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. 2003 Jul 2;23(13):5607-16.
doi: 10.1523/JNEUROSCI.23-13-05607.2003.

Categorically distinct acute stressors elicit dissimilar transcriptional profiles in the paraventricular nucleus of the hypothalamus

Teresa M Reyes  1 , John R Walker, Casey DeCino, John B Hogenesch, Paul E Sawchenko
Affiliations

Categorically distinct acute stressors elicit dissimilar transcriptional profiles in the paraventricular nucleus of the hypothalamus

Teresa M Reyes et al. J Neurosci. .

Abstract

The paraventricular hypothalamic nucleus (PVH) is a key site for integrating neuroendocrine, autonomic, and behavioral adjustments to diverse homeostatic challenges, including "physiological" (e.g., infection or hemorrhage) and "emotional" [e.g., restraint (RST) or footshock] stresses. Both types of challenges ultimately converge to activate common response systems represented in PVH, including the hypothalamo-pituitary-adrenal axis and the sympathoadrenal system. Oligonucleotide microarrays (U74A; Affymetrix, Santa Clara, CA) were used to compare and contrast gene expression profiles in the PVH elicited at 1 and 3 hr after acute exposure to representative physiological [intraperitoneal injection of 10 microg lipopolysaccharide (LPS)] and emotional (30 min RST) stressors. In general, the two challenges recruited relatively few genes in common, with the degree of overlap varying across functional classes of genes. The greatest degree of commonality was seen among signaling molecules and neuropeptides, whereas transcription factors upregulated by RST and LPS were largely distinct. Unexpectedly, RST induced a number of immune-related molecules, which were not regulated by LPS. Hybridization histochemical analyses localized a subset of responsive transcripts to the PVH and/or immediately adjoining regions. Immunerelated molecules in particular distributed broadly to vascular and other barrier-associated cell types. These global transcriptional profiles inform the search for early (transcription factors) and late (target genes) mechanisms in the modulation of PVH, and generalized CNS, responses to categorically distinct stressors.

