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Intravenous 2-Deoxy-D-Glucose Injection Rapidly Elevates Levels of the Phosphorylated Forms of p44/42 Mitogen-Activated Protein Kinases (Extracellularly Regulated Kinases 1/2) in Rat Hypothalamic Parvicellular Paraventricular Neurons

Program in Neural, Informational and Behavioral Sciences, University of Southern California, Los Angeles, California 90089

Address all correspondence and requests for reprints to: Dr. Arshad M. Khan, Department of Biological Sciences, Hedco Neuroscience Building, MC 2520, 3641 Watt Way, University of Southern California, Los Angeles, California 90089-2520. E-mail: arshadk{at}usc.edu.

CRH neurons within the medial parvicellular part of the hypothalamic paraventricular nucleus (PVHmp) can respond to afferent inputs encoding stress-related information by initiating peptide synthesis (signaling cascades, transcription, and translation) and/or peptide release. However, understanding these cellular events is hampered by three outstanding issues: 1) neural inputs that activate CRH neurons remain incompletely identified; 2) the identity and temporal dynamics of signaling pathways within CRH neurons are poorly understood; and 3) the precise coupling of the first two issues has not been established. Here, we report that the phosphorylated forms of p44/p42 MAPKs (pERK1/2) are rapidly detected in PVHmp cells after iv infusion of the antimetabolite, 2-deoxy-D-glucose (2-DG). Combined immunocytochemistry and in situ hybridization revealed that pERK1/2 immunoreactivity is detectable 10 min after 2-DG infusion not only within most PVHmp neurons containing CRH mRNA (78.6% of mean total CRH cells counted) but also in many non-CRH neurons (45.5% of mean total sampled cells). In contrast, Fos protein in the PVHmp was not detected within this time period, consistent with the known time course for its translation. Stress associated with halothane exposure also robustly elevated pERK1/2 levels in PVHmp neurons approximately 10 min after exposure. Our results implicate pERK1/2 in stress-induced activation of CRH neurosecretory cells and underscore their utility as indices of rapid cellular activation. Because 2-DG-induced activation of CRH gene transcription in these neurons requires a catecholaminergic input, our data also suggest that pERK1/2 could couple afferent catecholaminergic signals with CRH gene expression in these neurons.

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