Re-purposing of histological tissue sections for corroborative western blot analysis of hypothalamic metabolic neuropeptide expression following delineation of transactivated structures by Fos immuno-mapping

Neuropeptides
Fahaad S H AlenaziKaren P Briski

Abstract

Fos immunocytochemistry is a valuable anatomical mapping tool for distinguishing cells within complex tissues that undergo genomic activation, but it is seldom paired with corroborative molecular analytical techniques. Due to preparatory requirements that include protein cross-linking for specimen sectioning, histological tissue sections are regarded as unsuitable for those methods. Our studies show that pharmacological activation of the hindbrain energy sensor AMPK by AICAR elicits estradiol (E)-dependent patterns of Fos immunolabeling of hypothalamic metabolic loci. Here, Western blotting was applied to hypothalamic tissue removed from histological sections of E- versus oil (O)-implanted ovariectomized (OVX) female rat brain to measure levels of metabolic transmitters associated with Fos-positive structures. In both E and O rats, AICAR treatment elicited alterations in pro-opiomelanocortin, neuropeptide Y, SF-1, and orexin-A neuropeptide expression that coincided with patterns of Fos labeling of structures containing neurons that synthesize these neurotransmitters, e.g. arcuate and ventromedial nuclei and lateral hypothalamic area. O, but not E animals also exhibited parallel augmentation of tissue corticotropin-releasing hor...Continue Reading

References

Jan 1, 1992·Neuroscience and Biobehavioral Reviews·G N Wade, J E Schneider
Jan 10, 2006·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·Bjarne KrebsHans A Kretzschmar
May 7, 2009·Journal of Neurochemistry·Gabriele V RonnettSusan Aja
Mar 21, 2012·Journal of Neuroscience Research·Ajeesh Koshy Cherian, Karen P Briski

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