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Figures

Figure 1.
Figure 1.
Dissection procedure. A photograph of a coronal brain slice to illustrate the dissection procedure. A series of six cuts were performed using a razor blade. Viewing the ventral surface of the brain, two coronal cuts were made to isolate a hypothalamic block using the apex of the optic chiasm and the rostral margin of the mammillary bodies as landmarks. This slab was then placed flat and the first two cuts were placed on either side of the chiasm. The third cut was placed just dorsal to the third ventricle. Finally, this last block was bisected horizontally with the dorsal half representing the PVH-enriched region (a) and the ventral half comprising the ARH-enriched region (b). Magnification, 7×.
Figure 2.
Figure 2.
Induced Fos expression in response to LPS injection or restraint. Expression of the immediate early gene product, Fos, in the PVH of control (saline-injected), LPS-challenged (10 μg, i.p.), and acutely restrained animals (30 min). At 2 hr after stress, both treatments led to comparable patterns of Fos induction in PVH, over and above the low basal levels of expression seen in saline-injected controls, with LPS provoking a somewhat stronger response. Magnification, 130×.
Figure 3.
Figure 3.
Overlap in the sets of genes regulated by the two stressors. A depiction of the extent of overlap between the genes that met the following criteria: significant change from saline control (p < 0.05) and a fold change of at least 2.5. Numbers of genes that met these criteria are indicated within each box. There was minimal overlap between the sets of genes upregulated in response to either stressor at both time points, with values ranging between 4 and 16% (LPS, white; Shared, gray; RST, black). A similar pattern is observed in the genes that are downregulated in response to either stressor; however, there is substantially greater overlap at the 1 hr time point (24–35%) versus 3 hr (8–9%).
Figure 4.
Figure 4.
LPS-induced expression of the chemokine IP-10. In situ hybridization was used to confirm the expression of IP-10 in the PVH. Top, Chemokine expression was not detected in saline-treated animals (left) but was rapidly induced in response to LPS (middle; magnification, 70×). Immunolocalization for NeuN to identify neurons (right, top; magnification, 440×) or CD31 to identify blood vessels (BV) (right, bottom; magnification 280×) was combined with in situ hybridization for IP-10 (black grains) in tissue from LPS-treated animals. A NeuN/IP-10 doubly labeled cell (arrowhead) is apparent, but the bulk of IP-10 expression appears to be non-neuronal. Extensive codistribution of CD31 and IP-10 confirms the presence of this transcript on vascular-associated cells. IP-10 was also induced by LPS in other barrier-related areas (bottom), including BV (left), the choroid plexus (Chp) and SFO (middle), and AP (right). Small, discretely labeled cells, possibly glia, are also apparent throughout the brains of LPS-treated animals (magnification, 35×). v3, Third ventricle.
Figure 5.
Figure 5.
LPS-induced expression of additional chemokines, MCP-1 and Gro 1. Other chemokines showed induced patterns of expression that were similar, although not as dramatic as that exhibited by CXCL10, including MCP-1 (top) and Gro 1 (bottom). Dark-field images show expression of mRNA for both chemokines within or immediately adjacent to PVH, as well as in barrier-related areas, including SFO and choroid plexus (MCP-1, top right) and blood vessels (Gro 1, bottom right). Magnification: left, 45×; right, 90×.
Figure 6.
Figure 6.
Expression of the LPS-responsive transcription factor C/EBPδ. Dark-field images illustrating the LPS-induced expression pattern of mRNA encoding the transcription factor C/EBPδ. This shows a distribution in barrier-related structures similar to that of the chemokines (CXCL 10, MCP-1, and Gro1). Little if any expression is apparent in control animals (left). After injection of LPS, there is a dramatic upregulation of this transcript in a number of areas, including the SFO, choroid plexus (Chp), PVH, blood vessels (BVs), and meninges (Men). Magnification, 70×.
Figure 7.
Figure 7.
Orexin induction in response to RST. The left panel shows the distribution of orexin mRNA (black grains) within the LHA. The boxed area indicates the approximate region that was quantified. Orexin mRNA is significantly upregulated in response to 30 min RST. Representative images from the brains of control and acutely restrained animals are shown in dark field in the middle and right panels. The upregulation of orexin mRNA is statistically significant (p < 0.003). Magnification, 70×.
Figure 8.
Figure 8.
Neuropeptides that change similarly in response to both stressors. NPY, ppENK, and CCK are similarly affected by acute exposure to systemic LPS or restraint. The bar graphs show the fold change for each neuropeptide at 1 hr (left) and 3 hr (right). In situ hybridization was used to confirm the changes in ppENK mRNA at 2 hr after LPS administration or 30 min RST. Whereas increased signal is apparent within the PVH proper, the upregulation is primarily localized to the region just lateral to the PVH and medial to the fornix. Magnification, 75×.
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References

    1. Ando T, Brown RF, Berg RD, Dunn AJ ( 2000) Bacterial translocation can increase plasma corticosterone and brain catecholamine and indoleamine metabolism. Am J Physiol Regul Integr Comp Physiol 279: R2164–R2172. - PubMed
    1. Baeuerle PA ( 1998) IkappaB-NF-kappaB structures: at the interface of inflammation control. Cell 95: 729–731. - PubMed
    1. Bajetto A, Bonavia R, Barbero S, Florio T, Schettini G ( 2001) Chemokines and their receptors in the central nervous system. Front Neuroendocrinol 22: 147–184. - PubMed
    1. Bluthe RM, Bristow A, Lestage J, Imbs C, Dantzer R ( 2001) Central injection of interleukin-13 potentiates LPS-induced sickness behavior in rats. NeuroReport 12: 3979–3983. - PubMed
    1. Broberger C, de Lecea L, Sutcliffe JG, Hokfelt T ( 1998) Hypocretin/orexin- and melanin-concentrating hormone-expressing cells form distinct populations in the rodent lateral hypothalamus: relationship to the neuropeptide Y and agouti gene-related protein systems. J Comp Neurol 402: 460–474. - PubMed

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