Radioautographic Studies of the Incorporation of [35s]Cystine into Wool

Downes, AM; Lyne, A. G.; Clarke, WH

doi:10.1071/bi9620713

Cited by 17 publications

(9 citation statements)

References 8 publications

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“…This is at a higher level above the follicle bulb than the appearance of binding of antibodies to the other groups of keratin proteins studied to date Kemp and Rogers 1970) and suggests that the highsulfur proteins are synthesized later than the other fibre proteins. These results are in agreement with the observations of Frater (1976), obtained using polyclonal antibodies to high-sulfur protein preparations, and support the conclusions reached from biochemical and ultrastructural studies of the wool follicle (Downes et at. 1962;L.…”

Section: Discussionsupporting

confidence: 82%

“…2A). Surprisingly, only one (KB 2E2) of the monoclonal antibodies exhibited binding to the wool fibre cortex, in spite of the known presence of the proteins in the cortex (Downes et al 1962). As the high-sulfur proteins readily form extensive disulfide cross-links (Joubert et al 1968), it was thought that the structures into which they are incorporated in the developing fibre might rapidly become inaccessible to antibody penetration.…”

Section: Resultsmentioning

confidence: 99%

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Immunolocalization Studies of the High-sulfur Protein Fraction of Merino Wool using Monoclonal Antibodies

French¹,

Hewish²

1987

Aust. Jnl. Of Bio. Sci.

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Monoclonal antibodies raised to the high-sulfur fraction of solubilized wool protein were used to study the location of these proteins in sections of Merino sheep skin, using indirect immunofluorescence. Poor antibody binding was observed to untreated wool follicles, but mild trypsin digestion of the sections allowed penetration of the antibodies and specific antibody localization was observed. The high-sulfur protein monoclonal antibodies bound to the cells of the wool cortex in the keratogenous zone, and to the cells of the inner root sheath and fibre cuticle regions. The specificities of the antibodies were determined by their binding to high-sulfur proteins separated using high-performance liquid chromatography.

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Section: Discussionsupporting

confidence: 82%

Section: Resultsmentioning

confidence: 99%

Immunolocalization Studies of the High-sulfur Protein Fraction of Merino Wool using Monoclonal Antibodies

French¹,

Hewish²

1987

Aust. Jnl. Of Bio. Sci.

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“…This estimate agrees well with that of 0.92 m m / 2 4 hours made by Fleischer, Vidaver, and Haurowitz (1959). The rate of hair growth in sheep (Downes, Lyne, and Clarke, 1962), and man (Myers and Hamilton, 1951), is about 0.35 m m / 2 4 hours. The faster rate in the rat is associated with the distribution of growing hairs in a wave, while the slower rate in sheep and man is associated with the distribution of growing hairs in a mosaic.…”

Section: Migration Of Label With Hair Growthmentioning

confidence: 95%

“…The incorporation of S~-cysteine into the keratogenous zone of hair cortex has been demonstrated in rats, mice, lambs, and sheep (B61anger, 1956;Harkness and Bern, 1957;Ryder, 1959;Downes, Lyne, and Clarke, 1962). The cysteine could be incorporated into precursor proteins that are formed in the hair bulb and carried up to the keratogenous zone as suggested by Harkness and Bern (1957), or into new proteins as they are synthesized in the keratogenous zone.…”

Section: Introductionmentioning

confidence: 99%

The Incorporation and Fate of H3-Tyrosine in the Hair Cortex of Rats Observed by Radioautography

