Localization of serotonin in the central nervous system by immunohistochemistry: Description of a specific and sensitive technique and some applications

Steinbusch, Harry W.M.; Verhofstad, A.A.J.; Joosten, H.W.J.

doi:10.1016/0306-4522(78)90033-7

Cited by 525 publications

(106 citation statements)

References 40 publications

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“…Somatodendritic 5HT1A autoreceptors are found in both the dorsal (DRN) and median (MRN) raphé nuclei, which send topographically distinct, as well as overlapping, serotonin projections to glutamatergic nuclei that innvervate the accumbens (eg Azmitia and Segal, 1978;Steinbusch et al, 1978;Herve et al, 1987;Vertes, 1991;Vertes et al, 1999). Whereas DRN and MRN neurons dually innervate some aspects of the frontal cortex (eg cingulate, infralimbic, and prelimbic cortices), the hippocampus, the mediodorsal nucleus of the thalamus, and the basolateral and central nuclei of the amygdala (Vertes, 1991;Vertes et al, 1999), the DRN innervates strongly the majority of thalamic and amygdalar nuclei, as well as other cortical regions (incl., parietal, occipital, frontal orbital, and piriform cortices) (Vertes, 1991;Vertes et al, 1999).…”

Section: Introductionmentioning

confidence: 99%

Dissociable Roles for the Dorsal and Median Raphé in the Facilitatory Effect of 5-HT1A Receptor Stimulation upon Cocaine-Induced Locomotion and Sensitization

Szumlinski

Frys

Kalivas

2004

Neuropsychopharmacol

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A distinct role for serotonin transmission from the dorsal and median raphé nuclei (DRN and MRN, respectively) was identified in regulating the behavioral and neurochemical effects of acute and repeated cocaine administration. Serotonin 1A (5-hydroxytryptophan (5-HT)1A) receptors were stimulated by intraraphé microinjection of 8-hydroxy-2-(di-n-propylamino)tetralin (DPAT; 5 or 10 mg) and behavior, as well as extracellular neurotransmitter content in the nucleus accumbens was measured. Pretreatment of the DRN with DPAT caused a sensitization-like potentiation of acute cocaine-induced motor activity and an elevation in extracellular dopamine and glutamate. In contrast, DPAT microinjection into the MRN did not alter acute cocaine-induced motor activity or extracellular levels of dopamine or glutamate. Acutely, DPAT microinjection into either raphé nucleus reduced the basal and acute cocaine-stimulated levels of extracellular serotonin. Pretreatment with DPAT before systemic cocaine administration was continued for 5 days, and 3 weeks after the last injection, all rats were administered a cocaine challenge injection. The sensitized behavioral and neurochemical response produced by repeated cocaine in control subjects was unaffected by the intra-DRN administration of DPAT. However, in animals administered DPAT into the MRN, both the sensitized motor response and the increase in glutamate were augmented, while the sensitized serotonin response was blocked, without altering dopamine sensitization. These data show a differential role for 5-HT1A receptors in the DRN and MRN in the acute and sensitized effects of cocaine. While the DRN is involved in the acute effects of cocaine, neuroadaptations in the MRN may regulate the long-term consequences of repeated cocaine exposure.

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Section: Introductionmentioning

confidence: 99%

Dissociable Roles for the Dorsal and Median Raphé in the Facilitatory Effect of 5-HT1A Receptor Stimulation upon Cocaine-Induced Locomotion and Sensitization

Szumlinski

Frys

Kalivas

2004

Neuropsychopharmacol

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“…The GFA antiserum has been used previously in this laboratory to specifically localize astrocytes in tissue sections (8,9). The specificity ofthese antisera to 5-HT and GFA antisera is well documented (10)(11)(12) The publication costs ofthis article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U. S. C. §1734 solely to indicate this fact.…”

mentioning

confidence: 99%

Immunocytochemical demonstration of monoamine oxidase B in brain astrocytes and serotonergic neurons.

Levitt

Pintar

Breakefield

1982

Proc. Natl. Acad. Sci. U.S.A.

