Residual effect of organic zinc complexes in acid and calcareous soils on navy bean (phaseolus vulgaris l.) and flax (linum usitatissimum l.) crops under greenhouse conditions. Influence of the moisture conditions = Efecto residual de complejos orgánicos de zinc en suelos ácido y calizo en cultivos de judía (phaseolus vulgaris l.) y lino (linum usitatissimum l.) Realizados en condiciones de invernadero. Influencia de las diferentes condiciones de humedad

El Zn es un elemento esencial para el crecimiento saludable y reproducción de plantas, animales y humanos. La deficiencia de Zn es una de las carencias de micronutrientes más extendidas en muchos cultivos, afectando a grandes extensiones de suelos en diferentes áreas agrícolas. La biofortificación agronómica de diferentes cultivos, incrementando la concentración de micronutriente Zn en la planta, es un medio para evitar la deficiencia de Zn en animales y humanos. Tradicionalmente se han utilizado fertilizantes de Zn inorgánicos, como el ZnSO4, aunque en los últimos años se están utilizado complejos de Zn como fuentes de este micronutriente, obteniéndose altas concentraciones de Zn soluble y disponible en el suelo. Sin embargo, el envejecimiento de la fuente en el suelo puede causar cambios importantes en su disponibilidad para las plantas. Cuando se añaden al suelo fuentes de Zn inorgánicas, las formas de Zn más solubles pierden actividad y extractabilidad con el paso del tiempo, transformándose a formas más estables y menos biodisponibles. En esta tesis se estudia el efecto residual de diferentes complejos de Zn de origen natural y sintético, aplicados en cultivos previos de judía y lino, bajo dos condiciones de riego distintas (por encima y por debajo de la capacidad de campo, respectivamente) y en dos suelos diferentes (ácido y calizo). Los fertilizantes fueron aplicados al cultivo previo en tres dosis diferentes (0, 5 y 10 mg Zn kg-1 suelo). El Zn fácilmente lixiviable se estimó con la extracción con BaCl2 0,1M. Bajo condiciones de humedad por encima de la capacidad de campo se obtuvieron mayores porcentajes de Zn lixiviado en el suelo calizo que en el suelo ácido. En el caso del cultivo de judía realizado en condiciones de humedad por encima de la capacidad de campo se compararon las cantidades extraídas con el Zn lixiviado real. El análisis de correlación entre el Zn fácilmente lixiviable y el estimado sólo fue válido para complejos con alta movilidad y para cada suelo por separado. Bajo condiciones de humedad por debajo de la capacidad de campo, la concentración de Zn biodisponible fácilmente lixiviable presentó correlaciones positivas y altamente significativas con la concentración de Zn disponible en el suelo. El Zn disponible se estimó con varios métodos de extracción empleados habitualmente: DTPA-TEA, DTPA-AB, Mehlich-3 y LMWOAs. Estas concentraciones fueron mayores en el suelo ácido que en el calizo. Los diferentes métodos utilizados para estimar el Zn disponible presentaron correlaciones positivas y altamente significativas entre sí. La distribución del Zn en las distintas fracciones del suelo fue estimada con diferentes extracciones secuenciales. Las extracciones secuenciales mostraron un descenso entre los dos cultivos (el anterior y el actual) en la fracción de Zn más lábil y un aumento en la concentración de Zn asociado a fracciones menos lábiles, como carbonatos, óxidos y materia orgánica. Se obtuvieron correlaciones positivas y altamente significativas entre las concentraciones de Zn asociado a las fracciones más lábiles (WSEX y WS+EXC, experimento de la judía y lino, respectivamente) y las concentraciones de Zn disponible, estimadas por los diferentes métodos. Con respecto a la planta se determinaron el rendimiento en materia seca y la concentración de Zn en planta. Se observó un aumento del rendimiento y concentraciones con el efecto residual de la dosis mayores (10 mg Zn kg-1) con respecto a la dosis inferior (5 mg Zn 12 kg-1) y de ésta con respecto a la dosis 0 (control). El incremento de la concentración de Zn en todos los tratamientos fertilizantes, respecto al control, fue mayor en el suelo ácido que en el calizo. Las concentraciones de Zn en planta indicaron que, en el suelo calizo, serían convenientes nuevas aplicaciones de Zn en posteriores cultivos para mantener unas adecuadas concentraciones en planta. Las mayores concentraciones de Zn en la planta de judía, cultivada bajo condiciones de humedad por encima de la capacidad de campo, se obtuvieron en el suelo ácido con el efecto residual del Zn-HEDTA a la dosis de 10 mg Zn kg-1 (280,87 mg Zn kg-1) y en el suelo calizo con el efecto residual del Zn-DTPA-HEDTA-EDTA a la dosis de 10 mg Zn kg-1 (49,89 mg Zn kg-1). En el cultivo de lino, cultivado bajo condiciones de humedad por debajo de la capacidad de campo, las mayores concentraciones de Zn en planta ese obtuvieron con el efecto residual del Zn-AML a la dosis de 10 mg Zn kg-1 (224,75 mg Zn kg-1) y en el suelo calizo con el efecto residual del Zn-EDTA a la dosis de 10 mg Zn kg-1 (99,83 mg Zn kg-1). El Zn tomado por la planta fue determinado como combinación del rendimiento y de la