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Immunohistochemical localization of two types of choline acetyltransferase in neurons and sensory cells of the octopus arm

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Abstract

Cholinergic structures in the arm of the cephalopod Octopus vulgaris were studied by immunohistochemistry using specific antisera for two types (common and peripheral) of acetylcholine synthetic enzyme choline acetyltransferase (ChAT): antiserum raised against the rat common type ChAT (cChAT), which is cross-reactive with molluscan cChAT, and antiserum raised against the rat peripheral type ChAT (pChAT), which has been used to delineate peripheral cholinergic structures in vertebrates, but not previously in invertebrates. Western blot analysis of octopus extracts revealed a single pChAT-positive band, suggesting that pChAT antiserum is cross-reactive with an octopus counterpart of rat pChAT. In immunohistochemistry, only neuronal structures of the octopus arm were stained by cChAT and pChAT antisera, although the pattern of distribution clearly differed between the two antisera. cChAT-positive varicose nerve fibers were observed in both the cerebrobrachial tract and neuropil of the axial nerve cord, while pChAT-positive varicose fibers were detected only in the neuropil of the axial nerve cord. After epitope retrieval, pChAT-positive neuronal cells and their processes became visible in all ganglia of the arm, including the axial and intramuscular nerve cords, and in ganglia of suckers. Moreover, pChAT-positive structures also became detectable in nerve fibers connecting the different ganglia, in smooth nerve fibers among muscle layers and dermal connective tissues, and in sensory cells of the suckers. These results suggest that the octopus arm has two types of cholinergic nerves: cChAT-positive nerves from brain ganglia and pChAT-positive nerves that are intrinsic to the arm.

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Abbreviations

ACh:

Acetylcholine

AChE:

Acetylcholinesterase

ChAT:

Choline acetyltransferase

cChAT:

Common type of choline acetyltransferase

pChAT:

Peripheral type of choline acetyltransferase

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Acknowledgments

The authors thank the late Professor Tindaro G. Renda (University la Sapienza, Roma) for performance of preliminary immunohistochemical staining using pChAT antiserum in the octopus arm. J-P. B was supported by a Grant-in-Aid for KAKENHI from the Japan Society for the Promotion of Science (No. 24592334).

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Authors and Affiliations

  1. Molecular Neuroscience Research Center, Shiga University of Medical Science, Seta Tsukinowa-cho, Otsu, Shiga, 520-2192, Japan

    Yuko Sakaue, Jean-Pierre Bellier, Shin Kimura & Hiroshi Kimura

  2. Department of Pediatrics, Shiga University of Medical Science, Seta Tsukinowa-cho, Otsu, Shiga, 520-2192, Japan

    Yuko Sakaue & Yoshihiro Takeuchi

  3. Laboratory of Immunohistochemistry Tindaro G. Renda, Department of Anatomic, Histologic, Forensic and Locomotor Apparatus Sciences, Sapienza University of Rome, 00161, Rome, Italy

    Loredana D’Este

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  1. Yuko Sakaue
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  2. Jean-Pierre Bellier
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  3. Shin Kimura
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  4. Loredana D’Este
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  5. Yoshihiro Takeuchi
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Correspondence to Jean-Pierre Bellier.

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Sakaue, Y., Bellier, JP., Kimura, S. et al. Immunohistochemical localization of two types of choline acetyltransferase in neurons and sensory cells of the octopus arm. Brain Struct Funct 219, 323–341 (2014). https://doi.org/10.1007/s00429-012-0502-6

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