mM Of Taurine Research Articles

Taurine Prevents Angiotensin II-Induced Human Endocardial Endothelium Morphological Remodeling and the Increase in Cytosolic and Nuclear Calcium and ROS.

Endocardial endothelium (EE) is a layer of cells covering the cardiac cavities and modulates cardiomyocyte function. This cell type releases several cardioactive factors, including Angiotensin II (Ang II). This octopeptide is known to induce cardiac hypertrophy. However, whether this circulating factor also induces EE hypertrophy is not known. Taurine is known to prevent cardiac hypertrophy. Whether this endogenous antioxidant prevents the effect of Ang II on human EE (hEE) will be verified. Using quantitative fluorescent probe imaging for calcium and reactive oxygen species (ROS), our results show that Ang II induces (10-7 M, 48 h treatment) an increase in hEE cell (hEEC) volume and its nucleus. Pretreatment with 20 mM of taurine prevents morphological remodeling and increases intracellular calcium and ROS. These results suggest that the reported Ang II induces cardiac hypertrophy is associated with hEEC hypertrophy. This later effect is prevented by taurine by reducing intracellular calcium and ROS overloads. Thus, taurine could be an excellent tool for preventing Ang II-induced remodeling of hEECs.

EFFECTS OF ADDITIN OF TAURINE AND MANGANESE IN EXTENDER ON POST THAW SEMEN QUALITY OF SAHIWAL BULLS

Effect of two antioxidants (Taurine and Manganese) in extender on different spermparameters (sperm motility, live sperm percentage, membrane and acrosome integrity) of Sahiwal bullswas evaluated. Semen was collected twice a week for 8 weeks from six adult, regular donor andclinically healthy Sahiwal bulls. A total of 96 ejaculates were collected and pooled after initialevaluation. The pooled semen samples were diluted at with tris-citric acid,fructose, egg yolk andglycerol extender either containing different level of taurine (20.0, 40.0 and 60.0 mM), manganese(100.0, 150.0 and 200.0 µM), or combinations of taurine and manganese (20.0 mM + 100.0 µM, 40.0mM + 150.0 µM and 60 mM + 200 µM) at a temperature of 37oCand without antioxidants as control.Semen samples were loaded in 0.5 mL French straws and cooled to 4°C. Samples were frozen andstored in liquid nitrogen. These samples were thawed and anlayzed for different parameters. The postthaw sperm motility (%) was significantly (P<0.01) higher in treatment groups containing 20.0 mM oftaurine, 100.0 and 150.0 µM of manganese and 20.0 mM + 100.0 µM of taurine and manganeserespectively. Likewise, the live sperm percentage was higher (P<0.01) in 20.0 mM of taurine and100.0, 150.0 µM of manganese supplemented groups. The maximal sperm acrosome integrity of 20.0mm was observed in taurine group. A higher proportion (P<0.01) of sperm with intact membrane wasobserved in 20.0 mM of taurine and 150.0 µM in manganese containing groups. It was concluded thataddition of taurine and manganese in cryodiluents with a concentration of 20 mM and 150 µMrespectively could improve the post-thaw sperm quality in Sahiwal bulls.

Extender Supplementation with Glutathione (GSH) and Taurine Improves In Vitro Sperm Quality and Antioxidant Status of New Zealand Rabbits during Chilled Storage for up to 72 hours.