concentración en planta. Bajo condiciones de humedad por encima de capacidad de campo, con lixiviación, el Zn tomado por la judía disminuyó en el cultivo actual con respecto al cultivo anterior. Sin embargo, en el cultivo de lino, bajo condiciones de humedad por debajo de la capacidad de campo, se obtuvieron cantidades de Zn tomado superiores en el cultivo actual con respecto al anterior. Esta tendencia también se observó, en ambos casos, con el porcentaje de Zn usado por la planta. Summary Zinc is essential for healthy growth and reproduction of plants, animals and humans. Zinc deficiency is one of the most widespread micronutrient deficiency in different crops, and affect different agricultural areas. Agronomic biofortification of crops produced by an increased of Zn in plant, is one way to avoid Zn deficiency in animals and humans Sources with inorganic Zn, such as ZnSO4, have been used traditionally. Although, in recent years, Zn complexes are used as sources of this micronutrient, the provide high concentrations of soluble and available Zn in soil. However, the aging of the source in the soil could cause significant changes in their availability to plants. When an inorganic source of Zn is added to soil, Zn forms more soluble and extractability lose activity over time, transforming into forms more stable and less bioavailable. This study examines the residual effect of different natural and synthetic Zn complexes on navy bean and flax crops, under two different moisture conditions (above and below field capacity, respectively) and in two different soils (acid and calcareous). Fertilizers were applied to the previous crop in three different doses (0, 5 y 10 mg Zn kg-1 soil). The easily leachable Zn was estimated by extraction with 0.1 M BaCl2. Under conditions of moisture above field capacity, the percentage of leachable Zn in the calcareous soil was higher than in acid soil. In the case of navy bean experiment, performed in moisture conditions of above field capacity, amounts extracted of easily leachable Zn were compared with the real leachable Zn. Correlation analysis between the leachable Zn and the estimate was only valid for complex with high mobility and for each soil separately. Under moisture conditions below field capacity, the concentration of bioavailable easily leachable Zn showed highly significant positive correlations with the concentration of available soil Zn. The available Zn was estimated with several commonly used extraction methods: DTPA-TEA, AB-DTPA, Mehlich-3 and LMWOAs. These concentrations were higher in acidic soil than in the calcareous. The different methods used to estimate the available Zn showed highly significant positive correlations with each other. The distribution of Zn in the different fractions of soil was estimated with different sequential extractions. The sequential extractions showed a decrease between the two crops (the previous and current) at the most labile Zn fraction and an increase in the concentration of Zn associated with the less labile fractions, such as carbonates, oxides and organic matter. A positive and highly significant correlation was obtained between the concentrations of Zn associated with more labile fractions (WSEX and WS + EXC, navy bean and flax experiments, respectively) and available Zn concentrations determined by the different methods. Dry matter yield and Zn concentration in plants were determined in plant. Yield and Zn concentration in plant were higher with the residual concentrations of the higher dose applied (10 mg Zn kg-1) than with the lower dose (5 mg Zn kg-1), also these parameters showed higher values with application of this dose than with not Zn application. The increase of Zn concentration in plant with Zn treatments, respect to the control, was greater in the acid soil than in the calcareous. The Zn concentrations in plant indicated that in the calcareous soil, new applications of Zn are desirable in subsequent crops to maintain suitable concentrations in plant. 15 The highest concentrations of Zn in navy bean plant, performed under moisture conditions above the field capacity, were obtained with the residual effect of Zn-HEDTA at the dose of 10 mg Zn kg-1 (280.87 mg Zn kg-1) in the acid soil, and with the residual effect of Zn- DTPA-HEDTA-EDTA at a dose of 10 mg Zn kg-1 (49.89 mg Zn kg-1) in the calcareous soil. In the flax crop, performed under moisture conditions below field capacity, the highest Zn concentrations in plant were obtained with the residual effect of Zn-AML at the dose of 10 mg Zn kg-1 (224.75 Zn mg kg-1) and with the residual effect of Zn-EDTA at a dose of 10 mg Zn kg-1 (99.83 mg Zn kg-1) in the calcareous soil. The Zn uptake was determined as a combination of yield and Zn concentration in plant. Under moisture conditions above field capacity, with leaching, Zn uptake by navy bean decreased in the current crop, respect to the previous crop. However, in the flax crop, under moisture conditions below field capacity, Zn uptake was higher in the current crop than in the previous. This trend is also observed in both cases, with the percentage of Zn used by the plant

