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Total RNA was extracted from goldfish brain using the TRIzol® RNA isolation reagent (Invitrogen, Canada).
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Using reverse-transcriptase polymerase chain reaction (RT-PCR), Choi and Habibi (2003) also detected both ERs in goldfish brain.
In the present study, on the basis of multiple goldfish microarray datasets, we used comprehensive normalizations, differential gene expression identification, multivariate and gene ontology (GO) analyses to characterize the season-related expression patterns in the female goldfish brain.
As expected, the antibody targeted full-length NUCB2 in rat and goldfish brain samples (Figure 5C).
Anatomical localization and nomenclature were based on the goldfish brain atlas [26].
A sagittal view of the goldfish brain stained with nesfatin-1 antiserum, revealed a large number of ir cell bodies within the goldfish hypothalamus (Figure 6A F).
To date, many other well established neuropeptides with mammalian orthologues have been characterized within the NLT of the goldfish brain.
Following bioinformatics analysis, we collected goldfish brain samples at six seasonal time points for experimental verification of gene expression patterns.
On the basis of this meta-type dataset, we identified four clear seasonal gene expression patterns in female goldfish brain.
No band representing the processed nesfatin-1 (expected size: ∼9.5 kDa) was detected in either rat or goldfish brain samples.
Although further studies are required to elucidate the mechanisms of actions and interactions of nesfatin-1 in the goldfish brain, our results clearly indicate an anorexigenic effect for the native form of nesfatin-1 in goldfish.
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