Immunohistochemical localization of taurine in the rat ovary, oviduct, and uterus.

The distribution of the amino acid taurine in the female reproductive organs has not been previously analyzed in detail. The aim of this study was to determine taurine localization in the rat ovary, oviduct, and uterus by immunohistochemical methods. Taurine was localized in the ovarian surface epithelium. The granulosa cells and oocytes of primordial follicles were immunonegative. In primary and antral follicles, taurine was found mainly in theca cells and oocytes, whereas the zona pellucida, antrum, and most granulosa cells were unstained. However, taurine immunoreactivity in theca cells and oocytes decreased during follicular atresia. During corpora lutea development, the number of immunopositive theca lutein cells increased as these cells invaded the granulosa-derived region. Therefore, most luteal cells from the mature corpora lutea were stained. In the regressing corpora lutea, however, taurine staining in luteal cells decreased. In the fimbriae, infundibulum, and uterotubal junction, taurine was localized in most epithelial cells. In the ampullar and isthmic segments, taurine was found in the cilia of most ciliated cells and in the apical cytoplasm of some non-ciliated cells. In the uterus, most epithelial cells were immunopositive during diestrus and metestrus, whereas most of them were immunonegative during estrus and proestrus. Moreover, taurine immunoreactivity in the oviduct and uterus decreased with pregnancy. (J Histochem Cytochem 49:1133-1142, 2001)

Taurine activates GABA(A) but not GABA(B) receptors in rat hippocampal CA1 area.

We investigated if taurine, an endogenous GABA analog, could mimic both hyperpolarizing and depolarizing GABA(A)-mediated responses as well as pre- and postsynaptic GABA(B)-mediated actions in the CA1 region of rat hippocampal slices. Taurine (10 mM) perfusion induced changes in membrane potential and input resistance that are compatible with GABA(A) receptor activation. Local pressure application of taurine and GABA from a double barrel pipette positioned along the dendritic shaft of pyramidal cells revealed that taurine evoked a very small change of membrane potential and resistance compared with the large changes induced by GABA in these parameters. Moreover, in the presence of GABA(A) antagonists, local application of GABA on the dendrites evoked a GABA(B)-mediated hyperpolarization while taurine did not induce any change. Taurine neither mimicked baclofen inhibitory actions on presynaptic release of glutamate and GABA as judging by the lack of taurine effect on paired-pulse facilitation ratio and slow inhibitory postsynaptic potentials, respectively. These results show that taurine mainly activates GABA(A) receptors located on the cell body, indicating therefore that if taurine has any action on the dendrites it will not be mediated by either GABA(A) or GABA(B) receptors activation.

Immunohistochemical localization of taurine in the male reproductive organs of the rat.

The amino acid taurine has been implicated in several aspects of reproductive system physiology. However, its localization in these organs has not been previously analyzed. The aim of this study was to characterize its distribution in male rat reproductive organs by immunohistochemical methods. Taurine was localized in the smooth muscle cells of the tissues studied and in the skeletal fibers of the cremaster muscle. In the testis, taurine was found in Leydig cells, vascular endothelial cells, and other interstitial cells. No immunoreactivity was observed in the cells of the seminiferous tubules, either in germ cells at all spermatogenic stages or in Sertoli cells. However, peritubular myoid cells were immunostained. Most epithelial cells of the efferent ducts were immunolabeled, whereas the epithelial cells of the rete testis (extratesticular segments), epididymis (caput, corpus, and cauda regions), and ductus deferens were unstained. However, most epithelial cells from the intratesticular segments of the rete were immunopositive. Some cells identified as intraepithelial macrophages and lymphocytes, apical cells, and narrow cells were intensely immunolabeled. Regional differences in the distribution of these cell types along the ducts studied were also noted. The possible functional roles for taurine in these cells are discussed.

Weak organic acids induce taurine release through an osmotic-sensitive process in in vivo rat hippocampus.

Isotonic media containing sodium salts from weak organic acids induce cell swelling in several experimental preparations (Grinstein et al., 1984; Jakubovicz et al., 1987). In vivo perfusion of rat dentate gyrus, using a microdialysis probe, with modified Krebs-Ringer bicarbonate solutions in which 50 mM NaCl was isotonically substituted by the sodium salts from organic acids with a pKa value of greater than 2 (acetate, propionate, or pyruvate), induced a reversible increase in the extracellular taurine concentration. By contrast, similar NaCl substitutions with sodium salts from the stronger organic acids isethionate and methane-sulfonate did not change extracellular taurine levels. Extracellular taurine increases evoked by acetate, propionate, or pyruvate were almost completely abolished when the perfusion liquid was made hypertonic by adding sucrose (50 mM). A 30% reduction of the acetate-induced extracellular taurine increase was observed both when amiloride was present or when the [Na+]0 was lowered. Both conditions are known to inhibit Na+/H+ exchange. These results are compatible with the hypothesis that acid load-induced taurine release is stimulated by an osmotic sensitive mechanism, part of which is dependent on activation of the Na+/H+ exchange.

Neurochemical findings in neuroacanthocytosis.