Origin and patterns of distribution of trace elements in street dust. Unleaded petrol and urban lead

The elemental composition, patterns of distribution and possible sources of street dust are not common to all urban environments, but vary according to the peculiarities of each city. The common features and dissimilarities in the origin and nature of street dust were investigated through a series of studies in two widely different cities, Madrid (Spain) and Oslo (Norway), between 1990 and 1994. The most comprehensive sampling campaign was carried out in the Norwegian capital during the summer of 1994. An area of 14 km2, covering most of downtown Oslo and some residential districts to the north of the city, was divided into 1 km2 mapping units, and 16 sampling increments of approximately 150 g were collected from streets and roads in each of them. The fraction below 100 μm was acid-digested and analysed by ICP-MS. Statistical analyses of the results suggest that chemical elements in street dust can be classified into three groups: “urban” elements (Ba, Cd, Co, Cu, Mg, Pb, Sb, Ti, Zn), “natural” elements (Al, Ga, La, Mn, Na, Sr, Th, Y) and elements of a mixed origin or which have undergone geochemical changes from their original sources (Ca, Cs, Fe, Mo, Ni, Rb, Sr, U). Soil resuspension and/or mobilisation appears to be the most important source of “natural” elements, while “urban” elements originate primarily from traffic and from the weathering and corrosion of building materials. The data for Pb seem to prove that the gradual shift from leaded to unleaded petrol as fuel for automobiles has resulted in an almost proportional reduction in the concentration of Pb in dust particles under 100 μm. This fact and the spatial distribution of Pb in the city strongly suggest that lead sources other than traffic (i.e. lead accumulated in urban soil over the years) may contribute as much lead, if not more, to urban street dust.

Structural distribution of stability in a thermophilic enzyme

Stability parameters for individual residues in Thermus thermophilus cysteine-free RNase H were determined by native state hydrogen exchange, thus providing a unique comparison of regional thermodynamics between thermophilic and mesophilic homologues. The general distribution of stability in the thermophilic protein is similar to that of its mesophilic homologue, with a proportional increase in stability for almost all residues. As a consequence, the residue-specific stabilities of the two proteins are remarkably similar under conditions where their global stabilities are the same. These results indicate that T. thermophilus RNase H is stabilized in a delocalized fashion, preserving a finely tuned balance of stabilizing interactions throughout the structure. Therefore, although protein stability can be altered by single amino acid substitution, evolution for optimal function may require more subtle and delocalized mechanisms.