This study assessed the influence of supplementing the rabbit semen extender with various concentrations of glutathione (GSH) and taurine at 24, 48, and 72 h postchilling at 5°C. Semen samples were collected from 20 New Zealand bucks, and ejaculates with standard color, motility (>85%), about 0.5 mL volume, and ∼400 × 106/mL concentration were used and diluted with extenders supplemented with 0.5, 1, and 2 mM of GSH and 1, 5, and 10 mM of taurine and chilled at 5°C. Nonsupplemented samples were used as a control. Sperm's progressive motility, acrosome reaction, and extracellular oxidative stress biomarkers such as MDA contents and GPx, SOD, and CAT concentrations and intracellular transcriptomic levels of SOD and CAT genes were assessed. GSH and taurine supplementation improved the sperm's kinetics by reducing cooling-associated stress, which was ascertained by lowering MDA concentration and increasing SOD, CAT, and GPx concentrations (P < 0.05). Increasing the levels of antioxidant enzymes in the extender was due to the increasing mRNA copies of the SOD and CAT genes (P < 0.05). Furthermore, GSH and taurine maintained the fructose levels in the extender and lowered the GPT levels, which implies sperm membrane stability is maintained through GSH and taurine supplementation. GSH and taurine supplementation to the extender had protective influences on the in vitro rabbit semen quality during chilled storage for up to 72 h, which were remarkable with increasing supplementation dose and cooling time at 5°C.

Synergistic enhancement of rat intestinal alkaline phosphatase activity by taurine and sodium butyrate protects against endotoxin-induced bowel inflammation.

The effect of sodium butyrate (SB) and taurine on rat intestinal alkaline phosphatase (RIA) and the effect of the interaction of taurine and/or SB with bacterial lipopolysaccharides on ALP activity were investigated. In vitro analysis of the activity of RIA was carried out using various concentrations of SB and/or taurine. Substrate concentration-dependent kinetic study was performed at 1-10mM of taurine and SB at 5.17mM of p-nitrophenyl phosphate (p-NPP). The in vivo effect of lipopolysaccharide (LPS) in the presence and absence of taurine and SB on the activity of RIA was also evaluated. LPS was administered to rats intraperitoneally and 20min after; this was followed by oral administration of SB and/or taurine. The hydrolysis of p-NPP by RIA was enhanced by taurine and SB at different concentrations. The in vivo kinetic study revealed that RIA activity was greater (588.23×10-3 μmol/min/ml) when taurine and SB were co-administered with bacterial LPS, yielding a low Km (0.12mM) value. This suggested an increased affinity for the substrate by the enzyme. The degree of activation was highest when SB and taurine were administered together with LPS. The study concluded that SB and taurine are activators of RIA and their positive synergistic interaction in the presence of bacterial LPS may further emphasize the role of both activators in attenuating bacterial LPS-mediated diseases. PRACTICAL APPLICATIONS: The development and progression of a myriad of diseases such as inflammatory bowel disease, atherosclerosis, sepsis, multiple sclerosis, and rheumatoid arthritis have been linked to bacterial endotoxin. Taurine is an amino acid derived from cysteine, while sodium butyrate is a short-chain fatty acid. Consumption of food and food supplement rich in taurine and sodium butyrate can help protect against endotoxemic injury and aid tissue repair in the small intestine, digestibility, growth, and overall health of animals.

Taurine in semen extender modulates post-thaw semen quality, sperm kinematics and oxidative stress status in mithun (Bos frontalis) spermatozoa

Objective: To assess the effect of taurine on post-thaw semen quality parameters, sperm kinematics, antioxidant and oxidative stress profiles and sperm cholesterol efflux in mithun (Bos frontalis). Methods: A total of 50 ejaculates (n=25 samples) were selected based on biophysical parameters. Each sample was split into four equal aliquots after dilution with the Tris-citrate-glycerol extender. Group I, II, III and IV contained 0 mM (the control), 25 mM, 50 mM and 100 mM of taurine, respectively. Frozen-thawed samples were analysed for motility parameters (progressive forward and in bovine cervical mucus penetration test), kinetic and velocity parameters by computer-assisted sperm analyzer, viability, sperm and nuclear abnormalities, acrosome integrity, plasma membrane and nuclear integrities, sperm enzymatic leakage and biochemical (sperm cholesterol and oxidative stress) profiles. Results: The extender containing 50 mM taurine led to a significant enhancement in viability, acrosomal integrity, plasma membrane integrity, motility (progressive and in cervical mucus), and sperm cholesterol content and notably reduced sperm morphological and nuclear abnormalities, and leakage of intracellular enzymes compared to other taurine treated and untreated control groups (P&lt;0.05). Moreover, in addition to significant improvement in kinetic and velocity profiles, 50 mM taurine protected the integrity of acrosome and biochemical membranes than in the untreated control and other taurine treated groups. Inclusion of 50 mM taurine held a clear advantage over the control or 25 mM or 100 mM taurine in cryopreservation of mithun semen. Conclusions: Taurine (50 mM) supplementation in semen extender can be effectively utilized to reduce oxidative stress and improve post-thaw semen quality in mithun.

