The quantitative estimation of desoxyribose nucleic acid (DNA) in isolated individual animal nuclei by the caspersson ultraviolet method

Leuchtenberger, Cecilie; Leuchtenberger, Rudolf; Vendrely, C; Vendrely, R

doi:10.1016/0014-4827(52)90045-1

Cited by 71 publications

(12 citation statements)

References 3 publications

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“…For example, some changes in the sperm cell membrane during aging might interfere progressively with the entrance of the leucofuchsin, as iron ions have been shown to interfere in Tetrahymena and Acanthamoeba (7), or might shift the point of o p t i m u m hydrolysis, thus reducing the intensity of the staining reaction. This possibility, among others, is being tested, and independent techniques such as ultraviolet microspectrophotometry (14) and chemical determinations (8) are to be employed to determine the identity of the material which is depleted on aging of spermatozoa.…”

Section: Discussionmentioning

confidence: 99%

DECREASE IN NUCLEAR FEULGEN-POSITIVE MATERIAL (DNA) UPON AGING IN IN VITRO STORAGE OF BOVINE SPERMATOZOA

Salisbury

Birge

Torre

et al. 1961

The Journal of Cell Biology

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The Feulgcn-DNA content of sperm ceUs from 5 bulls was studied by means of microspectrophotomctry after storage at 5°C for 2, 3, 5, and l0 days in a yolk-citrate diluent permitting slow aerobic metabolism. A subsample of sperm cells from each bull was subjected to the Feulgen technique on each of the storage days selected. The cells sampled on each of these days received a standard 12 minute, 60°C hydrolysis. Absorption measurements at 546 m# of the individual cells indicated a marked progressive decrease in the Feulgen-DNA content of the stored spermatozoa. The loss of 30 per cent of the initial DNA at the end of 5 days' storage was highly significant statistically. This decrease approximately parallels the known decrease in fertility of stored sperm cells, as well as the increase in apparent embryonic mortality resulting from the use of similarly aged spermatozoa for artificial insemination.The relative constancy of the deoxyribonucleic acid (DNA) in the resting cell nucleus is a basic tenet of modern biology (8). This tenet for the male gametes is implicit in the recommendation that Feulgen-stained bull spermatozoa could provide a standard reference material for the microphotometric quantitation of DNA in other cell nuclei (14).The assumption of constancy in spermatozoa arises from a number of observations. Among these is the correlation, first established in the bovine (34), between the DNA content of haploid spermatozoan nuclei and the DNA content and the number of chromosome sets to be found in spermatogenic and somatic tissues (10,17,21,34).A second supporting fact is that in the absence of chromosome change the DNA content of cell nuclei appears to be relatively stable (1), for, except in the case of some secretory cells (16), it does not become metabolically so involved in the activity of cells as do other cellular components. This fact has lent support to the commonly held belief that the spermatozoon serves primarily as a convenient mechanism for transport of genetic information via DNA. During this transport it has been assumed that the DNA is wholly passive and is not involved in energy exchange mechanisms.No one seems to have seriously challenged the concept of constancy of the DNA in living spermatozoa, except Salisbury, who did so recently (22) on the basis of an increase in apparent early embryonic mortality in cattle resulting from the aging of spermatozoa by storage at 5°C before insemination (23). The evidence involving the aging of spermatozoa in the incidence of early embryonic mortality in cattle is supported by evidence on aging of spermatozoa in the female reproductive tract for other mammalian species--the rat and the guinea pig (28, 29)--and for poultry (9,20)

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Section: Discussionmentioning

confidence: 99%

DECREASE IN NUCLEAR FEULGEN-POSITIVE MATERIAL (DNA) UPON AGING IN IN VITRO STORAGE OF BOVINE SPERMATOZOA

Salisbury

Birge

Torre

et al. 1961

The Journal of Cell Biology

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“…Measurement of integrated absorbance at a wave-length near the nucleic acid absorption peak has provided reproducible measurements of the quantity of nucleic acid in individual cells which were closely correlated with biochemical estimations performed upon large numbers of cells (Leuchtenberger, Leuchtenberger, Vendrely & Vendrely, 1952;Walker & Yates, 1952;Leuchtenberger, Klein & Klein, 1952). Lindstrom, Zetterberg & Carlson (1966) showed that the absorption spectra of RNA solutions and of concentrated dried microdroplets are practically identical.…”

mentioning

confidence: 99%

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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“…The mean DNA content of the resting somatic cells is nearly constant for any species (Leuchtenberger et al,1952). It would be of interesting to know how the nucleic acid content is related to the cell size or the cell number.…”

mentioning

confidence: 99%

Nucleic Acid Contents of Various Organs in Rats During Postnatal Growth

Fujii

Koyama

1962

Endocrinol Japon

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The quantitative estimation of desoxyribose nucleic acid (DNA) in isolated individual animal nuclei by the caspersson ultraviolet method

Cited by 71 publications

References 3 publications

DECREASE IN NUCLEAR FEULGEN-POSITIVE MATERIAL (DNA) UPON AGING IN IN VITRO STORAGE OF BOVINE SPERMATOZOA

DECREASE IN NUCLEAR FEULGEN-POSITIVE MATERIAL (DNA) UPON AGING IN IN VITRO STORAGE OF BOVINE SPERMATOZOA

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

Nucleic Acid Contents of Various Organs in Rats During Postnatal Growth

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