Neuroethological analysis of song learning mechanism by using antisense DNA method in birds

2001 Fiscal Year Final Research Report Summary

• Project/Area Number: 11640677
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: 動物生理・代謝
• Research Institution: Shizuoka University
• Principal Investigator: TAKEUCHI Hiro-aki Shizuoka University, Faculty of Science, Associate Professor, 理学部, 助教授 (90216854)
• Project Period (FY): 1999 – 2001
• Keywords: learning / song / bird / Bengalese finch / zenk / immediate early gene / antisense DNA / brain

Research Abstract

In the present study, as a step to understand the brain mechanisms of song learning in Bengalese finch, I attempted to elucidate the functions of intracerebral zenk using the antisense DNA method. For designing oligodeoxynucleotide (ODN), I decided 1549bp of sequence in zenk cDNA using 5'/3'RACE and direct sequencing. The sequence of Bengalese finch's zenk showed 68.1 %, 67.1 % and 66.3% homology with those of human, mouse and zebra fish, respectively. To investigate the localization of applied ODN, FITC-labeled antisense ODN was enwrapped in HVJ-liposome complex of cation type, and then injected into the brain of bird. The result exhibited that sustained fluorescence localized in the nucleus for at least 1 week, and that FITC-labeled cells as well as ZENK-immunonegative cells was distributed within a distance of about 100-200μm from the injection site. The distribution of ZENK-immunonegative cells was too narrow to judge wether the cells were suppressed zenk expression by injected antisense ODN or by surgical damage. Several methodological trials, concerning the chronic injection of antisense, ODN into the brain of behaving bird and the behavioral quantification of the effects, failed to obtain the quantitative data to clear the causal relationship between intracerebral zenk expression and specific behavior