Fertilization Rate and Number of Embryos on Day 2 after Intrauterine and Deep Intrauterine Insemination Using Frozen-Thawed Boar Semen in Multiparous Sows

The present study determines fertilization rate and number of embryos on Day 2 after intrauterine insemination (IUI) and deep intrauterine insemination (DIUI) using frozen-thawed (FT) boar semen in multiparous sows. Twelve crossbred Landrace × Yorkshire multiparous sows were included. The sows were inseminated at 24 h after oestrus detection and reinseminated every 12 h until ovulation took place. The inseminations were conducted using IUI with 2 × 109 FT sperm per dose (n = 6) and DIUI with 1 × 109 FT sperm per dose (n = 6). The sows were slaughtered at 45.1 ± 7.2 h after ovulation. Embryos and unfertilized oocytes were flushed from the oviducts. IUI yielded a better fertilization rate than DIUI (66.0% versus 31.0%, P < .001). The number of embryos was 13.5 ± 2.7 and 6.6 ± 3.2 embryos/sow in IUI and DIUI groups, respectively (P = .08). The proportion of sows having unilateral fertilization was higher in the DIUI (3/5) than the IUI group (1/6). In conclusion, IUI with at least 2 × 109 total number of FT boar spermatozoa is recommended

Double cloprostenol administration during mid luteal phase of oestrous cycle does not modify the interoestrous interval in gilts

The present study was undertaken to test the effect of two vulva injections of D-cloprostenol on day 7, 9 and 10 of oestrous cycle on the duration of the interestrous interval in gilts. Following a pre-treatment oestrous cycle, 87 gilts were assigned to receive vulva injections of 75 μg D-cloprostenol at 08:00 and 14:00 h on day 7 (D7; n=30), day 9 (D9; n=29) or day 10 (D10; n=28) of their second observed oestrous cycle. Across the treatments, the duration of the oestrous cycle with D-cloprostenol treatment (19.1±0.1 d) was not different from that of the previous oestrous cycle (20.1±0.4 days). Plasma progesterone concentrations were evaluated 6 h before and 24 and 72 h after D-cloprostenol treatment in the D9 group. Compared to pretreatment levels (9.6±0.4 ng/mL), plasma progesterone concentrations were reduced (P<0.05) at 24 h (6.3±1.0 ng/mL) and 72 h after treatment but complete luteolysis did not occur. These data indicate that in gilts double vulva administration of D-cloprostenol is not able to induce a complete luteolisys and hence the duration of the oestrous cycle is not modified

Cell cycle analysis and interspecies nuclear transfer of cat cells treated with chemical inhibitors

This study investigated the effect of chemical inhibitors on the cell-cycle synchronisation in cat fibroblast cells and evaluated the development of interspecies embryos reconstructed from cat donor cells and enucleated bovine oocytes. Cat fibroblast cells were treated with 15 μg/mL roscovitine or 0.05 μg/mL deme-colcine prior to cell cycle analysis and nuclear transfer. The percentage of cat fibroblast cells arrested at the G0/G1 phase in the roscovitine group was similar to that in the control group without any treatment. The percentage of cells arrested at the G2/M phase was significantly higher in the demecolcine group than in the control group. The fusion rate of interspecies couplets was significantly greater in the roscovitine group than in the control group. Most embryos stopped the development at the 2- or 4-cell stage, and none developed into blastocysts. Chemical inhibitor-induced donor cell cycle synchronisation did not overcome developmental arrest in interspecies cloned embryos

Establishment of a rabbit induced pluripotent stem cell (RbiPSC) line using lentiviral delivery of human pluripotency factors

Rabbit Embryonic Fibroblast (RbEF) cells (from Hycole hybrid rabbit foetus) were reprogrammed by lentiviral delivery of a self-silencing hOKSM polycistronic vector. The pluripotency of the newly generated RbiPSC was verified by the expression of pluripotency-associated markers and by in vitro spontaneous differentiation towards the 3 germ layers. Furthermore, the spontaneous differentiation potential of the iPSC was also tested in vivo by teratoma assay. The iPSC line showed normal karyotype. The advantages of using RbiPSC are the easy access to primary material and the possibility to study the efficacy and safety of the iPSC-based therapies on a non-rodent animal model

HISTONE H3 MODIFICATION OF ISCNT EMBRYOS

This study aimed to determine the acetylation patterns on histone H3K9/18/23 and the dimethylation pattern on histone H3K9 during early embryogenesis among 50 nM Trichostatin A (TSA)-treated iSCNT cat-cow embryos, untreated iSCNT cat-cow embryos (control) and bovine in vitro fertilisation (IVF) embryos, because TSA-treated iSCNT embryos are able to develop into blastocysts. The results show that the acetylation levels of H3K9/18/23 in the TSA-treated iSCNT and bovine IVF embryos were higher than those in the control embryos at almost all of the examined stages (2 h post-fusion / post-insemination (PF/PI), pronuclear (PN), two-cell, four-cell and eight-cell stages). At 6 h PF/PI the acetylation levels on H3K9/23 in the TSA-treated iSCNT and bovine IVF embryos were lower than those in the control, and there was no difference in the acetylation levels of H3K18 among the three groups. The acetylation levels of H3K9/23 increased either in the TSA-treated iSCNT or and bovine IVF embryos increased when those embryos developed to the PN and two-cell stages. The dimethylation level of H3K9 in the TSA-treated iSCNT embryos resembled that of the bovine IVF embryos at all examined stages (2h PF/PI, 6 h PF/PI and PN stages), and these levels were greater than those of the control. This result suggests that treatment of iSCNT embryos with TSA modifies the patterns of histone acetylation and dimethylation at certain lysine residues in a manner that is comparable with that seen in IVF embryos during early embryogenesis

Different DNA methylation patterns detected by the Amplified Methylation Polymorphism Polymerase Chain Reaction (AMP PCR) technique among various cell types of bulls

Background: The purpose of this study was to apply an arbitrarily primed methylation sensitive polymerase chain reaction (PCR) assay called Amplified Methylation Polymorphism Polymerase Chain Reaction (AMP PCR) to investigate the methylation profiles of somatic and germ cells obtained from Holstein bulls

Etude comparee du developpement in vivo des embryons de lapine cultives, congeles et cultives-congeles apres transfert synchrone et asynchrone

SIGLECNRS T 58795 / INIST-CNRS - Institut de l'Information Scientifique et TechniqueFRFranc