Antibody Staining And In Situ Hybridization Reveals That A Subset Of Neurons In The Rabbit Neonatal Brain Produce Insulin View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

1987-04

AUTHORS

Ruben Schechter, Lynn Karyckl, Farouk Sadig, Thomas Hilliard, Arnold Kahn, Sherin Devaskar

ABSTRACT

We have previously shown that insulin is present in whole brain extracts of fetal/neonatal rabbits (BBRC 136:208, 1986). However, the source of this hormone remains controversial. To resolve this issue, glia and neurons from 10 day old newborn rabbits were grown in tissue culture and subsequently analyzed for hormone synthesis by the peroxidase-antiperoxidase (PAP) technique and in situ HYB. Glia and neurons were identified on the basis of morphology and positive staining with enolase (neurons) and fibrillary acidic protein (glia). The monoclonal antibody (1:100) used in the PAP reaction was specific for insulin. In situ HYB was conducted at 37°C under stringent conditions using a biotin-labeled rat insulin cDNA probe (Dr. A. Permutt). Biotinylated pBR322 DNA served as a control. HYB was detected using avidin-Peroxidase. The data show that insulin is present in only ∼5% of the neurons but not glia. Pretreating the cells with the ionophore monensin, to block hormone secretion, augmented the PAP reaction but did not increase the number of positive cells. Similarly, in situ HYB revealed insulin mRNA in ∼5% of the neurons, with no such transcripts identified in glia. We conclude that insulin detected in fetal/neonatal brain extracts is synthesized locally by a select subset of neurons. The biologic function of this neuronal insulin remains to be determined, but possibilities include neurotransmitter activity and/or growth promotion. More... »

PAGES

221-221

Identifiers

URI

http://scigraph.springernature.com/pub.10.1203/00006450-198704010-00330

DOI

http://dx.doi.org/10.1203/00006450-198704010-00330

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1035601701


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