Une formation nucléaire paracristalline dans les cellules tubulaires proximales rénales de chien

Résumé Des inclusions intranucléaires paracristallines ont été mises en évidence dans les tubes proximaux de reins de chien. Ces inclusions sont constituées par une répétition régulière d'unités élémentaires microtubulaires de 85±15 Å de diamètre avec une périodicité de 120±20 Å. Elles ne sont jamais en contact ni avec le nucléole ni avec la membrane nucléaire. L'étude histochimique montre qu'il s'agit d'inclusions de nature protéique. Ces inclusions ont été observées à la fois dans les reins greffés (autogreffes) après conservation dans un perfusat artificiel (selon «Collins» ou «Perfudex») pendant 24 heures à 0°C et dans les reins normaux. N'étant jamais rencontrées au niveau de cellules rénales lésées, il ne semble pas que ces inclusions expriment une dégénérescence cellulaire.

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Paracrystalline nuclear inclusions in the proximal tubule cells of the canine kidney

Summary The occurrence of intranuclear paracrystalline inclusions in proximal tubules of the canine kidney is described. These inclusions are composed of a regular repetition of microtubular elementary units of 85±15 Å thickness with a periodicity of 120±20 Å. They have no contact with the nucleolus or the nuclear membrane. Histochemical findings suggest that these inclusions may be proteinaceous. They are both observed in grafted kidneys (autografts) after conservation during 24 hours at 0°C in a synthetic solution (according to Collins or Perfudex) and in normal kidneys. There were no signs of degeneration in cells containing such inclusions; therefore, the possibility that these structures are of degenerative nature seems less probable.

     

Stream power influence on southern Californian riparian vegetation

Abstract. Mechanical damage by floodwaters is frequently invoked to explain the distribution of riparian plant species, but data have been lacking to relate vegetation to specific estimates of flood damage potential. This research uses detailed estimates of unit stream power (an appropriate measure of the potential for mechanical damage) in conjunction with vegetation cover data to test this relationship at 37 valley-bottom sites in the Transverse Ranges of Southern California. A computer program, HEC-2, was used to model the slope and the variation in flow depth and velocity of the 20-yr flood across the sites. Regression models tested the influence of stream power (and of height above the water table) on the woody species composition of 393 4-m cross-section segments of the valley-bottom sites. Results indicate that unit stream power does have a significant effect on the riparian vegetation, but that the amount of that influence and its importance relative to the influence of height above the water table varies between watersheds. Some species are found primarily in locations of high stream power, while others are limited to portions of the valley bottom that experience only low stream power.     

Leaf Anatomy of Fourteen Species in Carex Subgenus Indocarex (Cyperaceae)

Among the subgenera of the genus Carex, the subgenus Indocarex has been seldom studied in any respects, Its systematic position and its subdivision are still disputable. Leaf anatomy of 14 species in the subgenus lndocarex from China was studied. The anatomical characters are proved to be systematically valuable. (1) Characters of lamina transverse section: All leaves of these 14 species are dorsiventral. The outline mostly V-shaped, occasionally flat or nearly flat, with adaxial lateral rib in each half of lamina and some of them flanged. The cells of adaxial surface larger than those of abaxial surface, and the epidermal cells over veins usually smaller than others. Air-cavities between vascular bundles are well developed, and bulliform cells also well developed in most taxa. The vascular bundles are collaterai, bundle sheaths double-layered, and the outer sheath parenchymatous and the inner sheath fibrous. (2) Characters of lamina epidermis: The shape of the cell on both surfaces is generally rectangular, and the anticlinal wall of epidermal cell sinuous; stomata is paracytic, elliptic to oblong, rarely sub-circular; prickles occur on adaxial surfaces of certain species; papillae are only obvious on abaxial surface of C. moupinensis Franch. The characters of transverse section and epidermis of leaf blades of these 14 species differ from each other to certain degree, and closely related species are similar in anatomical characters. The anatomical characters of lamina are of value for classification at specific and sectional level of the subgenus Indocarex. Despite of the variation of these characters among species, a certain num ber of characters appears to be shared by the members of the subgenus, and some of the common characters are primitive. In addition, some gross morphological characters are common and primitive also. Therefore, the subgenus Indocarex may be primitive in the genus Carex. The anatomical and morphological characters of C. scaposa C. B. Clakre and C. densifimbriata Tang et Wang ex S. Y. Liang are distinct. The two species and their allies should be treated as section instead of subsection. The three species in the sectionPolystachyae share some anatomical characters and comprise a coherent group.     

