Expression of genes encoding antimicrobial peptides in the Harderian gland of the bullfrog Lithobates catesbeianus

Itaru Hasunuma; Shawichi Iwamuro; Tetsuya Kobayashi; Kazuhiko Shirama; J. Michael Conlon; Sakae Kikuyama

doi:10.1016/j.cbpc.2010.05.005

Expression of genes encoding antimicrobial peptides in the Harderian gland of the bullfrog Lithobates catesbeianus

Itaru Hasunuma
, Shawichi Iwamuro
, Tetsuya Kobayashi
, Kazuhiko Shirama
, J. Michael Conlon
, Sakae Kikuyama

School of Biomedical Sciences

Research output: Contribution to journal › Article › peer-review

9 Citations (Scopus)

Abstract

The Harderian gland is an orbital gland found in many tetrapod species that possess a nictitating membrane. While the main role of the Harderian gland is lubrication of the eyeballs, numerous other functions are attributed to this gland. In amphibians, mast cells have been detected in the Harderian gland, suggesting that the gland is involved in the host's system of innate immunity defending against microbial invasions. Using reverse-transcription polymerase chain reaction, we cloned from the bullfrog Harderian gland total RNA preparations, cDNAs encoding biosynthetic precursors for the antimicrobial peptides temporin-CBa (FLPIASLLGKYL-NH₂), previously isolated from an extract of bullfrog skin, and chensirin-2CBa (IIPLPLGYFAKKP) that contained the amino acid substitution Thr¹³→Pro compared with chensirin-2 from the Chinese brown frog, Rana chensinensis. By means of in situ hybridization using digoxigenin-labeled cRNA probes for preprotemporin-CBa and preprochensirin-2CBa, we have demonstrated for the first time in an amphibian the presence of mRNAs encoding these two precursors in the cytoplasm of the glandular cells in the bullfrog Harderian gland.

Original language	English
Pages (from-to)	301-305
Number of pages	5
Journal	Comparative Biochemistry and Physiology - C Toxicology and Pharmacology
Volume	152
Issue number	3
DOIs	https://doi.org/10.1016/j.cbpc.2010.05.005
Publication status	Published (in print/issue) - Sept 2010

Funding

The authors thank Dr. Okada and Mr. Nakano in Saitama University for collection of frog HG samples. This work was supported by Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science to I.H. (22770060) and to S.I., T.K., and S.K. (21570068) and by Toho University Nukada Memorial Scholarship Fund to S.I.

Funders	Funder number
Japan Society for the Promotion of Science	22770060, 21570068

Keywords

Antimicrobial peptide
Bullfrog
Chensirin-2
Harderian gland
Temporin

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10.1016/j.cbpc.2010.05.005

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title = "Expression of genes encoding antimicrobial peptides in the Harderian gland of the bullfrog Lithobates catesbeianus",

abstract = "The Harderian gland is an orbital gland found in many tetrapod species that possess a nictitating membrane. While the main role of the Harderian gland is lubrication of the eyeballs, numerous other functions are attributed to this gland. In amphibians, mast cells have been detected in the Harderian gland, suggesting that the gland is involved in the host's system of innate immunity defending against microbial invasions. Using reverse-transcription polymerase chain reaction, we cloned from the bullfrog Harderian gland total RNA preparations, cDNAs encoding biosynthetic precursors for the antimicrobial peptides temporin-CBa (FLPIASLLGKYL-NH2), previously isolated from an extract of bullfrog skin, and chensirin-2CBa (IIPLPLGYFAKKP) that contained the amino acid substitution Thr13→Pro compared with chensirin-2 from the Chinese brown frog, Rana chensinensis. By means of in situ hybridization using digoxigenin-labeled cRNA probes for preprotemporin-CBa and preprochensirin-2CBa, we have demonstrated for the first time in an amphibian the presence of mRNAs encoding these two precursors in the cytoplasm of the glandular cells in the bullfrog Harderian gland.",

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author = "Itaru Hasunuma and Shawichi Iwamuro and Tetsuya Kobayashi and Kazuhiko Shirama and Conlon, \{J. Michael\} and Sakae Kikuyama",

year = "2010",

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doi = "10.1016/j.cbpc.2010.05.005",

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AU - Hasunuma, Itaru

AU - Iwamuro, Shawichi

AU - Kobayashi, Tetsuya

AU - Shirama, Kazuhiko

AU - Conlon, J. Michael

AU - Kikuyama, Sakae

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N2 - The Harderian gland is an orbital gland found in many tetrapod species that possess a nictitating membrane. While the main role of the Harderian gland is lubrication of the eyeballs, numerous other functions are attributed to this gland. In amphibians, mast cells have been detected in the Harderian gland, suggesting that the gland is involved in the host's system of innate immunity defending against microbial invasions. Using reverse-transcription polymerase chain reaction, we cloned from the bullfrog Harderian gland total RNA preparations, cDNAs encoding biosynthetic precursors for the antimicrobial peptides temporin-CBa (FLPIASLLGKYL-NH2), previously isolated from an extract of bullfrog skin, and chensirin-2CBa (IIPLPLGYFAKKP) that contained the amino acid substitution Thr13→Pro compared with chensirin-2 from the Chinese brown frog, Rana chensinensis. By means of in situ hybridization using digoxigenin-labeled cRNA probes for preprotemporin-CBa and preprochensirin-2CBa, we have demonstrated for the first time in an amphibian the presence of mRNAs encoding these two precursors in the cytoplasm of the glandular cells in the bullfrog Harderian gland.

AB - The Harderian gland is an orbital gland found in many tetrapod species that possess a nictitating membrane. While the main role of the Harderian gland is lubrication of the eyeballs, numerous other functions are attributed to this gland. In amphibians, mast cells have been detected in the Harderian gland, suggesting that the gland is involved in the host's system of innate immunity defending against microbial invasions. Using reverse-transcription polymerase chain reaction, we cloned from the bullfrog Harderian gland total RNA preparations, cDNAs encoding biosynthetic precursors for the antimicrobial peptides temporin-CBa (FLPIASLLGKYL-NH2), previously isolated from an extract of bullfrog skin, and chensirin-2CBa (IIPLPLGYFAKKP) that contained the amino acid substitution Thr13→Pro compared with chensirin-2 from the Chinese brown frog, Rana chensinensis. By means of in situ hybridization using digoxigenin-labeled cRNA probes for preprotemporin-CBa and preprochensirin-2CBa, we have demonstrated for the first time in an amphibian the presence of mRNAs encoding these two precursors in the cytoplasm of the glandular cells in the bullfrog Harderian gland.

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KW - Temporin

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IS - 3

ER -

Expression of genes encoding antimicrobial peptides in the Harderian gland of the bullfrog Lithobates catesbeianus

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