Multiple protein domains determine the cell type-specific nuclear distribution of the catalytic subunit required for apolipoprotein B mRNA editing

Apolipoprotein B (apoB) mRNA editing catalyzed by apoB mRNA editing catalytic subunit 1 (APOBEC-1) has been proposed to be a nuclear process. To test this hypothesis, the subcellular distribution of hemagglutinin-(HA) tagged APOBEC-1 expressed in transiently transfected hepatoma cells was determined by indirect immunofluorescence microscopy. HA-APOBEC-1 was detected in both the nucleus and cytoplasm of rat and human hepatoma cells. Mutagenesis of APOBEC-1 demonstrated that the N-terminal 56 amino acids (1–56) were necessary for the nuclear distribution of APOBEC-1, but this region did not contain a functional nuclear localization signal (NLS). However, we identified a 24-amino acid domain in the C terminus of APOBEC-1 with characteristics of a cytoplasmic retention signal (CRS) or a nuclear export signal (NES). These data suggest, therefore, that the nuclear import of APOBEC-1 may not be mediated by a positive NLS; rather, it may be achieved by overcoming the effect of a CRS/NES. We also demonstrated that the nuclear distribution of APOBEC-1 occurred only in cell lines that were capable of editing apoB RNA. We propose that the cellular distribution of APOBEC-1 is determined by multiple domains within this protein, and a nuclear localization of the enzyme may be regulated by cell type-specific factors that render these cells uniquely editing competent.

Retinoic acid alters the intracellular trafficking of the mannose-6-phosphate/insulin-like growth factor II receptor and lysosomal enzymes

Previously, we showed that retinoic acid (RA) binds to the mannose-6-phosphate/insulin-like growth factor II receptor (M6P/IGF2R) with high affinity, suggesting that M6P/IGF2R may be a receptor for RA. Here, we show that RA, after 2–3 h of incubation with cultured neonatal-rat cardiac fibroblasts, dramatically alters the intracellular distribution of M6P/IGF2R as well as that of cathepsin B (a lysosomal protease bearing M6P). Immunofluorescence techniques indicate that this change in intracellular distribution is characterized by a shift of the proteins from the perinuclear area to cytoplasmic vesicles. The effect of RA was neither blocked by an RA nuclear receptor antagonist (AGN193109) nor mimicked by a selective RA nuclear-receptor agonist (TTNPB). Furthermore, the RA-induced translocation of cathepsin B was not observed in M6P/IGF2R-deficient P388D1 cells but occurred in stably transfected P388D1 cells expressing the receptor, suggesting that the effect of RA might be the result of direct interaction with M6P/IGF2R, rather than the result of binding to the nuclear receptors. These observations not only support the idea that M6P/IGF2R mediates an RA-response pathway but also indicate a role for RA in control of intracellular trafficking of lysosomal enzymes. Therefore, our observations may have important implications for the understanding of the diverse biological effects of retinoids.

Mitochondrial carbonic anhydrase CA VB: Differences in tissue distribution and pattern of evolution from those of CA VA suggest distinct physiological roles

A cDNA for a second mouse mitochondrial carbonic anhydrase (CA) called CA VB was identified by homology to the previously characterized murine CA V, now called CA VA. The full-length cDNA encodes a 317-aa precursor that contains a 33-aa classical mitochondrial leader sequence. Comparison of products expressed from cDNAs for murine CA VB and CA VA in COS cells revealed that both expressed active CAs that localized in mitochondria, and showed comparable activities in crude extracts and in mitochondria isolated from transfected COS cells. Northern blot analyses of total RNAs from mouse tissues and Western blot analyses of mouse tissue homogenates showed differences in tissue-specific expression between CA VB and CA VA. CA VB was readily detected in most tissues, while CA VA expression was limited to liver, skeletal muscle, and kidney. The human orthologue of murine CA VB was recently reported also. Comparison of the CA domain sequence of human CA VB with that reported here shows that the CA domains of CA VB are much more highly conserved between mouse and human (95% identity) than the CA domains of mouse and human CA VAs (78% identity). Analysis of phylogenetic relationships between these and other available human and mouse CA isozyme sequences revealed that mammalian CA VB evolved much more slowly than CA VA, accepting amino acid substitutions at least 4.5 times more slowly since each evolved from its respective human–mouse ancestral gene around 90 million years ago. Both the differences in tissue distribution and the much greater evolutionary constraints on CA VB sequences suggest that CA VB and CA VA have evolved to assume different physiological roles.

