A one-step cobalt-ferrocyanide method for histochemical demonstration of acetylcholinesterase activity in central nervous system tissue.

Schätz, Christoph R.; Geula, Changiz; Morecraft, Robert J.; Mesulam, Marsel

doi:10.1177/40.3.1552180

Cited by 9 publications

(8 citation statements)

References 10 publications

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“…Acteylcholinesterase (AChE) activity was visualized by using a modified "Karnowski-protocol" (Schatz and Veh, 1987;Schatz et al, 1992). Briefly, cryostat or Vibratome sections were rinsed in 0.1 M maleate buffer, pH 6.3, and preincubated for 30 minutes in a solution containing 3.15 mM cobalt chloride and 5 mM sodium citrate in 0.1 M maleate buffer.…”

Section: Acetylcholinesterase Cytochemistrymentioning

confidence: 99%

Morphologic and cytochemical criteria for the identification and delineation of individual subnuclei within the lateral habenular complex of the rat

Geisler

Andres

Veh

2003

J of Comparative Neurology

117

142

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The lateral habenular complex is part of the habenular nuclei, a distinct structure in the dorsal diencephalon of all vertebrates. In contrast to the bewildering diversity of behaviors, in which the lateral habenular complex is thought to be involved, there is an astonishing lack of information concerning its cellular organization, its neuronal circuits, and the neurophysiological mechanisms, which may provide the physiological and molecular basis for its diverse biological functions. This problem may be due to an unexpected heterogeneity of the lateral habenular complex. Recently, a detailed subnuclear organization has been described (Andres et al. [1999] J Comp Neurol 407:130-150), which provides the base for a subsequent physiological and behavioral analysis of this area. Available criteria, however, can be applied to semithin sections only. To facilitate further investigations, the present work aimed to elaborate novel morphologic and immunocytochemical criteria that can be applied to conventional cryostat or Vibratome sections to allow identification and delineation of subnuclei of the lateral habenular complex. Consequently, the regional, cellular, and subcellular localization of approximately 30 different neuroactive molecules was investigated. Of these candidate molecules, gamma-aminobutyric acid-B receptor protein, Kir3.2 potassium channel protein, tyrosine hydroxylase, and neurofilament heavy chain proved to be suitable markers. Our observation suggests that the habenular subnuclei express distinct immunocytochemical characteristics. These features may be used to identify and delineate the subnuclei on conventional cryostat or Vibratome sections. From our results, it is expected that the further functional analysis of the lateral habenular complex will be facilitated considerably.

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Section: Acetylcholinesterase Cytochemistrymentioning

confidence: 99%

Morphologic and cytochemical criteria for the identification and delineation of individual subnuclei within the lateral habenular complex of the rat

Geisler

Andres

Veh

2003

J of Comparative Neurology

117

142

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“…Morphological analysis of the vertebrate central nervous system conventionally requires the visualization of nerve cell bodies and myelinated fibers, more recently complemented by staining for acetylcholinesterase reactivity (Koelle and Friedenwald 1949;Schätz et al 1992).…”

Section: Discussionmentioning

confidence: 99%

“…While classic neuroanatomical investigations (Brodmann 1914;Vogt and Vogt 1919) were predominantly based on the study of Nissl-stained sections (Nissl 1894), in recent times acetylcholine esterase histochemistry is increasingly appreciated (Koelle and Friedenwald 1949;Schätz and Veh 1987;Schätz et al 1992). When the acetylcholine esterase staining is applied to the truncothalamic complex distinct nuclei can be easily distinguished (Paxinos and Watson 1998).…”

Section: Introductionmentioning

confidence: 98%

An optimized method for simultaneous demonstration of neurons and myelinated fiber tracts for delineation of individual trunco- and palliothalamic nuclei in the mammalian brain

Geisler

Heilmann

Veh

2001

Histochem Cell Biol

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The truncothalamic complex has long been considered to be a nuclear group with "non-specific" projections. More recently, it is suggested that these thalamic nuclei play an important role in regulating distinct basal ganglia circuits. To further analyze the exact biological function of individual nuclei of the truncothalamic complex a simple and reliable technique for an exact delineation of distinct nuclei is desirable. Therefore, we evaluated and optimized several potential procedures for a combined visualization of neurons and myelinated fibers. Fiber staining with gold toning or immunocytochemical visualization of myelin basic protein shows high contrast and precision but precludes sufficient demonstration of neuronal cell bodies. When the most common technique for the simultaneous visualization of both structures, the Kluver-Barrera procedure, is used, demonstration of myelinated fibers is restricted when the technique is applied to cryostat or vibratome sections. In the present report this limitation was abolished. The final protocol includes lipid extraction prior to the incubation with Luxol Fast Blue and uses carefully characterized staining conditions for Luxol Fast Blue and cresyl violet rendering microscopically controlled differentiation steps unnecessary. The optimized Kluver-Barrera technique results in high precision localization of individual axons and cell bodies and thus permits an exact and simple delineation of individual nuclei in the vertebrate thalamus.

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“…Another shortcoming of the present method is that when DFP pretreatment is not possible (e.g., human postmortem specimens), visualization of AChE-positive somata may be obscured by non-somatal staining. Under these circumstances, immunohistochemical (Mesulam et al 1994) or alternative enzymatic methods (Kujat et al 1993;Schatz et al 1992;Kugler 1987;Tago et al 1986;Hedreen et al 1985) for AChE staining may be preferable when coupled with other methods of MAO staining (e.g., Nakos and Gossrau 1993;Gossrau and Richter 1992).…”

Section: Discussionmentioning

confidence: 99%

A Simple Enzyme Histochemical Method for the Simultaneous Demonstration of Acetylcholinesterase and Monoamine Oxidase in Fixed-Frozen Sections

Dunning

McHaffie

Stein

1997

J Histochem Cytochem.

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SUMMARYWe describe an enzyme histochemical technique for the simultaneous demonstration of acetylcholinesterase (AChE) and monoamine oxidase (MAO) (Types A, B, or A ϩB) in fixed-frozen sections. Several regions in the mesencephalon and brainstem were examined for both somatic and neuropil labeling. The results obtained are equivalent or superior to those obtained using previous methods for the individual localization of these enzymes. The simultaneous visualization of AChE and MAO in the same section allows the relationship of the two enzymes to be easily assessed with brightfield microscopy.

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A one-step cobalt-ferrocyanide method for histochemical demonstration of acetylcholinesterase activity in central nervous system tissue.

Cited by 9 publications

References 10 publications

Morphologic and cytochemical criteria for the identification and delineation of individual subnuclei within the lateral habenular complex of the rat

Morphologic and cytochemical criteria for the identification and delineation of individual subnuclei within the lateral habenular complex of the rat

An optimized method for simultaneous demonstration of neurons and myelinated fiber tracts for delineation of individual trunco- and palliothalamic nuclei in the mammalian brain

A Simple Enzyme Histochemical Method for the Simultaneous Demonstration of Acetylcholinesterase and Monoamine Oxidase in Fixed-Frozen Sections

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