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Synchronous development of Eimeria tenella in chicken caeca and utility of laser microdissection for purification of single stage schizont RNA

Published online by Cambridge University Press:  20 August 2012

M. MATSUBAYASHI ,
T. HATTA ,
T. MIYOSHI ,
M. A. ALIM ,
K. YAMAJI ,
K. SHIMURA ,
T. ISOBE  and
N. TSUJI
M. MATSUBAYASHI
Affiliation:
National Institute of Animal Health, National Agricultural and Food Research Organization, Tsukuba, Ibaraki 305-0856, Japan
T. HATTA
Affiliation:
National Institute of Animal Health, National Agricultural and Food Research Organization, Tsukuba, Ibaraki 305-0856, Japan
T. MIYOSHI
Affiliation:
National Institute of Animal Health, National Agricultural and Food Research Organization, Tsukuba, Ibaraki 305-0856, Japan
M. A. ALIM
Affiliation:
National Institute of Animal Health, National Agricultural and Food Research Organization, Tsukuba, Ibaraki 305-0856, Japan
K. YAMAJI
Affiliation:
National Institute of Animal Health, National Agricultural and Food Research Organization, Tsukuba, Ibaraki 305-0856, Japan
K. SHIMURA
Affiliation:
National Institute of Animal Health, National Agricultural and Food Research Organization, Tsukuba, Ibaraki 305-0856, Japan
T. ISOBE
Affiliation:
National Institute of Animal Health, National Agricultural and Food Research Organization, Tsukuba, Ibaraki 305-0856, Japan
N. TSUJI*
Affiliation:
National Institute of Animal Health, National Agricultural and Food Research Organization, Tsukuba, Ibaraki 305-0856, Japan Department of Global Agricultural Sciences, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo-ku, Tokyo 113-8657, Japan
*
*Corresponding author: Laboratory of Parasitic Diseases, National Institute of Animal Health, National Agricultural and Food Research Organization, 3-1-5, Kannondai, Tsukuba, Ibaraki 305-0856, Japan. Tel: +81 29 838 7749. Fax: +81 29 838 7749. E-mail: tsujin@affrc.go.jp.
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Summary

Eimeria tenella is recognized worldwide as a significant pathogen in the poultry industry. However, a lack of methods for isolating developing schizonts has hindered the use of transcriptome analyses to discover novel and developmentally regulated genes. In the present study, we characterized the long-term successive development of E. tenella in infected chicken caeca and assessed the utility of laser microdissection (LMD) for the isolation of schizont RNA. Developmental stages, including those of the first, second, and third-generation schizonts and gametocytes, were synchronous. Using LMD, only the mature second-generation schizonts were successfully excised from the lamina propria, and non-degraded RNA was purified from the schizonts. E. tenella-specific genes were amplified by reverse transcription polymerase chain reaction (RT-PCR). These results augment our understanding of the E. tenella life cycle, and reveal LMD as a potentially useful tool for gene expression analyses of the intracellular stages of E. tenella.

Keywords

Eimeria tenellalaser microdissectionschizontsynchronous development

Information

Type
Research Article
Information
Parasitology , Volume 139 , Issue 12 , October 2012 , pp. 1553 - 1561
Copyright
Copyright © Cambridge University Press 2012

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