Application of the silver-gold intensified 3,3′ -diaminobenzidine chromogen to the light and electron microscopic detection of the luteinizing hormone-releasing hormone system of the rat brain

Liposits, Zsolt; Sétáló, G.; Flerkó, Béla

doi:10.1016/0306-4522(84)90245-8

Cited by 198 publications

(84 citation statements)

References 35 publications

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“…After a brief rinse in Tris-buffered saline, pH 7.60, Hcrt-immunoreactive (-ir) elements were visualized using a nickel-diaminobenzidine chromogen that revealed Hcrt fibers in black. The nickel-diaminobenzidine precipitate was then silver-gold intensified, as described previously (Liposits et al, 1984), to increase the contrast in electron density between Hcrt and cholinergic structures, but the thyoglycolic acid pretreatment was omitted from the original protocol.…”

Section: Methodsmentioning

confidence: 99%

Hypocretin/Orexin Innervation and Excitation of Identified Septohippocampal Cholinergic Neurons

Wu¹,

Záborszky²,

Hajszán³

et al. 2004

J. Neurosci.

131

101

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Hypothalamic fibers containing the wake-promoting peptides, hypocretins (Hcrts) or orexins, provide a dense innervation to the medial septum-diagonal band of Broca (MSDB), a sleep-associated brain region that has been suggested to show intense axonal degeneration in canine narcoleptics. The MSDB, via its cholinergic and GABAergic projections to the hippocampus, controls the hippocampal theta rhythm and associated learning and memory functions. Neurons of the MSDB express very high levels of the Hcrt receptor 2, which is mutated in canine narcoleptics. In the present study, we investigated the electrophysiological effects of Hcrt peptides on septohippocampal cholinergic neurons that were identified in living brain slices of the MSDB using a selective fluorescent marker. Hcrt activation of septohippocampal cholinergic neurons was reversible, reproducible, and concentration dependent and mediated via a direct postsynaptic mechanism. Both Hcrt1 and Hcrt2 activated septohippocampal cholinergic neurons with similar EC 50 values. The Hcrt effect was dependent on external Na ϩ , reduced by external Ba 2ϩ , and also reduced in recordings with CsCl-containing electrodes, suggesting a dual underlying ionic mechanism that involved inhibition of a K ϩ current, presumably an inward rectifier, and a Na ϩ -dependent component. The Na ϩ component was dependent on internal Ca 2ϩ , blocked by replacing external Na ϩ with Li ϩ , and also blocked by bath-applied Ni 2ϩ and KB-R7943, suggesting involvement of the Na ϩ -Ca 2ϩ exchanger. Using double-immunolabeling studies at light and ultrastructural levels, we also provide definitive evidence for a hypocretin innervation of cholinergic neurons. Thus Hcrt effects within the septum should increase hippocampal acetylcholine release and thereby promote hippocampal arousal.

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Section: Methodsmentioning

confidence: 99%

Hypocretin/Orexin Innervation and Excitation of Identified Septohippocampal Cholinergic Neurons

Wu¹,

Záborszky²,

Hajszán³

et al. 2004

J. Neurosci.

131

101

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“…The colour reaction was developed by incubating sections with 0.05% 3′3-diaminobenzidine tetrahydrochloride chromogen (Sigma) and 0.001% hydrogen peroxide in 0.05 M Tris buffer. Selected hypothalamic material was additionally stained by the silver intensification method of Liposits et al (1984).…”

Section: Immunohistochemistry (Ihc)mentioning

confidence: 99%

The effect of intracerebroventricular infusions of ghrelin on the secretory activity of the GnRH/LH system in peripubertal ewes

Wójcik-Gładysz¹,

Wańkowska²,

Gajewska³

et al. 2014

J. Anim. Feed Sci.

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“…To control for nonspecific silver intensification of the peroxidase deposit (Liposits et al, 1984(Liposits et al, , 1986) generated by the ABC reaction, two sections of postcommissural putamen that contained dense anterograde labeling from CM were processed according to the same protocol as described above, except that the immunogold localization of mGluR1a was performed first, followed by the histochemical ABC reaction to visualize BDA. The overall pattern of single and double labeling generated by this approach was the same as that resulting from the original protocol, which demonstrates the reliability and high degree of specificity of this method.…”

mentioning

confidence: 99%

Group I Metabotropic Glutamate Receptors in the Monkey Striatum: Subsynaptic Association with Glutamatergic and Dopaminergic Afferents

2003

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Group I metabotropic glutamate receptors (mGluRs) are involved in long-term synaptic plasticity and neuroprotection in the striatum, but the specific role(s) of mGluR1 and mGluR5 remain poorly understood. In this study, we used electron-microscopic immunocytochemistry to compare the pattern of subsynaptic and subcellular distribution of mGluR1a and mGluR5 in relation to putative glutamatergic and dopaminergic inputs to the monkey striatum. At the light-microscopic level, both group I mGluRs are expressed in the striatal neuropil. In addition, numerous perikarya of striatal output neurons are immunostained for mGluR5, but much less frequently for mGluR1a. At the electron-microscopic level, immunoreactivity for both receptor subtypes is primarily expressed postsynaptically in dendrites and spines, although presynaptic mGluR1a labeling of glutamatergic thalamostriatal boutons and, less frequently, dopaminergic and corticostriatal terminals is also seen. In contrast to mGluR1a, mGluR5 immunoreactivity is rarely encountered presynaptically. In postsynaptic elements, 40 -70% of immunoreactivity for both receptor subtypes is expressed intracellularly, whereas 30 -60% is apposed to the plasma membrane. More than 80% of the labeling apposed to the plasma membrane is extrasynaptic. The remaining 20% is located at the edges of putative glutamatergic synapses or in the active zone of symmetric synapses. In mGluR5-, but not mGluR1a-immunostained sections, ϳ70% of dopaminergic symmetric synapses are labeled perisynaptically.These data emphasize the differential pattern of subsynaptic localization of the two group I mGluRs and provide various presynaptic and postsynaptic sites whereby mGluR1 and mGluR5 could mediate different, but complementary, effects on glutamatergic and dopaminergic transmission in the primate striatum.

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Application of the silver-gold intensified 3,3′ -diaminobenzidine chromogen to the light and electron microscopic detection of the luteinizing hormone-releasing hormone system of the rat brain

Cited by 198 publications

References 35 publications

Hypocretin/Orexin Innervation and Excitation of Identified Septohippocampal Cholinergic Neurons

Hypocretin/Orexin Innervation and Excitation of Identified Septohippocampal Cholinergic Neurons

The effect of intracerebroventricular infusions of ghrelin on the secretory activity of the GnRH/LH system in peripubertal ewes

Group I Metabotropic Glutamate Receptors in the Monkey Striatum: Subsynaptic Association with Glutamatergic and Dopaminergic Afferents

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