Isolation and ultrastructure of human synaptic complexes

Wannamaker, Braxton B.; Kornguth, Steven; Scott, Grayson; Dudley, Alden W.; Kelly, Alma

doi:10.1002/neu.480040607

Cited by 10 publications

(3 citation statements)

References 17 publications

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“…In the current investigation, we have observed also in both the rat and human hypothalamus that like the tissue concentration of TRH (Okon and Koch, 1976;Parker and Porter, 1982), the subcellular compartmentalization of TRH remains stable for many hours after death. The apparent postmortem stability of the subcellular compartmentalization of TRH in the rat and human is similar to that observed for several other neural substances in human and animal brain tissues (Swanson et al, 1973;Wannamaker et al, 1973;Garey et al, 1974;Klemm and Kuhar, 1979). Thus, it seems likely that by utilizing postmortem brain tissues it will be possible to investigate mechanisms controlling release, receptor binding, degradation, and synaptosomal uptake of TRH or its metabolites (Parker et al, 1977) in the human.…”

Section: Discussionsupporting

confidence: 74%

Regional Localization and Subcellular Compartmentalization of Thyrotropin‐Releasing Hormone in Adult Human Brain

Parker

Porter²

1983

Journal of Neurochemistry

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In the current study, we sought to define the subcellular compartmentalization of thyrotropin-releasing hormone (TRH) in adult human brain tissues. Upon evaluating tissues (3-24 h post mortem) from 62 humans, ranging in age from 5 to 75 years, we found that TRH was widely distributed throughout the brain. The highest TRH concentration (ng/mg protein) was in the stalk-median eminence region of the hypothalamus (19.3 +/- 3.3, mean +/- SE); the TRH concentration in the hypothalamus, exclusive of the stalk-median eminence, was much lower (1.7 +/- 0.2). Substantial quantities of TRH also were detected in the medulla oblongata (0.26 +/- 0.08), mammillary bodies (0.33 +/- 0.25), and optic chiasm (0.14 +/- 0.07). Lower levels of TRH were found in the amygdala (0.060 +/- 0.015) and the corpus striatum (0.033 +/- 0.010). TRH was near or below the limits of detection in tissues of the cerebral and cerebellar cortices, the olfactory bulbs, the pons, and the hippocampus. When homogenates of medial basal hypothalamic tissue (prepared in 0.32 M sucrose-10 microM CaCl2) were fractionated by means of differential centrifugation, most of the TRH was recovered in subcellular particles which were pelleted at 10,000 X g and which contained the highest amounts of occluded LDH activity. When the nuclei-free supernatant fluid (900 X g S) was fractionated on discontinuous sucrose density gradients or continuous sucrose density gradients, most of the TRH was recovered in subcellular fractions containing synaptosomes. The subcellular distribution of TRH appeared to be stable for up to 24 h post mortem in rat and human brain tissue.(ABSTRACT TRUNCATED AT 250 WORDS)

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Section: Discussionsupporting

confidence: 74%

Regional Localization and Subcellular Compartmentalization of Thyrotropin‐Releasing Hormone in Adult Human Brain

Parker

Porter²

1983

Journal of Neurochemistry

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“…7,8 However, human postmortem brain samples can be associated with PMIs of hours to days, and perfusion-fixation of human brain is exceptionally rare. Despite these challenges, qualitative and quantitative EM studies of postmortem human brain tissue have been performed in immersion-fixed samples with PMIs ranging from 4 to 100 hr, 9,[10][11][12][13][14][15][16] though the presence of postsynaptic densities (PSD), myelin sheaths, and mitochondria appear to be the measures most resilient to extended PMIs. 9,[12][13][14][15]17 Moreover, other pre-and postmortem factors can affect tissue and ultrastructure quality other than PMI.…”

Section: Introductionmentioning

confidence: 99%

“…Despite these challenges, qualitative and quantitative EM studies of postmortem human brain tissue have been performed in immersion-fixed samples with PMIs ranging from 4 to 100 hr, 9,[10][11][12][13][14][15][16] though the presence of postsynaptic densities (PSD), myelin sheaths, and mitochondria appear to be the measures most resilient to extended PMIs. 9,[12][13][14][15]17 Moreover, other pre-and postmortem factors can affect tissue and ultrastructure quality other than PMI. 3,[18][19][20] For example, experimental models show that acidic pH values are associated with swelling of neuronal processes and mitochondria, 18 and acidic brain pH may reflect hypoxia or lactic acidosis processes occurring pre-and perimortem [20][21][22] which can negatively impact general tissue preservation.…”

Section: Introductionmentioning

confidence: 99%

Factors Affecting Ultrastructural Quality in the Prefrontal Cortex of the Postmortem Human Brain

Glausier

Konanur

Lewis

2018

J Histochem Cytochem.

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Electron microscopy (EM) studies of the postmortem human brain provide a level of resolution essential for understanding brain function in both normal and disease states. However, processes associated with death can impair the cellular and organelle ultrastructural preservation required for quantitative EM studies. Although postmortem interval (PMI), the time between death and preservation of tissue, is thought to be the most influential factor of ultrastructural quality, numerous other factors may also influence tissue preservation. The goal of the present study was to assess the effects of pre-and postmortem factors on multiple components of ultrastructure in the postmortem human prefrontal cortex. Tissue samples from 30 subjects were processed using standard EM histochemistry. The primary dependent measure was number of identifiable neuronal profiles, and secondary measures included presence and/or integrity of synapses, mitochondria, and myelinated axonal fibers. Number of identifiable neuronal profiles was most strongly affected by the interaction of PMI and pH, such that short PMIs and neutral pH values predicted the best preservation. Secondary measures were largely unaffected by preand postmortem factors. Together, these data indicate that distinct components of the neuropil are differentially affected by PMI and pH in postmortem human brain. (J Histochem Cytochem 67:185-202, 2019)

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Slice preparation of rat medulla and pons maintained for five hours in vitro

Hinrichsen¹

1980

J. Neurobiol.

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Slices of rat medulla and pons were incubated in bicarbonate-buffered medium and their electrical activity was monitored for five hours with microelectrodes. The morphology of these slices was compared with that of the same region of rats of the same age using prior perfusion or immersion in fixatives before incubation. Many neurons in incubated slices show shrinkage necrosis (apoptosis) but not dilatation of the endoplasmic reticulum seen in most neurons fixed immediately after slicing. In incubated slices, some processes but not somata of glia appeared swollen: to a lesser extent some dendritic and axonal processes were swollen. Glia showed no cytoplasmic reaction after five hours to indicate that they might phagocytose damaged tissue components. Synapses appeared morphologically normal after the period of incubation and there was an apparent increase in numbers of profiles resembling growth cones.

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Isolation and ultrastructure of human synaptic complexes

Cited by 10 publications

References 17 publications

Regional Localization and Subcellular Compartmentalization of Thyrotropin‐Releasing Hormone in Adult Human Brain

Regional Localization and Subcellular Compartmentalization of Thyrotropin‐Releasing Hormone in Adult Human Brain

Factors Affecting Ultrastructural Quality in the Prefrontal Cortex of the Postmortem Human Brain

Slice preparation of rat medulla and pons maintained for five hours in vitro

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