John B. Lombardini

Taurine: retinal function.

The status and potential functions of taurine in the retina have been reviewed. Taurine is present in high concentrations in the retina of all species tested, while the retinal concentrations of the enzymes necessary to synthesize taurine are presumed to vary among those species. The documented low activity of cysteinesulfinic acid decarboxylase, a key enzyme in taurine biosynthesis, in the livers of the cat, monkey and human possibly reflect low activity in their retinas, indicating reliance on the diet as an important source of taurine. Both high- and low-affinity binding proteins and uptake systems have been described for taurine in retinal tissue. Evoked release of taurine by light and other depolarizing stimuli have been well documented. Retinal pathologies including diminished ERGs and morphologic changes have been reported for animals and man deficient in taurine. Possible functions for taurine in the retina include: (1) protection of the photoreceptor - based on the shielding effects of taurine on rod outer segments exposed to light and chemicals; (2), regulation of Ca2+ transport - based on the modulatory effects of taurine on Ca2+ fluxes in the presence and absence of ATP; and (3) regulation of signal transduction - based on the inhibitory effects of taurine on protein phosphorylation.

Effects of Taurine on Calcium Ion Uptake and Protein Phosphorylation in Rat Retinal Membrane Preparations

Abstract: The effects of taurine on ATP‐dependent calcium ion uptake and protein phosphorylation of rat retinal membrane preparations were investigated. Taurine (20 mM) stimulates ATP‐dependent calcium ion uptake by twofold in crude retinal homogenates. In contrast, it inhibits the phosphorylation of specific membrane proteins as shown by acrylamide gel electrophoresis and autoradiography. The close structural analogue of taurine, 2‐aminoethylhydrogen sulfate, demonstrates similar effects in both systems, i.e., stimulation of ATP‐dependent calcium ion uptake and inhibition of protein phosphorylation, whereas isethionic acid and guanidinoethanesulfonate have no effect on either system. A P1 subcellular fraction of the retinal membrane preparation that contains photoreceptor cell synaptosomes has a higher specific activity for the uptake of calcium ions. Phosphorylation of specific proteins in the P1 fraction is also inhibited by the addition of 20 mM taurine. Taurine has no effect on retinal ATPase activities or on phosphatase activity, thus suggesting that it directly affects a kinase system.

Is taurine a hypothalamic neurotransmitter? : a model of the differential uptake and compartmentalization of taurine by neuronal and glial cell particles from the rat hypothalamus

Although taurine has been postulated to be a neurotransmitter or neuromodulator in the mammalian CNS, little is known concerning its role in brain function. Evidence suggesting that taurine may influence endocrine and homeostatic mechanisms via the hypothalamus resulted in our investigations into its function in this brain region. The main objectives of the research were to characterize the specific binding, uptake, and release of taurine in the hypothalamus. A specific aim was to examine the proposed neurotransmitter role for taurine in the hypothalamus. This was accomplished by comparing the characteristics and properties of the binding, uptake, and release of taurine with those for the classical neurotransmitters which satisfy the criteria for a neurotransmitter. On such a comparative basis, the characteristics of taurine uptake satisfy the neurotransmitter criterion of inactivation of taurine in the hypothalamus. However, the observed characteristics of taurine binding and release in the hypothalamus do not satisfy the respective neurotransmitter criteria of specific receptors and Ca2+-dependent evoked release. Therefore, solely on the basis of the experimental observations reported herein, we must conclude that taurine apparently does not function as a neurotransmitter in the hypothalamus. Two uptake systems were found in the P2 fraction, a high affinity uptake system and a low affinity uptake system. Uptake systems for taurine have previously been reported in glial and nerve cell homogenates, and therefore, because of the known contamination of crude synaptosomal preparations with glial particles, we sought to determine the cellular origin of the two taurine uptake systems in our crude preparation. Using a variety of diverse biochemical techniques such as hypo-osmotic shock, release experiments and Arrhenius plots, we determined that physical changes of the media or depolarizing stimuli which would influence neuronal and glial cell particles differently, also had differing effects on high and low affinity taurine uptake or its release from the respective uptake compartments. We conclude that the high affinity taurine uptake system/compartment is located on/in neuronal membranes/particles/particles and that the low affinity taurine uptake system/compartment is located on/in neuronal membranes/particles and that model for the differential cellular transport and compartmentalization of taurine into neuronal and glial cells has important implications concerning its possible role in the CNS.

Inhibition by taurine of the phosphorylation of specific synaptosomal proteins in the rat cortex: effects of taurine on the stimulation of calcium uptake in mitochondria and inhibition of phosphoinositide turnover.

