Expression of AP-1 family transcription factors in the amygdala during conditioned taste aversion learning: role for Fra-2.

Conditioned taste aversion (CTA) learning occurs after the pairing of a novel taste with a toxin (e.g. sucrose with LiCl). The immediate early gene c-Fos is necessary for CTA learning, but c-Fos alone cannot be sufficient for consolidation. The expression of other AP-1 proteins from the Fos- and Jun-families may also be required shortly after conditioning for CTA consolidation. To screen for the expression of AP-1 transcription factors within small subregions, RT-PCR analysis was used after laser capture microdissection of the amygdala. Rats were infused intraorally with 5% sucrose (6 ml/6 min) or injected with LiCl (12 ml/kg, 0.15 M, i.p.) or given sucrose paired with LiCl (sucrose/LiCl), or not treated; 1 h later their brains were dissected. The lateral (LA), basolateral (BLA), and central (CeA) subnuclei of the amgydala of single 5 microm sections from individual rats were dissected using the Arcturus PixCell II system. Semi-quantitative RT-PCR showed the consistent presence of c-Fos, Fra-2, c-Jun, and JunD in the amygdala. In situ hybridization confirmed that c-Fos and Fra-2 mRNA expression was increased in the CeA after LiCl and sucrose/LiCl treatment. Immunohistochemistry for Fra-2 revealed high baseline levels of Fra-2 protein in the BLA and CeA, but also an increase in Fra-2 in the BLA and CeA after LiCl and sucrose/LiCl treatment. The similarity of response in LiCl and sucrose/LiCl treated groups might reflect activation by LiCl in both groups. To control for the effects of LiCl, rats were tested in a learned safety experiment. Fra-2 and c-Fos were examined in response to sucrose/LiCl in rats with prior familiarity with sucrose compared to rats without prior exposure to sucrose. The familiar (pre-exposure) group showed a significantly decreased number of Fra-2-positive cells compared with the novel group in the BLA, but not in the CeA. Because pre-exposure to sucrose attenuates CTA learning, a decreased cellular response in pre-exposed rats suggests a specific correlation with CTA learning. Changes in Fra-2 and c-Fos expression in the BLA and CeA at the time of conditioning, together with constitutive expression of c-Jun and JunD, may contribute to CTA learning.

Alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate receptor subunit expression in rat olfactory bulb.

The alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) receptors (AMPARs) mediate rapid responses at most central excitatory synapses, including those in the olfactory bulb (OB). These receptors are composed of the glutamate subunits GluR1-4, which each has two splice variant (flip/flop) forms. We recently showed that AMPARs on OB neurons are kinetically and pharmacologically diverse. Here, we explored whether this functional heterogeneity reflects a diverse expression of AMPAR subunits and/or splice variants. Total RNA from rat OBs was amplified by RT-PCR. Digestion of the panGluR PCR product with subunit-specific restriction enzymes revealed that the OB expresses mRNAs for GluR1-4 but in different relative amounts i.e., GluR2 (61 +/- 2.4%), GluR1 (31 +/- 3.5%), GluR4 (6.3 +/- 1.4%), GluR3 (1.4 +/- 0.7%). Furthermore, GluR2 and GluR4 transcripts were composed of similar amounts of flip and flop, whereas GluR1 and GluR3 transcripts consisted mostly of flip. If similar to other brain regions, this heterogeneity in patterns of expression may facilitate information processing.

The maize Single myb histone 1 gene, Smh1, belongs to a novel gene family and encodes a protein that binds telomere DNA repeats in vitro.

We screened maize (Zea mays) cDNAs for sequences similar to the single myb-like DNA-binding domain of known telomeric complex proteins. We identified, cloned, and sequenced five full-length cDNAs representing a novel gene family, and we describe the analysis of one of them, the gene Single myb histone 1 (Smh1). The Smh1 gene encodes a small, basic protein with a unique triple motif structure of (a) an N-terminal SANT/myb-like domain of the homeodomain-like superfamily of 3-helical-bundle-fold proteins, (b) a central region with homology to the conserved H1 globular domain found in the linker histones H1/H5, and (c) a coiled-coil domain near the C terminus. The Smh-type genes are plant specific and include a gene family in Arabidopsis and the PcMYB1 gene of parsley (Petroselinum crispum) but are distinct from those (AtTRP1, AtTBP1, and OsRTBP1) recently shown to encode in vitro telomere-repeat DNA-binding activity. The Smh1 gene is expressed in leaf tissue and maps to chromosome 8 (bin 8.05), with a duplicate locus on chromosome 3 (bin 3.09). A recombinant full-length SMH1, rSMH1, was found by band-shift assays to bind double-stranded oligonucleotide probes with at least two internal tandem copies of the maize telomere repeat, TTTAGGG. Point mutations in the telomere repeat residues reduced or abolished the binding, whereas rSMH1 bound nonspecifically to single-stranded DNA probes. The two DNA-binding motifs in SMH proteins may provide a link between sequence recognition and chromatin dynamics and may function at telomeres or other sites in the nucleus.