Cholecystokinin-containing and nociceptive neurons in rat edinger-westphal nucleus

doi:10.1016/0006-8993(86)91008-5

Brain Research

Volume 363, Issue 2, 22 January 1986, Pages 230-238

https://doi.org/10.1016/0006-8993(86)91008-5 Get rights and content

Abstract

The putative peptide neurotransmitter cholecystokinin (CCK) is co-localized witsubstance P (SP) in a dense cluster of neurons located in the rat Edinger-Westphal (EW) nucleus. In an attempt to record electrophysiologically from these CCK-containing neurons, we have identified a group of nociceptive neurons located within the confines of the EW nucleus. The firing pattern of these nociceptive neurons is erratic, sometimes wita bursting pattern and at other times witfairly regular rates whicvary generally from 1 to 7 Hz. These neurons respond to noxious stimuli, sucas toe pinch, witan increase in rate and sometimes enter an apparent depolarization blockade (preceded by an increase in the duration and a decrease in the amplitude of the action potential). Systemically administered morphine suppresses botthe spontaneous firing rate and the toe pinch-induced increase in firing rate. Naloxone is able to reverse the effects of morphine. Althougwe have identified in the EW area a moderate density of terminals containing enkephalin-like immunoreactivity, morphine locally applied via microiontophoresis is largely without effect on the firing of these neurons. We hypothesize that the opiate-induced suppression of these nociceptive neurons is not mediated directly on the EW cells, but rather indirectly througafferent systems.

Reference (26)

AghajanianG.K. et al.
Serotonergic and non-serotonergic neurons of the dorsal raphe: reciprocal changes in firing induced by peripheral nerve stimulation
Brain Research
(1978)
AkertK. et al.
The Edinger-Westphal nucleus in the monkey. A retrograde tracer study
Brain Research
(1980)
HaysS.E. et al.
Demonstration of a putative receptor site for cholecystokinin in rat brain
Neuropeptides
(1980)
ItohS. et al.
Caerulein and cholecystokinin suppress β-endorphin-induced analgesia
Eur. J. Pharmacol.
(1982)
LoewyA.D. et al.
Re-evaluation of the efferent projections of the Edinger-Westphal nucleus in the cat
Brain Research
(1978)
LoewyA.D. et al.
Edinger-Westphal nucleus: projections to the brain stem and spinal cord in the cat
Brain Research
(1978)
PhippsB.S. et al.
Edinger-Westphal neurons that project to spinal cord contain Substance P
Neurosci. Lett.
(1983)
PingetM. et al.
Release of cholecystokinin peptides from a synaptosome-enriched fraction of rat cerebral cortex
Life Sci.
(1979)
SkirbollL.R. et al.
Peptide-monoamine coexistence: studies of the action of cholecystokinin-like peptide on the electrical activity of midbrain dopamine neurons
Neuroscience
(1981)
SkirbollL. et al.
Coexistence of substance P and cholecystokinin-like immunoreactivity in neurons of the mesencephalic periaqueductal central gray
Neurosci. Lett.
(1982)

ToyoshimaK. et al.

On the neuronal origin of the afferents to the ciliary ganglion in cat

Brain Research

(1980)

AghajanianG.K. et al.

Intracellular identification of central noradrenergic and serotonergic neurons by a new double labeling procedure

J. Neurosci.

(1982)

DoddS. et al.

The action of cholecystokinin and related peptides on pyramidal neurons of the mammalian hippocampus

Brain Research

(1981)

Cited by (50)

