title: Seminal plasma and seminal plasma proteins added to bulk sorted sperm do not alter the mRNA expression of in vitro produced bovine embryos.
abstract: Although sex-sorted sperm have been used for AI and IVF for over a decade there is still need to improve the technology as the results are highly variable. The goal of the present study was to assess the effect of seminal plasma and seminal plasma proteins as a supplement to sorted sperm on subsequent embryonic development, as a beneficial effect of these substances has been reported. In vitro matured oocytes were fertilized in vitro with either unsorted sperm (n = 215; Group 1), bulk sorted sperm (n = 226; Group 2), bulk sorted sperm extended in the presence of 1% seminal plasma (n = 185; Group 3) or bulk sorted sperm supplemented with seminal plasma proteins (4 mg mL(-1); n = 254; Group 4). An additional group of oocytes (n = 307; Group 5) was fertilized with the semen of another bull routinely used for IVF and served as a laboratory standard control. Subsequently, the presumptive zygotes were cultured for 8 days under standard conditions (SOFaa, 39 °C, 5% CO(2), 5% N(2)). Cleavage rates were assessed on day 3 p.i. (post insemination; group 1: 30.5 ± 14.7%; group 2: 28.8 ± 9.8%; group 3: 20.8 ± 14.9%; group 4: 25.7 ± 8.2%; group 5: 54.8 ± 11.5%). Development rates were documented on days 7 p.i. (group 1: 7.3 ± 6.6%; group 2: 5.6 ± 3.1%, group 3: 6.2 ± 7.7%, group 4: 6.7 ± 5.9%, group 5: 20.2 ± 6.9%) and 8 p.i. (group 1: 8.9 ± 7.0%; group 2: 6.0 ± 2.9%; group 3: 8.6 ± 11.3%; group 4: 7.8 ± 6.2%; group 5: 23.3 ± 7.8%), respectively. Significant differences among cleavage and development rates could only be seen for Group 5 compared to all other groups. However, this difference between Groups 1-4 vs. Group 5 regarding the development rates on Day 8 could not be detected when assessing the development rates on base of the number of cleaved embryos instead of the number of oocytes fertilized (group 1: 31.4 ± 17.2%; group 2: 26.0 ± 21.0%; group 3: 33.3 ± 19.05%; group 4: 26.6 ± 17.8%; group 5: 42.6 ± 11.3%). The relative abundance of six different developmentally important gene transcripts (G6PD, HSP1A1, SLC2A3, BAX, BCL2L1, DNMT3A) was determined using single Day 8 expanded blastocysts of all five groups. No significant differences were seen among the embryos of the five groups. Our results show that neither the bulk sorting procedure nor the addition of seminal plasma or seminal plasma proteins, respectively, affected cleavage and development rates when sperm from a specific bull was used. Additionally, sorting and subsequent exposure of sperm to either seminal plasma or seminal plasma proteins did not influence mRNA expression in bovine IVP embryos.
authors.affiliation: Clinic for Cattle, University of Veterinary Medicine, Hanover, Germany. Hanna.stinshoff@tiho-hannover.de
authors.forename: H
authors.initials: H
authors.lastname: Stinshoff
authors.forename: M
authors.initials: M
authors.lastname: Krienke
authors.forename: M
authors.initials: M
authors.lastname: Ekhlasi-Hundrieser
authors.forename: S
authors.initials: S
authors.lastname: Wilkening
authors.forename: A
authors.initials: A
authors.lastname: Hanstedt
authors.forename: D
authors.initials: D
authors.lastname: Frese
authors.forename: D
authors.initials: D
authors.lastname: Rath
authors.forename: H
authors.initials: H
authors.lastname: Bollwein
authors.forename: C
authors.initials: C
authors.lastname: Wrenzycki
mesh_terms: D000818:Animals; D002417:Cattle / Q000196:embryology / Q000235:genetics / Q000378:metabolism; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D002970:Cleavage Stage, Ovum / Q000166:cytology / Q000187:drug effects; D046149:Embryo Culture Techniques; D004622:Embryo, Mammalian / Q000187:drug effects* / Q000378:metabolism; D047108:Embryonic Development / Q000187:drug effects / Q000235:genetics; D005260:Female; D005307:Fertilization in Vitro / Q000187:drug effects / Q000379:methods / Q000662:veterinary; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D059471:In Vitro Oocyte Maturation Techniques / Q000662:veterinary; D008297:Male; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D012661:Semen / Q000502:physiology*; D029589:Seminal Plasma Proteins / Q000494:pharmacology*; D013094:Spermatozoa / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism
chemical_list: D012333:RNA, Messenger; D029589:Seminal Plasma Proteins