Absolute quantification of Atlantic salmon and rainbow trout vitellogenin by the 'signature peptide' approach using electrospray ionization QqToF tandem mass spectrometry.

In this work, we present a simple method for absolute quantification of plasma vitellogenin from both rainbow trout and Atlantic salmon. Plasma samples obtained from control and beta-estradiol induced fish were digested with trypsin. A characteristic 'signature peptide' was selected and analyzed by high performance liquid chromatography (HPLC) coupled to an electrospray quadrupole-time-of-flight tandem mass spectrometer, using a deuterated homolog peptide as an internal standard. The hybrid tandem mass spectrometer was operated in a 'pseudo' selected reaction-monitoring mode in which three diagnostic product ions were monitored for identification and quantification purposes. The reproducibility (coefficient of variation approximately 5%) and sensitivity (limit of quantification (LOQ) of 0.009 mg/ml) achieved by this simple assay allow it to be considered as an alternative to immunological assays.

'De novo' sequencing of Atlantic cod vitellogenin tryptic peptides by matrix-assisted laser desorption/ionization quadrupole time-of-flight tandem mass spectrometry: similarities with haddock vitellogenin.

Vitellogenin is a protein produced by the liver of oviparous animals in response to circulating estrogens. The amino acid sequence of vitellogenin from Atlantic cod (Gadus morhua) has not yet been determined. In this study Atlantic cod vitellogenin was characterized using a 'bottom-up' mass spectrometric approach. Vitellogenin synthesis was induced 'in vivo' with beta-estradiol, and subjected to trypsin digestion for characterization by matrix-assisted laser desorption/ionization quadrupole time-of-flight tandem mass spectrometry. A peptide mass fingerprint was obtained and 'de novo' sequencing of the most abundant tryptic peptides was performed by low-energy collision-induced dissociation tandem mass spectrometry. As a result of these experiments, the sequences of various tryptic peptides have been elucidated. The database search has shown that Atlantic cod vitellogenin shares a series of common peptides with the two different known vitellogenin sequences of haddock, a closely related species. These findings allow us to propose that Atlantic cod might also co-express at least two distinct forms of vitellogenin.

Novel objects in a holeboard probe the role of the locus coeruleus in curiosity: support for two modes of attention in the rat.

Idazoxan, an alpha 2 adrenoceptor antagonist (2 mg/kg), enhanced novel object investigation in a holeboard in rats as previously reported (V. Devauges & S. J. Sara, 1990). Two weeks of 10 min/day in 37 degrees C water increased dopamine-beta-hydroxylase staining density in the locus coeruleus but did not enhance novel object investigation. In contrast to idazoxan, however, the warm water treatment increased rearing, center entries, and activity, a pattern previously described during tonic infusion of norepinephrine into the hippocampus. Correlations among dopamine-beta-hydroxylase measures and behavior reinforced these tonic norepinephrine/behavior associations. The behavioral effects across the idazoxan and warm water experiments support G. Aston-Jones et al.'s (1999) 2 modes of attention hypothesis for locus coeruleus function: Phasic locus coeruleus activity promotes focused attention; tonic locus coeruleus activity promotes scanning attention.