The structure and biochemistry of the vacuolar H+ ATPase in proximal and distal urinary acidification

Vacuolar H⁺ ATPases participate in renal hydrogen ion secretion in both the proximal and distal nephron. These plasma membrane forms of the vacuolar H⁺ ATPase are regulated physiologically to maintain the acid-base balance of the organism. Proton transporting renal cells have requirements for constitutive acidification of intracellular compartments for normal endocytic and secretory functions. Recent experiments have begun to reveal how the kidney regulates these proton pumps independently. Vacuolar H⁺ ATPases are a family of structurally similar enzyme which differ in the composition of specific subunits. Cytosolic regulatory enzymes are present in renal cells which may affect vacuolar H⁺ ATPases in certain membrane compartments selectively. The vacuolar H⁺ ATPase in the plasma membrane of intercalated cells resides in a specialized proton-transporting apparatus that translocates the enzyme between an intracellular membrane pool and the plasma membrane in response to physiologic stimuli.This review will focus on the structure, enzymology, and regulation of the vacuolar H⁺ ATPase in the mammalian kidney. Because of space limitations, it will cover predominantly work from our laboratory. However, a number of investigators, including Brown (Brownet al., 1987, 1988a,b, 1989), Burckhardt (Sabolicet al., 1985; Turriniet al., 1989; Simon and Burckhardt, 1990), Madsen and Tisher (Madsen and Tisher, 1985; Verlanderet al., 1987, 1989). Steinmetz (Steinmetz, 1986; Stetson and Steinmetz, 1986), Schwartz (Scwartzet al., 1985, 1988; Satlin and Schwartz, 1989), Sabatini and Kurtzman (Sabatiniet al., 1990a,b), DuBose (Diaz-Diazet al., 1986; Gurich and DuBose, 1989), Al-Awqati (Van Adelsberg and Al-Awqati, 1986), and their coworkers, and many other investigators have made important contributions to this field.     

阎甲采集方法的比较及应用

结合阎甲的生活习性介绍了野外采集阎甲科昆虫的方法,主要分为客观采集法(包括引诱物法、土网筛法、巴氏罐诱法和灯诱法)和特殊生境(树皮下、动物巢穴和储藏物)主观采集法;并比较各种方法的特性和优缺点,认为引诱物法采集效率最高。通过分析各种方法所采集的主要阎甲种类,说明主观采集法的类群针对性强。探讨各种采集方法在不同研究方向的现状和潜在的应用前景。     

Characterization of Whole Cell Chloride Conductances in a Mouse Inner Medullary Collecting Duct Cell Line mIMCD-3

The chloride conductance of inner medullary collecting duct cells (mIMCD-3 cell line) has been investigated using the whole cell configuration of the patch clamp technique. Seventy-seven percent of cells were chloride selective when measured with a NaCl-rich bathing solution and a TEACl-rich pipette solution. Seventy-five percent of chloride-selective cells (90/144) had whole cell currents which exhibited an outwardly-rectifying (OR) current-voltage (I/V) relationship, while the remaining cells exhibited a linear (L) I/V relationship. The properties of the OR and L chloride currents were distinct. OR currents (mean current densities at ±60 mV of 66 ± 5 pA/pF and 44 ± 3 pA/pF), were time- and voltage-independent with an anion selectivity (from calculated permeability ratios) of SCN⁻ (2.3), NO⁻ ₃ (1.8), ClO⁻ ₄ (1.7), Br⁻ (1.7), I⁻ (1.6), Cl⁻ (1.0), HCO⁻ ₃ (0.5), gluconate⁻ (0.2). Bath additions of NPPB, flufenamate, glibenclamide (all 100 μm) and DIDS (500 μm) produced varying degrees of block of OR currents with NPPB being the most potent (IC₅₀ of approximately 50 μm) while DIDS was the least effective. Linear chloride currents had similar current densities to the OR chloride currents and were also time- and voltage-independent. The anion selectivity sequence was SCN⁻ (2.5), NO⁻ ₃ (1.9), Br⁻ (1.4), I⁻ (1.1), Cl⁻ (1.0), ClO⁻ ₄ (0.5), HCO⁻ ₃ (0.5), gluconate⁻ (0.3). In contrast to the OR conductance, glibenclamide was the most potent and DIDS the least potent blocker of L currents. An IC₅₀ of >100 μm was observed for NPPB block. Neither OR of L chloride currents were affected by acutely or chronically increased intracellular cAMP and were not affected when intracellular Ca²⁺ levels were increased or decreased. The molecular identity and physiological role of OR and linear currents in mIMCD-3 cells are discussed. Received: 13 June 1995/Revised: 15 September 1995     

Morphologic response of the rabbit cortical collecting tubule to peritubular hypotonicity: Quantitative examination with differential interferene contrast microscopy

Summary The isolated and perfused cortical collecting tubule of the rabbit was examined by differential interference contrast microscopy in order to characterize the morphologic response of this nephron segment to peritubular hypotonicity. Computer-assisted, morphometric procedures were developed to obtain measurements of cell volume and lateral intercellular space geometry from interference contrast images of perfused nephron segments. Following dilution of the bath from 290 to 190 mOsm in the absence of antidiuretic hormone (T=25°C), the cells swelled rapidly to a new steady-state volume which was maintained for at least 20 to 30 min and which was about 90% of that predicted for ideal osmometric behavior. The increase in cell volume was accomplished entirely by bulging of the cells into the lumen; lateral space width and outside tubule diameter were unaffected by peritubular hypotonicity. In addition, the swelling of the cells was associated with an apparent swelling of intracellular organelles, e.g., nuclei and mitochondria. Our results indicate that cells of the mammalian collecting tubule swell without the capacity for significant volume regulation at 25°C and without the cytoplasmic vacuolation and dilation of the lateral intercellular spaces observed following the onset of antidiuretic hormone-dependent volume reabsorption (E. Ganote, J. Grantham, H. Moses, M. Burg and J. Orloff,J. Cell Biol. 36:355, 1968).     

