Characterization and visualization of cholecystokinin receptors in rat brain using [3H]pentagastrin

doi:10.1016/0196-9781(83)90032-3

Peptides

Volume 4, Issue 5, September–October 1983, Pages 755-762

https://doi.org/10.1016/0196-9781(83)90032-3 Get rights and content

Abstract

[³H]Pentagastrin binds specifically to an apparent single class of CCK receptors on slide-mounted sections of rat brain (K_D=5.6 nM; B_max=36.6 fmol/mg protein). This specific binding is temperature-dependant and regulated by ions and nucleotides. The relative potencies of C-terminal fragments of CCK-8(SO₃H), benzotript and proglumide in inhibiting specific [³H]pentagastrin binding to CCK brain receptors reinforce the concept of different brain and pancreas CCK receptors. CCK receptors were visualized by using tritium-sensitive LKB film analyzed by computerized densitometry. CCK receptors are highly concentrated in the cortex, dentate gyrus, granular and external plexiform layers of the olfactory bulb, anterior olfactory nuclei, olfactory tubercle, claustrum, accumbens nucleus, some nuclei of the amygdala, thalamus and hypothalamus.

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Cited by (113)

Somatodendritic Release of Cholecystokinin Potentiates GABAergic Synapses Onto Ventral Tegmental Area Dopamine Cells
2023, Biological Psychiatry
Citation Excerpt :
Might CCK release occurring during depolarization cause LTP? CCK acts via 2 G protein–coupled receptors, CCK1R and CCK2R (61,62); CCK2R is predominant in the brain (63–66). We therefore bath applied the CCK2R antagonist, LY225910, and tested whether depolarization was still capable of inducing LTP.
Neuropeptides are contained in nearly every neuron in the central nervous system and can be released not only from nerve terminals but also from somatodendritic sites. Cholecystokinin (CCK), among the most abundant neuropeptides in the brain, is expressed in the majority of midbrain dopamine neurons. Despite this high expression, CCK function within the ventral tegmental area (VTA) is not well understood.
We confirmed CCK expression in VTA dopamine neurons through immunohistochemistry and in situ hybridization and detected optogenetically induced CCK release using an enzyme-linked immunosorbent assay. To investigate whether CCK modulates VTA circuit activity, we used whole-cell patch clamp recordings in mouse brain slices. We infused CCK locally in vivo and tested food intake and locomotion in fasted mice. We also used in vivo fiber photometry to measure Ca²⁺ transients in dopamine neurons during feeding.
Here we report that VTA dopamine neurons release CCK from somatodendritic regions, where it triggers long-term potentiation of GABAergic (gamma-aminobutyric acidergic) synapses. The somatodendritic release occurs during trains of optogenetic stimuli or prolonged but modest depolarization and is dependent on synaptotagmin-7 and T-type Ca²⁺ channels. Depolarization-induced long-term potentiation is blocked by a CCK₂ receptor antagonist and mimicked by exogenous CCK. Local infusion of CCK in vivo inhibits food consumption and decreases distance traveled in an open field test. Furthermore, intra-VTA–infused CCK reduced dopamine cell Ca²⁺ signals during food consumption after an overnight fast and was correlated with reduced food intake.
Our experiments introduce somatodendritic neuropeptide release as a previously unknown feedback regulator of VTA dopamine cell excitability and dopamine-related behaviors.
Cholecystokinin acts in the dorsomedial hypothalamus of young male rats to suppress appetite in a nitric oxide-dependent manner
2021, Neuroscience Letters
Citation Excerpt :
We have demonstrated that CCK decreases food intake in young rats when administered directly into the DMH. CCK acts through two receptors, CCK1 and CCK2 receptors, both of which are expressed in the DMH [2,12]. Previous research has demonstrated that CCK acts through CCK2 receptors to modulate synaptic transmission in the DMH in young rats [9].
Cholecystokinin (CCK) is an appetite-suppressing hormone that acts in the dorsomedial hypothalamus (DMH) in adult rats to suppress food intake. It remains unknown, however, whether CCK has the same affect in young animals, despite the rising prevalence of childhood obesity and drastic need for research in this area. At the synaptic level, CCK has been shown to inhibit putative orexigenic DMH neurons in young male rats by increasing GABA release onto these neurons via a CCK2 receptor and nitric oxide-dependent pathway. Whether this pathway leads to appetite suppression in young rats is not known. We therefore investigated whether intra-DMH administration of CCK, with or without inhibitors of the CCK2 receptor and nitric oxide signaling pathways, affects food intake in young, male, fasted Sprague-Dawley rats. We implanted bilateral guide cannulas into the DMH and allowed animals to recover from the surgery. Animals were then fasted for 24 h, following which they received intra-DMH microinjections of vehicle, CCK-8S, or CCK-8S combined with either LY-225910 (CCK2 receptor antagonist), L-NAME (a nitric oxide synthase inhibitor), or ODQ (a soluble guanylate cyclase inhibitor) and were given access to regular chow. Following a two hour refeeding period during which food intake, latency to feed, and body weight were measured, brains were subsequently removed to confirm cannula placement in the DMH. The effect of CCK on these parameters in rats given a high fat diet were also measured. Here we show that intra-DMH administration of CCK suppresses food intake and body weight in young rats. This effect requires activation of CCK2 receptors and nitric oxide signaling. Finally, CCK has no effect on consumption of a high fat diet when administered into the DMH. Overall, these findings demonstrate a potential pathway through which CCK might suppress appetite in young rats.
