Studies of the direct coloring thiocholine method for localizing cholinesterase activity

Broderson, Stevan H.; Westrum, Lesnick E.; Sutton, Alexander E.

doi:10.1007/bf00490269

Cited by 16 publications

(7 citation statements)

References 15 publications

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“…Reaction product was developed with sodium sulfide in 0.2 M acetic acid after incubation times of up to 48 hr. Some tissue was processed by the "direct coloring" method of Karnovsky and Roots (1964) with the incubation medium and inhibitors suggested by Broderson et al (1974). In selected sections, staining was intensified with 0.1 to 1.0% silver nitrate (Geneser-Jensen and Blackstad, 1971).…”

Section: Methodsmentioning

confidence: 99%

Development of prestriate visual projections in the monkey and human fetal cerebrum revealed by transient cholinesterase staining

Kostović¹,

Rakić²

1984

J. Neurosci.

255

148

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Cholinesterase (ChE) staining was used to reveal the timing and pattern of development of afferents to the prestriate visual cortex (areas 18, 19, 20, and 21 of Brodmann) in a series of developing human and monkey fetal brains. This investigation was possible because the nucleus pulvinaris of the thalamus, the main source of subcortical projections to the prestriate cortex, displays positive reactivity after thiocholine incubation during the last three quarters of gestation, while neighboring thalamic nuclei that project to the adjacent neocortical areas are unstained. Staining of the pulvinar and its prestriate projections passes through six broad stages. Stage I begins in both species at the end of the first third of gestation. Positively stained fibers originate from the pulvinar and enter but do not extend beyond the hemispheric stalk. During stage II, pulvinar axons gradually invade the intermediate zone of the occipital lobe, and in stage III they reach the level of the subplate zone. In stage IV, which occurs around mid-gestation in both species, cholinesterase-positive fibers accumulate within the subplate zone subjacent to the developing prestriate cortex. During stage V, ChE-positive fibers penetrate the prospective prestriate cortex but do not yet form the alternating columnar pattern characteristic of pulvinar input to this area in the adults. Rather, ChE activity is concentrated in two continuous bands situated within prospective layers III-IV and VI; also a narrow band is visible in upper layer I. In stage V a clear histochemical border forms between prestriate and striate areas with ChE activity in prospective area 17 limited mostly to the superficial strata of layers I and II. This histochemical differentiation precedes the emergence of cytoarchitectonic landmarks. During stage VI, which begins in the last fifth of gestation in both species, the pulvinar become progressively less stainable and its projections can no longer be traced by ChE histochemistry.

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Section: Methodsmentioning

confidence: 99%

Development of prestriate visual projections in the monkey and human fetal cerebrum revealed by transient cholinesterase staining

Kostović¹,

Rakić²

1984

J. Neurosci.

255

148

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“…As eserine inhibited all staining, whereas isoOMPA had little effect, this indicated that the reaction obtained is due to the true AChE. The reaction product was developed with sodium sulfide in 0.2 M acetic acid after incubation for up to 24 h. Some sections were also processed by the Karnovsky-Roots "direct coloring" method (Karnovsky-Roots, 1964) as described by Broderson et al (1974) and modified by Tago et al (1986).…”