It has been previously observed that taurine inhibits PKC-activated phosphorylation of specific proteins including a approximately 20k Mr protein in rat cortical synaptosomes. In the present study, the mechanism of the above effects of taurine were investigated. In an intrasynaptosomal cytosol fraction obtained by subcellular fractionation, taurine did not have inhibitory effects on protein phosphorylation. However, taurine did inhibit the phosphorylation of the approximately 20k Mr protein in a reconstituted preparation containing intrasynaptosomal cytosol and mitochondria. Experiments measuring calcium uptake demonstrated that taurine increased the accumulation of 45Ca2+ in the mitochondrial fraction in incubation systems both in the absence and presence of added ATP. In addition, taurine inhibited the accumulation of 32P-labeled phosphatidic acid in synaptosomes indicative of a reduction in the levels of diacylglycerol. These results suggest that taurine may inhibit specific protein phosphorylation both by reducing cytosolic calcium levels and by inhibiting the turnover of phosphoinositides. These effects of taurine on the signal transduction cascade involving PKC and phosphoinositide metabolism indicate a potential biological role for taurine in the nervous system.

The role of taurine in the pathogenesis of the cardiomyopathy of insulin-dependent diabetes mellitus

The cellular and molecular physiology and pathology of insulin-dependent diabetes mellitus (IDDM) and non-insulin-dependent diabetes mellitus (NIDDM) are mostly studied and understood through the use of animal models. Fundamental differences between the IDDM and NIDDM animal models may help to explain the etiology behind diabetic cardiomyopathy, one of the most severe complications of IDDM. Experimental rat models of IDDM exhibit a characteristic increase in tissue levels of taurine in the heart, a change that is not seen in NIDDM rats. This article deals with the causes and possible consequences of this observation which may contribute to the development of diabetic cardiomyopathy. Modulation of pyruvate dehydrogenase (lipoamide) (PDH; EC 1.2.4.1) activity was found to be a possible mode for taurine involvement. PDH is a mitochondrial protein and is the rate-limiting step in the generation of acetyl CoA from glycolysis. In IDDM, PDH activity is decreased through a mechanism that includes the stimulation of the de novo synthesis of a kinase activator protein (KAP) which phosphorylates PDH and inactivates the enzyme. This lesion does not occur in NIDDM rat hearts. Taurine is known to inhibit the phosphorylation of PDH in vitro, and in taurine-depleted rats PDH phosphorylation is known to increase. Thus, the increased levels of taurine in the diabetic heart may be inhibiting this phosphorylation which in turn may be stimulating the synthesis of KAP through a negative feedback process. The main argument for this theory would be the lack of change in both the taurine levels and the activity of PDH in the NIDDM rat model.

The inhibitory effects of taurine on protein phosphorylation: comparison of various characteristics of the taurine-affected phosphoproteins present in rat retina, brain and heart.

The role of taurine in the animal kingdom has been questioned for many years since its discovery in ox bile in 1827 by Tiedemann and Gmelin (20). Eventually, the seminal review by Jacobsen and Smith published in 1968 (1) provided the impetus to a small but determined segment of the scientific community to study this β-amino sulfonic acid systematically for the past 25 years. Since 1975 some 10 international symposia have been devoted to taurine (18). Nevertheless, the basic question as to the function of taurine remains unanswered. In the early years of the study of taurine phenomenology was the norm, while in recent years many of the questions posed are directed towards the mechanisms of action of taurine at the molecular level. Consequently, my laboratory has pursued the understanding, on a mechanistic basis, of the effects of taurine on Ca2+ uptake and protein phosphorylation in excitable tissues. This chapter summarizes the recent and ongoing studies of the effects of taurine on protein phosphorylation.

Taurine Inhibits Protein Kinase C‐Catalyzed Phosphorylation of Specific Proteins in a Rat Cortical P2 Fraction

