Behavioural neuroscienceEarly socio-emotional experience induces expression of the immediate-early gene Arc/arg3.1 (activity-regulated cytoskeleton-associated protein/activity-regulated gene) in learning-relevant brain regions of the newborn chick
Section snippets
RNA isolation and cDNA synthesis
Total RNA was isolated from the brains of 1-day-old chicks (Gallus gallus domesticus) and adult rats (Rattus norvegicus) and treated with RNase-free DNase I (Promega, Mannheim, Germany). First-strand cDNA was then synthesized using either random nonamers (Stratagene, Amsterdam Zuidoost, The Netherlands) or the primer RoRidT17 (5′-ATCGATGGTCGACGCATGCGGATCCAAAGCTTGAATTCGAGCTCT17-3′; Harvey et al., 1991) and Moloney murine leukemia virus reverse transcriptase (Promega).
Amplification of a rat Arc/arg3.1 partial cDNA
An ∼1.2 kb cDNA fragment was
Results
By screening a chicken brain library with a rat Arc/arg3.1 cDNA probe, and the subsequent application of a nested RACE protocol (Frohman and Martin 1989, Harvey et al 1991, Stühmer et al 1996), we have isolated a full-length cDNA that encodes a polypeptide of 404 amino acids (Fig. 1), with a predicted molecular weight of 46,307 Da. This polypeptide exhibits 74%, 74% and 75% identity to the mouse (EMBL accession number AF162777), rat (Link et al., 1995; EMBL accession number Z46925) and human (
Discussion
We describe here the isolation of a full-length cDNA that encodes a 404 amino-acid protein that displays ∼75% sequence identity to mammalian Arc/arg3.1 proteins, defining it as the chicken orthologue. The sequence of this clone has been used to design a specific 45-base oligonucleotide, which has been used in in situ hybridization experiments to investigate changes in the expression of the Arc/arg3.1 gene in response to a short (30-min) acoustic imprinting paradigm in newborn chicks. This
Acknowledgments
This work was supported in part by the Deutsche Forschungsgemeinschaft (SFB444/A5 to M.G.D.), an award from the Fonds der Chemischen Industrie (to M.G.D.), and the VolkswagenStiftung (to K.B.). We thank Agata Blaszcyk-Wewer and Sönke Harder (Hamburg) for DNA sequencing and oligonucleotide synthesis, respectively.
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2018, Seminars in Cell and Developmental BiologyCitation Excerpt :Other forms of synaptic stimulation have also been shown to upregulate Arc transcription [20] and behavioural studies have revealed that Arc expression is increased by a number of tasks involving learning and memory formation. These include auditory imprinting in chicks [21], novel object recognition tasks in rodents [22], spatial memory recall [23] and fear conditioning paradigms [24]. It has been established that these increased levels of Arc mRNA are required for the synaptic plasticity underlying the subsequent learning and memory, as Arc knockout mice are deficient in spatial learning, aversive conditioning and long-term object recognition [16].
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