On the Isolation of Nerve Endings and Synaptic Vesicles

Robertis, E. De; Iraldi, Amanda Pellegrino de; Rodríguez, G B Germán; Gómez, César Cinesi

doi:10.1083/jcb.9.1.229

Cited by 164 publications

(31 citation statements)

References 11 publications

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“…These small vesicles had similar characteristics, as judged from their distribution in density gradients, and electron microscopic appearance, to the ACh containing vesicles isolated from guinea-pig brain by Whittaker (1959) and from rat brain by de Robertis et al (1961). The finding that ACh occurred in the neural lobe in small vesicles did not, however, clarify the basic question, namely whether the small vesicles, occurring in the same nerve as the hormone granules, contained ACh, or whether the AChcontaining vesicles occurred in separate (cholinergic) nerve endings.…”

Section: Discussionmentioning

confidence: 73%

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Neuronal and subcellular localization of acetylcholine in the posterior pituitary of the rabbit

Lederis¹,

Livingston²

1970

The Journal of Physiology

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SUMMARY1. Acetylcholine and neurohypophysial hormones were measured in subcellular fractions of rabbit neurohypophysis.2. Differential anddensity gradient centrifugation, followed by bio-assay and electron microscopy were used to separate and characterize the subcellular particles, or isolated nerve endings.3. Acetylcholine in the rabbit neurohypophysis was found to occur in small vesicles of about 40 nm diameter which had similar characteristics to the synaptic vesicles from central nervous tissue.4. Examination of intact nerve endings isolated from the rabbit neurohypophysis indicated that at least some acetylcholine and neurohypophysial hormones occurred in separate nerve endings.

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Section: Discussionmentioning

confidence: 73%

“…1. This procedure was based on the methods of Whittaker (1959) and de Robertis et al (1961), for the preparation of a pellet containing nerve endings and mitochondria from brain tissue. All centrifugation experiments were done on an MSE superspeed 40 refrigerated ultracentrifuge at 20 C using a 3 x 5 ml.…”

Section: Differential Centrifugationmentioning

confidence: 99%

Neuronal and subcellular localization of acetylcholine in the posterior pituitary of the rabbit

Lederis¹,

Livingston²

1970

The Journal of Physiology

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“…Nonmitochondrial organelles originally detected in the mitochondrial fraction of rat brain (259) were eventually purified by sedimentation on sucrose density gradients and largely identified as pinched-off nerve terminals (synaptosomes) on the basis of their content of synaptic vesicles and mitochondria (67,127). Since then, mammalian brain synaptosomes purified with different protocols have been found to contain other minor particulates such as free mitochondria and fragments of axons, dendrites, and glial processes (54, 105, 124, 136, 234, 264, 310; for a review, see Ref.…”

Section: B Vertebrate Nerve Endingsmentioning

confidence: 99%

Local Gene Expression in Axons and Nerve Endings: The Glia-Neuron Unit

Giuditta¹,

Chun²,

Eyman³

et al. 2008

Physiological Reviews

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Neurons have complex and often extensively elongated processes. This unique cell morphology raises the problem of how remote neuronal territories are replenished with proteins. For a long time, axonal and presynaptic proteins were thought to be exclusively synthesized in the cell body, which delivered them to peripheral sites by axoplasmic transport. Despite this early belief, protein has been shown to be synthesized in axons and nerve terminals, substantially alleviating the trophic burden of the perikaryon. This observation raised the question of the cellular origin of the peripheral RNAs involved in protein synthesis. The synthesis of these RNAs was initially attributed to the neuron soma almost by default. However, experimental data and theoretical considerations support the alternative view that axonal and presynaptic RNAs are also transcribed in the flanking glial cells and transferred to the axon domain of mature neurons. Altogether, these data suggest that axons and nerve terminals are served by a distinct gene expression system largely independent of the neuron cell body. Such a local system would allow the neuron periphery to respond promptly to environmental stimuli. This view has the theoretical merit of extending to axons and nerve terminals the marginalized concept of a glial supply of RNA (and protein) to the neuron cell body. Most long-term plastic changes requiring de novo gene expression occur in these domains, notably in presynaptic endings, despite their intrinsic lack of transcriptional capacity. This review enlightens novel perspectives on the biology and pathobiology of the neuron by critically reviewing these issues.

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“…Synaptosomes were prepared by a modification of the techniques of Gray and Whittaker [13] and De Robert is et al [10]. Tissue frag ments were homogenized in cold 0.32 M sucrose with eight up and down strokes of a Teflon pestle in a smooth glass homogenizer.…”

Section: Tissue Preparationmentioning

confidence: 99%

Characteristics of Dopamine Uptake and 3,4-Dihydroxyphenylacetic Acid (DOPAC) Formation in the Dopaminergic Terminals of the Neurointermediate Lobe of the Pituitary Gland

Annunziato¹,

Weiner²

1980

Neuroendocrinology

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The kinetics of dopamine (DA) uptake were studied in synaptosomal preparations of the neurointermediate lobe (NIL) of the pituitary gland, a region containing the terminals of the tuberoinfundibular dopaminergic neurons. Lineweaver-Burk plots of the initial velocity of DA uptake versus the concentration of DA yielded a single straight line in the NIL. The K_m values in the NIL of the rat and the steer were 2.2 ± 0.5 × 10^–6M and 2.0 ± 0.1 × 10^–6M, respectively. The uptake was predominantly into dopaminergic terminals since preincubation with desipramine did not affect the V_max or K_m of DA uptake. Oberved uptake was predominantly due to transport across the neuronal membrane and not into storage granules, since reserpine caused only a small decrease in uptake. The concentration of the DA metabolite 3,4-dihydroxyphenylacetic acid (DOPAC) in the NIL was 3.6 ng/mg protein. The ratio of DA to DOPAC in the NIL (3.73) was similar to that obtained in the medial basal hypothalamus, another region innervated by the tuberoinfundibular dopaminergic neurons. The kinetics of DA uptake in the nerve terminals of the NIL are similar to those observed in the DA terminals in the median eminence. The affinity of uptake in the terminal fields of the tuberoinfundibular system is considerably lower than in terminals of the mesolimbic and nigrostriatal dopaminergic terminals.

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On the Isolation of Nerve Endings and Synaptic Vesicles

Cited by 164 publications

References 11 publications

Neuronal and subcellular localization of acetylcholine in the posterior pituitary of the rabbit

Neuronal and subcellular localization of acetylcholine in the posterior pituitary of the rabbit

Local Gene Expression in Axons and Nerve Endings: The Glia-Neuron Unit

Characteristics of Dopamine Uptake and 3,4-Dihydroxyphenylacetic Acid (DOPAC) Formation in the Dopaminergic Terminals of the Neurointermediate Lobe of the Pituitary Gland

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