Most Synaptic Vesicles Isolated from Rat Brain Carry Three Membrane Proteins, SV2, Synaptophysin, and p65

Floor, Erik; Feist, Blake E.

doi:10.1111/j.1471-4159.1989.tb09190.x

Cited by 59 publications

(27 citation statements)

References 39 publications

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“…Immunoprecipitation (Floor and Feist 1989) and electron microscopic (Jahn et al 1985;Navone et al 1986;Hiscock et al 2000) studies have localized synaptophysin to virtually all, or greater than 90%, of all vesicles. Synaptophysin immunoreactivity has been identified as a useful marker for synaptic density in many studies (Hamos et al 1989;Masliah et al 1990;.…”

Section: Author Manuscriptmentioning

confidence: 99%

Synaptophysin and postsynaptic density protein 95 in the human prefrontal cortex from mid-gestation into early adulthood

Glantz¹,

Gilmore²,

Hamer³

et al. 2007

Neuroscience

235

162

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Previous studies of postnatal synaptic development in human frontal cortex have shown that synaptic density rises after birth, reaches a plateau in childhood and then decreases to adult levels by late adolescence. A similar pattern has been seen in nonhuman primate cortex. These earlier studies in human cortex are limited, however, by significant age gaps in study subjects at critical inflection points of the developmental curve. Additionally, it is unclear if synaptic development occurs in different patterns in different cortical layers in prefrontal cortex (PFC). The purpose of this study was to examine synaptic density in human PFC across development by measuring two synaptic marker proteins: synaptophysin (presynaptic), and postsynaptic density-95 (PSD-95; postsynaptic). Western blotting was used to assess the relative levels of synaptophysin and PSD-95 in dorsolateral PFC of 42 subjects, distributed in age from 18 weeks gestation to 25 years. In addition, synaptophysin immunoreactivity was examined in each layer of areas 9 and 46 of PFC in 24 subjects, ranging in age from 0.1 to 25 years. Synaptophysin levels slowly increased from birth until age 5 and then increased more rapidly to peak in late childhood around age 10. Synaptophysin subsequently decreased until the adult level was reached by mid-adolescence, around age 16. PSD-95 levels increased postnatally to reach a stable plateau by early childhood with a slight reduction in late adolescence and early adulthood. The pattern of synaptophysin immunoreactivity seen with immunohistochemistry was similar to the Western experiments but the changes across age were more subtle, with little change by layer within and across age. The developmental patterns exhibited by these synaptic marker proteins expand upon previous studies of developmental synaptic changes in human frontal cortex; synaptic density increases steadily from birth to late childhood, then decreases in early adolescence to reach adult levels by late adolescence. Keywordsadolescence; development; human; synapses; pruning; synaptogenesisThe human brain has the most protracted period of development of any species, with neurodevelopment continuing well into early adulthood. Imaging studies have shown that in the human, the overall volume of the brain increases during childhood and adolescence (Giedd Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. NIH Public Access Author ManuscriptNeuroscience. Author manuscript; available in PMC 2008 November 9. Sowell et al. 2002;Gilmore et al. 2007). Specifically, the cortical gray matter volume, particularly the f...

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Section: Author Manuscriptmentioning

confidence: 99%

Synaptophysin and postsynaptic density protein 95 in the human prefrontal cortex from mid-gestation into early adulthood

Glantz¹,

Gilmore²,

Hamer³

et al. 2007

Neuroscience

235

162

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“…Synaptophysin is an established marker of synaptic vesicles (Floor and Feist, 1989). Immunofluorescence staining with synaptophysin revealed that YFP-GABARAP was expressed in dendrites that are making synaptic contact (Fig.…”

Section: Localization Of Yfp-gabarap In Hippocampal Neuronsmentioning

confidence: 99%

GABA_AReceptor-Associated Protein Traffics GABA_AReceptors to the Plasma Membrane in Neurons

Leil

Chen²,

Chang³

et al. 2004

J. Neurosci.

166

171

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The trafficking of GABA A receptors is an important component of the pathway that regulates plasticity of inhibitory synapses. The 17 kDa GABA A receptor-associated protein (GABARAP) has been implicated in the trafficking of GABA A receptors because of its ability to interact not only with the ␥2 subunit of the receptor but also with microtubules and the N-ethylmaleimide-sensitive factor (NSF). To elucidate the role of GABARAP in the trafficking of GABA A receptors, we have constructed a yellow fluorescent protein (YFP) fusion protein of GABARAP and expressed it in neurons using adenovirus, so that its function may be examined. YFP-GABARAP colocalized with ␥2 subunit-containing GABA A receptors and NSF to the perinuclear cytoplasm in cultured hippocampal neurons and to the proximal regions of dendrites that are making synaptic contact. Expression of YFP-GABARAP in Cos7 cells and cultured hippocampal neurons was able to increase the level of GABA A receptors detected at the plasma membrane, even at low levels of YFP-GABARAP expression. This effect is specific to the function of GABARAP on GABA A receptor trafficking, because point mutations in the ␥2-binding domain of YFP-GABARAP interfered with the ability of YFP-GABARAP to increase GABA A receptor surface levels. These mutations also disrupted the colocalization of YFP-GABARAP with the ␥2 subunit and with NSF in hippocampal neurons. The results of this study show for the first time that GABARAP has a functional effect on the trafficking of GABA A receptors and provide decisive evidence for the role of GABARAP in transporting GABA A receptors to the plasma membrane in neurons.