Antioxidants improve the semen quality following cryopreservation in Indian yak bulls

The present study was conducted to elucidate the effect of different antioxidants on the semen quality of cryopreserved yak semen. The ejaculates found suitable were extended (1:10) with 4 different Tris extenders by split sample technique containing different antioxidants in each of 3 extenders, viz. Taurine @ 50 mM, Trehalose @ 100 mM or vitamin E @ 2 mM and one control (without additive). The sperm motility, live sperm, HOSTreacted sperm, total incidence of acrosomal changes and extracellular release of ALT and AST differed significantly between antioxidants. Total sperm motility, live sperm and HOST-reacted sperm were significantly higher and total incidence of acrosomal changes and extracellular release of AST were significantly lower in extender containing antioxidants than that in control. The difference between taurine, trehalose and vitamin E was not significant for parameters like sperm motility, HOST-reacted sperm and extracellular release of ALT. The per cent live sperm was significantly higher and total incidence of acrosomal changes was significantly lower for taurine than for trehalose and vitamin E. The extracellular release of ALT was significantly lower in extender containing taurine or trehalose than in control while the extracellular release of AST was significantly lower in the extender containing taurine than that in containing trehalose, vitamin E and control. The findings of the present study revealed that addition of taurine, trehalose and vitamin E in Tris extender significantly improved the post thaw quality of yak semen. Among the antioxidants evaluated, 50 mM of taurine in Tris extender could be effectively used to obtain better quality of frozen thawed yak semen.

Compatible osmolytes modulate mitochondrial function in a marine osmoconformer Crassostrea gigas (Thunberg, 1793)

Salinity is an important environmental factor affecting physiology of marine organisms. Osmoconformers such as marine mollusks maintain metabolic function despite changes of the osmolarity and composition of the cytosol during salinity shifts. Currently, metabolic responses to the salinity-induced changes of the intracellular milieu are not well understood. We studied the effects of osmolarity (450 vs. 900 mOsm) and compatible osmolytes (70–590 mM of taurine or betaine) on isolated gill mitochondria of a marine osmoconformer, the Pacific oyster Crassostrea gigas. Physiological concentrations of taurine enhanced mitochondrial ATP synthesis and electron transport system (ETS) capacity, increased mitochondrial coupling and stimulated the forward flux through the Complex I. Notably, the stimulatory effects of taurine were more pronounced at 900 mOsm compared to 450 mOsm. In contrast, betaine proportionally increased the rates of the mitochondrial proton leak, oxidative phosphorylation and ETS flux (with no net effect on the mitochondrial coupling) and suppressed the activity of cytochrome c oxidase in oyster mitochondria. However, the effective concentration of betaine (590 mM) was higher than typically found in bivalves, and thus betaine is not likely to affect oyster mitochondria under the physiological conditions in vivo. Our findings indicate that taurine may support the mitochondrial bioenergetics during hyperosmotic stress in oysters. Compatibility of taurine with the metabolic functions and its beneficial effects on mitochondria may have contributed to its broad distribution as an osmolyte in marine osmoconformers and might explain the earlier reports of the positive effects of taurine supplementation on energy metabolism of other organisms, including mammals.

A Novel Cysteine Sulfinic Acid Decarboxylase Knock-Out Mouse: Immune Function (II).