Dopamine receptor subtypes modulate olfactory bulb γ-aminobutyric acid type A receptors

The γ-aminobutyric acid type A (GABAA) receptor is the predominant Cl− channel protein mediating inhibition in the olfactory bulb and elsewhere in the mammalian brain. The olfactory bulb is rich in neurons containing both GABA and dopamine. Dopamine D1 and D2 receptors are also highly expressed in this brain region with a distinct and complementary distribution pattern. This distribution suggests that dopamine may control the GABAergic inhibitory processing of odor signals, possibly via different signal-transduction mechanisms. We have observed that GABAA receptors in the rat olfactory bulb are differentially modulated by dopamine in a cell-specific manner. Dopamine reduced the currents through GABA-gated Cl- channels in the interneurons, presumably granule cells. This action was mediated via D1 receptors and involved phosphorylation of GABAA receptors by protein kinase A. Enhancement of GABA responses via activation of D2 dopamine receptors and phosphorylation of GABAA receptors by protein kinase C was observed in mitral/tufted cells. Decreasing or increasing the binding affinity for GABA appears to underlie the modulatory effects of dopamine via distinct receptor subtypes. This dual action of dopamine on inhibitory GABAA receptor function in the rat olfactory bulb could be instrumental in odor detection and discrimination, olfactory learning, and ultimately odotopic memory formation.

γ-Aminobutyric acid type B receptors are expressed and functional in mammalian cardiomyocytes

γ-Hydroxybutyrate (GHB), an anesthetic adjuvant analog of γ-aminobutyrate (GABA), depresses cell excitability in hippocampal neurons by inducing hyperpolarization through the activation of a prominent inwardly rectifying K+ (Kir3) conductance. These GABA type B (GABAB)-like effects are clearly shown at high concentrations of GHB corresponding to blood levels usually reached during anesthesia and are mimicked by the GABAB agonist baclofen. Recent studies of native GABAB receptors (GABABRs) have favored the concept that GHB is also a selective agonist. Furthermore, cloning has demonstrated that GABABRs assemble heteromeric complexes from the GABABR1 and GABABR2 subtypes and that these assemblies are activated by GHB. The surprisingly high tissue content, together with anti-ischemic and protective effects of GHB in the heart, raises the question of a possible influence of GABAB agonists on excitable cardiac cells. In the present study, we provide electrophysiological evidence that GHB activates an inwardly rectifying K+ current in rat ventricular myocytes. This effect is mimicked by baclofen, reversibly inhibited by GABAB antagonists, and prevented by pertussis toxin pretreatment. Both GABABR1 and GABABR2 are detected in cardiomyocytes by Western blotting and are shown to coimmunoprecipitate. Laser scanning confocal microscopy discloses an even distribution of the two receptors in the sarcolemma and along the transverse tubular system. Hence, we conclude that GABABRs are distributed not only in neuronal tissues but also in the heart, where they can be activated and induce electrophysiological alterations through G-protein-coupled inward rectifier potassium channels.