Lumbo-sacral neural crest derivatives fate mapped with the aid of Wnt-1 promoter integrate but are not essential to kidney development

Neural crest (NC) cells may be involved in kidney organogenesis by providing inductive signals and contributing to cells of the renal stroma. We show here that the lumbo-sacral NC cells fate mapped with the aid of Wnt-1 promoter in the mouse migrate close to the metanephros at the initiation of organogenesis but these cells remain superficial to the condensed Pax2-expressing mesenchymal cells. NC-derived cells enter later into the kidney proper from the midline region. The NC cells contribute also to development of the extra-adrenal para-aortic bodies, Zuckerkandl's bodies and the nerve cord of the sympathetic nervous system. Splotch (Sp^2H/Sp^2H) embryos, having a NC defect in the lumbo-sacral region, develop a normal metanephros even though the kidney does not express the NC markers Sox10, Phox2b and tyrosine hydroxylase. Consistent with the histological findings, the kidneys of Sp^2H/Sp^2H embryos also express the stromal genes Foxd1, Hoxa10 and RARβ normally. Wnt-1 promoter-marked wild-type LacZ NC cells migrate intensely from the heterologous inducer tissue of the embryonic dorsal spinal cord (SPC) to the kidney mesenchyme, but tubule induction does not depend on NC migration, since the Sp^2H/Sp^2H SPC also induces tubulogenesis. The Sp^2H/Sp^2H mesenchyme also remains competent for tubulogenesis. We conclude that the NC cells fate mapped with the aid of Wnt-1 promoter migrate to the close to the metanephros and form later derivatives integrating with the kidney, but they may not be essential to the development of the stromal cells nor they may provide critical morphogenetic signals to regulate early kidney development in vivo.     

Molecular and functional characterization of choline transporter in rat renal tubule epithelial NRK-52E cells

Homeostatic regulation of the plasma choline concentration depends on the effective functioning of a choline transporter in the kidney. However, the nature of the choline transport system in the kidney is poorly understood. In this study, we examined the molecular and functional characterization of choline uptake in the rat renal tubule epithelial cell line NRK-52E. Choline uptake was saturable and mediated by a single transport system, with an apparent Michaelis-Menten constant (K_m) of 16.5 μM and a maximal velocity (V_max) of 133.9 pmol/mg protein/min. The V_max value of choline uptake was strongly enhanced in the absence of Na⁺ without any change in K_m values. The increase in choline uptake under Na⁺-free conditions was inhibited by Na⁺/H⁺ exchanger (NHE) inhibitors. Choline uptake was inhibited by the choline uptake inhibitor hemicholinium-3 (HC-3) and organic cations, and was decreased by acidification of the extracellular medium and by intracellular alkalinization. Collapse of the plasma membrane H⁺ electrochemical gradient by a protonophore inhibited choline uptake. NRK-52E cells mainly express mRNA for choline transporter-like proteins (CTL1 and CTL2), and NHE1 and NHE8. CTL1 protein was recognized in both plasma membrane and mitochondria. CTL2 protein was mainly expressed in mitochondria. The biochemical and pharmacological data indicated that CTL1 is functionally expressed in NRK-52E cells and is responsible for choline uptake. This choline transport system uses a directed H⁺ gradient as a driving force, and its transport functions in co-operation with NHE8. Furthermore, the presence of CTL2 in mitochondria provides a potential site for the control of choline oxidation.     

Adenosine deamination to inosine in isolated basolateral membrane from kidney proximal tubule: Implications for modulation of the membrane-associated protein kinase A

In this work, the metabolism of adenosine by isolated BLM associated-enzymes and the implications of this process for the cAMP-signaling pathway are investigated. Inosine was identified as the major metabolic product, suggesting the presence of adenosine deaminase (ADA) activity in the BLM. This was confirmed by immunoblotting and ADA-specific enzyme assay. Implications for the enzymatic deamination of adenosine on the receptor-modulated cAMP-signaling pathway were also investigated. We observed that inosine induced a 2-fold increase in [³⁵S] GTPγS binding to the BLM and it was inhibited by 10⁻⁶ M DPCPX, an A₁ receptor-selective antagonist. Inosine (10⁻⁷ M) inhibited protein kinase A activity in a DPCPX-sensitive manner. Molecular association between ADA and G_αi-3 protein-coupled A₁ receptor was demonstrated by co-immunoprecipitation assay. These data show that adenosine is deaminated by A₁ receptor-associated ADA to inosine, which in turn modulates PKA in the BLM through A₁ receptor-mediated inhibition of adenylyl cyclase.     

