Requirement of new protein synthesis of a transcription factor for memory consolidation: Paradoxical changes in mRNA and protein levels of C/EBP

Dai Hatakeyama, Hisayo Sadamoto, Takayuki Watanabe, Akiko Wagatsuma, Suguru Kobayashi, Yutaka Fujito, Masakane Yamashita, Manabu Sakakibara, György Kemenes, Etsuro Ito

研究成果: Article

39 引用 (Scopus)

抄録

Some specific transcription factors are essential for memory consolidation across species. However, it is still unclear whether only the activation of constitutively expressed forms of these conserved transcription factors is involved in memory consolidation or their de novo synthesis also occurs after learning. This question has remained unanswered partly because of the lack of an efficient method for the determination of copy numbers of particular mRNAs in single neurons, which allows the detection of new transcription at the cellular level. Here we applied a newly developed protocol of single-cell quantitative real-time polymerase chain reaction (qRT-PCR) to single neurons playing an important role in associative learning. Specifically, we examined the changes in the mRNA and protein expression levels of a highly conserved transcription factor, CCAAT/enhancer binding protein (C/EBP), in the paired B2 motoneurons of the pond snail Lymnaea stagnalis. These buccal neurons are involved in the motor control of feeding behavior, with a potentially important role in conditioned taste aversion (CTA). Single-cell qRT-PCR revealed a significant decrease in LymC/EBP mRNA copy numbers in the B2 motoneurons during memory consolidation after CTA training. By contrast, isoelectric focusing and immunoblotting of extracts of the buccal ganglia showed that translation and phosphorylation levels of LymC/EBP significantly increased during memory consolidation. The C/EBP-like immunoreactivity in the B2 motoneurons, which are the major immunopositive component in the buccal ganglia, also significantly increased during memory consolidation, suggesting that the main source of increase in the level of protein in the buccal ganglia are the B2 motoneurons. Thus, early memory consolidation after CTA learning in L. stagnalis involves both the rapid synthesis and phosphorylation of LymC/EBP as well as the rapid breakdown of LymC/EBP mRNA in the neural network controlling feeding, suggesting that all of these processes play a role in the function of C/EBP in memory consolidation.

元の言語English
ページ(範囲)569-577
ページ数9
ジャーナルJournal of Molecular Biology
356
発行部数3
DOI
出版物ステータスPublished - 2006 2 24
外部発表Yes

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CCAAT-Enhancer-Binding Proteins
Transcription Factors
Cheek
Messenger RNA
Motor Neurons
Ganglia
Proteins
Learning
Neurons
Real-Time Polymerase Chain Reaction
Phosphorylation
Lymnaea
Snails
Isoelectric Focusing
Feeding Behavior
Memory Consolidation
Immunoblotting

ASJC Scopus subject areas

  • Virology

これを引用

Requirement of new protein synthesis of a transcription factor for memory consolidation : Paradoxical changes in mRNA and protein levels of C/EBP. / Hatakeyama, Dai; Sadamoto, Hisayo; Watanabe, Takayuki; Wagatsuma, Akiko; Kobayashi, Suguru; Fujito, Yutaka; Yamashita, Masakane; Sakakibara, Manabu; Kemenes, György; Ito, Etsuro.

:: Journal of Molecular Biology, 巻 356, 番号 3, 24.02.2006, p. 569-577.

研究成果: Article

Hatakeyama, D, Sadamoto, H, Watanabe, T, Wagatsuma, A, Kobayashi, S, Fujito, Y, Yamashita, M, Sakakibara, M, Kemenes, G & Ito, E 2006, 'Requirement of new protein synthesis of a transcription factor for memory consolidation: Paradoxical changes in mRNA and protein levels of C/EBP', Journal of Molecular Biology, 巻. 356, 番号 3, pp. 569-577. https://doi.org/10.1016/j.jmb.2005.12.009
Hatakeyama, Dai ; Sadamoto, Hisayo ; Watanabe, Takayuki ; Wagatsuma, Akiko ; Kobayashi, Suguru ; Fujito, Yutaka ; Yamashita, Masakane ; Sakakibara, Manabu ; Kemenes, György ; Ito, Etsuro. / Requirement of new protein synthesis of a transcription factor for memory consolidation : Paradoxical changes in mRNA and protein levels of C/EBP. :: Journal of Molecular Biology. 2006 ; 巻 356, 番号 3. pp. 569-577.
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abstract = "Some specific transcription factors are essential for memory consolidation across species. However, it is still unclear whether only the activation of constitutively expressed forms of these conserved transcription factors is involved in memory consolidation or their de novo synthesis also occurs after learning. This question has remained unanswered partly because of the lack of an efficient method for the determination of copy numbers of particular mRNAs in single neurons, which allows the detection of new transcription at the cellular level. Here we applied a newly developed protocol of single-cell quantitative real-time polymerase chain reaction (qRT-PCR) to single neurons playing an important role in associative learning. Specifically, we examined the changes in the mRNA and protein expression levels of a highly conserved transcription factor, CCAAT/enhancer binding protein (C/EBP), in the paired B2 motoneurons of the pond snail Lymnaea stagnalis. These buccal neurons are involved in the motor control of feeding behavior, with a potentially important role in conditioned taste aversion (CTA). Single-cell qRT-PCR revealed a significant decrease in LymC/EBP mRNA copy numbers in the B2 motoneurons during memory consolidation after CTA training. By contrast, isoelectric focusing and immunoblotting of extracts of the buccal ganglia showed that translation and phosphorylation levels of LymC/EBP significantly increased during memory consolidation. The C/EBP-like immunoreactivity in the B2 motoneurons, which are the major immunopositive component in the buccal ganglia, also significantly increased during memory consolidation, suggesting that the main source of increase in the level of protein in the buccal ganglia are the B2 motoneurons. Thus, early memory consolidation after CTA learning in L. stagnalis involves both the rapid synthesis and phosphorylation of LymC/EBP as well as the rapid breakdown of LymC/EBP mRNA in the neural network controlling feeding, suggesting that all of these processes play a role in the function of C/EBP in memory consolidation.",
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AU - Watanabe, Takayuki

AU - Wagatsuma, Akiko

AU - Kobayashi, Suguru

AU - Fujito, Yutaka

AU - Yamashita, Masakane

AU - Sakakibara, Manabu

AU - Kemenes, György

AU - Ito, Etsuro

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