37a66236da52b5efed13f4c0bc0d47cf pmc_35203238.pdf b4f1fcb8f2aea576bdfbfd4e7a3fa694bf39657e pmc_35203238.pdf 945e6cc3f64c01b3e7cc40f55dca6148624f2b0820c81ff1a031ac182bcfe2f1 pmc_35203238.pdf Title: The Role of Aquaporin 7 in the Movement of Water and Cryoprotectants in Bovine In Vitro Matured Oocytes Subject: Aquaglyceroporins are known as channel proteins, and are able to transport water and small neutral solutes. In this study, we evaluate the effect of exposure of in vitro matured bovine oocytes to hyperosmotic solutions containing ethylene glycol (EG), dimethyl sulfoxide (Me2SO) or sucrose on the expression levels of AQP3, AQP7 and AQP9. Moreover, we studied whether artificial protein expression of AQP7 in bovine oocytes increases their permeability to water and cryoprotectants. Exposure to hyperosmotic solutions stimulated AQP3 and AQP7 but not AQP9 expression. Oocytes exposed to hyperosmotic Me2SO solution exhibited upregulated AQP3 expression, while AQP7 expression was upregulated by EG hyperosmotic exposure. Microinjection of oocytes at the germinal vesicle stage with enhanced green fluorescent protein (EGFP) or EGFP+AQP7 cRNAs resulted in the expression of the corresponding proteins in 86% of the metaphase-II stage oocytes. AQP7 facilitated water diffusion when bovine MII oocytes were in presence of Me2SO solution but not EG or sucrose solution. However, the overexpression of this aquaporin did not increase membrane permeability to Me2SO or EG. In summary, cryoprotectant-induced increase of AQP3 and AQP7 expression could be one of the mechanisms underlying oocyte tolerance to hyperosmotic stress. Water diffusion appears to be improved when AQP7 overexpressed oocytes are exposed to Me2SO, shortening the time required for oocytes to achieve osmotic balance with cryoprotectant solutions. Keywords: cell membrane permeability; cryoprotective agents; artificial protein expression; aquaporin 3; aquaporin 9; cryopreservation; dimethyl sulfoxide; ethylene glycol; facilitated diffusion Author: Tania García-Martínez, Iris Martínez-Rodero, Joan Roncero-Carol, Meritxell Vendrell-Flotats, Jaume Gardela, Alfonso Gutiérrez-Adán, Priscila Ramos-Ibeas, Adam Z. Higgins and Teresa Mogas Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Tue Feb 22 10:45:04 2022 CET ModDate: Tue Feb 22 10:47:43 2022 CET Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 16 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 2382491 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- IBTYZX+VnURWPalladioL Type 1 Custom yes yes no 10 0 UMUMPQ+URWPalladioL-Bold Type 1 Custom yes yes no 15 0 MQLSSW+URWPalladioL-Roma Type 1 Custom yes yes no 20 0 SUIGJF+URWPalladioL-Ital Type 1 Custom yes yes no 24 0 DAEAVA+CMSY10 Type 1 Builtin yes yes no 28 0 IOJCNK+VnURWPalladioL-Bold Type 1 Custom yes yes no 32 0 MCBCCI+SourceSansRoman-Regular Type 1C Custom yes yes no 40 0 MCBCCM+SourceSansRoman.613wght Type 1C Custom yes yes no 44 0 MCBCCO+SourceSansRoman-Bold Type 1C Custom yes yes no 48 0 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no 196 0 EEMNDD+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 203 0 EEMNDF+PalatinoLinotype TrueType WinAnsi yes yes no 206 0 EEMNDH+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 209 0 EEMNDD+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 219 0 EEMNDF+PalatinoLinotype TrueType WinAnsi yes yes no 222 0 EEMNDH+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 225 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2022-06-03 02:24:15 CEST RepresentationInformation: pmc_35203238.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2022-02-27 18:35:56 CET Size: 2382491 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 446 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4923ab24 Item: Title: Materials and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@44c8afef Children: Item: Title: Chemicals Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7b69c6ba Item: Title: Oocyte Collection and In Vitro Maturation Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@46daef40 Item: Title: Analysis of AQP3, AQP7 and AQP9 Expression Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@12f41634 Item: Title: Preparation of AQP7 cRNA Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@13c27452 Item: Title: Microinjection