Increased cholecystokinin labeling in the hippocampus of a mouse model of epilepsy maps to spines and glutamatergic terminals
2012, Neuroscience
Citation Excerpt :
The conditions for, and function of, CCK release from glutamatergic axon boutons and the reason for its increase after pilocarpine injection remains to be established. In normal conditions, the sulfated octapeptide is postulated to be released under strong, repetitive stimulation (Verhage et al., 1991; Karson et al., 2008; Deng et al., 2010), activating predominantly G-protein-coupled CCK2 receptors in the brain (Gaudreau et al., 1983; Hill et al., 1987; Carlberg et al., 1992; Harro et al., 1993; Wang et al., 2011; Lee et al., 2011). In general, the functional role of CCK is not entirely understood, although that is not due to lack of investigation (for review see Lee and Soltesz, 2011).
The neuropeptide cholecystokinin (CCK) is abundant in the CNS and is expressed in a subset of inhibitory interneurons, particularly in their axon terminals. The expression profile of CCK undergoes numerous changes in several models of temporal lobe epilepsy. Previous studies in the pilocarpine model of epilepsy have shown that CCK immunohistochemical labeling is substantially reduced in several regions of the hippocampal formation, consistent with decreased CCK expression as well as selective neuronal degeneration. However, in a mouse pilocarpine model of recurrent seizures, increases in CCK-labeling also occur and are especially striking in the hippocampal dendritic layers of strata oriens and radiatum. Characterizing these changes and determining the cellular basis of the increased labeling were the major goals of the current study. One possibility was that the enhanced CCK labeling could be associated with an increase in GABAergic terminals within these regions. However, in contrast to the marked increase in CCK-labeled structures, labeling of GABAergic axon terminals was decreased in the dendritic layers. Likewise, cannabinoid receptor 1-labeled axon terminals, many of which are CCK-containing GABAergic terminals, were also decreased. These findings suggested that the enhanced CCK labeling was not due to an increase in GABAergic axon terminals. The subcellular localization of CCK immunoreactivity was then examined using electron microscopy, and the identities of the structures that formed synaptic contacts were determined. In pilocarpine-treated mice, CCK was observed in dendritic spines and these were proportionally increased relative to controls, whereas the proportion of CCK-labeled terminals forming symmetric synapses was decreased. In addition, CCK-positive axon terminals forming asymmetric synapses were readily observed in these mice. Double labeling with vesicular glutamate transporter 1 and CCK revealed colocalization in numerous terminals forming asymmetric synapses, confirming the glutamatergic identity of these terminals. These data raise the possibility that expression of CCK is increased in hippocampal pyramidal cells in mice with recurrent, spontaneous seizures.
Overlapping regional distribution of CCK and TPPII mRNAs in Cynomolgus monkey brain and correlated levels in human cerebral cortex (BA 10)
2006, Brain Research
Citation Excerpt :
Intracellularly, CCK is degraded in the endosomal/lysosomal system after internalization of CCK receptor-ligand complex (Roettger et al., 1997; Tarasova et al., 1997), the lysosomal enzyme TPPI being essential for this degradation (Warburton and Bernardini, 2002). In this study, we concentrate on TPPII as it has the potential to inactivate extracellular CCK-8 and CCK-5, a fragment known to have an affinity comparable to CCK-8 to the CCK B receptor (Gaudreau et al., 1983) (predominant in the brain). Distribution of TPPII has been mostly studied in rat brain (Facchinetti et al., 1999; Rose et al., 1996; Tomkinson and Nyberg, 1995), while distribution and gene expression of CCK (Burgunder and Young, 1990; Lindefors et al., 1991, 1993; Schiffmann and Vanderhaeghen, 1991) and CCK B receptor (Cross et al., 1988; Dietl et al., 1987; Durieux et al., 1988; Mercer et al., 1996; Van Dijk et al., 1984) are also studied in primate and human brain.