Taurine deficient mice lacking cysteine sulfinic acid decarboxylase (CSAD KO) were developed for investigating the various physiological roles of taurine including the development of the brain and eye as well as immune function. Due to severe abnormalities of immune function in a taurine deficient cat, the immune function including adoptive and innate immunity in taurine-deficient mice have been studied. Previously we demonstrated that B cell function in CSAD KO was reduced in both females and males. However, T cell function was significantly reduced only in females. In this study, we have examined innate immunity using macrophage activation with LPS or/and IFN-γ and polymorphonuclear leukocytes (PMN) activation with phorbol myristate acetate (PMA). Pro- and anti-inflammatory cytokines including IL-6, TNF-α and IL-10 as well as nitric oxide (NO) were determined using ELISA and Griess reagent, respectively. Peritoneal macrophages were activated with 1 μg/mL of lipopolysaccharide (LPS) and/or 50 U/mL of IFN-γ. In addition, superoxide anion was measured using peritoneal PMN activated with PMA in the presence and absence of superoxide dismutase. Superoxide anion production in activated PMN from CSAD KO homozygotes (HO) was not significantly different from wild-type (WT) with and without 25 mM taurine. IL-10 and TNF-α production in both female and male CSAD KO were not significantly different. IL-6 and NO were significantly lower only in females as previously observed in Con A-activated cellular proliferation of splenocytes. Cytokine production with 10 mM of taurine was not different, indicating the reduction of NO and IL-6 in females may be due to the absence of the CSAD gene, not due to low taurine concentrations.These data indicate that some measures of innate immunity were altered in female CSAD mice.

Effect of Addition of Taurine on the Liquid Storage (5°C) of Mithun (Bos frontalis) Semen

The present study was undertaken to assess the effect of taurine on sperm motility, viability, total sperm abnormalities, acrosomal and plasma membrane integrity, enzymatic profiles such as reduced glutathione (GSH), glutathione peroxidase (GPX), superoxide dismutase (SOD), and catalase (CAT), and biochemical profiles such as cholesterol efflux and malondialdehyde (MDA) production. A total of 50 ejaculates were collected twice a week from 8 mithun bulls, and semen was split into 4 equal aliquots and diluted with the TEYC extender. Group 1: semen was without additives (control); groups 2 to 4: semen was diluted with 25 mM, 50 mM, and 100 mM of taurine, respectively. Seminal parameters and enzymatic and biochemical profiles were assessed at 5°C. Inclusion of taurine into diluent resulted in significant (P < 0.05) decreases in percentages of dead spermatozoa, abnormal spermatozoa, and acrosomal abnormalities after liquid storage compared with the control group. Additionally, taurine at 50 mM has significant improvement in quality of mithun semen than taurine at 25 or 100 mM stored in in vitro at 5°C. It was concluded that the possible protective effects of taurine on sperm parameters are from enhancing the function of antioxidant enzymes, preventing efflux of cholesterol from cell membranes and decreased MDA production.

Taurina no desenvolvimento de embriões bovinos fecundados in vitro

O objetivo deste trabalho foi avaliar o efeito de diferentes concentrações de taurina no desenvolvimento de embriões bovinos fecundados in vitro em meio de cultivo com diferentes fontes de soro. No experimento 1, zigotos (n=440) fecundados in vitro foram distribuídos aleatoriamente nos tratamentos com 0, 3, 7 ou 14 mM de taurina em meio de cultivo acrescido de 10% de soro fetal bovino (SFB) e 3g/l de albumina sérica bovina (BSA). No experimento 2, os zigotos (n=940) foram divididos nos tratamentos com 0, 3 ou 14 mM de taurina em meio acrescido de 10% de SFB ou 3g/l de BSA. No experimento 3, os zigotos (n=191) foram divididos nos tratamentos com 0 ou 3 mM de taurina em meio de cultivo sem fonte de soro, porém adicionado de 3 g/l de álcool polivinil. Nos experimentos 1 e 2 não se observou diferença (P&gt;0,05) na taxa de clivagem, na produção de blastocistos e no número de células entre as concentrações de taurina avaliadas. No experimento 3 encontraram-se maior (P&lt;0,05) taxa de clivagem (68,5% vs. 16,9%) e produção de blastocistos (8,3% vs. 0%) na presença de taurina. O cultivo de zigotos em meio adicionado de SFB produziu maior (P&lt;0,01) taxa de blastocistos no sétimo (25,6% vs. 6,7%) e oitavo (30,8% vs. 13,9%) dia pós-fecundação e número de células/blastocistos (104,8± 2,63 vs. 84,7± 3,86) do que no cultivo com BSA, apesar de menor (P&lt;0,01) taxa de clivagem (58,1% vs. 71,3%). Conclui-se que o efeito benéfico da taurina no desenvolvimento embrionário somente é observado na ausência de SFB e BSA. O SFB produz menor taxa de clivagem mas melhora o desenvolvimento embrionário após as primeiras divisões celulares.