Abstract: We previously reported that taurine inhibits the phosphorylation of specific proteins in a P2 synaptosomal fraction prepared from the rat cortex. In the present study, the regulation of the phosphorylation of an ∼20K Mr protein whose phosphorylation is inhibited by taurine was further investigated. The phosphorylation of the ∼20K Mr protein in a hypo‐osmotically shocked P2 fraction from rat cortex was dependent on the free Ca2+ in the reaction medium. Depolarization induced by 30 mM K+ stimulated the phosphorylation of the ∼20K Mr protein in an intact synaptosomal P2 preparation by 30‐fold. This stimulation was inhibited 35% by taurine, whereas guanidinoethanesulfonic acid, a taurine analogue, did not have any effect, thereby indicating the specificity of taurine. Addition of phorbol 12‐myristate 13‐acetate, a phorbol ester, together with phosphatidylserine, stimulated the phosphorylation of the ∼20K Mr protein in the hypo‐osmotically shocked P2 synaptosomal fraction by fivefold, whereas cyclic AMP, cyclic GMP, and calmodulin did not have any effect on the phosphorylation of this particular protein. Phorbol 12‐myristate 13‐acetate–stimulated phosphorylation of the ∼20K Mr protein is blocked 30% by taurine. Taurine also inhibited phorbol 12‐myristate 13‐acetate‐activated phosphorylation of two other proteins that were similar in molecular weight and isoelectric point to the ∼20K Mr protein on two‐dimensional gels. These results suggest that taurine modulates the phosphorylation of specific proteins regulated by the signal transduction system in the brain. Thus, taurine may modulate neuroactivity by inhibiting the phosphorylation of specific proteins involved in regulatory function.

Dietary taurine supplementation: Hypolipidemic and antiatherogenic effects

Abstract Taurine supplementation may prove to be a safe and convenient method to reverse high blood cholesterol and the associated rise in atherosclerosis. Although human studies are limited, experiments using animal models provide extensive proof of the hypolipidemic and antiatherogenic effects of taurine. Examples of these animal models involve feeding with high-fat diets, genetically determined or heritable disease conditions, and artificially induced or genetic diabetes. Most importantly, the addition of taurine to the diet clearly has effects against pathological increases in serum and liver cholesterol and triglycerides. Another consistent and noteworthy effect of taurine is the simulation of cholesterol 7α-hydroxylase activity, the enzyme that is responsible for the catabolism of cholesterol into bile acids. Taurine also exhibited considerable effects on atherosclerotic lipid accumulation, perhaps through an antioxidative mechanism and through the elevation of HDL cholesterol levels. Data from animal model systems support the specific cardiovascular benefits of taurine, and hopefully, this research will be continued in human studies in the future.

Effects of ATP and taurine on calcium uptake by membrane preparations of the rat retina.

Abstract: The effects of ATP and taurine on the kinetics of calcium uptake in rat retinal membrane preparations were determined. ATP increased calcium uptake at low calcium ion concentrations. Addition of ATP plus taurine further increased calcium uptake. Cooperative relationships were observed for calcium uptake in the absence of ATP and taurine. In the presence of phosphate ions reciprocal plots demonstrated upward deflections from linear ty, while in the absence of phosphate ions downward deflections were noted. Addition of ATP plus taurine to the incubation system appeared to obliterate the cooperativity. Two uptake systems for calcium were observed.

Inhibition of rat vascular smooth muscle cell proliferation by taurine and taurine analogues

The growth of rat aorta vascular smooth muscle cells (VSMCs) was measured in the presence and absence of taurine. Concentrations of taurine as low as 0.3 mM in the culture medium inhibited the proliferation of the cells, as monitored by measuring cell count, and also inhibited the rate of DNA synthesis, as examined by measuring [3H]thymidine incorporation into DNA. However, even at the highest concentration of taurine (30 mM), the doubling time of the VSMCs was only increased by 38%. Protein content of the VSMCs was decreased by 30 mM taurine. [3H]Leucine incorporation into newly synthesized protein was not affected by the highest concentration of taurine tested (30 mM), indicating that taurine did not inhibit protein synthesis but rather decreased total protein content by inhibiting cellular proliferation. The effects of other amino acids such as alanine, glycine, and serine and of various taurine analogues such as beta-alanine, guanidinoethanesulfonic acid (GES), and isethionic acid also were tested at a concentration of 20 mM for their effects on the growth of the VSMCs. Alanine, glycine, and serine had only a minimal effect or no effect on cell count, quantity of protein, and incorporation of [3H]thymidine into DNA. GES, beta-alanine, and isethionic acid had a significant effect on cell count, protein content, and incorporation of [3H]thymidine into DNA. Beta-alanine was the only analogue tested that significantly depressed [3H]leucine incorporation into newly synthesized protein. It is concluded that taurine, GES, and isethionic acid inhibited proliferation of VSMCs but did not alter normal protein synthesis or survivability of VSMCs. In contrast, other amino acids, alanine, glycine and serine, had minimal effects on VSMC proliferation and protein synthesis, whereas beta-alanine appeared to be toxic, inhibiting both VSMC synthesis and de novo protein synthesis.