Fluorescent imaging is widely available and is a standard method of analysis of gene function in zebrafish embryos; image acquisition is rapid, but is of limited use in adult wild-type zebrafish. Our data indicate that the useful range of the stereomicroscope can be considerably extended to the adult fish, in both the visible and fluorescent ranges, due to the optical clarity of our model. When combined with confocal microscopy, single cell resolution is easily achievable, and the use of dual-photon confocal microscopy is likely to yield much greater depth of image resolution in the casper line. Crossing the casper line to the numerous transgenic lines that label vasculature (i.e. fli-GFP, shown in ) or internal organs with fluorescent tags (i.e. pdx-GFP for pancreas, LFABP-GFP for liver) may be of considerable use to those interested in in vivo analysis of angiogenesis, organ homeostasis and regeneration after injury. However, direct injection of transgenic constructs in the casper background may be advantageous since it eliminates the need for complex breeding strategies to maintain double homozygosity of roy and nacre.
Early warning signs of endocrine disruption in adult fish from ..
PCR products of , or were subjected to a T7E1 assay (Methods) and confirmed by gel electrophoresis. a, The result of the or T7E1 assay for ten adult fish. ‘M.’ is a 100 bp DNA ladder marker (NEB). In the gel picture, 810 bp (black arrowhead) is the wild-type band as observed in cont. lane (wild type without gRNA injection). All ten fish showed smaller and bottom shifted products (red arrowheads) compared to negative control fish, indicating that all fish have mutations in the target region of . In the gel picture, 1,089 bp is the wild-type band. All ten fish into which gRNAs were injected showed smaller and bottom shifted products compared to negative control fish, indicating that all fish have mutations in the target region of . In the gel picture, 823 bp is the wild-type band. Eight of ten fish showed smaller and bottom shifted products, indicating that 80% of fish have mutations in the target region of b, The efficiency of CRISPR/Cas9 deletion for in zebrafish. Almost all adult fish into which gRNAs and mRNA were injected have mutations at the target positions. c, The efficiency of germline transmission of CRISPR/Cas9 mutant fish. Identified mutant fish were outcrossed to wild-type fish to obtain embryos and confirmed germline transmission. Obtained embryos were lysed individually at 48 hpf, genotyped by T7E1 assay and sequenced. Because of CRISPR/Cas9 mosaicism, some different mutation patterns, which result in a non-frameshift or frameshift mutation, were observed.
Optic nerve regeneration in adult fish and apolipoprotein A-I. - NCBI
Clown loaches are often sold as tiny fish, only 3cm long or so. These little babies won’t stay that size for long though, clown loaches can easily reach 30cm or more in length. The little fellow at the front is the size often sold in shops, his companions are on their way to adult size but still have a way to go.
Optic nerve regeneration in adult fish and apolipoprotein A-I