An investigation on skin wound healing in mice with a taurine-chitosan gel formulation.

The process of wound healing begins immediately following surface lesions or just after exposure to radiation, chemical agents or extreme temperatures. Taurine (2-aminoethane sulfonic acid), an amino acid containing sulfur, is found in almost all tissues in mammals, playing various important physio-logical roles in each organ. Taurine exhibits an antioxidant effect and is also known to have effects on cell proliferation, inflammation and collagenogenesis. Many antioxidants have been used to eliminate the negative effects of oxygen free radicals on wound healing. The objective of the present study was to examine the wound healing effect in mice of taurine-chitosan gel, which releases taurine slowly over a long time period. Fifty mM of taurine in 1.5% chitosan polymer (TAU-GEL) and 1.5% chitosan polymer (CHI-GEL) were applied to full thickness skin wounds of mice once a day for seven days. After seven days of treatment, lipid peroxide formation-malondialdehyde (MDA) and hydroxyproline (HPX) levels and the tensile strength of wound tissues were measured. All results were compared with those of the untreated control group (CONT). The structural alterations in the skin layers were also histologically investigated. It was found that locally administered TAU-GEL form significantly increased wound tensile strength by decreasing the MDA and increasing HPX levels. These results were supported by histological findings. All observations suggest that taurine gel may be effective in wound healing.

Predominance of human versus rat phenotype in the metabolic pathways for bile acid synthesis by hybrid WIF-B9 cells

The rat hepatoma–human fibroblast hybrid cell line WIF-B9 stably exhibits the structural and functional characteristics of normal differentiated hepatocytes. The abilities of these cells to synthesize bile acids and amidate them with glycine and taurine were investigated. The release of bile acids into the culture media over 72 h was assessed by gas chromatography-mass spectrometry. WIF-B9 cells were able to synthesize bile acids (1.10±0.17 nmol/mg protein) but less efficiently than rat hepatocytes in primary culture (2.19±0.19 nmol/mg protein; P<0.01). The patterns of major bile acid species produced by both types of cells were also different. Cholic acid (CA; 72%) and β-muricholic acid (19%) were the major bile acids produced by rat hepatocytes, while chenodeoxycholic acid (CDCA) accounted for only 4.5% of total bile acids. In contrast, muricholic acids were absent, while CA (62%) and CDCA (34%) were the most abundant bile acids synthesized by WIF-B9 cells. Using reverse transcription-polymerase chain reaction and gene- and species-specific primers for key enzymes involved in bile acid synthesis, the expression of human, but not rat, orthologues of CYP7A1, CYP27, CYP8B and CYP7B1 was found in WIF-B9 cells. Induction of cell stress by serum deprivation did not change the amount of total bile acids synthesized by these cells, but an inversion of the CA-to-CDCA ratio from 1.8 to 0.3 together with a marked increase in the proportion of intermediate metabolites related to the acidic pathway was found. Using 500 μM radiolabeled CA and 2 mM of taurine or glycine, the ability to amidate CA over 48 h was determined by high performance liquid chromatography. Rat hepatocytes conjugated more than 90% CA with either amino acid, whereas this ability was very poor (<2%) in WIF-B9 cells. Regarding the expression of enzymes and the products of bile acid synthesis, it may be concluded that the human phenotype predominates over that of the rat in WIF-B9 cells. Moreover, these cells are almost completely unable to further conjugate primary bile acids, which facilitates the manipulation of these steroids in analytical procedures. These characteristics make WIF-B9 cells a suitable in vitro model to carry out studies on bile acid synthesis by ‘human-like’ metabolic pathways.