Sims

1964

The Journal of Cell Biology

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The incorporation of H3-tyrosine into the protein of the cells in the cortex of rat hair has been investigated by radioautography. In growing hairs, radioactivity is found in the matrix, the upper bulb, and the whole of the keratogenous zone up to the fully keratinized part of the shaft, 10 and 30 minutes after an injection of labelled tyrosine. This is unequivocal evidence of protein synthesis at these sites. There is a very precise relationship between the end of protein synthesis and the hardening of the cortical cells at the top of the keratogenous zone. The way in which the silver grains of the radioautographs are clustered indicates that at 30 minutes after the injection the isotope is distributed more evenly in the matrix and upper bulb than in the top of the keratogenous zone. Possibly this reflects a difference, at these sites, in the cell components engaged in protein synthesis, or in the proteins being synthesized. The fully keratinized and hardened part of the hair was not radioactive at 10 and 30 minutes after the injection of H3-tyrosine. The rate at which the radioactivity moves into this region shows that the hair of rats grows 0.9 mm/24 hours. Comparison of the degree of radioactivity along the growing hair in the 30-minute, 12-hour, and 36-hour materials shows conclusively that protein accumulates in the cortical cells during their keratinization. An injection of a labelled amino acid does not behave as an ideal pulse dose; consequently, the grain density over the hair cortex at 36 hours is 100 per cent larger than would be expected if an ideal pulse dose situation existed.

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“…12, the mode of incorporation of [ 35 S]cystine in wool follicles differs from that of [3H]thymidine. The [ 35 S]cystine is incorporated into the keratogenous zone of wool fibres (Ryder 1958;Downes et at. 1962;Chapman and Gemmell 1973) and, in contrast to [3H]thymidine, is retained in the fibres as indicated by the increase in the specific activity of the petroleum-washed fibres with time (Fig.…”

Section: Effects Of Nutritional Changementioning

confidence: 99%

Migration and Keratinization of Cells in Wool Follicles

Chapman¹,

Downes²,

Pa³

1980

Aust. Jnl. Of Bio. Sci.

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. .Migration of' cells in wool follicles of an adult Merino sheep was studied autoradiographically in skin samples taken at intervals after an intravenous injection of [3Hjthymidine. Fibre arid inner root sheath cells incorporated [3Hjthymidine in a cone-shaped region of the follicle bulb. Labelled inner sheath cells migrated out of the bulb ahead of contemporaneous cells in the fibre and remained in advance, although to a progressively lesser extent, until the inner sheath cells sloughed into the follicle lumen. Outer root sheath cells incorporated [3Hjthymidine along the length of the follicle. Cells in the proximal half of the outer sheath migrated inwards and distally and sloughed into the follicle lumen before contemporaneous inner sheath cells. Other cells in the distal half of the outer sheath migrated past the level where cells from the proximal population were shed and also sloughed into the lumen. In the most distal part of the outer sheath, which formed the epidermis-like lining of the follicle canal, little migration of cells was observed during 8 days of observation.The specific activity of tritium in fibres plucked from the same sheep at intervals after the intravenous injection of [3Hjthymidine was determined by scintillation counting and assessed in terms of cell migration and hardening of the fibres. The time at which the specific activity of solventdegreased fibres reached a maximum was found to give an estimate of the time for cells in the fibre to migrate to the upper limit of the keratogenous zone. When the plucked fibres were extracted with 8 M urea the times of the maximum specific activities of the urea-dispersible and urea-insoluble material provided respectively estimates of the times at which hardening of the fibres began and ended.The effects of different planes of nutrition were examined in two other Merino sheep by radioassay· of fibres plucked after intravenous injections of [3Hjthymidine given after an equilibration period of at least 2 months on each level of feeding. A high plane of nutrition increased the rate of cell migration and hastened the onset of hardening of the fibres, but prolonged the hardening process. The prolongation of the hardening process was confirmed by the specific activities of fibres plucked after intravenous inje,

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Radioautographic Studies of the Incorporation of [35s]Cystine into Wool

Cited by 17 publications

References 8 publications

Immunolocalization Studies of the High-sulfur Protein Fraction of Merino Wool using Monoclonal Antibodies

Immunolocalization Studies of the High-sulfur Protein Fraction of Merino Wool using Monoclonal Antibodies

The Incorporation and Fate of H3-Tyrosine in the Hair Cortex of Rats Observed by Radioautography

Migration and Keratinization of Cells in Wool Follicles

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