Creatinine, arachidonic acid (AA), docosahexaenoic acid and uric acid were obtained from the National Institutes for Food and Drug Control. L-valine was purchased from Amresco Company. Kynurenic acid, hypotaurine were purchased from Sigma Company or Aladdin Company. Antibodies against transforming growth factor-β 1 (TGF-β1), TGF-β RII, TGF-β RI, Smad2, Smad3, p-Smad2, p-Smad3, Smad4, Smad7, α-SMA, collagen I, fibronectin, E-cadherin, FSP1, vimentin, collagen III, and GAPDH were purchased from Santa Cruz Biotechnology, Cell Signaling Technology, or Abcam Company. Ultra purity water was prepared using a Milli-Q water purification system (Billerica, MA, United States). Other chemicals were of analytical grade, and their purity was above 99.5%.
Hypotaurine
Manufactured by Merck Group
Sourced in United States
Hypotaurine is a chemical compound used in laboratory equipment. It serves as a core functional component in various applications, but a detailed description without interpretation or extrapolation cannot be provided.
Automatically generated - may contain errors
Lab products found in correlation
Penicillamine, by Merck Group (4 mentions)
Epinephrine, by Merck Group (4 mentions)
Tamoxifen, by Merck Group (2 mentions)
Bsa fraction 5, by Merck Group (2 mentions)
Fibronectin, by Santa Cruz Biotechnology (1 mentions)
Vimentin, by Santa Cruz Biotechnology (1 mentions)
Collagen 1, by Santa Cruz Biotechnology (1 mentions)
Collagen 3, by Santa Cruz Biotechnology (1 mentions)
α sma, by Santa Cruz Biotechnology (1 mentions)
E cadherin, by Santa Cruz Biotechnology (1 mentions)
Kynurenic acid, by Merck Group (1 mentions)
Gapdh, by Santa Cruz Biotechnology (1 mentions)
Heparin, by Merck Group (1 mentions)
Mineral oil, by Merck Group (1 mentions)
Hybond ecl nitrocellulose membrane, by Cytiva (1 mentions)
Hank s medium, by Merck Group (1 mentions)
Rabbit anti mouse secondary antibody, by Santa Cruz Biotechnology (1 mentions)
β oestradiol, by Merck Group (1 mentions)
Goat anti rabbit secondary antibody, by Santa Cruz Biotechnology (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
10 protocols using hypotaurine
1
Transforming Growth Factor-β Signaling Pathway
2
In Vitro Fertilization of Bovine Oocytes
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After 20–24 h of IVM, COCs were washed twice in TL medium before being transferred in groups of 5–48 μl droplets under mineral oil. The IVF droplets consisted of modified TL medium supplemented with fatty-acid-free BSA (0.6% w/v), pyruvic acid (0.2 mM), heparin (2 μg/mL) and gentamycin (50 mg/mL). COCs were transferred to IVF droplets 15 min prior to adding the spermatozoa. To stimulate sperm motility, penicillamine (2 mM; Sigma-Aldrich), hypotaurine (1 mM; Sigma-Aldrich) and epinephrine (250 mM; Sigma-Aldrich) were added to each droplet. The selected spermatozoa were counted using a hemocytometer and diluted with IVF medium to obtain a final concentration of 1 × 106 sperm/mL. Finally, 2 μL of the sperm suspension was added to the droplets containing the matured COCs. The fertilization medium was incubated at 38.5°C for 18 h in a humidified atmosphere of 95% air and 5% CO2. Presumptive zygotes were denuded by treatment with 0.1% bovine testicular hyaluronidase.
3
In vitro bovine embryo production
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In vitro-matured COCs were fertilized with frozen-thawed bovine sperm from a batch that had previously been used in our laboratory. COCs (10/drop) were transferred to 50 μL FERT-TALP medium (IVL02, Caisson Laboratories, 836 South 100 East Smithfield, UT 84335) supplemented with 0.1 mg/mL Heparin (Sigma H0519), 1 mM Hypotaurine (Sigma H1384), 250 mM Epinephrine (Sigma E4642), 2 mM Penicillamine (Sigma P4875), and 1x antibiotic solution (ICN 1670049 MP Biomedicals, Fisher Scientific Company, Ottawa, ON, USA). Semen was thawed at 36 °C for 1 min, and motile spermatozoa were separated using percoll gradients (Allgrad® 90% and 45% LifeGlobal group, 393 Soundview Rd, Guilford, CT 06437) by centrifugation at 780× g for 5 min at room temperature. The pellet was diluted with 300 μL of FERT-TL media, and the final concentration was adjusted at 2 × 106 sperm/mL, and added in each drop containing COCs [11 (link)]. COCs and semen were incubated for 18 to 20 h at 38.5 °C in a humidified atmosphere of 5% CO2 in air [23 (link),41 (link)].
