Anti-Histone H3 (full length) polyclonal antibody

Successful fertilization by intracytoplasmic sperm injection (ICSI) is possible as long as the sperm genome is intact, even in the context of defective sperm or sustained adverse treatment. However, there are few reports on rescuing gene-modified mouse lines after accidental death. To investigate whether sperm from a dead transgenic mouse can fertilize an oocyte and enable embryo development into a pup, Nestin-GFP transgenic male mice were sacrificed, and sperm was collected 14 h, 24 h, and 48 h after death. The collected sperm was injected into oocytes from hybrid B6D2F1 or inbred C57BL/6 N mice. The results showed that the sperm in the three groups activated oocytes from B6D2F1 and supported embryo development to the blastocyst stage. For ICSI embryos derived from B6D2F1 mice, the cleavage and blastocyst rates were significantly lower in the three experimental groups than in the control group (0 h) (P < 0.05), and the birth rate in the 24 h and 48 h groups was significantly lower than that in the 14 h and control groups (0 h). For C57BL/6N-derived ICSI embryos, the cleavage rates were significantly lower at 24 h and 48 h than at 14 h and 0 h (control group), and the birth rate in the three experimental groups was significantly lower than that in the control group (0 h). The F0 mice derived from B6D2F1 and C57BL/6 N oocytes had normal reproductive ability, and F1 mice were successfully obtained. The characteristics of the GFP gene were preserved and inherited. The histone H2AX phosphorylation assay showed that the proportion of focus-negative embryos was markedly and significantly lower in the 14 h, 24 h, and 48 h groups than in the control group (0 h). The proportion of focus-negative embryos was significantly lower at 48 h than at 14 h or 24 h. The number of foci was significantly higher in the three experimental groups than in the control group (0 h), indicating that sperm DNA sustained more damage after death and that few sperm had an intact genome. In summary, sperm obtained from mice 14 h, 24 h, and 48 h after death is capable of activating an oocyte and supporting complete embryo development into a pup. This study provides an effective way to rescue accidently died mouse strains. (C) 2020 Elsevier Inc. All rights reserved.

Pre-eclampsia (PE) is a major cause of maternal and perinatal death. Previous research has indicated the role of histone deacetylase 2 (HDAC2) in the pathogenesis of PE but the relevant molecular mechanisms are unknown. However, there is hitherto little information concerning the molecular mechanism behind HDAC2 in PE. Herein, we hypothesized that HDAC2 promotes trophoblast cell proliferation and this requires the involvement of microRNA-183 (miR-183), forkhead box protein A1 (FOXA1), and interleukin 8 (IL-8). We collected placental specimens from 30 PE affected and 30 normal pregnant women. HDAC2 and FOXA1 were poorly expressed while miR-183 and IL-8 were highly expressed in placental tissues in PE. In vitro, HDAC2 overexpression enhanced the proliferation, migration, and invasion of human trophoblast cells HTR-8/SVNEO. HDAC2 inhibited the expression of miR-183 by diminishing H4 acetylation in the miR-183 promoter region. miR-183 inhibition by its specific inhibitor increased the expression of FOXA1 and thus enhanced HTR-8/SVNEO cell proliferation, migration, and invasion. FOXA1, a transcriptional factor, enhanced HTR-8/SVNEO cell proliferation, migration, and invasion by inhibiting the transcription of IL-8. We also observed HDAC2 knockdown was lost upon FOXA1 overexpression, suggesting that HDAC2 could promote HTR-8/SVNEO proliferation, migration, and invasion through the miR-183/FOXA1/IL-8 pathway. In summary, the results highlighted the role of the HDAC2/miR-183/FOXA1/IL-8 pathway in PE pathogenesis and thus suggest a novel molecular target for PE.