Oxytocin Receptors in the Mouse Centrally-projecting Edinger-Westphal Nucleus and their Potential Functional Significance for Thermoregulation
2022, Neuroscience
The centrally-projecting Edinger-Westphal nucleus (EWcp) has been shown to contribute to regulation of multiple functions, including responses to stress and fear, attention, food consumption, addiction, body temperature and maternal behaviors. However, receptors involved in regulation of these behaviors through EWcp remain poorly characterized. On the other hand, the oxytocin peptide (OXT) is also known to regulate a substantial number of physiological responses and behaviors. Here we show that mRNA encoding OXT receptors (Oxtr) is expressed in EWcp of male and female C57BL/6J mice. These receptors are present on urocortin 1 (Ucn) mRNA-containing neurons and, to a lesser extent, on neurons in EWcp expressing the vesicular glutamate transporter 2 (Vglut2) mRNA of EWcp. Using RNAscope in situ hybridization, we show that neurons containing Ucn and Vglut2 mRNAs are two intermingled, but independent subpopulations in EWcp and characterize their relationship with other populations of neurons in the vicinity of this nucleus. Using immunohistochemistry, we show that intraperitoneal (IP) administration of OXT can induce FOS in Oxtr-containing neurons, suggesting that these receptors on EWcp neurons are functional. A follow up study showed that injection of OXT (2.3 or 7.7 mg/kg, IP) is accompanied by a decrease in body temperature. Since EWcp is known to be involved in regulation of body temperature, we hypothesize that OXT’s effects on body temperature could be mediated through the EWcp. The contribution of OXTR in EWcp to regulation of various functions of EWcp and OXT needs to be deciphered.
Vesicular glutamate transporter 2-containing neurons of the centrally-projecting Edinger-Westphal nucleus regulate alcohol drinking and body temperature
2021, Neuropharmacology
Citation Excerpt :
In addition, it is increasingly recognized that the EWcp is not a uniform structure but rather consists of multiple cellular subpopulations (Zuniga and Ryabinin, 2020). Specifically, while the predominant population of neurons within the EWcp co-expresses peptides Ucn1, cocaine- and amphetamine-regulated transcript (CART) and nesfatin-1 (Kozicz, 2003; Okere et al., 2010), a smaller subpopulation expresses tyrosine hydroxylase (Bachtell et al., 2002), whereas a third subpopulation co-express cholecystokinin (CCK) and substance P (Innis and Aghajanian, 1986). Our own analysis of the Allen Brain Atlas (Lein et al., 2007) also suggested the presence of Vglut2-positive neurons within the EWcp.
Previous studies in rodents have repeatedly demonstrated that the centrally-projecting Edinger-Westphal nucleus (EWcp) is highly sensitive to alcohol and is also involved in regulating alcohol intake and body temperature. Historically, the EWcp has been known as the main site of Urocortin 1 (Ucn1) expression, a corticotropin-releasing factor-related peptide, in the brain. However, the EWcp also contains other populations of neurons, including neurons that express the vesicular glutamate transporter 2 (Vglut2). Here we transduced the EWcp with adeno-associated viruses (AAVs) encoding Designer Receptors Exclusively Activated by Designer Drugs (DREADDs) to test the role of the EWcp in alcohol drinking and in the regulation of body temperature. Activation of the EWcp with excitatory DREADDs inhibited alcohol intake in a 2-bottle choice procedure in male C57BL/6J mice, whereas inhibition of the EWcp with DREADDs had no effect. Surprisingly, analysis of DREADD expression indicated Ucn1-containing neurons of the EWcp did not express DREADDs. In contrast, AAVs transduced non-Ucn1-containing EWcp neurons. Subsequent experiments showed that the inhibitory effect of EWcp activation on alcohol intake was also present in male Ucn1 KO mice, suggesting that a Ucn1-devoid population of EWcp regulates alcohol intake. A final set of chemogenetic experiments showed that activation of Vglut2-expressing EWcp neurons inhibited alcohol intake and induced hypothermia in male and female mice. These studies expand on previous literature by indicating that a glutamatergic, Ucn1-devoid subpopulation of the EWcp regulates alcohol consumption and body temperature.
Melanocortin 4 receptor ligands modulate energy homeostasis through urocortin 1 neurons of the centrally projecting Edinger-Westphal nucleus
