An autoradiographic study of the incorporation of nucleic‐acid precursors by neurones and glia during nerve stimulation.

Watson, W. E.

doi:10.1113/jphysiol.1965.sp007729

Cited by 91 publications

(5 citation statements)

References 37 publications

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Section: Discussionsupporting

confidence: 90%

“…Our data are consistent with in vivo [ 3 H]uridine labelling studies showing rapid accumulation of radiolabeled RNA in neuronal nuclei following intracranial, intravenous or subcutaneous injection, reaching a plateau within 30-40 min after injection [16,66,75]. In large neurons, nuclear [ 3 H]uridine labelling persisted for 24 h and showed a gradual reduction in subsequent days, while in other cell types (choroid plexus, liver) labelling disappeared more quickly [16,75]. In our study, we found different temporal EU staining profiles within the same neuronal tissue depending on cell type, and varying from disappearance of detectable staining within 6 h postinjection in cerebellar granule cells to staining that persisted for longer than 2 weeks in Purkinje cells, Bergman glia and molecular layer interneurons.…”

Section: Discussionsupporting

confidence: 90%

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In vivo 5-ethynyluridine (EU) labelling detects reduced transcription in Purkinje cell degeneration mouse mutants, but can itself induce neurodegeneration

Sant

White

Hoeijmakers

et al. 2021

acta neuropathol commun

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Fluorescent staining of newly transcribed RNA via metabolic labelling with 5-ethynyluridine (EU) and click chemistry enables visualisation of changes in transcription, such as in conditions of cellular stress. Here, we tested whether EU labelling can be used to examine transcription in vivo in mouse models of nervous system disorders. We show that injection of EU directly into the cerebellum results in reproducible labelling of newly transcribed RNA in cerebellar neurons and glia, with cell type-specific differences in relative labelling intensities, such as Purkinje cells exhibiting the highest levels. We also observed EU-labelling accumulating into cytoplasmic inclusions, indicating that EU, like other modified uridines, may introduce non-physiological properties in labelled RNAs. Additionally, we found that EU induces Purkinje cell degeneration nine days after EU injection, suggesting that EU incorporation not only results in abnormal RNA transcripts, but also eventually becomes neurotoxic in highly transcriptionally-active neurons. However, short post-injection intervals of EU labelling in both a Purkinje cell-specific DNA repair-deficient mouse model and a mouse model of spinocerebellar ataxia 1 revealed reduced transcription in Purkinje cells compared to controls. We combined EU labelling with immunohistology to correlate altered EU staining with pathological markers, such as genotoxic signalling factors. These data indicate that the EU-labelling method provided here can be used to identify changes in transcription in vivo in nervous system disease models.

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Section: Discussionsupporting

confidence: 90%

Section: Discussionsupporting

confidence: 90%

In vivo 5-ethynyluridine (EU) labelling detects reduced transcription in Purkinje cell degeneration mouse mutants, but can itself induce neurodegeneration

Sant

White

Hoeijmakers

et al. 2021

acta neuropathol commun

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“…The changes occur at about the same rate as changes in RNA metabolism observed autoradiographically when the functional state of neurones is changed (Watson, 1965b).…”

mentioning

confidence: 58%

“…After injuring the left hypoglossal nerve the whole tongue must be controlled by the right nerve, and it is possible that the changes in the uninjured neurones are those of work hypertrophy. The changes occur at about the same rate as changes in RNA metabolism observed autoradiographically when the functional state of neurones is changed (Watson, 1965b …”

mentioning

confidence: 63%

Observations on the nucleolar and total cell body nucleic acid of injured nerve cells

Watson¹

1968

The Journal of Physiology

259

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SUMMARY1. The nucleic acid content of neuronal nucleoli and the total cell body nucleic acid content of neurones of the hypoglossal nucleus were measured by ultraviolet absorption microspectrography.2. After nerve injury both the nucleolar nucleic acid and the total cell body nucleic acid increased: nucleolar changes preceded those of the cell body.3. The closer to the nerve cell body that the axon was injured the earlier was the onset and the decline of the nucleolar response.4. Actinomycin D was given to prevent DNA-primed RNA synthesis, and the rate of 'decay' of nucleolar RNA was measured. This rate varied after nerve injury and was closely related to the nucleolar nucleic acid content.5. The apparent rate of transfer of labelled RNA from the neuronal nucleus into the cytoplasm changed after nerve injury in a manner closely related to the changes in nucleolar nucleic acid content.6. It was demonstrated by making consecutive nerve injuries or by preventing or delaying nerve regeneration, that the nucleic acid changes were not induced by removal of contact between the neurone and its motor end-plate, and were not repressed by the restoration of such

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“…This method was considered suitable to yield correlates for the comparison of protein-synthetic activity between the neurosecretory cells of the SON and of the PVN, under different types of stimulation which were known to result in preponderant secretion of one or the other of the neurohypo physial hormones, namely dehydration and lactation. Watson [1965b] compared RNA transfer rates from nucleus to cytoplasm between the SON and the PVN of mice subjected to varying degrees of dehydration. He did not study the lactating animal.…”

Section: Introductionmentioning

confidence: 99%

Localization of functions in the magnocellular neurosecretory nuclei of the mammalian hypothalamus by autoradiography

Bandaranayake¹

1976

Cells Tissues Organs

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Nucleo-cytoplasmic transfer rates of RNA were used as indices of protein-synthetic activity in mammalian supraoptic and paraventricular neurones. Normal, dehydrated and lactating rats were exposed, for varying periods of time, to tritiated uridine, and autoradiographs obtained from hypothalamic blocks. Grain count and cytometry enabled a cytoplasmic/nuclear ratio of grain densities to be obtained for each neurosecretory nucleus. Comparison of these ratios, as well as the regression slopes for their increase with time of exposure to the isotope, indicated the presence of a differential response between the supraoptic and paraventricular nuclei, with the former responding more to the stimulus of dehydration, and the latter to that of lactation. The observations substantiate the view that a partial division of labour exists between the two nuclei in the elaboration of each of the mammalian neurohypophysial octapeptides, vasopressin and oxytocin.

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An autoradiographic study of the incorporation of nucleic‐acid precursors by neurones and glia during nerve stimulation.

Cited by 91 publications

References 37 publications

In vivo 5-ethynyluridine (EU) labelling detects reduced transcription in Purkinje cell degeneration mouse mutants, but can itself induce neurodegeneration

In vivo 5-ethynyluridine (EU) labelling detects reduced transcription in Purkinje cell degeneration mouse mutants, but can itself induce neurodegeneration

Observations on the nucleolar and total cell body nucleic acid of injured nerve cells

Localization of functions in the magnocellular neurosecretory nuclei of the mammalian hypothalamus by autoradiography

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