MiR-202 controls female fecundity by regulating medaka oogenesis.

Female gamete production relies on coordinated molecular and cellular processes that occur in the ovary throughout oogenesis. In fish, as in other vertebrates, these processes have been extensively studied both in terms of endocrine/paracrine regulation and protein expression and activity. The role of small non-coding RNAs in the regulation of animal reproduction remains however largely unknown and poorly investigated, despite a growing interest for the importance of miRNAs in a wide variety of biological processes. Here, we analyzed the role of miR-202, a miRNA predominantly expressed in male and female gonads in several vertebrate species. We studied its expression in the medaka ovary and generated a mutant line (using CRISPR/Cas9 genome editing) to determine its importance for reproductive success with special interest for egg production. Our results show that miR-202-5p is the most abundant mature form of the miRNA and that it is expressed in granulosa cells and in the unfertilized egg. The knock out (KO) of mir-202 gene resulted in a strong phenotype both in terms of number and quality of eggs produced. Mutant females exhibited either no egg production or produced a dramatically reduced number of eggs that could not be fertilized, ultimately leading to no reproductive success. We quantified the size distribution of the oocytes in the ovary of KO females and performed a large-scale transcriptomic analysis approach to identified dysregulated molecular pathways. Together, cellular and molecular analyses indicate that the lack of miR-202 impairs the early steps of oogenesis/folliculogenesis and decreases the number of large (i.e. vitellogenic) follicles, ultimately leading to dramatically reduced female fecundity. This study sheds new light on the regulatory mechanisms that control the early steps of follicular development, including possible targets of miR-202-5p, and provides the first in vivo functional evidence that a gonad-predominant microRNA may have a major role in female reproduction.

Incidence and predictive factors of atrial fibrillation in paced patients.

We have designed a prospective observational study to analyze the incidence and predictive factors of atrial fibrillation (AF) during a long follow-up, in a large population. Atrial fibrillation episodes were documented by the fallback mode switch (FMS) provided by implanted pacemakers. We have included 377 patients (61% men). The pacing indications were atrioventricular (AV) block (49%), sinus node disease (SND, 16%), bradycardia-tachycardia syndrome (BTS, 5%), AV block + SND (19%), AV block + BTS (6%), and BTS + SND (5%). The mean age at implant was 75 +/- 12 (range 28-95). Atrial fibrillation before inclusion was documented in 10% of patients. Drug therapy at first follow-up included beta-adrenergic blockers (17% of the patients), amiodarone (13%), and others (16%). The mean follow-up was 30 +/- 24 weeks. At least one AF episode was stored during follow-up in the memory of 169 pacemakers (45%). Among patients without history of AF at implant, 46% had documented FMS during follow-up. Patients with AF received more antiplatelet medications than patients without AF (P = 0.03). In patients with AF, New York Heart Association functional class was slightly higher, amiodarone and sotalol were more often prescribed, and the proportion of hypertension was higher than in patients without AF. However, these trends were not statistically significant. A significant higher incidence of premature atrial beats was observed in patients with AF than patients without AF (P < 0.0002). Patients with AF had a lower atrial percentage of paced events (55%) than patients without AF (63%, P < 0.02). These preliminary results confirm the high incidence of AF in paced patients and suggest a preventive effect of atrial pacing. The effects of other clinical variables may be confirmed with a longer follow-up in a larger population.