Proliferating cell nuclear antigen in gonad and associated storage tissue of the Pacific oyster Crassostrea gigas: seasonal immunodetection and expression in laser microdissected tissues |
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Authors: | Alban Franco Aude Jouaux Michel Mathieu Pascal Sourdaine Christophe Lelong Kristell Kellner Clothilde Heude Berthelin |
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Institution: | 1. UMR 100 Ifremer, Physiologie et Ecophysiologie des Mollusques Marins, IFR 146 ICORE, Université de Caen Basse-Normandie, 14 032, Caen Cedex, France 2. UMR 100?M Ifremer, IFR 146 ICORE, Université de Caen Basse-Normandie, 14 032, Caen Cedex, France
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Abstract: | To understand the processes involved in tissue remodeling associated with the seasonal reproductive cycle of the oyster Crassostrea gigas, we used immunodetection and expression measurements of proliferating cell nuclear antigen (PCNA). The expression of the
PCNA gene was measured by real-time polymerase chain reaction in the whole gonadal area compared with laser microdissected
gonad and storage tissue. Results underlined the advantage of the laser microdissection approach to detect expression, mainly
for early stages of spermatogenesis. In the storage tissue, PCNA expression was reduced in the gonadal tubules, but immunolabeled
hemocytes and vesicular cells were detected when the storage tissue was being restored. In the gonadal tubules, the PCNA gene
was more highly expressed in males than in females. As soon as spermatogenesis was initiated, PCNA expression showed a high
and constant level. In females, the expression level increased gradually until the ripe stage. The immunological approach
established the involvement of peritubular cells in gonadal tubule expansion during early gametogenesis. In both sexes, gonial
mitosis was immunodetected throughout the reproductive cycle. In males, the occurrence of two types of spermatogonia was ascertained
by differential immunolabeling, and intragonadal somatic cell proliferation was noted. As expected, immunolabeling was never
observed from stage II spermatocytes to spermatozoa. In females, positively stained cells were detected from oogonia to growing
oocytes with various labeled intracellular locations. |
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