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First published online May 24, 2004
Journal of Experimental Biology 207, 2255-2265 (2004)
Published by The Company of Biologists 2004
doi: 10.1242/jeb.01004

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Dietary influences over proliferating cell nuclear antigen expression in the locust midgut

E. Zudaire¹, S. J. Simpson², I. Illa¹ and L. M. Montuenga^1,*

¹ Department of Histology and Pathology, Schools of Medicine and Sciences, University of Navarra, E-31080 Pamplona, Spain
² Department of Zoology and University Museum of Natural History, University of Oxford, South Parks Road, Oxford OX1 3PS, UK

^* Author for correspondence (e-mail: lmontuenga{at}unav.es)

Accepted 29 March 2004

We have studied the influence of variations in dietary protein (P) and digestible carbohydrate (C), the quantity of food eaten, and insect age during the fifth instar on the expression of the proliferating cell nuclear antigen (PCNA) in the epithelial cells of the midgut (with special reference to the midgut caeca) in the African migratory locust, Locusta migratoria. Densitometric analysis of PCNA-immunostained cells was used as an indirect measure of the levels of expression of PCNA, and a PCNA cellular index (PCNA-I) was obtained. Measurements of the DNA content of the cells have also been carried out by means of microdensitometry of Feulgen-stained, thick sections of midgut. A comparison between the PCNA nuclear level and the DNA content was performed. The PCNA levels were significantly different among the cells of the five regions studied: caeca, anterior ventricle, medial ventricle, posterior ventricle and ampullae of the Malpighian tubules. We have studied in more detail the region with highest PCNA-I, i.e. the caeca. The quality and the quantity of food eaten under ad libitum conditions were highly correlated with both the PCNA and DNA levels in the caeca cells. Locusts fed a diet with a close to optimal P:C content (P 21%, C 21%) showed the highest PCNA and DNA content. In locusts fed a food that also contained a 1:1 ratio of P to C but was diluted three-fold by addition of indigestible cellulose (P 7%, C 7%), a compensatory increase in consumption was critical to maintaining PCNA levels. Our measurements also showed that the nuclear DNA content of the mature and differentiated epithelial cells was several-fold higher than the levels in the undifferentiated stem cells of the regenerative nests. These results, combined with the low number of mitotic figures found in the regenerative nests of the caeca and the marked variation in PCNA levels among groups, suggest that some type of DNA endoreduplication process may be taking place. Our data also indicate that the DNA synthetic activity in the midgut is related to feeding in locusts. The possible dietary and nutritional regulatory mechanisms and the significance of the differences found are discussed.

Key words: proliferating cell nuclear antigen, PCNA, Locusta migratoria, Insecta, BrdU incorporation, DNA synthesis, protein, carbohydrate, midgut, feeding behaviour, diet, nutritional balance

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