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Circulating small non-coding RNAs provide new insights into vitamin K nutrition and reproductive physiology in teleost fish
Authors:Ignacio Fernández  Jorge M.O. Fernandes  Vânia P. Roberto  Martina Kopp  Catarina Oliveira  Marta F. Riesco  Jorge Dias  Cymon J. Cox  M. Leonor Cancela  Elsa Cabrita  Paulo Gavaia
Affiliation:1. Centro de Ciências do Mar (CCMAR), Universidade do Algarve, Campus de Gambelas, 8005-139 Faro, Portugal;2. Center for Aquaculture Research, Agro-Technological Institute of Castilla y Leon, Ctra. Arévalo, s/n. 40196 Zamarramala, Segovia, Spain;3. Marine Genomics Group, Faculty of Biosciences and Aquaculture, Nord University, 8049 Bodø, Norway;4. Planta de Cultivos el Bocal, Spanish Institute of Oceanography (IEO), Barrio Corbanera, s/n 39012, Monte, Santander, Spain;5. SPAROS Ltd, Área Empresarial de Marim, Lote C, 8700-221 Olhão, Portugal;6. Department of Biomedical Sciences and Medicine, University of Algarve, Campus de Gambelas, 8005-139 Faro, Portugal;7. Algarve Biomedical Center (ABC), Universidade do Algarve, Campus de Gambelas, 8005-139 Faro, Portugal
Abstract:

Background

Vitamin K (VK) is a fat-soluble vitamin known for its essential role in blood coagulation, but also on other biological processes (e.g. reproduction, brain and bone development) have been recently suggested. Nevertheless, the molecular mechanisms behind its particular function on reproduction are not yet fully understood.

Methods

The potential role of VK on reproduction through nutritional supplementation in Senegalese sole (Solea senegalensis) was assessed by gonadal maturation and 11-ketosterone, testosterone and estriol plasma levels when fed with control or VK supplemented (1250?mg?kg?1 of VK1) diets along a six month trial. At the end, sperm production and quality (viability and DNA fragmentation) were evaluated. Circulating small non-coding RNAs (sncRNAs) in blood plasma from males were also studied through RNA-Seq.

Results

Fish fed with dietary VK supplementation had increased testosterone levels and lower sperm DNA fragmentation. SncRNAs from blood plasma were found differentially expressed when nutritional and sperm quality conditions were compared. PiR-675//676//4794//5462 and piR-74614 were found up-regulated in males fed with dietary VK supplementation. Let-7g, let-7e(18nt), let-7a-1, let-7a-3//7a-2//7a-1, let-7e(23nt) and piR-675//676//4794//5462 were found to be up-regulated and miR-146a and miR-146a-1//146a-2//146a-3 down-regulated when fish with low and high sperm DNA fragmentation were compared. Bioinformatic analyses of predicted mRNAs targeted by sncRNAs revealed the potential underlying pathways.

Conclusions

VK supplementation improves fish gonad maturation and sperm quality, suggesting an unexpected and complex regulation of the nutritional status and reproductive performance through circulating sncRNAs.

General significance

The use of circulating sncRNAs as reliable and less-invasive physiological biomarkers in fish nutrition and reproduction has been unveiled.
Keywords:Fat soluble vitamins  Phylloquinone  Small non-coding RNAs  Sperm quality  Reproduction  Corresponding author at: Center for Aquaculture Research, Ctra. Arévalo, s/n. 40196 Zamarramala, Spain.
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