9cf3cb23ac0a7f5c72556cb21c86ae77 animals_a2022v12n8p1012.pdf bb299d868364fe29667415317b7677e5934818cf animals_a2022v12n8p1012.pdf 228d44a29b625caa6087f5c5871ce56985ba6fd79e970a5b2675c99a1153385c animals_a2022v12n8p1012.pdf Title: Bicarbonate-Triggered In Vitro Capacitation of Boar Spermatozoa Conveys an Increased Relative Abundance of the Canonical Transient Receptor Potential Cation (TRPC) Channels 3, 4, 6 and 7 and of CatSper- Subunit mRNA Transcripts Subject: Sperm capacitation is a stepwise complex biochemical process towards fertilization. It includes a crucial early calcium (Ca2+) transport mediated by CatSper channels and Canonical Transient Potential Channels (TRPC). We studied the relative abundance of mRNA transcripts changes of the CatSper , and subunits and TRPC-channels 1, 3, 4, 6 and 7 in pig spermatozoa, after triggering in vitro capacitation by bicarbonate ions at levels present in vivo at the fertilization site. For this purpose, we analyzedfive5 ejaculate pools (from three fertile adult boars) before (control-fresh samples) and after in vitro exposure to capacitation conditions (37 mM NaHCO3, 2.25 mM CaCl2, 2 mM caffeine, 0.5% bovine serum albumin and 310 mM lactose) at 38 C, 5% CO2 for 30 min. In vitro capacitation using bicarbonate elicits an increase in the relative abundance of mRNA transcripts of almost all studied Ca2+ channels, except CatSper- and TRPC1 (significantly reduced). These findings open new avenues of research to identify the specific role of each channel in boar sperm capacitation and elucidate the physiological meaning of the changes on sperm mRNA cargo. Keywords: calcium channels; catSper subunits; TRPC; pig; capacitation Author: Estíbaliz Lacalle, César Consuegra, Cristina A. Martínez, Manuel Hidalgo, Jesús Dorado, Felipe Martínez-Pastor, Manuel Álvarez-Rodríguez and Heriberto Rodríguez-Martínez Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Thu Apr 14 07:13:32 2022 CEST ModDate: Thu Apr 14 07:16:16 2022 CEST 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: 1172959 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- AKXRSZ+EURB10 Type 1 Builtin yes yes no 10 0 BVJSWV+VnURWPalladioL-Bold Type 1 Custom yes yes no 14 0 PWPXSM+VnURWPalladioL Type 1 Custom yes yes no 19 0 CCBNGP+EURM10 Type 1 Builtin yes yes no 23 0 SFEUXR+URWPalladioL-Bold Type 1 Custom yes yes no 27 0 DVHOUL+URWPalladioL-Roma Type 1 Custom yes yes no 32 0 BKISQV+URWPalladioL-Ital Type 1 Custom yes yes no 36 0 YFFKLH+CMSY10 Type 1 Builtin yes yes no 40 0 RUWDWW+EURM7 Type 1 Builtin yes yes no 44 0 MCBCCI+SourceSansRoman-Regular Type 1C Custom yes yes no 52 0 MCBCCM+SourceSansRoman.613wght Type 1C Custom yes yes no 56 0 MCBCCO+SourceSansRoman-Bold Type 1C Custom yes yes no 60 0 SZMRXJ+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 72 0 YDGZHF+PazoMath-Italic Type 1 Builtin yes yes no 97 0 MPUDFL+PazoMath Type 1 Builtin yes yes no 107 0 BCDEEE+PalatinoLinotype-Bold TrueType WinAnsi yes yes no 122 0 BCDFEE+PalatinoLinotype-Roman TrueType WinAnsi yes yes no 126 0 BCDGEE+PalatinoLinotype-Italic TrueType WinAnsi yes yes no 130 0 BCDIEE+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 134 0 BCDEEE+PalatinoLinotype-Bold TrueType WinAnsi yes yes no 159 0 BCDFEE+PalatinoLinotype-Roman TrueType WinAnsi yes yes no 163 0 BCDGEE+PalatinoLinotype-Italic TrueType WinAnsi yes yes no 167 0 BCDKEE+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 171 0 WABLSD+SFRM1000 Type 1 Custom yes yes no 186 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2022-04-27 02:23:43 CEST RepresentationInformation: animals_a2022v12n8p1012.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2022-04-20 11:27:20 CEST Size: 1172959 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 472 FreeObjects: 3 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7b69c6ba Item: Title: Materials and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@46daef40 Children: Item: Title: Ethics Statement Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@12f41634 Item: Title: Sperm Handling and In Vitro Capacitation Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@13c27452 Item: Title: Experimental Design Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@262b2c86 Item: Title: Sperm Evaluation Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@371a67ec Item: Title: Capacitation Status Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@5ed828d Children: Item: Title: Membrane Fluidity Assessment Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@50d0686 Item: Title: cAMP and PKA Assessment Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7a3d45bd Item: Title: RNA Extraction and qPCR Analyses of Calcium Channels Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1e7c7811 Item: Title: Statistical Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77ec78b9 Item: Title: Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1a3869f4 Children: Item: Title: In Vitro Capacitation Increases the Live-Capacitated Sperm Ratio Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@a38d7a3 Item: Title: In Vitro Capacitation for 30 min Increases the Sperm Protein Kinase A Content Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77f99a05 Item: Title: mRNA Expression of Catsper Subunits and TRPC Channels Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@63440df3 Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@3aeaafa6 Item: Title: Conclusions Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@76a3e297 Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4d3167f4 Info: Title: Bicarbonate-Triggered In Vitro Capacitation of Boar Spermatozoa Conveys an Increased Relative Abundance of the Canonical Transient Receptor Potential Cation (TRPC) Channels 3, 4, 6 and 7 and of CatSper- Subunit mRNA Transcripts Author: Estíbaliz Lacalle, César Consuegra, Cristina A. Martínez, Manuel Hidalgo, Jesús Dorado, Felipe Martínez-Pastor, Manuel Álvarez-Rodríguez and Heriberto Rodríguez-Martínez Subject: Sperm capacitation is a stepwise complex biochemical process towards fertilization. It includes a crucial early calcium (Ca2+) transport mediated by CatSper channels and Canonical Transient Potential Channels (TRPC). We studied the relative abundance of mRNA transcripts changes of the CatSper , and subunits and TRPC-channels 1, 3, 4, 6 and 7 in pig spermatozoa, after triggering in vitro capacitation by bicarbonate ions at levels present in vivo at the fertilization site. For this purpose, we analyzedfive5 ejaculate pools (from three fertile adult boars) before (control-fresh samples) and after in vitro exposure to capacitation conditions (37 mM NaHCO3, 2.25 mM CaCl2, 2 mM caffeine, 0.5% bovine serum albumin and 310 mM lactose) at 38 C, 5% CO2 for 30 min. In vitro capacitation using bicarbonate elicits an increase in the relative abundance of mRNA transcripts of almost all studied Ca2+ channels, except CatSper- and TRPC1 (significantly reduced). These findings open new avenues of research to identify the specific role of each channel in boar sperm capacitation and elucidate the physiological meaning of the changes on sperm mRNA cargo. 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