We performed a neurochemical study of the brain of two unrelated patients, living in different continents, with neuroacanthocytosis. The levels of monoamines and their metabolites, gamma-aminobutyric acid and substance P, were measured in several brain areas and the monoamine metabolites in cerebrospinal fluid. The binding of 3H-spiperone to striatal membranes and to lymphocytes was also measured. Both patients had a progressive neurological disorder with onset in the third decade of life and characterized by a complex movement disorder, epilepsy, muscular wasting, and changes in behavior. The movement disorder initially manifested with oromandibular dystonia and limb chorea, but at the time of death was characterized by a severe dystonic syndrome. The chemical changes were similar in the two patients. The most important neurochemical findings were a depletion of dopamine and its metabolites in most brain areas, most notably in the striatum, and elevation of norepinephrine levels in the putamen and globus pallidus. Substance P was markedly reduced in the striatum and substantia nigra. Our findings may provide clues to the neurochemical mechanisms underlying dystonia.

Placental formation of lactate from transferred L-alanine and its impairment by aminooxyacetate in the late-pregnant rat.

After 20 min infusion of L-[U-14C]alanine through the left uterine artery in 21-day-pregnant rats, the radioactivity in the plasma of fetuses from the left uterine horn was much higher than in their mothers and was composed of approximately equal parts of [14C]alanine and [14C]lactate, with a minor percentage of [14C]glucose. Radioactivity in fetal plasma was much lower when the mothers were infused with alpha-amino[14C]isobutyric acid. The simultaneous infusion of aminooxyacetate decreased materno-fetal transfer of radioactivity from [14C]alanine but not from alpha-amino[14C]isobutyric acid, and this effect corresponded to a complete disappearance of the [14C]lactate in fetal plasma without affecting [14C]alanine levels or alanine concentration in the fetuses. Placenta slices in vitro metabolized L-[U-14C]alanine into [14C]lactate and 14CO2 at the rate of 7 nmol/g per min, and this process was inhibited by the presence of 1 mM aminooxyacetate in the medium. Placental uptake of alpha-amino[14C]isobutyric acid was half that of [U-14C]alanine, and aminooxyacetate did not affect this parameter with either of the labelled compounds. Results indicate that the lower transfer to the rat fetus of the 14C atoms from alpha-amino[14C]isobutyric acid as compared to that from [14C]alanine is due not only to the diminished placental carrier system of the former but also to its non-metabolyzable condition. It is proposed that the capacity of the placenta to metabolyze L-alanine to lactate and the subsequent release of lactate to the fetus constitute important factors for the fetal metabolic economy.

Electrophysiological evidence that nipecotic acid can be used in vivo as a false transmitter.

Dentate gyrus of the rat hippocampal formation was perfused with the potent inhibitor of GABA uptake, nipecotic acid, by means of an implanted dialytrode. Evoked population spikes in dentate gyrus were decreased in amplitude and often abolished during perfusion. However, multiple (2-4) population spikes developed shortly after nipecotic acid withdrawal. This excitability increase, which presented a pattern of repetitive discharge resembling that following blocking of GABAergic transmission was interpreted as electrophysiological evidence that nipecotic acid can act as a false transmitter 'in vivo', as previously postulated from uptake and release 'in vitro' studies.

Aminobutyric acid greatly increases the in vivo extracellular taurine in the rat hippocampus.

The effects of gamma-aminobutyric acid (GABA) on the extracellular levels of taurine and on excitability in the dentate gyrus were studied in anesthetized rats by the dialytrode technique. The dentate gyrus was perfused by means of a dialytrode with Krebs-Ringer-bicarbonate or GABA solutions. Amino acid contents in perfusates and dentate field potentials evoked by electrical stimulation of the perforant pathway were evaluated. GABA drastically elevated the levels of extracellular taurine in a dose-dependent manner, decreasing the amplitude of the population spike. This result indicates that GABA stimulates taurine release, probably by a counter-transport process. It is suggested that in physiological conditions an increase in extracellular taurine may be produced by synaptically released GABA.

In vivo effects of nipecotic acid on levels of extracellular GABA and taurine, and hippocampal excitability.

The effect of nipecotic acid on the extracellular levels of the putative hippocampal inhibitory transmitters, gamma-aminobutyric acid (GABA) and taurine, and also excitability in the dentate gyrus, were studied in anaesthetised rats using the dialytrode technique. The dentate gyrus was perfused by means of a dialytrode , using Krebs-Ringer bicarbonate or nipecotic acid solutions. Dentate field potentials, evoked by electrical stimulation of the perforant pathway, were recorded. Nipecotic acid drastically elevated the levels of extracellular GABA and taurine and the amplitude of population spikes in the dentate was concomitantly decreased. These results indicate that changes in extracellular levels of endogenous GABA and/or taurine influence the excitability of the hippocampus.

Chemical characterization of the selenoprotein component of clostridial glycine reductase: identification of selenocysteine as the organoselenium moiety.

A small, heat-stable selenoprotein, one of the components of the glycine reductase complex, was labeled with 75Se by growth of Clostridium sticklandii in the presence of Na2 75SeO3. The selenium-containing moiety, which is essential for the biological activity of the protein, was shown to be a selenocysteine residue. It was isolated as its Se-carboxymethyl, Se-carboxyethyl, and Se-aminoethyl derivatives from digests of the pure 75Se-labeled protein that had been reduced and treated with the various alkylating agents prior to hydrolysis. In each instance the 75Se-labeled moiety obtained from an alkylated protein sample and the corresponding alkyl derivative of authentic selenocysteine were indistinguishable. Several studies of the native selenoprotein detected a chromophore (UVmax 238nm) that appeared upon reduction of the protein with KBH4 and rapidly disappeared upon exposure to oxygen. This oxygen-labile chromophore is thought to be the ionized -SeH group of the selenocysteine residue.