4
Sperm Entry Point Mapping in Bovine Oocytes
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To investigate whether sperm entry point
(SEP) is random or preferential in bovine,
oocytes (n=311) were submitted to in vitrofertilization (IVF) as described previously (10 (link)).
In brief, frozen-thawed and washed sperm from
a single Holstein sire of proven in vitro fertility
were used for fertilization after capacitation by
the swim-up procedure. Spermatozoa (1×106/
ml sperm) and matured COCs (40-45 COCs/200
μl) were co-incubated in modified fertilization-
Tyrode’s albumin lactate pyruvate (TALP,
handmade) medium containing 0.01 mM
heparin (Sigma, USA), 0.2 mM penicillamine
(Sigma, USA) and 0.1 mM hypotaurine
(Sigma, USA) for 18 hours at 39.5˚C, 6%
CO2 in humidified air. Fertilized oocytes at
4-5 hours post fertilization (hpf) were fixed,
stained with Hoechst-33342 and visualized
with a fluorescence-assisted micromanipulator
microscope (Olympus, Japan). Schurmann et al.
(14 (link)) demonstrated 4-6 hpf as the optimum timepoint
of maximum early fertilization. To map
the SEP, eggs were rotated under constant UVlight
until MII-spindle (the approved reference
point of SEP) was positioned to 3 O’clock.
Then, the spatial relationship between SEP and MII-spindle was measured as described
by Motosugi et al. (Fig .2 ) (15 (link)). In this scheme,
zones I and IV are the closest and the furthest
from the MII-spindle, respectively.
(SEP) is random or preferential in bovine,
oocytes (n=311) were submitted to in vitrofertilization (IVF) as described previously (10 (link)).
In brief, frozen-thawed and washed sperm from
a single Holstein sire of proven in vitro fertility
were used for fertilization after capacitation by
the swim-up procedure. Spermatozoa (1×106/
ml sperm) and matured COCs (40-45 COCs/200
μl) were co-incubated in modified fertilization-
Tyrode’s albumin lactate pyruvate (TALP,
handmade) medium containing 0.01 mM
heparin (Sigma, USA), 0.2 mM penicillamine
(Sigma, USA) and 0.1 mM hypotaurine
(Sigma, USA) for 18 hours at 39.5˚C, 6%
CO2 in humidified air. Fertilized oocytes at
4-5 hours post fertilization (hpf) were fixed,
stained with Hoechst-33342 and visualized
with a fluorescence-assisted micromanipulator
microscope (Olympus, Japan). Schurmann et al.
(14 (link)) demonstrated 4-6 hpf as the optimum timepoint
of maximum early fertilization. To map
the SEP, eggs were rotated under constant UVlight
until MII-spindle (the approved reference
point of SEP) was positioned to 3 O’clock.
Then, the spatial relationship between SEP and MII-spindle was measured as described
by Motosugi et al. (
zones I and IV are the closest and the furthest
from the MII-spindle, respectively.
5
Bovine Oocyte Maturation and Evaluation
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The following reagents were used in the study: Hank’s medium (SIGMA, USA), gentamicin (SIGMA, USA), penicillin (SIGMA, USA), streptomycin (SIGMA, USA), PBS (SIGMA, USA), TCM-199 HEPES (SIGMA, USA), foetal bovine serum (FBS) (GIBCO, Scotland), mineral oil (SIGMA, USA), FSH (SIGMA, USA), β-oestradiol (SIGMA, USA), sodium pyruvate (MERCK, France), BSA fraction V (SIGMA, USA), BSA fraction FAF (SIGMA, USA), penicillamine (SIGMA, USA), hypotaurine (SIGMA, USA), epinephrine (SIGMA, USA), High Pure RNA Isolation Kit (Roche Diagnostics), SYBR Green (Roche Applied Science), Hybond-ECL nitrocellulose membranes (Amersham Biosciences), Trypan blue (SIGMA) antibody against HSP70 (Abcam, ab6535), antibody against OVGP1 (Abcam, ab74544), rabbit anti-mouse secondary antibodies (Santa Cruz Biotechnology, sc-2005), goat anti-rabbit secondary antibodies (Santa Cruz Biotechnology, sc-2004), ECL Plus (Amersham Life Sciences), Fluoromount (Sigma USA).
