I. the origin of species by means of natural selection

Ecological implications of genetically modified fishes in freshwater fisheries, with a focus on salmonids

Fluorescence microscopy and RT-PCR technique were used to detect the expression of exogenous gene in the genetic strains of white cloud mountain minnow Tanichthys albonubes successfully constructed and modified by recombinant plasmid pDsRed-mylz2 from the zebrafish inserted myosin light chain 2 promoter and red fluorescent protein gene.The results showed that the expression of the red fluorescent gene was initially observed 19 days after fertilization in F3,and expressed mostly in the 25-30 days,the latest expression of the genetically modified fish appeared in the 35 days.The red fluorescent protein was expressed in muscle,heart,hepatopancrease,spleen,gill,swim bladder,intestines and gonad,and the expression was much stronger in the muscles,hepatopancrease,and gill than that in other tissues,but the fluorescence was not detected in epidermis and fin ray.Although postponement of the red fluorescent protein expression and the change in specificity was appeared occasionally in transgenic offspring of the minnow,the genetic expression of the exogenous gene in the offspring was stable in general.So it is expected to produce a stable strain of red fluorescent protein transgenic white cloud mountain minnow.

The expression of the red fluorescent protein gene in transgenic white cloud mountain minnow Tanichthys albonubes

In laboratory fish research, the zebrafish Danio rerio (Cyprinidae) represents the equivalent of the mouse in mammalian research. This species has become a major model for studies in developmental and behavioural genetics, neurophysiology, biomedicine, ecotoxicology, and behavioural and evolutionary ecology. To meet the need for accurate and reproducible data in both fundamental and applied sciences, it is of primary importance to be able to tag and/or recognize individual zebrafish. However, classic methods used in fish ecology and aquaculture are generally difficult to apply to such small fish. Recently, various new tagging methods have been developed. This paper presents a first review of current identification and marking methods applied to zebrafish, from external observation methods (such as skin pattern recognition, fin clipping, scale regeneration, colour and transgenic methods) to the most advanced technological developments in electronic (low- and high- radio-frequencies PIT tags, microchip) and image analysis methods (video tracking). This review aims to help researchers and zebrafish facility managers select the identification method (ID) best adapted to their needs. The main characteristics of each ID method are examined (including detection range, durability, speed and repetitiveness, ID code combination, size dependence and ethical considerations), and their pros and cons are summarized in a decision table to help select the most appropriate option for a research or management program. Finally, contextual applications of these ID methods and future developments are discussed.

/pdf/individual-identification-and-marking-techniques-for-38kzzjbdpt.pdf

Individual identification and marking techniques for zebrafish

Demonstrating relationships between sexual selection mechanisms and trait evolution is central to testing evolutionary theory. Using zebrafish, we found that wild-type males possessed a significant advantage in mate competition over transgenic RFP Glofish® males. In mating trials, wild-type males were aggressively superior to transgenic males in male-male chases and male-female chases; as a result, wild-type males sired 2.5× as many young as did transgenic males. In contrast, an earlier study demonstrated that female zebrafish preferred transgenic males as mates when mate competition was excluded experimentally. We tested the evolutionary consequence of this conflict between sexual selection mechanisms in a long-term study. The predicted loss of the transgenic phenotype was confirmed. More than 18,500 adults collected from 18 populations across 15 generations revealed that the frequency of the transgenic phenotype declined rapidly and was eliminated entirely in all but one population. Fitness component data for both sexes indicated that only male mating success differed between wild-type and transgenic individuals. Our predictive demographic model based on fitness components closely matched the rate of transgenic phenotype loss observed in the long-term study, thereby supporting its utility for studies assessing evolutionary outcomes of escaped or released genetically modified animals.

Mate competition and evolutionary outcomes in genetically modified zebrafish (Danio rerio)

Shoaling and sexual behaviour of wild-type male and female white cloud mountain minnow Tanichthys albonubes were measured in the presence of the red fluorescent transgenic conspecifics under laboratory conditions. Wild-type female test fish showed no significant preference, whereas wild-type male test fish preferred to be near a shoal of red transgenic fish rather than wild-type fish. When placed in a potentially reproductive context, wild-type males had a higher competitive ability over transgenic males; wild-type females spent more time with wild-type males in visually mediated experiments, but wild-type males performed more courtship displays towards transgenic females. These results suggest that the red body colouration does not appear to disturb signal communication between wild-type and transgenic T. albonubes in shoaling behaviour; transgenic males have no mating advantage over wild-type males, but the red body colouration of transgenic females may affect mate choice of wild-type males.

Shoaling and mate choice of wild-type Tanichthys albonubes in the presence of the red fluorescent transgenic conspecifics.

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Shoaling and mate choice of wild-type Tanichthys albonubes in the presence of the red fluorescent transgenic conspecifics.