Tripeptidyl peptidase II (TPPII) is a high molecular weight exopeptidase important in inactivating extracellular cholecystokinin (CCK). Our aims were to study the anatomical localization of TPPII and CCK mRNA in the Cynomolgus monkey brain as a basis for a possible functional anatomical connection between enzyme (TPPII) and substrate (CCK) and examine if indications of changes in substrate availability in the human brain might be reflected in changes of levels of TPPII mRNA. Methods: mRNA in situ hybridization on postmortem brain from patients having had a schizophrenia diagnosis as compared to controls and on monkey and rat brain slices. Results: overlapping distribution patterns of mRNAs for TPPII and CCK in rat and monkey. High amounts of TPPII mRNA are seen in the neocortex, especially in the frontal region and the hippocampus. TPPII mRNA is also present in the basal ganglia and cerebellum where CCK immunoreactivity and/or CCK B receptors have been found in earlier studies, suggesting presence of CCK-ergic afferents from other brain regions. Levels of mRNAs for CCK and TPPII show a positive correlation in postmortem human cerebral cortex Brodmann area (BA) 10. TPPII mRNA might be affected following schizophrenia. Discussion: overall TPPII and CCK mRNA show a similar distribution in rat and monkey brain, confirming and extending earlier studies in rodents. In addition, correlated levels of TPPII and CCK mRNA in human BA 10 corroborate a functional link between CCK and TPPII in the human brain.
Atypical behavioural responses to CCK-B receptor ligands in Fawn-Hooded rats
2003, Life Sciences
At present there is an increasing literature demonstrating heterogeneity of the CCK-B receptor. Recent reports by our laboratory have demonstrated that the Fawn-Hooded rat demonstrates atypical neurochemical responses to CCK₄, in vitro. Since the ability of CCK-B receptor ligands to modulate affective state is dependent on the putative receptor subtype activated, the aim of the present study was to examine the behavioural effects of the CCK-B receptor agonist, t-boc-CCK₄, and the CCK-B receptor antagonist, Ci-988 in Fawn-Hooded and Wistar Kyoto rats. Interestingly, both t-boc-CCK₄ and Ci-988 produced an anxiolytic profile in FH rats as determined by an increased time spent on the open arms of an elevated plus maze, while both drugs were devoid of any behavioural effect in WKY rats, lending further support to the theory that the FH rat strain has an atypical relative proportion of these putative subtypes apparently resulting in a predominantly CCK-B₂ receptor effect.
Neuroendocrine pharmacology of stress
2003, European Journal of Pharmacology
Exposure to hostile conditions initiates responses organized to enhance the probability of survival. These coordinated responses, known as stress responses, are composed of alterations in behavior, autonomic function and the secretion of multiple hormones. The activation of the renin–angiotensin system and the hypothalamic–pituitary–adrenocortical axis plays a pivotal role in the stress response. Neuroendocrine components activated by stressors include the increased secretion of epinephrine and norepinephrine from the sympathetic nervous system and adrenal medulla, the release of corticotropin-releasing factor (CRF) and vasopressin from parvicellular neurons into the portal circulation, and seconds later, the secretion of pituitary adrenocorticotropin (ACTH), leading to secretion of glucocorticoids by the adrenal gland. Corticotropin-releasing factor coordinates the endocrine, autonomic, behavioral and immune responses to stress and also acts as a neurotransmitter or neuromodulator in the amygdala, dorsal raphe nucleus, hippocampus and locus coeruleus, to integrate brain multi-system responses to stress. This review discussed the role of classical mediators of the stress response, such as corticotropin-releasing factor, vasopressin, serotonin (5-hydroxytryptamine or 5-HT) and catecholamines. Also discussed are the roles of other neuropeptides/neuromodulators involved in the stress response that have previously received little attention, such as substance P, vasoactive intestinal polypeptide, neuropeptide Y and cholecystokinin. Anxiolytic drugs of the benzodiazepine class and other drugs that affect catecholamine, GABA_A, histamine and serotonin receptors have been used to attenuate the neuroendocrine response to stressors. The neuroendocrine information for these drugs is still incomplete; however, they are a new class of potential antidepressant and anxiolytic drugs that offer new therapeutic approaches to treating anxiety disorders. The studies described in this review suggest that multiple brain mechanisms are responsible for the regulation of each hormone and that not all hormones are regulated by the same neural circuits. In particular, the renin–angiotensin system seems to be regulated by different brain mechanisms than the hypothalamic–pituitary–adrenal system. This could be an important survival mechanism to ensure that dysfunction of one neurotransmitter system will not endanger the appropriate secretion of hormones during exposure to adverse conditions. The measurement of several hormones to examine the mechanisms underlying the stress response and the effects of drugs and lesions on these responses can provide insight into the nature and location of brain circuits and neurotransmitter receptors involved in anxiety and stress.

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Characterization and visualization of cholecystokinin receptors in rat brain using [3H]pentagastrin

Abstract

Brain Res

Neuropeptides

Neuropeptides

Life Sci

Life Sci

Science

Eur J Pharmacol

Can J Physiol Pharmacol

Ann Neurol

Peptides

J Neurosci

Brain Res

Acta Physiol Scand

Life Sci

Nature

Neuroscience

Biochem Biophys Res Commun

Peptides

Characterization and visualization of cholecystokinin receptors in rat brain using [³H]pentagastrin