6
Reagents and Chemicals Procurement
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Hypotaurine, (–)epinephrine, 17α-estradiol (17αE2), 17βE2, fluorescein isothiocyanate and bovine serum albumin (BSA)-conjugated 17βE2 (BSA-17βE2), GABA, melatonin, sodium taurocholate, sodium metabisulfite, and tamoxifen were purchased from Sigma-Aldrich (St Louis, MO, USA). BSA fraction V was purchased from Merck KGaA (Darmstadt, Germany). Other reagent-grade chemicals were purchased from Wako Pure Chemical Industries (Osaka, Japan).
7
Antioxidant Enzyme Gene Expression in V. alginolyticus
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Genes encoding antioxidant-related enzymes of SOD, GR and CAT in V. alginolyticus were cloned and sequenced in our previous work, and qPCR primers were designed according to the sequenced results (Table 1 ), referenced by 16S rDNA gene. Primers were synthesized by Sangon Biological Engineering Technology & Services Co., Ltd. (China). The kits of RNA extraction and reverse transcription were from Beijing TransGen Biotech Co., Ltd. (China). Sodium hydrosulfide (NaSH, H2S donor), hypotaurine (HT, H2S scavenger), sodium nitroprusside (SNP, NO donor) and 2-phenyl-4, 4, 5, and 5-tetramethylimidazoline-1-oxyl 3-oxide (PTIO, NO scavenger) came from Sigma-Aldrich Co. LLC (USA), while NOR came from Guangzhou Technology Company Limited (China). ELISA kits for microorganism CBS and NOS, H2S and NO were provided by Shanghai Jianglai Industrial Ltd (China). Hydrogen Peroxide Assay Kit and Total Protein Quantification Assay Kit were from Nanjing Jiancheng Bioengineering Institute (China).
8
Musk Shrew Zygote Culture
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Pronuclear zygotes (n = 24) were collected from the oviducts of female musk shrews (n = 9) 26–27 h after mating with a stud male. The zygotes were cultured for 20 h in KSOM, TYH, m-KRB with
1 mM hypotaurine (Sigma-Aldrich Inc., MO, USA), or m-PZM at 37°C in a 5% CO2 atmosphere. The developmental stages of embryos were confirmed under a light microscope.
1 mM hypotaurine (Sigma-Aldrich Inc., MO, USA), or m-PZM at 37°C in a 5% CO2 atmosphere. The developmental stages of embryos were confirmed under a light microscope.
9
Chromatographic Resin Purification Protocol
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Chromatographic resins were purchased from GE Healthcare (Italy). Chemicals including Luria Bertani (LB), tryptone, agar, ampicillin, Isopropyl-beta-D-thiogalactopyranoside (IPTG), phenylmethanesulfonylfluoride (PMSF), riboflavin, flavin adenine dinucleotide (FAD), βmercaptoethanol, glycerol, lysozyme, NADPH, NADP + , NADH, NAD + , hypotaurine, taurine, tamoxifen, fenthion, fenthion sulfoxide, phosphoric acid, acetonitrile, methanol, triethylamine and salts were all purchased from Sigma-Aldrich. tamoxifen N-oxide standard was purchased from Toronto Research Chemicals (Canada). All chemicals were of highest available purity and used without any further purification. All media, solutions and buffers were prepared with deionized Milli-Q water.
10
Fluorescent Probe Assay for Oxidants
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The molecular probe H 2 DCF-DA was obtained from Biotium (Hayward, CA, USA). 2-(N-morpholino) ethanesulfonic acid (MES), hypotaurine (HT), aminooxy acetic acid (AOA), hydroxylamine (NH 2 OH), potassium pyruvate (C 3 H 3 KO 3 ), ammonia (NH 3 ), ascorbic acid (ASA), catalase (CAT), diphenylene iodonium (DPI), salicylhydroxamic acid (SHAM), L-cysteine, D-cysteine, dimethyl sulfoxide (DMSO), N,Ndimethyl-p-phenylenediamine dihydrochloride and dithiothreitol (DTT) were obtained from Sigma-Aldrich (St Louis, MO, USA). Unless stated otherwise, the remaining chemicals were of the highest analytical grade available from various Chinese suppliers.
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