doi:10.3791/2126 (2010). A convenient method for chemically treating zebrafish is to introduce th... more doi:10.3791/2126 (2010). A convenient method for chemically treating zebrafish is to introduce the reagent into the tank water, where it will be taken up by the fish. However, this method makes it difficult to know how much reagent is absorbed or taken up per fish. Some experimental questions, particularly those related to metabolic studies, may be better addressed by delivering a defined quantity to each fish, based on weight. Here we present a method for intraperitoneal (IP) injection into adult zebrafish. Injection is into the abdominal cavity, posterior to the pelvic girdle. This procedure is adapted from veterinary methods used for larger fish. It is safe, as we have observed zero mortality. Additionally, we have seen bleeding at the injection site in only 5 out of 127 injections, and in each of those cases the bleeding was brief, lasting several seconds, and the quantity of blood lost was small. Success with this procedure requires gentle handling of the fish through several s...
This is an open-access article distributed under the terms of the Creative Commons Attribution-No... more This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial License
Raising larvae of Danio rerio Hamilton (zebrafish) is a challenging task that requires skill and ... more Raising larvae of Danio rerio Hamilton (zebrafish) is a challenging task that requires skill and a significant daily time investment. We have developed a simple nursery and a husbandry regimen that streamlines procedures and is feasible for small laboratories to carry out in the absence of support staff. The nursery is inexpensive to build and easy to maintain. The regimen uses a simple benchtop nursery that houses up to 300 larvae. Feeding is simplified by using defined volumes of microencapsulated feeds with only 1 type of live prey as a dietary supplement, Artemia franciscana Leach (brine shrimp). Tests of the regimen using wild-type lines showed that it supports timely entry into the metamorphic period and supports survival rates of at least 75%. Further, inexperienced users were able to raise larvae successfully. Here, we describe how to assemble the nursery and how to carry out the feeding and care regimen.
The anti-apoptotic protein Bcl-2 has been shown to function in roles unrelated to apoptosis in a ... more The anti-apoptotic protein Bcl-2 has been shown to function in roles unrelated to apoptosis in a variety of cell types. We have previously reported that loss of Bcl-2 expression alters chondrocyte morphology and modulates aggrecan expression via an apoptosis-independent pathway. Here we show that Bcl-2 is required for chondrocytes to maintain expression of a variety of cartilage-specific matrix proteins. Using quantitative, real-time PCR, we demonstrate that Bcl-2-deficient chondrocytes coordinately down-regulate genes coding for hyaline cartilage matrix proteins including collagen II, collagen IX, aggrecan, and link protein. The decrease in steady-state level of these mRNA transcripts results, in part, from decreased mRNA stability in Bcl-2-deficient chondrocytes. Transcriptional regulation is also likely involved because chondrocytes with decreased Bcl-2 levels show decreased expression of SOX9, a transcription factor necessary for expressing the major cartilage matrix proteins. I...
In recent years, there has been significant progress in understanding the detailed mechanisms of ... more In recent years, there has been significant progress in understanding the detailed mechanisms of pancreas development. These studies have in turn influenced research aimed at producing pancreatic islet cells from stem cells. Here, we review recent progress in both of these areas.
doi:10.3791/2126 (2010). A convenient method for chemically treating zebrafish is to introduce th... more doi:10.3791/2126 (2010). A convenient method for chemically treating zebrafish is to introduce the reagent into the tank water, where it will be taken up by the fish. However, this method makes it difficult to know how much reagent is absorbed or taken up per fish. Some experimental questions, particularly those related to metabolic studies, may be better addressed by delivering a defined quantity to each fish, based on weight. Here we present a method for intraperitoneal (IP) injection into adult zebrafish. Injection is into the abdominal cavity, posterior to the pelvic girdle. This procedure is adapted from veterinary methods used for larger fish. It is safe, as we have observed zero mortality. Additionally, we have seen bleeding at the injection site in only 5 out of 127 injections, and in each of those cases the bleeding was brief, lasting several seconds, and the quantity of blood lost was small. Success with this procedure requires gentle handling of the fish through several s...
This is an open-access article distributed under the terms of the Creative Commons Attribution-No... more This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial License
Raising larvae of Danio rerio Hamilton (zebrafish) is a challenging task that requires skill and ... more Raising larvae of Danio rerio Hamilton (zebrafish) is a challenging task that requires skill and a significant daily time investment. We have developed a simple nursery and a husbandry regimen that streamlines procedures and is feasible for small laboratories to carry out in the absence of support staff. The nursery is inexpensive to build and easy to maintain. The regimen uses a simple benchtop nursery that houses up to 300 larvae. Feeding is simplified by using defined volumes of microencapsulated feeds with only 1 type of live prey as a dietary supplement, Artemia franciscana Leach (brine shrimp). Tests of the regimen using wild-type lines showed that it supports timely entry into the metamorphic period and supports survival rates of at least 75%. Further, inexperienced users were able to raise larvae successfully. Here, we describe how to assemble the nursery and how to carry out the feeding and care regimen.
The anti-apoptotic protein Bcl-2 has been shown to function in roles unrelated to apoptosis in a ... more The anti-apoptotic protein Bcl-2 has been shown to function in roles unrelated to apoptosis in a variety of cell types. We have previously reported that loss of Bcl-2 expression alters chondrocyte morphology and modulates aggrecan expression via an apoptosis-independent pathway. Here we show that Bcl-2 is required for chondrocytes to maintain expression of a variety of cartilage-specific matrix proteins. Using quantitative, real-time PCR, we demonstrate that Bcl-2-deficient chondrocytes coordinately down-regulate genes coding for hyaline cartilage matrix proteins including collagen II, collagen IX, aggrecan, and link protein. The decrease in steady-state level of these mRNA transcripts results, in part, from decreased mRNA stability in Bcl-2-deficient chondrocytes. Transcriptional regulation is also likely involved because chondrocytes with decreased Bcl-2 levels show decreased expression of SOX9, a transcription factor necessary for expressing the major cartilage matrix proteins. I...
In recent years, there has been significant progress in understanding the detailed mechanisms of ... more In recent years, there has been significant progress in understanding the detailed mechanisms of pancreas development. These studies have in turn influenced research aimed at producing pancreatic islet cells from stem cells. Here, we review recent progress in both of these areas.
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