Kinetic study of acamprosate absorption in rat small intestine.

Acamprosate (calcium bis acetyl-homotaurine), a homotaurine derivative, a structural analogue of gamma-aminobutyric acid (GABA) and an upper homologue of taurine, is a relatively new drug used to prevent relapse in weaned alcoholics. When administered orally as enteric-coated tablets at relatively high doses, this drug has a bioavailability of about 11%; however, the intestinal absorption mechanism has not been studied in depth. The present study was therefore planned to characterize the intestinal transport of acamprosate in the rat and the effect of chronic alcohol treatment on this process, quantifying its kinetic parameters and investigating possible inhibitors. Using an in vitro technique, acamprosate absorption was measured in the rat intestine from three different groups: alcohol group [fed a liquid diet containing 5% (w/v) ethanol for 4 weeks], isocaloric pair-fed control, and a solid diet group. Intestinal acamprosate absorption was found to occur mainly by passive diffusion with a diffusive permeability of 0.213+/-0.004 cm/h in control pair-fed animals, 0.206+/-0.001 cm/h in animals receiving chronic alcohol treatment, and 0.193+/-0.001 cm/h in the solid diet group. Inhibition studies showed that at a 10(-3) M acamprosate concentration, some compounds such as GABA, taurine, proline, and glycine at 40 mM each did not affect acamprosate transport. Nevertheless, when a lower concentration of the drug (10(-4) M) was assayed, a significant reduction of acamprosate transport in the presence of taurine or GABA 40 mM was found. These results suggest that acamprosate in the rat jejunum, could be transported, in part, by a carrier system. Further experiments using different concentrations of taurine (10, 20, and 80 mM) showed that the maximum inhibition (32%) is achieved at 20 mM of taurine. These latter results suggest that acamprosate and taurine share, at least, an intestinal carrier system in rat jejunum. From the above results, it can be concluded that there are probably two pathways involved in the intestinal absorption of acamprosate: passive diffusion and mediated transport, with the former being predominant. Moreover, neither chronic ethanol intake nor the type of diet seems to alter the intestinal absorption of the drug.

Taurine blocks ATP-sensitive potassium channels of rat skeletal muscle fibres interfering with the sulphonylurea receptor.

Taurine is a sulphonic aminoacid present in high amounts in various tissues including cardiac and skeletal muscles showing different properties such as antioxidative, antimyotonic and anti-schaemic effects. The cellular mechanism of action of taurine is under investigation and appears to involve the interaction of the sulphonic aminoacid with several ion channels. Using the patch-clamp technique we studied the effects of taurine in rat skeletal muscle fibres on ATP-sensitive K(+) channel (K(ATP)) immediately after excision and on channels that underwent rundown. The cytoplasmic application of 20 mM of taurine reduced the K(ATP) current; this effect was reverted by washout of the drug solution. In this experimental condition the IC(50) was 20.1 mM. After rundown, taurine inhibited the K(ATP) current with similar efficacy. Competition experiments showed that taurine shifted the dose-response inhibition curve of glybenclamide to the left on the log-dose axis without significantly affecting those of ATP or Ca(2+) ion. Single channel recording revealed that taurine affects the close state of the channel prolonging it and reducing the bursts duration. Our data indicate that taurine inhibits the muscular K(ATP) channel interfering with the glybenclamide site on the sulphonylurea receptor of the channel or on the site allosterically coupled to it. During ischaemia and hypoxia, the skeletal and heart muscles undergo several changes; for example, the activation of K(ATP) channels and loss of the intracellular taurine content. The depletion of taurine during ischaemia would contribute to the early activation of K(ATP) channels and salvage the intracellular ATP content.