2017, Neuropharmacology
The role of the urocortin 1 (Ucn1) expressing centrally projecting Edinger-Westphal (EWcp) nucleus in energy homeostasis and stress adaptation response has previously been investigated. Morphological and functional studies have proven that orexigenic and anorexigenic peptidergic afferents and receptors for endocrine messengers involved in the energy homeostasis are found in the EWcp. The central role of the hypothalamic melanocortin system in energy homeostasis is well known, however, no data have been published so far on possible crosstalk between melanocortins and EWcp-Ucn1. First, we hypothesized that members of the melanocortin system [i.e. alpha-melanocyte stimulating hormone (alpha-MSH), agouti-related peptide (AgRP), melanocortin 4 receptor (MC4R)] would be expressed in the EWcp. Second, we put forward, that alpha-MSH and AgRP contents as well as neuronal activity and Ucn1 peptide content of the EWcp would be affected by fasting. Third, we assumed that the intra-EWcp injections of exogenous MC4R agonists and antagonist would cause food intake-related and metabolic changes.
Ucn1 neurons were found to carry MC4Rs, and they were contacted both by alpha-MSH and AgRP immunoreactive nerve fibers in the rat. The alpha-MSH immunosignal was reduced, while that of AgRP was increased upon starvation. These were associated with the elevation of FosB and Ucn1 expression. The intra-EWcp administration of MC4R blocker (i.e. HS024) had a similar, but enhanced effect on FosB and Ucn1. Furthermore, alpha-MSH injected into the EWcp had anorexigenic effect, increased oxygen consumption and caused peripheral vasodilation.
We conclude that the melanocortin system influences the EWcp that contributes to energy-homeostasis.
Postnatal developmental profile of urocortin 1 and cocaine- and amphetamine-regulated transcript in the perioculomotor region of C57BL/6J mice
2010, Brain Research
Citation Excerpt :
Moreover, finding that expression of CART and Ucn 1 appears in this brain region at such different ages, indicates differential regulation of genes expressed in pIII neurons during development. The pIIIu shows preferential expression of several additional genes involved in regulation of food consumption, stress and energy homeostasis, such as cholecystokinin, nesfatin and the growth hormone secretagogue receptor (also known as ghrelin receptor) (Brailoiu et al., 2007; Innis and Aghajanian, 1986; Maciewicz et al., 1984; Xu et al., 2009). It would be important to comparatively investigate developmental profiles of different behaviorally important genes in this brain region.
Urocortin 1 (Ucn 1) is an endogenous corticotropin releasing factor (CRF)-related peptide. Ucn 1 is most highly expressed in the perioculomotor urocortin containing neurons (pIIIu), previously known as the non-preganglionic Edinger-Westphal nucleus (npEW). Various studies indicate that these cells are involved in stress adaptation and the regulation of ethanol (EtOH) intake. However, the developmental trajectory of these neurons remained unexamined. Expression of the cocaine- and amphetamine-regulated transcript (CART), which co-localizes with Ucn 1 in the perioculomotor area (pIII) has been examined prenatally, but not postnatally. The goal of the current study was to characterize the ontogenetic profile of Ucn 1 and CART during postnatal development in C57BL/6J (B6) mice. B6 mice were bred, and brains were collected at postnatal days (PND) 1, 4, 8, 12, 16, 24 and 45. Brightfield immunohistochemical staining for Ucn 1 and CART showed that Ucn 1-immunoreactivity (ir) was absent at PND 1, while CART-ir was already apparent in pIIIu at birth, a finding indicating that although the pIIIu neurons have already migrated to their adult position, Ucn 1 expression is triggered in them at later postnatal stages. Ucn 1-ir gradually increased with age, approaching adult levels at PND 16. This developmental profile was confirmed by double-immunofluorescence, which showed that Ucn 1 was absent in CART-positive cells of pIII at PND 4 and that Ucn 1 and CART are strongly but not completely co-localized in pIII at PND 24. Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) analysis confirmed that Ucn 1 mRNA levels are significantly lower at PND 4 and PND 12 than in adult animals. The lack of brain Ucn 1 immunoreactivity at birth and the gradual postnatal increase in Ucn 1 in pIIIu suggests that this peptide plays a greater behavioral role in adulthood than during the early postnatal development of an organism.