445

214

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An antiserum to monoamine oxidase B (MAO-B) was used to define the distribution of this metabolic enzyme in the adult rat brain immunocytochemically. MAO-B is specifically located in two major central nervous system cell classes, astrocytes and serotonin-containing neurons. Double-immunofluorescence experiments using antisera to glial fibrillary acidic protein and MAO-B showed that both protoplasmic and fibrillary astrocytes throughout the brain contain MAO-B, whereas oligodendrocytes do not contain the enzyme. Areas lacking a blood-brain barrier, such as the specialized circumventricular organs, also contain MAO-B-positive cells. A double-immunofluorescence experiment using antisera to serotonin and MAO-B enabled the positive identification of neurons containing both molecules. The catecholamine-containing neurons of the brain did not contain detectable amounts of MAO-B. The specific distribution of MAO-B (1,2). MAO activity has been subdivided on a pharmacological basis into two types: MAO-A has a higher affinity for the substrates norepinephrine and serotonin (5-HT) and the inhibitor clorgyline, and MAO-B has a higher affinity for the substrates phenylethylamine and benzylamine and the inhibitor deprenyl. Recent biochemical evidence indicates that these activities are mediated by two distinct mitochondrial proteins that have different molecular weights (3-5). Both types of MAO activity can be measured in all CNS regions, but it has been difficult to elucidate the cellular distribution of MAO with current pharmacological and biochemical methods because of the diversity of neuronal and nonneuronal cell types in the brain. By using an antiserum directed against MAO-B and immunocytochemical methods, we have localized MAO-B Immunocytochemistry. Every third section, 10 ,Lm thick, through the entire brain was collected on a gelatin-coated slide and immunostained by using rabbit preimmune serum or antiserum prepared against pure bovine ; unpublished data; MAO-B was obtained from James Salach and Walter Weyler). Sections on slides were incubated for 5 min with 1% sodium borohydride in 0.1 M sodium phosphate (pH 7.2) and then for 3 hr at 370C with either MAO-B antiserum or preimmune serum diluted 1: 100 in a staining buffer of0.45 M NaCl/ 20 mM sodium phosphate, pH 7.4/0.3% Triton X-100/10% normal swine serum. The sections were washed for 30 min with the staining buffer and exposed to rhodamine-conjugated swine anti-rabbit IgG (Dako, Accurate Scientific) diluted 1: 40 in the staining buffer. Sections were washed for 30 min and placed under a coverslip with a glycerol/0. 1 M sodium phosphate, pH 7.2 (3:1), solution. The slides were examined on a Zeiss photomicroscope equipped with epifluorescence and photographed on Kodak Plus-X film using 1-to 2-min exposures. Some tissue sections were processed by using the peroxidaseantiperoxidase procedure (8,9). The cell cultures (see below) were fixed with 4% paraformaldehyde in 0.1 M sodium borate buffer (pH 11.0) for 90 min at 4°C, rinsed extensively, and then st...

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“…However, it is unlikely that such a general increase in excitability caused the increased responsiveness to 5-HT as responses to NA were not affected by dorsal root section and responses to ERP were not significantly larger. Although several authors found that the section of afferent pathways in the CNS failed to cause a specific supersensitivity (Krnjevic, Reiffenstein & Silver, 1970;Macon, 1979) some recent reports show that selective increases in sensitivity to a known neurotransmitter can occur (Wald, 1976;Haas, Wolf, Palacios, Garbarg, Barbin & Schwartz, 1978;McCall & Aghajanian, 1979 (Steinbusch, Verhofstad & Joosten, 1978). Although Lackovic (1978) gave a preliminary report that 5-HT may exist in primary afferents and Shibuya & Anderson (1968) showed that 25 % of 5-HT in the spinal cord survived total spinal transection, these reports have not yet been confirmed or explained.…”

Section: Discussionmentioning

confidence: 86%

The Effects of Chronic Section of Dorsal Roots on the Responsiveness of Motoneurones to 5‐hydroxytryptamine and a Substance P Analogue

Roberts¹,

Wright²

1981

British J Pharmacology

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1 Experiments were performed on rats 14-21 days after unilateral dorsal root section in order to determine if the effects on motoneurone excitability of a substance P analogue, 5-hydroxytryptamine (5-HT) and noradrenaline were altered by section of primary afferents.2 The effects of iontophoretic applications of these agents on motoneurone excitability were quantified by measuring the change in amplitude of the short latency field potentials evoked antidromically, by ventral root stimulation. 3 lontophoretic application of the substance P analogue (eledoisin-related peptide, ERP) always produced an increase in the amplitude of the field potential. These increases in amplitude were 25.9% larger on the sides of the cords with sectioned dorsal roots. This was not a statistically significant difference (P>0.05). 4 Section of dorsal roots did not alter responses to noradrenaline. 5 Responses to 5-HT were significantly larger following section of dorsal roots. There is very little evidence for the release of 5-HT by primary afferents and denervation supersensitivity is an improbable explanation. It is possible that descending 5-HT systems directly excite motoneurones and indirectly inhibit primary afferent transmission. Dorsal root section would alter the balance between these actions of 5-HT in favour of an excitatory effect.

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Localization of serotonin in the central nervous system by immunohistochemistry: Description of a specific and sensitive technique and some applications

Cited by 525 publications

References 40 publications

Dissociable Roles for the Dorsal and Median Raphé in the Facilitatory Effect of 5-HT1A Receptor Stimulation upon Cocaine-Induced Locomotion and Sensitization

Dissociable Roles for the Dorsal and Median Raphé in the Facilitatory Effect of 5-HT1A Receptor Stimulation upon Cocaine-Induced Locomotion and Sensitization

Immunocytochemical demonstration of monoamine oxidase B in brain astrocytes and serotonergic neurons.

The Effects of Chronic Section of Dorsal Roots on the Responsiveness of Motoneurones to 5‐hydroxytryptamine and a Substance P Analogue

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