A comparison of techniques for studying oogenesis in the European eel Anguilla anguilla
Ilaria
Mazzeo, Elisabetta Giorgini, Giorgia Gioacchini, Francesca Maradonna, M. Carmen
Vílchez, Sylvie Baloche, Sylvie Dufour, Luz Pérez, Oliana Carnevali, Juan F. Asturiano
Abstract
A multi-technique
approach was used to study the changes occurring in European eel Anguilla
anguilla ovaries during hormonally-induced vitellogenesis. Aside from
classic techniques used to monitor the vitellogenic process, such as ovary
histology, fat content analysis, sodium dodecyl sulfate–polyacrylamide gel
electrophoresis (SDS–PAGE), and vitellogenin enzyme linked immunosorbent assay
(ELISA), a new technique, Fourier Transform Infrared (FT-IR) Microspectroscopy,
was used for the first time to analyze A. anguilla ovaries. The results
from the different techniques provided different ways of approaching the same process.
Although it is considered a time consuming approach, of all the employed
techniques, histology provided the most direct evidences about vitellogenesis.
SDS-PAGE and ELISA were also useful for studying vitellogenesis, whereas fat
analysis cannot be used for this purpose. The FT-IR analysis provided a
representative IR spectrum for each ovarian stage (PV, EV, MV and LV), demonstrating
that it is a valid method able to illustrate the distribution of the oocytes
within the ovary slices. The obtained chemical maps confirmed changes in lipid
concentrations, and revealed their distribution within the oocytes at different
maturational stages. When the results and the accuracy of the FT-IR analysis were
compared to those of the traditional techniques commonly used to establish the
vitellogenic stage, it became evident that FT-IR is a useful and reliable tool,
with many advantages, including the fact that it requires little biological
material, the costs involved are low, analysis times are short, and, last but
not least, the fact that it offers the possibility of simultaneously analyzing
various biocomponents of the same oocyte.
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