Development of rabbit zygotes into blastocysts in defined protein-free medium and offspring born following culture and embryo transfer

We report here on an improved, completely defined culture system for producing embryos in vitro which mimics development in vivo. This system avoids the confounding effects of the many unknowns introduced by the multivariate components of the serum or by unknowns attached to bovine serum albumin (BSA). Zygotes were obtained from superovulated rabbits and cultured in modified defined RPMI 1640:Dulbecco's MEM, 1:1 (RD) medium. The effect of a novel and potentially ideal antioxidant, tempol, was tested (20 to 0.001 mM) but found to be either toxic or ineffective. In the presence of 20% O 2, 600 units of Superoxide dismutase or 2.5 mM of taurine increased embryo hatching after 72 h of culture in RD medium to 75 and 76%, respectively, compared with 46% in the control (P < 0.05). The need for antioxidants was reduced with 5% O 2. The beneficial effects of RD medium were demonstrated when 60 zygotes cultured for 48 h to the early blastocyst stage in this medium were transferred and resulted in 30 young (50%) compared with 35 60 (58%) young from uncultured control transfers. Only 12% of the young were obtained from slower developing morulae. Thus, high viability was established for rapidly growing embryos in culture, but fewer slow growing embryos survived after transfer. A further comparison of embryos cultured in RD medium with a high potassium simple, optimized, defined medium (KSOM), revealed that both yielded results approaching those of direct transfer without culture. Simple defined media may also be useful for the culture of embryos of other species.

Effect of taurine depletion on excitation-contraction coupling and C1 − conductance of rat skeletal muscle

The pharmacological action of taurine on skeletal muscle is to stabilize sarcolemma by increasing macroscopic conductance to Cl − ( G C1), whereas a proposed physiological role for the amino acid is to modulate excitation-contraction coupling mechanism via Ca 2+ availability. To get insight in the physiological role of taurine in skeletal muscle, the effects of its depletion were evaluated on voltage threshold for mechanical activation and G C1 with the two intracellular microelectrode method in ‘point’ voltage clamp mode and current clamp mode, respectively. The experiments were performed on extensor digitorum longus muscle fibers from rats depleted of taurine by a chronic 4 week treatment with guanidinoethane sulfonate, a known inhibitor of taurine transporter. The treatment significantly modified the mechanical threshold of striated fibers; i.e. at each pulse duration they needed significantly less depolarization to contract and the fitted rheobase voltage was more negative by 10 mV with respect to untreated muscle fibers. In parallel, the treatment with guanidinoethane sulfonate produced a significant 40% lowering of G C1. In vitro application of 60 mM of taurine to such depleted muscles almost completely restored the mechanical threshold and increased G C1 even above the value of untreated control. However, in vitro application of 60 mM of either taurine or guanidinoethane sulfonate to untreated control muscles did not cause any change of the mechanical threshold but increased G C1 by 40% and 21%, respectively. Furthermore, 100 μM of the S-(−) enantiomer of 2-( p-chlorophenoxy)propionic acid almost fully blocked G C1 but did not produce any change in the mechanical threshold of normal muscle fibers. The present results show that the large amount of intracellular taurine plays a role in the excitation-contraction coupling mechanism of striated muscle fibers. This action is independent from any effect involving muscle Cl − channels, but it is likely mediated by the proposed ability of taurine to modulate Ca 2+ availability through the interaction with the Ca 2+ transporters present on sarcoplasmic reticulum.

Effect of various extenders and taurine on survival of stallion sperm cooled to 5 °C

