Nuclear competence and genetic expression of growth differentiation factor‐9 (GDF-9) of canine oocytes in 3D culture
Articolo
Data di Pubblicazione:
2018
Citazione:
Nuclear competence and genetic expression of growth differentiation factor‐9 (GDF-9) of canine oocytes in 3D culture / M.G. Morselli, M. Loiacono, M. Colombo, M. Mortarino, G.C. Luvoni. - In: REPRODUCTION IN DOMESTIC ANIMALS. - ISSN 0936-6768. - 53:suppl. 3(2018 Nov), pp. 117-124. ((Intervento presentato al 20. convegno Congress of the European-Veterinary-Society-for-Small-Animal-Reproduction (EVSSAR) tenutosi a Wien nel 2017 [10.1111/rda.13336].
Abstract:
To evaluate the ability of a 3D culture system in improving the nuclear and molecular
competence of canine oocytes, barium alginate microcapsules were used for in vitro
maturation (IVM) and the expression profile of one selected oocyte‐secreted factor,
the growth differentiation factor‐9 (GDF‐9) was analysed. In Experiment I, canine
grade I cumulus–oocyte complexes (COCs) were in vitro matured in 3D microcapsules
in a controlled atmosphere for 72 hr, and meiosis resumption rates were compared
to those of oocytes cultured in traditional 2D microdrops of medium. In
Experiment II, a primer pair specific for canine GDF‐9 was designed, and preliminary
tested in conventional PCR on genomic DNA. Total RNA content was isolated from
oocytes at different time intervals (T0‐T24‐T48‐T72) during in vitro 3D culture, and a
reverse transcription to cDNA was performed. The expression of target gene was
assessed by quantitative Reverse Transcription Real‐Time PCR (qRT‐PCR), and the
obtained amplicons were sequenced to check the specificity of the analysis. Canine
COCs resumed meiosis at higher rates in 3D microcapsules than in 2D microdrops
(p < 0.05), even though no significant differences in the proportions of oocytes
achieving full maturational stages were obtained. A significant dynamic decrease in
GDF‐9 expression was recorded during culture: after 72 hr of IVM, the GDF‐9 transcription
significantly dropped (p = 0.018) compared to 24 and 48 hr. In conclusion, in
vitro 3D culture represents an efficient system for IVM of canine oocytes, and the
expression profile of GDF‐9 well reflects temporal dynamics for the acquisition of
developmental competence in this species.
Tipologia IRIS:
01 - Articolo su periodico
Keywords:
canine; GDF‐9; in vitro 3D culture; oocytes
Elenco autori:
M.G. Morselli, M. Loiacono, M. Colombo, M. Mortarino, G.C. Luvoni
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