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Developmental Biology

Using Chicken Embryo as a Powerful Tool in Assessment of Developmental Cardiotoxicities

Published: March 21st, 2021

DOI:

10.3791/62189

1School of Public Health, Qingdao University, 2Department of Pharmacology & Toxicology, Brody School of Medicine, East Carolina University

Chicken embryos, as a classical developmental model, are used in our lab to assess developmental cardiotoxicities following exposure to various environmental contaminants. Exposure methods and morphological/functional assessment methods established are described in this manuscript.

Chicken embryos are a classical model in developmental studies. During the development of chicken embryos, the time window of heart development is well-defined, and it is relatively easy to achieve precise and timely exposure via multiple methods. Moreover, the process of heart development in chicken embryos is similar to mammals, also resulting in a four-chambered heart, making it a valuable alternative model in the assessment of developmental cardiotoxicities. In our lab, the chicken embryo model is routinely used in the assessment of developmental cardiotoxicities following exposure to various environmental pollutants, including per- and polyfluoroalkyl substances (PFAS), particulate matter (PMs), diesel exhaust (DE) and nano materials. The exposure time can be freely selected based on the need, from the beginning of development (embryonic day 0, ED0) all the way to the day prior to hatch. The major exposure methods include air-cell injection, direct microinjection, and air-cell inhalation (originally developed in our lab), and the currently available endpoints include cardiac function (electrocardiography), morphology (histological assessments) and molecular biological assessments (immunohistochemistry, qRT-PCR, western blotting, etc.). Of course, the chicken embryo model has its own limitations, such as limited availability of antibodies. Nevertheless, with more laboratories starting to utilize this model, it can be used to make significant contributions to the study of developmental cardiotoxicities.

The chicken embryo is a classic developmental model, which has been used for over two hundred years1. The chicken embryo model has various advantages compared to traditional models. First of all, as early as over 70 years ago, the normal development of the chicken embryo had been illustrated very clearly in the Hamburger-Hamilton staging guide2, in which a total of 46 stages during chicken embryo development were defined with precise time and morphological characteristics, facilitating detections of abnormal development. Additionally, the chicken embryo model has other features such as being relatively low-cost and redun....

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All procedures described were approved by the Institutional Animal Care and Use Committee (IACUC) of Qingdao University. In our lab, the eggs were incubated in two incubators. Eggs were held upright in the incubator and randomly placed on the shelves. The incubation conditions for the eggs were as follows: incubation temperature started at 37.9 °C, and gradually decreased to 37.1 °C as incubation proceeded; the humidity started at 50% and gradually increased to 70%.

1. Exposure methods.......

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Exposure results
Air cell injection
Air cell injection can effectively expose developing chicken embryos to various agents, which may be subsequently detected in the collected samples (serum, tissue, etc.) of embryos/hatchling chickens. Here is an example, in which perfluorooctanoic acid (PFOA) was air-cell injected, and serum PFOA concentrations were then determined with Ultra-performance liquid chromatography-mass spectrometry. The serum concentrations corresponded with the injected doses, indicating the ef.......

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The chicken embryo has been a classical model in developmental studies for 200 years1. Our methods presented in this manuscript have been used in the assessment of several environmental contaminants, including perfluorooctanoic acid, particulate matter, and diesel exhaust with success5, 7,8,9,10,11

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This work was supported by National Natural Science Foundation of China (Grant No. 91643203, 91543208, 81502835).

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Name Company Catalog Number Comments
4% phosphate buffered formaldehydefixative Biosharp, Hefei, China REF: BL539A
75% ethanol Guoyao,Shanghai,China CAS:64-17-5
Biosignaling monitor BL-420E+ Taimeng, Chengdu, China BL-420E+
Candling lamp Zhenwei, Dezhou, China WZ-001
Disposable syringe Zhiyu, Jiangsu, China
Egg incubator Keyu,Dezhou, China KFX
Electrical balance OHAUS, Shanghai, China AR 224CN
Electro-thermal incubator Shenxian, Shanghai, China DHP-9022
Ethanol absolute Guoyao,Shanghai,China CAS:64-17-5
Fertile chicken egg Jianuo, Jining, China
Hematoxylin and Eosin Staining Kit Beyotime, Bejing, China C0105
Histology paraffin Aladdin, Shanghai, China P100928-500g Melt point 52~54°C
Histology paraffin Aladdin, Shanghai, China P100936-500g Melt point 62~64°C
IV catheter KDL, Zhejiang, China The catheters have to be soft, plastic ones.
Lentivirus Genechem, Shanghai, China The lentivirus were individually designed/synthesized by Genechem.
Masson's trichrome staining kit Solarbio, Beijing, China G1340
Metal probe Jinuotai, Beijing, China
Microinjector (5 uL) Anting,Shanghai, China
Microscope CAIKON, Shanghai, China XSP-500
Microtome Leica, Germany HistoCore BIOCUT
Microtome blade Leica,Germany Leica 819
Pentobarbitual sodium Yitai Technology Co. Ltd.,  Wuhan, China CAS: 57-33-0
Pipetter(10ul) Sartorius, Germany
Povidone iodide Longyuquan, Taian, China
Scissor Anqisheng,Suzhou, China
Sterile saline Kelun,Chengdu, China
Sunflower oil Mighty Jiage, Jiangsu, China Any commerical sunflower oil for human consumption should work
Tape M&G, Shanghai, China
Tedlar PVF Bag (5L) Delin, Dalian, China
Vortex mixer SCILOGEX, Rocky Hill, CT, US MX-F
Xylene Guoyao,Shanghai,China CAS:1330-20-7

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