of AQP7 cRNA into Bovine Oocytes Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@262b2c86 Item: Title: Expression of AQP7 in cRNA-Injected Oocytes Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@371a67ec Item: Title: Measurement of Permeability to Water and Cryoprotectants Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@5ed828d Item: Title: Experimental Design Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@50d0686 Item: Title: Statistical Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7a3d45bd Item: Title: Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1e7c7811 Children: Item: Title: Analysis of AQP3, AQP7 and AQP9 Expression in Oocytes after Treatment with Cryoprotection Solutions Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77ec78b9 Item: Title: Measurement of Permeability to Water and Cryoprotectants of In Vitro Matured Oocytes in Which AQP7 Was Artificially Overexpressed Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1a3869f4 Children: Item: Title: Expression of AQP7 in cRNA-Injected Oocytes Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@a38d7a3 Item: Title: Permeability to Water and Cryoprotectants of AQP7 cRNA-Injected Oocytes Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77f99a05 Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@63440df3 Item: Title: Conclusions Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@3aeaafa6 Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@76a3e297 Info: Title: The Role of Aquaporin 7 in the Movement of Water and Cryoprotectants in Bovine In Vitro Matured Oocytes Author: Tania García-Martínez, Iris Martínez-Rodero, Joan Roncero-Carol, Meritxell Vendrell-Flotats, Jaume Gardela, Alfonso Gutiérrez-Adán, Priscila Ramos-Ibeas, Adam Z. Higgins and Teresa Mogas Subject: Aquaglyceroporins are known as channel proteins, and are able to transport water and small neutral solutes. In this study, we evaluate the effect of exposure of in vitro matured bovine oocytes to hyperosmotic solutions containing ethylene glycol (EG), dimethyl sulfoxide (Me2SO) or sucrose on the expression levels of AQP3, AQP7 and AQP9. Moreover, we studied whether artificial protein expression of AQP7 in bovine oocytes increases their permeability to water and cryoprotectants. Exposure to hyperosmotic solutions stimulated AQP3 and AQP7 but not AQP9 expression. Oocytes exposed to hyperosmotic Me2SO solution exhibited upregulated AQP3 expression, while AQP7 expression was upregulated by EG hyperosmotic exposure. Microinjection of oocytes at the germinal vesicle stage with enhanced green fluorescent protein (EGFP) or EGFP+AQP7 cRNAs resulted in the expression of the corresponding proteins in 86% of the metaphase-II stage oocytes. AQP7 facilitated water diffusion when bovine MII oocytes were in presence of Me2SO solution but not EG or sucrose solution. However, the overexpression of this aquaporin did not increase membrane permeability to Me2SO or EG. In summary, cryoprotectant-induced increase of AQP3 and AQP7 expression could be one of the mechanisms underlying oocyte tolerance to hyperosmotic stress. Water diffusion appears to be improved when AQP7 overexpressed oocytes are exposed to Me2SO, shortening the time required for oocytes to achieve osmotic balance with cryoprotectant solutions. Keywords: cell membrane permeability; cryoprotective agents; artificial protein expression; aquaporin 3; aquaporin 9; cryopreservation; dimethyl sulfoxide; ethylene glycol; facilitated diffusion Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Tue Feb 22 10:45:04 CET 2022 ModDate: Tue Feb 22 10:47:43 CET 2022 ID: 0x49f942f4a25f28b3287363b9b363535e, 0x49f942f4a25f28b3287363b9b363535e Filters: FilterPipeline: FlateDecode Fonts: Type1: Font: BaseFont: IOJCNK+VnURWPalladioL-Bold FontSubset: true FirstChar: 161 LastChar: 189 FontDescriptor: FontName: IOJCNK+VnURWPalladioL-Bold Flags: Symbolic FontBBox: -152, -301, 1000, 935 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: OVNEXW+EURM10 FontSubset: true FirstChar: 22 LastChar: 22 FontDescriptor: FontName: OVNEXW+EURM10 Flags: Symbolic FontBBox: -32, -243, 1060, 720 FontFile: true Font: BaseFont: MQLSSW+URWPalladioL-Roma FontSubset: true FirstChar: 2 LastChar: 237 FontDescriptor: FontName: 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