Gonopodium development in normal male and 11-ketotestosterone-treated female mosquitofish (Gambusia affinis): a quantitative study using computer image analysis.

Mature male mosquitofish possess a highly modified anal fin called a gonopodium that develops from an unmodified female-like anal fin under the influence of endogenous androgens at sexual maturity. Although females do not normally develop gonopodia, their anal fins can be induced to develop into gonopodium-like structures by the administration of androgens. We used computer image analysis techniques to quantify the morphological changes in the anal fins of normal male and female mosquitofish during growth and development and the changes that occur in females exposed to an androgen, 11-ketotestosterone. The mosquitofish anal fin is a useful bioassay system for quantifying the effects of known or suspected environmental androgens.

Identification of androstenedione in a river containing paper mill effluent.

Effluent from a paper mill discharging into the Fenholloway River, Taylor County, Florida, USA, contains chemicals that masculinize females of the resident population of eastern mosquitofish (Gambusia holbrooki), as evidenced in females by elongated anal fins, which is normally a male-specific trait. To identify androgenic components in the effluent, water collected from the Fenholloway River and a control tributary was fractionated using solid-phase extraction and reverse-phase high-performance-liquid chromatography. Two Fenholloway River fractions induced androgen receptor-dependent transcriptional activity in transient transfection cell culture assays. Of these, androstenedione was confirmed by liquid chromatography-mass spectrometry with multiple reaction monitoring.

Mosquitofish (Gambusia affinis) vitellogenin: identification, purification, and immunoassay.

Vitellogenin is a phospholipoglycoprotein precursor of egg yolk. In mature female fish, vitellogenin is synthesized and secreted by the liver in response to circulating estrogens. Vitellogenin is normally undetectable in the blood of male fish, but can be induced by exposure to compounds possessing estrogenic activity. Thus, the presence of vitellogenin in blood of male fish can serve as a useful biomarker for assessing previous exposure to estrogenic compounds. In the present study, we report identification and purification of vitellogenin in the mosquitofish (Gambusia affinis). Anti-vitellogenin immune serum was generated and used to develop an immunoblot assay for detection of vitellogenin. A combination of immunoblotting and densitometric scanning was used to assess the time- and dose-dependent effects of 17alpha-ethynylestradiol on vitellogenesis in male G. affinis. The results indicate that changes in the level of vitellogenin in mosquitofish blood can be reliably detected by the immunoblot assay, and that the mosquitofish may be a useful bioindicator organism for detecting estrogenic contamination of the aquatic environment.

Quantification of the expression of a temperature-sensitive pigment allele in sailfin mollies (Poecilia latipinna) by image analysis.

Image analysis was used to quantify the activity of a temperature-sensitive macromelanophore-determining allele in sailfin mollies as the percentage of the body surface area covered by macromelanophores. Fish heterozygous for the macromelanophore-determining allele produced very few macromelanophores when raised at either 25 or 28 degrees C, even after more than 200 days. In contrast, the mean percent coverage for genetically identical fish raised at 22 degrees C increased steadily throughout the course of the experiment. Production of macromelanophores was sex influenced, with greater expressivity seen in males. At 22 degrees C, the mean percent coverages had significantly diverged between males and females by the age of 201 days. From that point on, the percent macromelanophore coverage of the males was consistently significantly higher than that of the females. The tendency to produce greater melanization at cooler temperatures is not the result of a heat-sensitive tyrosinase enzyme, as is the case in mammals carrying the Himalayan allele. In mollies, the activity of tyrosinase increases between 22 and 29 degrees C. We hypothesize that production of macromelanophores is under the control of a proto-oncogene.

Pigments and ultrastructures of pigment cells in xanthic sailfin mollies (Poecilia latipinna).

Electron micrographs of skin from xanthic (gold) sailfin mollies revealed numerous xanthophores, as well as scattered melanophores. The melanophores were seen to contain premelanosomes in various stages of development. This is consistent with the fact that xanthic mollies have been shown to be tyrosinase positive. Melanosomes in xanthic mollies appear to develop by one of two pathways: 1) from an endoplasmic reticulum-derived vesicle which develops an internal lamellar framework, and 2) by fusion of multiple Golgi-derived vesicles which lack an internal lamellar framework. Analysis of the pigments in the skin of the xanthic mollies identified four colorless pteridine pigments (xanthopterin, isoxanthopterin, neopterin, and pterin) and a carotenoid with an absorbance spectrum similar to beta-carotene. It appears that, unlike some other poeciliid fishes, sailfin mollies do not use pteridine pigments for orange coloration. Rather, they appear to rely primarily on carotenoids.

Population frequencies of carbonic anhydrase II (CA II), esterase D (EsD), and glyoxalase I (GLO) in the Metropolitan Birmingham, Alabama area.

Both black and white populations from Birmingham, Alabama were analyzed for the frequencies of carbonic anhydrase II (CA II), glyoxalase I (GLO) and esterase D (EsD) isoenzymes. The results compared favorably with published frequencies of these genetic markers in other populations.