Studies on the possible role of vitreous humor on protein synthesis and morphology of organ cultured adult rabbit lens. II. Epithelial cells

Banerjee, Amit Kumar; Richiert, Dawn M.; Emanuel, K; Singh, Aastha; Bagchi, M.

doi:10.1016/0167-4838(91)90472-c

Cited by 3 publications

(4 citation statements)

References 15 publications

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“…Figure 2 illustrates the DNA synthesis of epithelial cells of lenses cultured in 3% serum containing TC-199 medium. Control lenses were incubated with the 8 kD VH fraction (Banerjee et al, 1991). DNA synthesis of the experimental (no VH) epithelial cells of organ cultured lenses were considerably higher than control cells at all time periods examined, and this increase was most pronounced at 48 h of incubation.…”

Section: Figure L(i) Illustrates the Autoradiogram Of Iodinated Totalmentioning

confidence: 99%

“…Reports from our laboratory have clearly demonstrated that the rate of protein synthesis by epithelial cells of lenses incubated in the presence of total vitreous humor or an 8 kD VH factor, is significantly decreased when compared with epithelial cells of lenses cultured without VH (Singh and Bagchi, 1989;Banerjee et al, 1991). It has also been reported that the early events of cell proliferation are dependent on continuous protein synthesis and that a 50-70% inhibition of protein synthesis effectively inhibits cell growth (Pardee, 1989).…”

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confidence: 90%

“…factor was isolated according to the technique of Banerjee et al (1991). VH was dissected out from freshly isolated rabbit lenses and centrifuged €or 3 h at 100,OOOg.…”

Section: Isolation Of V H Protein the 8 Kd Vhmentioning

confidence: 99%

“…Experiments were performed in our laboratory to determine the effect of the 8 kD VH factor (Banerjee et al, 1991) as well as the total VH on the mitotic activity of the epithelial cells of the organ cultured lens.…”

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confidence: 99%

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Growth regulation of the mammalian ocular lens by vitreous humor

Banerjee

Parafina

Bagchi

1992

J of Cellular Biochemistry

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Experiments were performed in our laboratory to study the effects of a mammalian 8 kD vitreous humor (VH) factor on the DNA synthesis and mitosis of the epithelial cells of organ cultured rabbit lens. The 8 kD polypeptide factor was purified from mature rabbit vitreous humor by liquid chromatography. Proliferative activities of the epithelial cells of organ cultured lenses were stimulated by 3% rabbit serum. The data from our experiments depicted that the 8 kD VH factor effectively inhibits DNA synthesis and mitosis by the epithelial cells of the organ cultured lens. Our experiments also showed that this 8 kD VH factor can maintain its growth inhibitory activity even when heated for 3 min at 95 degrees C. The growth inhibitory effect of the 8 kD VH factor was dose dependent. Using iodinated vitreal proteins it was demonstrated that the VH proteins are able to enter or bind to lens epithelial cells. The growth inhibitory effect of the 8 kD VH factor was also tested on tissue cultured lens epithelial cells. These experiments showed that the 8 kD VH factor has no growth inhibitory effect on the tissue cultured lens epithelial cells. This experiment has been repeated many times using different concentrations of the factor. These observations suggest that the 8 kD VH factor may have receptors in the lens capsular material (extracellular matrix) and the factor-receptor binding is essential for the growth inhibitory effect.

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Section: Figure L(i) Illustrates the Autoradiogram Of Iodinated Totalmentioning

confidence: 99%

mentioning

confidence: 90%

“…factor was isolated according to the technique of Banerjee et al (1991). VH was dissected out from freshly isolated rabbit lenses and centrifuged €or 3 h at 100,OOOg.…”

Section: Isolation Of V H Protein the 8 Kd Vhmentioning

confidence: 99%

mentioning

confidence: 99%

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Growth regulation of the mammalian ocular lens by vitreous humor

Banerjee

Parafina

Bagchi

1992

J of Cellular Biochemistry

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The mammalian iris‐ciliary complex affects organization and synthesis of cytoskeletal protein of organ and tissue cultured lens epithelial cells

Banerjee¹,

Emanuel²,

Parafina³

et al. 1992

J of Cellular Biochemistry

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A water soluble growth inhibitor was isolated from the mammalian ocular iris-ciliary complex. The molecular weight of this protein is 10 kD or lower as determined by ultrafiltration fractionation. The iris-ciliary (IC) complex water soluble protein(s) significantly inhibits synthesis of lower molecular weight proteins of the epithelial cells of the organ cultured mammalian ocular lens. It was also found that this inhibitory effect of IC is mediated via the structural organization of the lens. Monolayer cultures of the lens epithelial cells exposed to IC did not manifest any inhibition of their protein synthesis. Moreover, these tissue cultured lens epithelial (TCLE) cells showed a significant increase in their protein synthetic activities in response to the presence of IC factors in the culture medium. It is postulated that the IC activity is modulated via either the lens capsule, an extracellular matrix, or due to the specific organization of the intact lens. The specific effects of IC on the cytoskeletal organization and synthesis in the organ cultured lens epithelial (OCLE) and TCLE cells were also examined. Both groups, treated with IC factors, manifested significant alterations in their protein synthetic activities and cytoskeletal architecture. The 3H-leucine incorporation experiments showed that alpha-actin and alpha-tubulin synthesis is partially inhibited by IC factors in OCLE cells but vimentin synthesis is not, whereas in TCLE cells all of them showed increased synthesis in response to IC factors. Turnover rates of these proteins in both OCLE and TCLE cells were also computed. The immunofluorescence and microscopic evaluation of OCLE and TCLE cells exposed to IC factors illustrated significant alteration in the cytoarchitecture of the filaments. We demonstrate that an inhibitor(s) molecule of 10 kD or lower size isolated from IC inhibited protein synthesis of OCLE cells and stimulated protein synthesis in TCLE cells. The IC factor also affects the synthesis and organization of cytoskeletal filaments of both the OCLE and TCLE cells.

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