in sensitivity to estrogen (CD-1 and CD10, resistant; CD3 and C57BL/6 responsive), and in F1 crosses. The order of decreasing resistance was CD-1 > or = CD10 > or = CD10 x CD3F1 > CD10 x B6F1 > CD3 > C57BL/6 and correlated with estrogen responsiveness in endocrine disruptor assays. Resistance to Candida vaginitis appears additive in CD10 x B6F1 animals and dominant in CD10 x CD3F1 animals.