Urocortin 1 microinjection into the mouse lateral septum regulates the acquisition and expression of alcohol consumption
2008, Neuroscience
Citation Excerpt :
Since our study included very low doses of Ucn1, this reasoning seems unlikely. As an alternative explanation, since pIIIu neurons contain other peptides besides Ucn1 as potential regulators of the rewarding properties of drugs of abuse [including CART, CCK and neuropeptide B (Maciewicz et al., 1984; Innis and Aghajanian, 1986; Couceyro et al., 1997; Kozicz, 2003; Tanaka et al., 2003)], it is possible that synergistic actions of these peptides are needed to increase the rewarding properties of alcohol. Lesions of the pIIIu or a study correlating the number of cells in the pIIIu with alcohol drinking cannot directly test this possibility, but can only point to an importance of the brain region in this behavior.
Previous studies using genetic and lesion approaches have shown that the neuropeptide urocortin 1 (Ucn1) is involved in regulating alcohol consumption. Ucn1 is a corticotropin releasing factor (CRF) –like peptide that binds CRF1 and CRF2 receptors. Perioculomotor urocortin-containing neurons (pIIIu), also known as the non-preganglionic Edinger-Westphal nucleus, are the major source of Ucn1 in the brain and are known to innervate the lateral septum. Thus, the present study tested whether Ucn1 could regulate alcohol consumption through the lateral septum. In a series of experiments Ucn1 or CRF was bilaterally injected at various doses into the lateral septum of male C57BL/6J mice. Consumption of 20% volume/volume ethanol or water was tested immediately after the injections using a modification of a 2-h limited access sweetener-free “drinking-in-the-dark” procedure. Ucn1 significantly suppressed ethanol consumption when administered prior to the third ethanol drinking session (the expression phase of ethanol drinking) at doses as low as 6 pmol. Ethanol intake was differentially sensitive to Ucn1, as equivalent doses of this peptide did not suppress water consumption. In contrast, CRF suppressed both ethanol and water intake at 40 and 60 pmol, but not at lower doses. Repeated administration of Ucn1 during the acquisition of alcohol consumption showed that 40 pmol (but not 2 or 0.1 pmol) significantly attenuated ethanol intake. Repeated administration of Ucn1 also resulted in a decrease of ethanol intake in sham-injected animals, a finding suggesting that the suppressive effect of Ucn1 on ethanol intake can be conditioned. Taken together, these studies confirm the importance of lateral septum innervation by Ucn1 in the regulation of alcohol consumption.
On the role of urocortin 1 in the non-preganglionic Edinger-Westphal nucleus in stress adaptation
2007, General and Comparative Endocrinology
The discovery of novel members of the CRF neuropeptide family, urocortin 1 (Ucn1), urocortin 2 and 3 has provided important insights into stress adaptation pathways, and predicted that stress adaptation involves more systems than the HPA-axis alone. This mini-review aims to summarize our recent data and research by others indicating that an important role is played by Ucn1 in the non-preganglionic Edinger–Westphal nucleus (npEW). These results point to an intriguing possibility that CRF/Ucn1 neuronal circuits comprise two separate, but functionally interrelated entities, which are coordinately regulated by acute stressors, but are inversely coupled during chronic stress. Such collaboration between the two systems would implicate a very important role of Ucn1 in adaptation to stress, and, as a consequence, in stress-related disorders like anxiety, major depression and use of drugs of abuse.

View all citing articles on Scopus

View full text

Cholecystokinin-containing and nociceptive neurons in rat edinger-westphal nucleus

Abstract

Brain Research

Brain Research

Neuropeptides

Eur. J. Pharmacol.

Brain Research

Brain Research

Neurosci. Lett.

Life Sci.

Neuroscience

Neurosci. Lett.

Brain Research

Intracellular identification of central noradrenergic and serotonergic neurons by a new double labeling procedure

J. Neurosci.

The action of cholecystokinin and related peptides on pyramidal neurons of the mammalian hippocampus

Brain Research