Section: Ache Histochemistrymentioning

confidence: 99%

Prenatal development of the human entorhinal cortex

Šimić

Krsnik

Knezović

et al. 2022

J of Comparative Neurology

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Little is known about the development of the human entorhinal cortex (EC), a major hub in a widespread network for learning and memory, spatial navigation, high‐order processing of object information, multimodal integration, attention and awareness, emotion, motivation, and perception of time. We analyzed a series of 20 fetal and two adult human brains using Nissl stain, acetylcholinesterase (AChE) histochemistry, and immunocytochemistry for myelin basic protein (MBP), neuronal nuclei antigen (NeuN), a pan‐axonal neurofilament marker, and synaptophysin, as well as postmortem 3T MRI. In comparison with other parts of the cerebral cortex, the cytoarchitectural differentiation of the EC begins remarkably early, in the 10th week of gestation (w.g.). The differentiation occurs in a superficial magnocellular layer in the deep part of the marginal zone, accompanied by cortical plate (CP) condensation and multilayering of the deep part of CP. These processes last until the 13–14th w.g. At 14 w.g., the superficial lamina dissecans (LD) is visible, which divides the CP into the lamina principalis externa (LPE) and interna (LPI). Simultaneously, the rostral LPE separates into vertical cell‐dense islands, whereas in the LPI, the deep LD emerges as a clear acellular layer. In the 16th w.g., the LPE remodels into vertical cell‐dense and cell‐sparse zones with a caudorostral gradient. At 20 w.g., NeuN immunoreactivity is most pronounced in the islands of layer II cells, whereas migration and differentiation inside‐out gradients are seen simultaneously in both the upper (LPE) and the lower (LPI) pyramidal layers. At this stage, the EC adopts for the first time an adult‐like cytoarchitectural organization, the superficial LD becomes discernible by 3T MRI, MBP‐expressing oligodendrocytes first appear in the fimbria and the perforant path (PP) penetrates the subiculum to reach its molecular layer and travels along through the Cornu Ammonis fields to reach the suprapyramidal blade of the dentate gyrus, whereas the entorhinal‐dentate branch perforates the hippocampal sulcus about 2–3 weeks later. The first AChE reactivity appears as longitudinal stripes at 23 w.g. in layers I and II of the rostrolateral EC and then also as AChE‐positive in‐growing fibers in islands of superficial layer III and layer II neurons. At 40 w.g., myelination of the PP starts as patchy MBP‐immunoreactive oligodendrocytes and their processes. Our results refute the possibility of an inside‐out pattern of the EC development and support the key role of layer II prospective stellate cells in the EC lamination. As the early cytoarchitectural differentiation of the EC is paralleled by the neurochemical development, these developmental milestones in EC structure and connectivity have implications for understanding its normal function, including its puzzling modular organization and potential contribution to consciousness content (awareness), as well as for its insufficiently explored deficits in developmental, psychiatric, and degenerative brain disorders.

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“…As described in other studies, there is a random, nonspecific scattering of crystalline precipitate over the tissue in all preparations. This is a n artifact of the tissue preparation and not associated with enzymatic activity or the distribution of reaction product (Broderson et al, 1974;Westrum and Broderson, 1976).…”

Section: Normalmentioning

confidence: 95%

“…Tissue was osmicated in the same buffer for %-1 h and prepared for electron microscopy by the usual methods (Westrum and Black, 1971). For additional details, see Broderson et al (1974) and Westrum and Broderson (1976). Semithin (1 pm) sections were made for orientation purposes; then the blocks trimmed for the specific nuclei and sublayers.…”

Section: Electron Microscopymentioning

confidence: 99%

“…Acetylcholinesterase (AChE), the glycoprotein enzyme that hydrolyses the neurotransmitter acetylcholine (ACh), has important functional implications in the nervous system and is particularly well known for its role in the termination of cholinergic transmission. The concentration and distribution of AChE in normal and injured nerves have been studied more or less along the entire neuronal axis, including peripheral endings, both sensory endings and motor end-plates (Gautron et al, 1983; McConnell and Simpson, 1976), pre-and postganglionic fibers (Malatova et al, 1985), cell bodies (Malatova et al, 1985; Pannese et al, 1974), and in terminal synaptic regions (Broderson et al, 1974; McDonald and Rasmussen, 1971; Westrum and Broderson, 1976).…”

Section: Introductionmentioning

confidence: 99%

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Ultrastructural changes in acetylcholinesterase activity in the deafferented spinal trigeminal nucleus

Wilson

Westrum

Broderson

1988

Synapse

Self Cite

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Acetylcholinesterase (AChE) activity was investigated in synaptic areas of the cat spinal trigeminal nucleus (pars interpolaris and pars caudalis) ipsilateral and contralateral to complete retrogasserian rhizotomy. Vibratome sections of tissue taken from animals of 1, 3, 6, 14, and 21 days survival were examined by electron microscopy following a histochemical reaction for AChE activity employing a method based on the Karnovsky-Roots technique for demonstrating reaction product. As degeneration progressed with survival time, enzymatic activity was initially reduced in synaptic clefts of injured afferent terminals and subsequently was enhanced throughout the extracellular space, including within synaptic clefts of possibly reinnervated sites. These changes in enzymatic activity with primary deafferentation are discussed in relation to the process of reinnervation, the development of neuronal hyperactivity, and possible noncholinergic functions of AChE.

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Studies of the direct coloring thiocholine method for localizing cholinesterase activity

Cited by 16 publications

References 15 publications

Development of prestriate visual projections in the monkey and human fetal cerebrum revealed by transient cholinesterase staining

Development of prestriate visual projections in the monkey and human fetal cerebrum revealed by transient cholinesterase staining

Prenatal development of the human entorhinal cortex

Ultrastructural changes in acetylcholinesterase activity in the deafferented spinal trigeminal nucleus

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