Newly raised anti‐VAChT and anti‐ChAT antibodies detect cholinergic cells in chicken embryos |
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Authors: | Tadayoshi Watanabe Takahiro Kiyomoto Ryosuke Tadokoro Yuta Takase Yoshiko Takahashi |
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Affiliation: | 1. Department of Zoology, Graduate School of Science, Kyoto University, Sakyo‐ku, Kyoto, Japan;2. AMED Core Research for Evolutional Science and Technology (AMED‐CREST), Japan Agency for Medical Research and Development (AMED), Chiyoda‐ku, Tokyo, Japan |
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Abstract: | The autonomic nervous system consists of sympathetic and parasympathetic nerves, which functionally antagonize each other to control physiology and homeostasis of organs. However, it is largely unexplored how the autonomic nervous system is established during development. In particular, early formation of parasympathetic network remains elusive because of its complex anatomical structure. To distinguish between parasympathetic (cholinergic) and sympathetic (adrenergic) ganglia, vesicular acetylcholine transporter (VAChT) and choline O‐acetyltransferase (ChAT), proteins associated with acetylcholine synthesis, are known to be useful markers. Whereas commercially available antibodies against these proteins are widely used for mammalian specimens including mice and rats, these antibodies do not work satisfactorily in chickens, although chicken is an excellent model for the study of autonomic nervous system. Here, we newly raised antibodies against chicken VAChT and ChAT proteins. One monoclonal and three polyclonal antibodies for VAChT, and one polyclonal antibody for ChAT were obtained, which were available for Western blotting analyses and immunohistochemistry. Using these verified antibodies, we detected cholinergic cells in Remak ganglia of autonomic nervous system, which form in the dorsal aspect of the digestive tract of chicken E13 embryos. The antibodies obtained in this study are useful for visualization of cholinergic neurons including parasympathetic ganglia. |
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Keywords: | choline O‐acetyltransferase cholinergic nerves parasympathetic ganglia remak ganglion vesicular acetylcholine transporter |
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