Localization of the Locus Coeruleus in the Mouse Brain

Schmidt, Katharina; Bari, Bilal A.; Ralle, Martina; Washington-Hughes, Clorissa; Muchenditsi, Abigael; Maxey, Evan; Lutsenko, Svetlana

doi:10.3791/58652

Cited by 15 publications

(12 citation statements)

References 34 publications

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“…The protocol reported by Schmidt et al [12] is very detailed; however, the operation is much more complicated: (1) The cutting plane is determined with an adult mouse brain slicer matrix and at Bregma -3 mm [12]; however, there are individual differences in mouse brain volume and the degree of dehydration also affects brain volume. At the same time, the brain tissue becomes hard after dehydration with sucrose and may not fit well with the slicer matrix, so there may be large individual differences for the measure of the positioning line.…”

Section: Discussionmentioning

confidence: 99%

“…Mice are widely used experimental animals, but mouse LC is small (about 500 μm in sagittal plane) [11] and lacks obvious localization landmarks, making it difficult for novices to accurately and quickly locate in brain slice sectioning. Schmidt et al [12] provided a step-by-step protocol to localize LC in mouse brain sectioning; however, the operations are complicated. Here we report an easy-to-follow method that quickly and stably determines the cutting plane and locates the LC, as well as the noradrenergic neuron most enriched planes.…”

Section: Introductionmentioning

confidence: 99%

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Fast Localization and Sectioning of Mouse Locus Coeruleus

Cao

Yuan

et al. 2020

BioMed Research International

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The locus nucleus (LC) is a multifunctional nucleus which is also the source of norepinephrine in the brain. To date, there is no simple and easy method to locate the small LC in brain sectioning. Here we report a fast, accurate, and easy-to-follow protocol for the localization of mice LC in frozen sectioning. After fixation and dehydration, the intact brains of adult mice were placed on a horizontal surface and vertically cut along the posterior margin of the bilateral cerebral cortex. In the coronal cutting plane, the aqueduct of midbrain can be seen easily with the naked eyes. After embedding the cerebellum part with optimal cutting temperature (OCT) compound, coronal brain slices were cut from the cutting plane, within 1 mm, the aqueduct of midbrain disappeared and the fourth ventricle appeared, then the brain slices contained LC and were collected. From the first collection, at~200 μm, the noradrenergic neurons' most enriched brain slices can be collected. The tyrosine hydroxylase immunofluorescence staining confirmed that the localization of LC with this method is accurate and the noradrenergic neuron most abundant slices can be determined with this method.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Fast Localization and Sectioning of Mouse Locus Coeruleus

Cao

Yuan

et al. 2020

BioMed Research International

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“…Immunocytochemistry was performed to confirm and extend the catecholamine fluorescence data, by staining the neuronal dendrite marker, microtubule-associated protein 2 (MAP2) and the noradrenergic marker, dopamine β-hydroxylase (DBH), the enzyme that synthesizes norepinephrine (Foote et al, 1983;Sanchez-Padilla et al, 2014;Schmidt et al, 2019). Dissociated neurons were fixed with 4% paraformaldehyde and 4% sucrose in the standard external solution at 4°C for 30 min.…”

Section: Characterization Of Lc Neurons By Immunocytochemistrymentioning

confidence: 99%

Calcium-induced calcium release in noradrenergic neurons of the locus coeruleus

et al. 2020

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The locus coeruleus (LC) is a nucleus within the brainstem that consists of norepinephrinereleasing neurons. It is involved in broad processes including autonomic regulation, and cognitive and emotional functions such as arousal, attention and anxiety. Understanding the mechanisms that control the excitability of LC neurons is important because they innervate widespread regions of the central nervous system. One of the key regulators is the cytosolic calcium concentration ([Ca 2+ ]c), the increases in which can be amplified by calcium-induced calcium release (CICR) from the intracellular calcium stores. Although the electrical activities of LC neurons are regulated by changes in [Ca 2+ ]c, the extent of CICR involvement in this regulation has remained unclear. Here we show that CICR hyperpolarizes acutely dissociated LC neurons of the rat brain and demonstrate the pathway whereby it does this. When CICR was activated by extracellular application of 10 mM caffeine, LC neurons were hyperpolarized in the current-clamp mode of the patch-clamp recording, and the majority of neurons showed an outward current in the voltage-clamp mode. This outward current was accompanied by an increase in membrane conductance, and its reversal potential was close to the K + equilibrium potential, indicating that it is mediated by the opening of K + channels. The outward current was generated in the absence of extracellular calcium and was blocked when the calcium stores were inhibited by applying ryanodine. Pharmacological experiments indicated that the outward current was mediated by Ca 2+ -activated K + channels of the nonsmall conductance type. Finally, the application of caffeine led to an increase in the [Ca 2+ ]c in these neurons, as visualized by fluorescence microscopy. These findings delineate a mechanism whereby CICR suppresses the electrical activity of LC neurons, and indicate that it could play a dynamic role in modulating the LC-mediated noradrenergic tone in the brain. RUNNING TITLECa 2+ release in locus coeruleus neurons KEYWORDS Ca 2+ -activated K + current, Ca 2+ -induced Ca 2+ release, caffeine, Ca 2+ imaging, dopamine βhydroxylase, intracellular Ca 2+ store, patch-clamp recording..

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“…Dopamine β-hydroxylase (DβH) is the key regulatory enzyme required to synthesize NE from DA (21), and it is also important to maintain the brain DA/NE balance. DβH is located in both central (catecholamine vesicles) and peripheral systems (sympathetic nerves and adrenal medulla) (22,23). Since DβH can be released from catecholamine vesicles, the protein can be determined in the plasma or serum (21).…”

Section: Introductionmentioning

confidence: 99%

Reduced Plasma Dopamine-β-Hydroxylase Activity Is Associated With the Severity of Bipolar Disorder: A Pilot Study

Sun

Mao

et al. 2021

Front. Psychiatry

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Dopamine-β-hydroxylase (DβH) is an enzyme converting dopamine to norepinephrine, a key neurotransmitter in mood disorders, such as major depressive disorder (MDD) and bipolar disorder (BD). Due to overlapping symptomology of unipolar and bipolar depression, the present study attempted to explorer if the plasma DβH activity could discriminate the depressive episodes of BD from MDD. The aim of this study was to compare the plasma DβH activity among MDD patients (n = 104), BD patients (n = 101), and healthy controls (n = 160). Clinical characteristics and cognitive function were assessed using the Young Mania Rating Scale (YMRS), Hamilton Depression Scale (HAM-D), Hamilton Anxiety Scale (HAM-A), Patient Health Questionnaire-9 (PHQ-9), and Repeatable Battery for the Assessment of Neuropsychological Status (RBANS). Our data showed a lower plasma DβH activity in patients with BD, not MDD, than that in controls. For the BD patients, the plasma DβH activities were negatively correlated with HAM-D scores and HAM-A scores. However, there was no significant correlation between plasma DβH activity and severity of depressive symptoms in MDD patients. No significant correlation between DβH activities and cognitive assessments neither in BD nor in MDD patients. The present study provides evidence that BD is associated with decreased circulating DβH activity.

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Localization of the Locus Coeruleus in the Mouse Brain

Cited by 15 publications

References 34 publications

Fast Localization and Sectioning of Mouse Locus Coeruleus

Fast Localization and Sectioning of Mouse Locus Coeruleus

Calcium-induced calcium release in noradrenergic neurons of the locus coeruleus

Reduced Plasma Dopamine-β-Hydroxylase Activity Is Associated With the Severity of Bipolar Disorder: A Pilot Study

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