e934addac466c9f3b4bef67165031dcb pmc_34068214.pdf 8a7ac18bcd49becd3e34456b7058ff25cbb9ebb9 pmc_34068214.pdf 10e7337e9446d9c32b9b857842cbd4621ef03924538c808293d97d01078d993e pmc_34068214.pdf Title: Specific Seminal Plasma Fractions Are Responsible for the Modulation of Sperm–PMN Binding in the Donkey Subject: While artificial insemination (AI) with frozen-thawed sperm results in low fertility rates in donkeys, the addition of seminal plasma, removed during cryopreservation, partially counteracts that reduction. Related to this, an apparent inflammatory reaction in jennies is induced following AI with frozen-thawed sperm, as a high amount of polymorphonuclear neutrophils (PMN) are observed within the donkey uterus six hours after AI. While PMN appear to select the sperm that ultimately reach the oviduct, two mechanisms, phagocytosis and NETosis, have been purported to be involved in that clearance. Remarkably, sperm interacts with PMN, but the presence of seminal plasma reduces that binding. As seminal plasma is a complex fluid made up of different molecules, including proteins, this study aimed to evaluate how different seminal plasma fractions, separated by molecular weight (<3, 3–10, 10–30, 30–50, 50–100, and >100 kDa), affect sperm–PMN binding. Sperm motility, viability, and sperm–PMN binding were evaluated after 0 h, 1 h, 2 h, 3 h, and 4 h of co-incubation at 38 C. Two seminal plasma fractions, including 30–50 kDa or 50–100 kDa proteins, showed the highest sperm motility and viability. As viability of sperm not bound to PMN after 3 h of incubation was the highest in the presence of 30–50 and 50–100 kDa proteins, we suggest that both fractions are involved in the control of the jenny’s post-breeding inflammatory response. In conclusion, this study has shown for the first time that specific fractions rather than the entire seminal plasma modulate sperm–PMN binding within the donkey uterus. As several proteins suggested to be involved in the control of post-AI endometritis have a molecular weight between 30 and 100 kDa, further studies aimed at determining the identity of these molecules and evaluating their potential effect in vivo are much warranted. Keywords: sperm; inflammatory reaction; polymorphonuclear neutrophils (PMN); sperm–PMN binding; donkey Author: Jordi Miró, Jaime Catalán, Henar Marín, Iván Yánez-Ortiz and Marc Yeste Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Thu May 13 12:25:09 2021 CEST ModDate: Thu May 13 12:25:09 2021 CEST Custom Metadata: yes Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 14 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 2463268 bytes Optimized: no PDF version: 1.5 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- FHNRXR+URWPalladioL-Ital Type 1 Custom yes yes no 139 0 KNXWCM+URWPalladioL-Bold Type 1 Custom yes yes no 140 0 PRJKGQ+VnURWPalladioL-Bold Type 1 Custom yes yes no 141 0 XXPQEC+URWPalladioL-Roma Type 1 Custom yes yes no 142 0 XEYJZU+CMSY10 Type 1 Builtin yes yes no 145 0 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ViewArea: CropBox ViewClip: CropBox PrintArea: CropBox PageClip: CropBox PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: ぉのぴひはつふっぴどはの〠 Item: Title: きちぴづひどちぬび〠ちのつ〠きづぴとはつび〠 Children: Item: Title: ぁのどねちぬび〠 Item: Title: くぢぴちどのどので〠こづねどのちぬ〠ぐぬちびねち〠こちねばぬづび〠 Item: Title: うひちっぴどはのどので〠こづねどのちぬ〠ぐぬちびねち〠こちねばぬづび〠 Item: Title: ぉびはぬちぴどはの〠はて〠ぐきぎ〠 Item: Title: こづねづの〠ぃはぬぬづっぴどはの〠ちのつ〠ぐひづばちひちぴどはの〠 Item: Title: ぃは〭どのっふぢちぴどはの〠はて〠こばづひね〠ちのつ〠ぐきぎ〠 Item: Title: ぅぶちぬふちぴどはの〠はて〠こばづひね〠ざどちぢどぬどぴべ〠ちのつ〠きはひばとはぬはでべ〠 Item: Title: ぅぶちぬふちぴどはの〠はて〠こばづひね〠きはぴどぬどぴべ〠 Item: Title: いづぴづひねどのちぴどはの〠はて〠こばづひね〓ぐきぎ〠あどのつどので〠 Item: Title: こぴちぴどびぴどっちぬ〠ぁのちぬべびづび〠 Item: Title: げづびふぬぴび〠 Children: Item: Title: ぅててづっぴ〠はて〠こづねどのちぬ〠ぐぬちびねち〠うひちっぴどはのび〠はの〠こばづひね〓ぐきぎ〠あどのつどので〠 Item: Title: ぅててづっぴ〠はて〠こづねどのちぬ〠ぐぬちびねち〠うひちっぴどはのび〠はの〠ぴとづ〠ざどちぢどぬどぴべ〠はて〠さのぢはふのつ〠ちのつ〠あはふのつ〭ぴは〭ぐきぎ〠こばづひね〠ぐはばふぬちぴどはのび〠 Item: Title: ぅててづっぴ〠はて〠こづねどのちぬ〠ぐぬちびねち〠うひちっぴどはのび〠はの〠ぴとづ〠きはぴどぬどぴべ〠はて〠さのぢはふのつ〠ちのつ〠あはふのつ〭ぴは〭ぐきぎ〠こばづひね〠ぐはばふぬちぴどはのび〠 Item: Title: いどびっふびびどはの〠 Item: Title: ぃはのっぬふびどはのび〠 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