Abstract
This study investigated the effect of Folliculinum 6 cH on the oocyte meiosis resumption and viability rates, progesterone production and mitochondrial activity after in vitro maturation of cumulus-oocyte complexes (COCs) in sheep. Sheep ovaries were collected at a local slaughterhouse and COCs were recovered by slicing technique. The selected COCs were maturated in TCM199 (Control treatment), or control medium supplemented with 0.05% ethanol (v/v) (the vehicle of the homeopathic preparation – Ethanol treatment) or with Folliculinum 6 cH. After 24 h of in vitro maturation (IVM), oocytes were mechanically denuded and incubated with Hoechst 33342 and MitoTracker (0.5 μM) Orange CMTMRos for analysis of viability and chromatin configuration, and mitochondrial activity, respectively. The results showed that Folliculinum 6 cH addition increased oocyte degeneration and reduced meiotic resumption compared to the control (P < 0.05). Interestingly, the percentages meiotic resumption and oocyte maturation were lower in the Folliculinum 6 cH treatment compared to its vehicle (Ethanol treatment) (P < 0.05). On the other hand, when the treatments were compared, higher mitochondrial activity was observed in the Ethanol treatment (P < 0.05). In conclusion, contrary to its vehicle, the addition of Folliculinum 6 cH to the IVM medium promoted oocyte degeneration and affected negatively the mitochondrial distribution, impairing meiosis resumption.
Highlights
Homeopathy is a therapeutic method based on the use of drugs in a minimal dosage during treatment (Teixeira et al, 2008)
The addition of ethanol to the control in vitro maturation (IVM) medium did not affect the percentage of degenerate, germinal vesicle (GV) and germinal vesicle breakdown (GVBD) and metaphase II (MII) oocytes after IVM (Table 1)
The percentages of meiotic resumption and MII-oocytes were lower in the Folliculinum 6 cH treatment compared to its vehicle (P < 0.05 - Table 1)
Summary
Homeopathy is a therapeutic method based on the use of drugs in a minimal dosage during treatment (Teixeira et al, 2008). Others argued that the supposed effect of homeopathic products is due to their vehicles, such as ethanol or even to a possible placebo effect (Moffett et al, 2006) In this sense, the in vitro models, for example the IVM, could represent an excellent tool to solve such intriguing issues. Concerning reproduction, few studies investigated the effects of homeopathic medicinal products on the in vitro preantral follicle survival and development (sheep – Lima et al, 2016 and pig - Lima et al, 2017). The originality of the present paper is to investigate for the first time the effect of Folliculinum 6 cH and its vehicle (ethanol) on the in vitro maturation, viability and mitochondrial activity of ovine oocytes
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