Autoradiographic Localization of Tritiated Thymidine in Whole-Mount Preparations of Lens Epithelium

Harding, Clifford V.; Hughes, Walter L.; Bond, V.P.; Schork, P.

doi:10.1001/archopht.1960.00950020060009

Cited by 62 publications

(11 citation statements)

References 8 publications

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“…In addition, there was no evidence of ectopic proliferation in the anterior subcapsular central zone. Harding et al (9), however, who used whole-mount preparations of the lens epithelium, recognized labelled cells also in the centroanterior subcapsular region. Their results were consistent with the ones reported here which showed considerable ectopic proliferation in the anterior subcapsular epithelium of three-weeks-old rats using whole-mount preparations.…”

Section: Resultsmentioning

confidence: 99%

The proliferative ability of the lens epithelium in rat galactose cataracts. Examination using whole-mount preparations.

Terubayashi

Tsuji

Okamoto

et al. 1991

Acta Histochem. Cytochem

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We gave three-weeks-old rats a diet which included galactose in three different concentrations (15%, 25%, 50%) producing sugar cataracts of three different degrees.Combining the 3H-thymidine autoradiographic method with the whole-mount preparations of total lens epithelial cells of one lens, we examined the proliferative activity of the lens epithelium in the early stages of cataract crisis (for 14 days). In the lenses of three-weeks-old normal rats, 3H-thymidine labelled cells were observed mainly in the proliferative zone at the lens equator, and at the same time a few were detected in other parts. On the first day after we started feeding the rats galactose-included diets, the very early stages of cataract formation and an increase in the number of labelled cells were observed in the proliferative zone when compared with the control group on a normal diet. On the fourth day, we found an increase in the number of labelled cells in the proliferative zone and a remarkable ectopic proliferation in other parts. However, in contrast to this, as the cataract progressed from the seventh day to the fourteenth day, we noticed a decrease in the number of labelled cells in the proliferative zone and in the other epithelial layers when compared with the control group. The peak in the number of labelled cells on the fourth day was observed regardless of the galactose concentrations in the diet, and was not proportional to the degree of the cataracts.In rat galactose cataracts, the cure phenomenon (Diet-Reversal), in which the lens opacity is removed by altering the galactose-including diet to the normal diet, was observed. This Diet-Reversal was proved through light microscopy (17). Moreover, we also reported that aldose reductase inhibitors (ART) had curative effects and preventive effects on galactose cataracts (1-3, 11, 12). On the basis of the above-mentioned results, we think it is essential that the cataract lens is repaired by the regenerated fibers at the lens equator. That is to say, the examination of the proliferative ability and the kinetics of the regenerated lens epithelium seems to be very important in order to develop the cataract therapeutic agents for the future, though it is impossible at present.We used the 3H-thymidine autoradiographic method on thin sections embedded with methacrylate, and proved that, in the developmental process of rat galactose cataracts, the lens epithelium will repeat the ectopic proliferation even in other parts than the lens equator, and play an important role in the repair of the regenerated lens fibers (13,14). In particular, on the fourth day after we started feeding rats the 50% galactose-including diet, a remarkable increase in the amount of ectopic proliferation was observed. But to give a conjecture on the whole layer of lens epithelium from only one thin section seemed to be unreasonable. Therefore, in this investigation we used the whole-mount preparations of the lens epithelial layer. In this study, we examined the ability of the rat lens epithelium to proliferate dur...

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Section: Resultsmentioning

confidence: 99%

The proliferative ability of the lens epithelium in rat galactose cataracts. Examination using whole-mount preparations.