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“…Markers for the 5-HT transporter included the specific binding of )H-imipramine (Marcusson et al, 1986(Marcusson et al, , 1989Graham et al, 1989) and 3H-paroxetine (Habert et al, 1985;DeSouza and Kuyatt, 1987;Hrdina et al, 1990), as well as the ability to take up 3H-5-HT. Synaptic vesicle markers included synaptophysin, a protein found in the membrane of nearly all synaptic vesicles (Jahn et al, 1985;Wiedenmann and Franke, 1985;Navone et al, 1986;Floor and Feist, 1989; Sudhof and Jahn, 199 1) and a form (-45 kDa) of serotonin-binding protein (SBP), which is found only in the synaptic vesicles of serotonergic neurons and the secretory vesicles of other 5-HT-storing cells that are derived from neuroectoderm (Jonakait et al, 1979;Tamir and Gershon, 1979;Gershon et al, 1983;Tamir, 1984, 1985;Barasch et al, 1987;Kirchgessner et al, 1988;Tamir et al, 1990). Finally, we investigated, as a function of age, the ability of reserpine, which blocks the transport of monoamines into synaptic vesicles (Carlsson, 1965;Giachetti et al, 1974;Shore and Giachetti, 19 78;Maron et al, 1979;Angelides, 19 80), to deplete 5-HT.…”

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confidence: 99%

Synaptic properties of serotonergic growth cones in developing rat brain

1994

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In order to gain insight into the events that take place when serotonergic growth cones are remodeled into synapses, we tested the hypothesis that neurotransmitter-related properties of presynaptic terminals are already present in these growth cones before synaptogenesis begins. The ontogeny of markers for the specific reuptake of 5-HT and for 5-HT-storing synaptic vesicles was studied in isolated growth cone (IGC) fractions from developing rat brain. High- affinity 3H-imipramine binding (a marker for the plasma membrane 5-HT transporter) was significantly enriched in IGC fractions prepared before the beginning of cortical synaptogenesis [embryonic day 15 (E15) and E20]. Radioautography with 3H-imipramine or 3H-paroxetine (another marker for the transporter) confirmed that the 5-HT transporter is present in the cerebral cortex when it contains serotonergic growth cones, but not serotonergic synapses. Specific uptake of 3H-5-HT was found in IGC fractions as early as E15; this uptake was inhibited by fluoxetine. Electron microscopic radioautography demonstrated directly that growth cones were the structures in IGC fractions that took up 3H- 5-HT. The synaptic vesicle protein synaptophysin and a 45 kDa protein found specifically in serotonergic synaptic vesicles, serotonin-binding protein (SBP), were each enriched in IGC fractions from E15 to postnatal day 5; SBP immunoreactivity increased approximately 10-fold between E15 and E20. Endogenous 5-HT was detected in IGC fractions at E15 and increased in amount as development proceeded. The ratio of 5-HT to 5-hydroxyindole acetic acid suggested that 5-HT within growth cones is protected from catabolism by monoamine oxidase. Reserpine-induced depletion of 5-HT, a marker for the vesicular carrier of 5-HT, was apparent in IGC fractions at E20, but not at E15. These data suggest that properties that characterize the presynaptic components of mature serotonergic synapses develop in growth cones before synapses are formed. The early development of these properties may permit neurotransmission to be established rapidly during synaptogenesis or, alternatively, enable 5-HT to play a role in ontogeny.

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Most Synaptic Vesicles Isolated from Rat Brain Carry Three Membrane Proteins, SV2, Synaptophysin, and p65

Cited by 59 publications

References 39 publications

Synaptophysin and postsynaptic density protein 95 in the human prefrontal cortex from mid-gestation into early adulthood

Synaptophysin and postsynaptic density protein 95 in the human prefrontal cortex from mid-gestation into early adulthood

GABA_AReceptor-Associated Protein Traffics GABA_AReceptors to the Plasma Membrane in Neurons

Synaptic properties of serotonergic growth cones in developing rat brain

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Most Synaptic Vesicles Isolated from Rat Brain Carry Three Membrane Proteins, SV2, Synaptophysin, and p65

Cited by 59 publications

References 39 publications

Synaptophysin and postsynaptic density protein 95 in the human prefrontal cortex from mid-gestation into early adulthood

Synaptophysin and postsynaptic density protein 95 in the human prefrontal cortex from mid-gestation into early adulthood

GABAAReceptor-Associated Protein Traffics GABAAReceptors to the Plasma Membrane in Neurons

Synaptic properties of serotonergic growth cones in developing rat brain

Contact Info

Product

Resources

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GABA_AReceptor-Associated Protein Traffics GABA_AReceptors to the Plasma Membrane in Neurons