Identification of two novel transmembrane γ-carboxyglutamic acid proteins expressed broadly in fetal and adult tissues

The proline-rich γ-carboxyglutamic acid (Gla) proteins (PRGPs) 1 and 2 are the founding members of a family of vitamin K-dependent single-pass integral membrane proteins characterized by an extracellular amino terminal domain of approximately 45 amino acids that is rich in Gla. The intracellular carboxyl terminal region of these two proteins contains one or two copies of the sequence PPXY, a motif present in a variety of proteins involved in such diverse cellular functions as signal transduction, cell cycle progression, and protein turnover. In this report, we describe the cloning of the cDNAs for two additional human transmembrane Gla proteins (TMG) of 20–24 kDa named TMG3 and TMG4. These two proteins possess extracellular Gla domains with 13 or 9 potential Gla residues, respectively, followed by membrane-spanning hydrophobic regions and cytoplasmic carboxyl terminal regions that contain PPXY motifs. This emerging family of integral membrane Gla proteins includes proline-rich Gla protein (PRGP) 1, PRGP2, TMG3, and TMG4, all of which are characterized by broad and variable distribution in both fetal and adult tissues. Members of this family can be grouped into two subclasses on the basis of their gene organization and amino acid sequence. These observations suggest novel physiological functions for vitamin K beyond its known role in the biosynthesis of proteins involved in blood coagulation and bone development. The identification and characterization of these proteins may allow a more complete understanding of the teratogenic consequences of exposure in utero to vitamin K antagonists, such as warfarin-based anticoagulants.

Hydrophobic moments of protein structures: Spatially profiling the distribution

It is generally accepted that globular proteins fold with a hydrophobic core and a hydrophilic exterior. Might the spatial distribution of amino acid hydrophobicity exhibit common features? The hydrophobic profile detailing this distribution from the protein interior to exterior has been examined for 30 relatively diverse structures obtained from the Protein Data Bank, for 3 proteins of the 30S ribosomal subunit, and for a simple set of 14 decoys. A second-order hydrophobic moment has provided a simple measure of the spatial variation. Shapes of the calculated spatial profiles of all native structures have been found to be comparable. Consequently, profile shapes as well as particular profile features should assist in validating predicted protein structures and in discriminating between different protein-folding pathways. The spatial profiles of the 14 decoys are clearly distinguished from the profiles of their native structures.

Shifts of Intracellular pH Distribution as a Part of the Signal Mechanism Leading to the Elicitation of Benzophenanthridine Alkaloids1 : Phytoalexin Biosynthesis in Cultured Cells of Eschscholtzia californica

Cultured cells of Eschscholtzia californica (Californian poppy) respond to a yeast elicitor preparation or Penicillium cyclopium spores with the production of benzophenanthridine alkaloids, which are potent phytoalexins. Confocal pH mapping with the probe carboxy-seminaphthorhodafluor-1-acetoxymethylester revealed characteristic shifts of the pH distribution in challenged cells: within a few minutes after elicitor contact a transient acidification of cytoplasmic and nuclear areas occurred in parallel with an increase of the vacuolar pH. The change of proton concentration in the vacuole and in the extravacuolar area showed a nearly constant relation, indicating an efflux of vacuolar protons into the cytosol. A 10-min treatment with 2 mm butyric or pivalic acid caused a transient acidification of the cytoplasm comparable to that observed after elicitor contact and also induced alkaloid biosynthesis. Experimental depletion of the vacuolar proton pool reversibly prevented both the elicitor-triggered pH shifts and the induction of alkaloid biosynthesis. pH shifts and induction of alkaloid biosynthesis showed a similar dependence on the elicitor concentration. Net efflux of K+, alkalinization of the outer medium, and browning of the cells were evoked only at higher elicitor concentrations. We suggest that transient acidification of the cytoplasm via efflux of vacuolar protons is both a necessary and sufficient step in the signal path toward biosynthesis of benzophenanthridine alkaloids in Californian poppy cells.