Megalin is downregulated via LPS-TNF-α-ERK1/2 signaling pathway in proximal tubule cells

Expression and function of megalin, an endocytic receptor in proximal tubule cells (PTCs), are reduced in diabetic nephropathy, involved in the development of proteinuria/albuminuria. Lipopolysaccharide (LPS) is chronically increased in diabetic sera, by the mechanism called metabolic endotoxemia. We investigated low-level LPS-mediated signaling that regulates megalin expression in immortalized rat PTCs (IRPTCs). Incubation of the cells with LPS (10 ng/ml) for 48 h suppressed megalin protein expression and its endocytic function. TNF-α mRNA expression was increased by LPS treatment, and knockdown of the mRNA with siRNA inhibited LPS-mediated downregulation of megalin mRNA expression at the 24-h time point. Incubation of IRPTCs with exogenous TNF-α also suppressed megalin mRNA and protein expression at the 24- and 48-h time points, respectively. MEK1 inhibitor PD98059 competed partially but significantly TNF-α-mediated downregulation of megalin mRNA expression. Collectively, low-level LPS-mediated TNF-α-ERK1/2 signaling pathway is involved in downregulation of megalin expression in IRPTCs.     

Modulation of ouabain-insensitive Na(+)-ATPase activity in the renal proximal tubule by Mg(2+), MgATP and furosemide

In addition to the (Na⁺+K⁺)ATPase another P-ATPase, the ouabain-insensitive Na⁺-ATPase has been observed in several tissues. In the present paper, the effects of ligands, such as Mg²⁺, MgATP and furosemide on the Na⁺-ATPase and its modulation by pH were studied in the proximal renal tubule of pig. The principal kinetics parameters of the Na⁺-ATPase at pH 7.0 are: (a) K_0.5 for Na⁺=8.9±2.2 mM; (b) K_0.5 for MgATP=1.8±0.4 mM; (c) two sites for free Mg²⁺: one stimulatory (K_0.5=0.20±0.06 mM) and other inhibitory (I_0.5=1.1±0.4 mM); and (d) I_0.5 for furosemide=1.1±0.2 mM. Acidification of the reaction medium to pH 6.2 decreases the apparent affinity for Na⁺ (K_0.5=19.5±0.4) and MgATP (K_0.5=3.4±0.3 mM) but increases the apparent affinity for furosemide (0.18±0.02 mM) and Mg²⁺ (0.05±0.02 mM). Alkalization of the reaction medium to pH 7.8 decreases the apparent affinity for Na⁺ (K_0.5=18.7±1.5 mM) and furosemide (I_0.5=3.04±0.57 mM) but does not change the apparent affinity to MgATP and Mg²⁺. The data presented in this paper indicate that the modulation of the Na⁺-ATPase by pH is the result of different modifications in several steps of its catalytical cycle. Furthermore, they suggest that changes in the concentration of natural ligands such as Mg²⁺ and MgATP complex may play an important role in the Na⁺-ATPase physiological regulatory mechanisms.     

New horizons in schizophrenia treatment: autophagy protection is coupled with behavioral improvements in a mouse model of schizophrenia

Autophagy plays a key role in the pathophysiology of schizophrenia as manifested by a 40% decrease in BECN1/Beclin 1 mRNA in postmortem hippocampal tissues relative to controls. This decrease was coupled with the deregulation of the essential ADNP (activity-dependent neuroprotector homeobox), a binding partner of MAP1LC3B/LC3B (microtubule-associated protein 1 light chain 3 β) another major constituent of autophagy. The drug candidate NAP (davunetide), a peptide fragment from ADNP, enhanced the ADNP-LC3B interaction. Parallel genetic studies have linked allelic variation in the gene encoding MAP6/STOP (microtubule-associated protein 6) to schizophrenia, along with altered MAP6/STOP protein expression in the schizophrenic brain and schizophrenic-like behaviors in Map6-deficient mice. In this study, for the first time, we reveal significant decreases in hippocampal Becn1 mRNA and reversal by NAP but not by the antipsychotic clozapine (CLZ) in Map6-deficient (Map6^+/−) mice. Normalization of Becn1 expression by NAP was coupled with behavioral protection against hyperlocomotion and cognitive deficits measured in the object recognition test. CLZ reduced hyperlocomotion below control levels and did not significantly affect object recognition. The combination of CLZ and NAP resulted in normalized outcome behaviors. Phase II clinical studies have shown NAP-dependent augmentation of functional activities of daily living coupled with brain protection. The current studies provide a new mechanistic pathway and a novel avenue for drug development.