Terubayashi

Tsuji

Okamoto

et al. 1991

Acta Histochem. Cytochem

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“…2A). In accordance with previous nomenclature (Harding et al, 1960) this region was called the germinative zone (GZ). At the equator, the reorganization of cell nuclei into meridional rows (MR) signified the onset of fiber cell differentiation, thus delineating the edge of the epithelium.…”

Section: Introductionmentioning

confidence: 86%

A stochastic model of eye lens growth

Šikić

Shi

Lubura

et al. 2015

Journal of Theoretical Biology

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The size and shape of the ocular lens must be controlled with precision if light is to be focused sharply on the retina. The lifelong growth of the lens depends on the production of cells in the anterior epithelium. At the lens equator, epithelial cells differentiate into fiber cells, which are added to the surface of the existing fiber cell mass, increasing its volume and area. We developed a stochastic model relating the rates of cell proliferation and death in various regions of the lens epithelium to deposition of fiber cells and lens growth. Epithelial population dynamics were modeled as a branching process with emigration and immigration between various proliferative zones. Numerical simulations were in agreement with empirical measurements and demonstrated that, operating within the strict confines of lens geometry, a stochastic growth engine can produce the smooth and precise growth necessary for lens function.

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“…Cataracts that develop in rats fed a galactose supplemented diet are thought to be useful as an animal model of diabetic cataracts [7,13,15]. We believe that the proliferation potency at the central part of the lens is strongly related to maturity onset cataracts or diabetic cataracts, because this proliferation potency decreases with aging [5,6,12,16] and increases in some types of human cataracts or galactose induced cataract in rats [4,14], while lens epithelial cells in normal rats grow vigorously at the equator. In the present study, the proliferation potency of lens epithelial cells at the central part of the lens was studied in rats fed 5%-25% galactose supplemented diets in order to examine the relationship between cataract development and epithelial cells.…”

Section: Discussionmentioning

confidence: 99%

“…Growth of lens epithelial cells, except for those at the lens equator, decreases after the juvenile growth spurt [5,6,12,16]. In general, while senile cataracts are caused by cortical changes such as swelling and liquefaction of lens fiber, subcapsular cataracts display abnormal growth and stratification of lens epithelial cells from the early stages of the disease, suggesting that the development of cataracts is profoundly influenced by epithelial cell growth.…”

Section: Introductionmentioning

confidence: 99%

Cell Growth of Rat Lens Epithelium in Galactose-induced Cataracts.

Kubo

Takayanagi

Tsuzuki

et al. 1997

Acta Histochem. Cytochem

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Cell growth of rat lens epithelium during galactose-induced cataract formation was examined in the present study using fluorescence cytophotometry or proliferating cell nuclear antigen (PCNA) immunohistochemistry.Six-week old male SpragueDawley rats were divided into 4 dietary groups according to galactose supplementation of the diet: 5%, 10%, 15% and 25% galactose diet groups. Whole-mount preparations of rat lens epithelium were prepared.The nuclear content of epithelial cells was measured using fluorescence cytophotometry, and cells undergoing DNA synthesis (PCNA labeled cells) were detected using the PCNA immunohistochemical staining method.The results of fluorescence cytophotometry showed that cells containing tetraploid nuclei (4C cells) gradually increased in number. In the 5% galactose diet fed rats, the number of 4C cells peaked on the 9th day after the start of dietary galactose supplementation, the 7th day in the 10% galactose diet fed rats and on the 5th day in the 15% and 25% galactose diet fed rats. On each day that 4C cells peaked, a large number of PCNA labeled cells were observed in the central zone of the epithelial cell layer. These results suggest that growth stimulation of lens epithelial cells depends upon the concentration of galactose in the diet and precedes changes in lens fiber.

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Autoradiographic Localization of Tritiated Thymidine in Whole-Mount Preparations of Lens Epithelium

Cited by 62 publications

References 8 publications

The proliferative ability of the lens epithelium in rat galactose cataracts. Examination using whole-mount preparations.

The proliferative ability of the lens epithelium in rat galactose cataracts. Examination using whole-mount preparations.

A stochastic model of eye lens growth

Cell Growth of Rat Lens Epithelium in Galactose-induced Cataracts.

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