A Two Emulsion Autoradiographic Technique and the Discrimination of the Three Different Types of Labelling After Double Labelling With 3h‐ and Wc‐thymidine

Schultze, B.; Maurer, W.; Hagenbusch, H.

doi:10.1111/j.1365-2184.1976.tb01272.x

Cited by 29 publications

(18 citation statements)

References 10 publications

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“…After drying, the gelatine was hardened in 20% formalin for 20 min. Gelatine layers prepared in this way were 5-0 0-6 pm thick in the dry state (Schultze, Maurer & Hagenbusch, 1976). This thickness ensures total absorption of 3H pparticles.…”

Section: A T E R I a L S A N D M E T H O D Smentioning

confidence: 99%

Cell population kinetics in pig epidermis: further studies

Morris

Hopewell

1987

Cell Proliferation

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Abstract. The cell population kinetics of the epidermis were studied in 4‐month‐old pigs. Mitotic figures were confined to the basal cell (L1) and the first suprabasal cell layer (L2). The mitotic index (MI) was 0.17 ± 0.04% for L1 and 0.08 ± 0.03% for L2. Labelled nuclei were distributed throughout the viable epidermis, the majority (79.1 ± 1.1%) were in L1 with 19.5 ± 1.2% in L2. The labelling indices (LI) in layers L1 and L2 were 7.1 ± 0.4% and 3.4 ± 0.1%, respectively. After labelling with two injections of tritiated thymidine [3H]TdR separated by 90 min, the LI increased to 8.2 ± 0.3% in L1 and to 4.0 ± 0.2% in L2. This increased labelling confirmed that cell proliferation occurs in both layers, L1 and L2, of the epidermis. The cell production rate (K) in L1 and L2 had an upper limit of 10.7 ± 1.0 and 6.2 + 1.8 cells per 1000 cells per hour respectively. The cell flow rate per hour (cell flux), into and out of the DNA synthesis phase (S), and the duration of DNA synthesis were determined from double‐labelling studies with [3H]TdR and [14C]TdR. The cell flux into and out of S was identical and was calculated as 0.6 ± 0.1%/hr (L1) and 0.5 ± 0.1%/hr (L2). Values for tS varied from 8 to 10 hr. The cell turnover times (tT) were in the range 89–129 hr and 180–261 hr for L1 and L2, respectively. Log normal curves were fitted to the fraction labelled mitoses data for L1 and L2. Values for tS for cells in L1 and L2 were 9.8 hr and 11.9 hr, respectively. tG2+ 1/2tM was 7.2 hr in L1 and 9.1 hr in L2.

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Section: A T E R I a L S A N D M E T H O D Smentioning

confidence: 99%

Cell population kinetics in pig epidermis: further studies

Morris

Hopewell

1987

Cell Proliferation

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“…In the case of labelling with [3H]TdR only, the slides were dipped into diluted Ilford K2 emulsion (emulsion : distilled water = 3 : 2), exposed for 1 4 d and developed in Amidol. In the case of double-labelling, two emulsion layer autoradiographs were prepared with minor changes by the method of Schultze, Maurer & Hagenbusch (1976). Briefly, a first thin emulsion layer (about 1 pm thick) was produced by dipping the smears into diluted Ilford K2 emulsion followed by a 1 d exposure at 4°C.…”

Section: Histology and Autoradiographymentioning

confidence: 99%

Tumour Cell Recruitment of the Jb‐1 and L 1210 Ascites Tumour Determined Directly By Double Labelling With [¹⁴C]‐ and [³H]‐Thymidine

1988

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Abstract. Tumour cell recruitment of the JB‐1 and L 1210 ascites tumour has been demonstrated directly by a double‐labelling method with [14C]‐ and [3H]‐thymidine (TdR). After [14C]‐labelling of all proliferating tumour cells by multiple injections of [14C]TdR, recruitment of resting cells was stimulated by removal of the majority of tumour cells, i.e. by maximum aspiration of ascitic fluid. the number of recruited resting cells in the remaining tumour that re‐enter the cell cycle after stimulation was demonstrated directly by a single injection of [3H]TdR given at different times after stimulation. the increase in the percentage of purely [3H]‐labelled cells, i.e. recruited cells, with increasing time after stimulation, shows that recruitment is not a synchronous but a continuous process, the maximum of which occurs earlier in the case of the L 1210 than the JB‐1 tumour. This suggests that there seems to be a relationship between the time required for maximum recruitment and the corresponding cell cycle parameters of the unperturbed tumour. There is a transitory increase of the growth fraction to about 100% and a considerable shortening of the cycle time at the maximum of recruitment.

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“…A second, thick layer of undiluted Ilford K2 emulsion (10-20 pm thick after development) was then spread onto the slides. After a 15-20-day exposure the slides were developed in diluted amidol developer (20 min) at 12OC, dried and covered with a cover slip (for details see Schultze, Maurer & Hagenbusch, 1976).…”

Section: Single-and Double-layer Autoradiography Of the Crypt Squashesmentioning

confidence: 99%

Influence of the Beta‐Adrenergic Antagonists Propranolol, Practolol and Oxprenolol On the Proliferation of Rat Jejunal Crypt Cells

et al. 1986

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Abstract. Beta‐adrenergic blockade by quite large doses of propranolol, practolol and oxprenolol, once or continuously applied, does not influence jejunal crypt‐cell proliferation in the rat. After a single i.p. injection of 20 mg/kg propranolol or practolol and even of 100 mg/kg practolol, the mitotic index, the labelling index and the duration of the S phase do not differ between treated and untreated control animals nor between animals treated with the different drugs. Continuous application of 30 mg/kg/d propranolol, practolol or oxprenolol for 7 or 14 days does not affect the mitotic and labelling indices either, nor does it change the duration of the cycle of the jejunal crypt cells and its phases as determined by the percent labelled mitoses method. These results are in contrast to those reported previously by Tutton & Helme (1974).

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A Two Emulsion Autoradiographic Technique and the Discrimination of the Three Different Types of Labelling After Double Labelling With 3h‐ and Wc‐thymidine

Cited by 29 publications

References 10 publications

Cell population kinetics in pig epidermis: further studies

Cell population kinetics in pig epidermis: further studies

Tumour Cell Recruitment of the Jb‐1 and L 1210 Ascites Tumour Determined Directly By Double Labelling With [¹⁴C]‐ and [³H]‐Thymidine

Influence of the Beta‐Adrenergic Antagonists Propranolol, Practolol and Oxprenolol On the Proliferation of Rat Jejunal Crypt Cells

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A Two Emulsion Autoradiographic Technique and the Discrimination of the Three Different Types of Labelling After Double Labelling With 3h‐ and Wc‐thymidine

Cited by 29 publications

References 10 publications

Cell population kinetics in pig epidermis: further studies

Cell population kinetics in pig epidermis: further studies

Tumour Cell Recruitment of the Jb‐1 and L 1210 Ascites Tumour Determined Directly By Double Labelling With [14C]‐ and [3H]‐Thymidine

Influence of the Beta‐Adrenergic Antagonists Propranolol, Practolol and Oxprenolol On the Proliferation of Rat Jejunal Crypt Cells

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Tumour Cell Recruitment of the Jb‐1 and L 1210 Ascites Tumour Determined Directly By Double Labelling With [¹⁴C]‐ and [³H]‐Thymidine