Cell type-specific glycoconjugates of collecting duct cells during maturation of the rat kidney

Summary The ontogeny of lectin-positive epithelial cell types and the maturation of polarized expression of the glycocalyx of the collecting ducts (CD) of the rat kidney were studied from samples of 18th-day fetal and neonatal kidneys of various ages. Lectins from Dolichos biflorus (DBA) and Vicia villosa (VVA), with preferential affinity to principal cells, stained virtually all CD cells of the fetal kidneys. However, within two days postnatally, the number of cells positive for DBA and VVA decreased to amounts found in the adult kidneys. Moreover, a characteristic change occurred rapidly after birth in the intracellular polarization of the reactive glycoconjugates, from a uniform plasmalemmal to a preferentially apical staining. In contrast, lectins from Arachis hypogaea (PNA), Maclura pomifera (MPA) and Lotus tetragonolobus (LTA), reacting indiscriminatively with principal and intercalated cells of adult kidneys, stained most CD cells in the fetal kidneys, and failed to show any postnatal change in the amount of positive cells or in the intracellular polarization.The immunocytochemical tests for (Na + K)-ATPase and carbonic anhydrase (CA II) revealed the characteristic postnatal decrease in the amount of principal cells and simultaneous increase in the amount of CA II rich intercalated cells. DBA and VVA reactive cells also decreased postnatally, paralleling the changes observed in the (Na + K)-ATPase positive principal cells. The present results suggest that the expression of the cell type-specific glycocalyx of principal and intercalated cells is developmentally regulated, undergoes profound changes during maturation, and is most likely associated with electrolyte transport phenomena.     

Changing attitudes to nature conservation: the botanical perspective

Concern to protect wild plants began possibly as early as the eighteenth century. The early-Victorian liberal tide brought the first protests at the over-collecting of rarities, but it seemingly needed the shock of the mid-century fern craze to swing botanical opinion to the cause of moderation. Protective legislation and the establishment of sanctuaries thereafter became the twin concerns, but progress was disappointing. Conservation and the creation of reserves, the fruit of ecological thinking, rose to be the dominant aim only after the Second World War. Developments since then have been rapid and substantial.     

Quantitative analysis of the structural events associated with antidiuretic hormone-induced volume reabsorption in the rabbit cortical collecting tubule

Summary We quantitatively examined the influence of antidiuretic hormone (ADH)-dependent volume reabsorption on the morphology of the rabbit cortical collecting tubule. Estimates of cell volume and the geometry of the lateral intercellular spaces were extracted from differential interference contrast images of perfused nephron segments using the morphometric procedures described in the preceding paper (K.L. Kirk, D.R. DiBona and J.A. Schafer,J. Membrane Biol. 79:53–64, 1984). The results indicate that ADH addition in the presence, but not absence, of a lumen-to-bath osmotic gradient (130 to 290 mOsm) stimulated transepithelial volume flow and simultaneously increased the volumes of both the cells (+28%) and the lateral intercellular spaces (+78%). In addition, the formation of cytoplasmic vacuoles could be observed during the latter stages of the swelling response, and vacuole formation continued well after new steadystate values for transepithelial water flow and cell volume had been reached. Two main conclusions can be drawn from these results. First, the cytoplasmic vacuoles comprise a slowly filling compartment that lies in parallel to the transepithelial pathway for ADH-stimulated volume reabsorption. Second, from the magnitude of the cell volume increase, we estimate that the hydraulic conductivities of the opposing cell membranes are nearly equal during maximal ADH stimulation.     

Convergent evolution of morphological specializations in Central European bee and honey wasp species as an adaptation to the uptake of pollen from nototribic flowers (Hymenoptera, Apoidea and Masaridae)

In the Central European apoid and masarid fauna, 13 bee species belonging to seven different taxonomic groups and one honey wasp species were found to be equipped with a specialized pollen-collecting apparatus on the face which is used to harvest pollen from nototribic flowers, viz. representatives of the Lamiaceae and the Scrophulariaceae. Pollen from these two plant families plays an important part in the larval nourishment of these hymenopteran species. The pollen-collecting apparatus consists of a peculiar facial pilosity composed of transformed hairs which are thickened at their base and either knobbed apically (Celonites abbreviatus) or extended into a thin tail which is either straight and bent at right angles ( Rophites spp., Anthophora borealis, A. furcata ) or wavy ( Anthidium spp., Anthocopa andrenoides, Osmia aurulenta, O. caerulescens ). Nototribic flowers are worked by pollen-collecting females by rubbing the facial area covered with the transformed hairs over the anthers (Celonites, Anthidium, Anthocopa, Osmia) , by buzzing the pollen amongst the transformed hairs (Anthophora Jurcata) or by a combination of rubbing movements and buzzing (Rophites) . Bee species lacking special morphological devices were observed to perform specialized behaviours when collecting pollen at nototribic flowers. Nototribic flowers, therefore, appear to be difficult to exploit for pollen. The raised position of the anthers in nototribic flowers as a possible means to reduce excessive pollen losses caused by pollen-collecting hymenopterans is discussed.