Mapping Intercellular CO2 Mole Fraction (Ci) in Rosa rubiginosa Leaves Fed with Abscisic Acid by Using Chlorophyll Fluorescence Imaging1 : Significance of Ci Estimated from Leaf Gas Exchange

Imaging of photochemical yield of photosystem II (PSII) computed from leaf chlorophyll fluorescence images and gas-exchange measurements were performed on Rosa rubiginosa leaflets during abscisic acid (ABA) addition. In air ABA induced a decrease of both the net CO2 assimilation (An) and the stomatal water vapor conductance (gs). After ABA treatment, imaging in transient nonphotorespiratory conditions (0.1% O2) revealed a heterogeneous decrease of PSII photochemical yield. This decline was fully reversed by a transient high CO2 concentration (7400 μmol mol−1) in the leaf atmosphere. It was concluded that ABA primarily affected An by decreasing the CO2 supply at ribulose-1,5-bisphosphate carboxylase/oxygenase. Therefore, the An versus intercellular mole fraction (Ci) relationship was assumed not to be affected by ABA, and images of Ci and gs were constructed from images of PSII photochemical yield under nonphotorespiratory conditions. The distribution of gs remained unimodal following ABA treatment. A comparison of calculations of Ci from images and gas exchange in ABA-treated leaves showed that the overestimation of Ci estimated from gas exchange was only partly due to heterogeneity. This overestimation was also attributed to the cuticular transpiration, which largely affects the calculation of the leaf conductance to CO2, when leaf conductance to water is low.

Occurrence of two somatostatin variants in the frog brain: characterization of the cDNAs, distribution of the mRNAs, and receptor-binding affinities of the peptides.

In tetrapods, only one gene encoding a somatostatin precursor has been identified so far. The present study reports the characterization of the cDNA clones that encode two distinct somatostatin precursors in the brain of the frog Rana ridibunda. The cDNAs were isolated by using degenerate oligonucleotides based on the sequence of the central region of somatostatin to screen a frog brain cDNA library. One of the cDNAs encodes a 115-amino acid protein (prepro-somatostatin-14; PSS1) that exhibits a high degree of structural similarity with the mammalian somatostatin precursor. The other cDNA encodes a 103-amino acid protein (prepro-[Pro2, Met13]somatostatin-14; PSS2) that contains the sequence of the somatostatin analog (peptide SS2) at its C terminus, but does not exhibit appreciable sequence similarity with PSS1 in the remaining region. In situ hybridization studies indicate differential expression of the PSS1 and PSS2 genes in the septum, the lateral part of the pallium, the amygdaloid complex, the posterior nuclei of the thalamus, the ventral hypothalamic nucleus, the torus semicircularis and the optic tectum. The somatostatin variant SS2 was significantly more potent (4-6 fold) than somatostatin itself in displacing [125I-Tyr0, D-Trp8] somatostatin-14 from its specific binding sites. The present study indicates that the two somatostatin variants could exert different functions in the frog brain and pituitary. These data also suggest that distinct genes encoding somatostatin variants may be expressed in the brain of other tetrapods.

Subcellular localization of gamma-aminobutyric acid type A receptors is determined by receptor beta subunits.

gamma-aminobutyric acid type A (GABAA) receptors are the major sites of fast synaptic inhibition in the brain. They are constructed from four subunit classes with multiple members: alpha (1-6), beta (1-4), gamma (1-4), and delta (1). The contribution of subunit diversity in determining receptor subcellular targeting was examined in polarized Madin-Darby canine kidney (MDCK) cells. Significant detection of cell surface homomeric receptor expression by a combination of both immunological and electrophysiological methodologies was only found for the beta 3 subunit. Expression of alpha/beta binary combinations resulted in a nonpolarized distribution for alpha 1 beta 1 complexes, but specific basolateral targeting of both alpha 1 beta 2 and alpha 1 beta 3 complexes. The polarized distribution of these alpha/beta complexes was unaffected by the presence of the gamma 2S subunit. Interestingly, delivery of receptors containing the beta 3 subunit to the basolateral domain occurs via the apical surface. These results show that beta subunits can selectively target GABAA receptors to distinct cellular locations. Changes in the spatial and temporal expression of beta-subunit isoforms may therefore provide a mechanism for relocating GABAA receptor function between distinct neuronal domains. Given the critical role of these receptors in mediating synaptic inhibition, the contribution of different beta subunits in GABAA receptor function, may have implications in neuronal development and for receptor localization/clustering.