Stallion semen was diluted in five different extenders (dimitro-poulus onze (Dimitro's), Kenney's modified tryode (Kenney's), modified INRA82 (INRA82), egg yolk-citrate-taurine (Citrate) and EZ-Mixin) and evaluated for motility after cooling and storage at 5 °C for 0, 24, 48, 72 and 96 h. EZ-Mixin extender was used as control while 70 and 100 mM of taurine were added to Dimitro's, Kenney's and INRA82 to study its effect under conditions of storage at 5 °C and varying processing modifications. Motility in INRA82 was 57.0, 58.4, 61.1, and 56.1% after 24, 48, 72 and 96 h, respectively and was higher (P < 0.05) than in other extenders after 48, 72, and 96 h. Motility decreased over time in Dimitro's (P < 0.05) and Kenney's (P < 0.01). Motility in INRA82 and EZ-Mixin decreased (P < 0.05) after 24 h and then remained unchanged until 96 h. In taurine (70 mM) containing extender Citrate, motility decreased throughout storage. Motility in INRA82 and Kenney's with taurine decreased (P < 0.05 and P < 0.01, respectively) after 48 h, and then remained stable until 96 h only in INRA82 with taurine. Motility in INRA82 with taurine was higher (P < 0.05) than in Citrate throughout incubation, whereas motility in INRA82 with taurine was higher than in EZ-Mixin and Kenney's with taurine after 24 and 48 h, respectively. Motility in INRA82 and Kenney's with taurine improved (P < 0.05) when osmotic pressure was increased with taurine (100 mM) but not when osmotic pressure was increased with Na ∗ and K ∗ salts. Motility was always higher (P < 0.01) in taurine (70 mM or 100 mM) containing extenders than in non-taurine extenders, Dimitro's, INRA82, and Kenney's, when sperm were incubated for 24 h at 5 °C in these extenders, then washed and incubated at 39 °C in Sp-TALP for 12 or 24 h. In conclusion 1) INRA82 was a better extender than the other extenders tested. 2) inclusion of taurine (100 mM) in INRA82 and Kenney's improved sperm survival until 96 and 48 h, respectively, and 3) sperm preincubation for 24 h in taurine containing extenders resulted in better sperm survival when washed and stored in Sp-TALP for further 12 or 24 h.

In vitro capacitation of boar ejaculated spermatozoa: effect of conditioned media prepared from preincubated sperm suspension.

The fertilizing ability of boar ejaculated spermatozoa was examined in vitro after preincubation at a concentration of 2.5 x 10(8)/ml for 4 hr in several conditioned media (CM). For preparation of CM, boar spermatozoa were incubated in a modified Krebs-Ringer bicarbonate solution (TYH) at concentrations of 20 to 40 x 10(8)/ml for several hours up to 4 hr; then their supernatant fluids were collected by centrifugation. When boar ejaculated spermatozoa were preincubated in TYH alone, 14.1% of oocytes were penetrated by them as we reported previously. On the other hand, preincubating them with CM, their fertilizing ability was elevated according as the incubation time of CM preparation was lengthened. The fertilization rate reached 75.0%, using 4 hr-incubated CM for the preincubation medium. The effect of CM was not deteriorated by heat treatments (56 degrees C, 30 min, or 100 degrees C, 5 min). The components of CM were separated at a molecular weight of 25,000 by ultrafiltration, and high fertilization rate (69.8%) was obtained when low molecular weight fraction was used for the preincubation medium. Sperm extracts prepared from directly frozen-thawed sperm suspension and 0.1-10 mM of taurine or hypotaurine had no effect on the fertilizing ability of boar spermatozoa. These results suggest that substances stimulating boar sperm capacitation were accumulated from viable spermatozoa into the medium during incubation and that the effective substances were heat-stable and of low molecular weight and were not taurine and hypotaurine.

Potentiation by taurine of inotropic effect of strophanthin-K on guinea-pig isolated auricles

Fresh guinea-pig auricles contain 39.4±4.2 mM of taurine per ml of intracellular water. When isolated, the auricles lose 15 per cent of their content in the Tyrode bath solution (30°C, pH 7.2–7.3, 97%O 2 +3% CO 2 ) during the first hour of incubation. After 5 hours of incubation taurine content is 70% of the initial value. At this time taurine content can be restored to its initial value by the addition of 8 mM taurine (final concentration) to the incubation fluid for 30 minutes; other times and concentrations have not been tested. Neither the taurine nor the homotaurine 8 mM concentrations affect the contractility or rate of the auricles. However, taurine, but not homotaurine, increases the contractileresponse to a fixed dose of strophanthin-K.