Maternal macho-1 is an intrinsic factor that makes cell response to the same FGF signal differ between mesenchyme and notochord induction in ascidian embryos

An extracellular signaling molecule acts on several types of cells, evoking characteristic and different responses depending on intrinsic factors in the signal-receiving cells. In ascidian embryos, notochord and mesenchyme are induced in the anterior and posterior margins, respectively, of the vegetal hemisphere by the same FGF signal emanating from endoderm precursors. The difference in the responsiveness depends on the inheritance of the posterior-vegetal egg cytoplasm. We show that macho-1, first identified as a localized muscle determinant, is also required for mesenchyme induction, and that it plays a role in making the cell response differ between notochord and mesenchyme induction. A zygotic event involving snail expression downstream of maternal macho-1 mediates the suppression of notochord induction in mesenchyme precursors.     

Exocytotic insertion of calcium channels constrains compensatory endocytosis to sites of exocytosis

Proteins inserted into the cell surface by exocytosis are thought to be retrieved by compensatory endocytosis, suggesting that retrieval requires granule proteins. In sea urchin eggs, calcium influx through P-type calcium channels is required for retrieval, and the large size of sea urchin secretory granules permits the direct observation of retrieval. Here we demonstrate that retrieval is limited to sites of prior exocytosis. We tested whether channel distribution can account for the localization of retrieval at exocytotic sites. We find that P-channels reside on secretory granules before fertilization, and are translocated to the egg surface by exocytosis. Our study provides strong evidence that the transitory insertion of P-type calcium channels in the surface membrane plays an obligatory role in the mechanism coupling exocytosis and compensatory endocytosis.     

Fusion of cell ghosts with intact cells in Dictyostelium discoideum: Differential response of opposite mating-type cells

The sexual cycle of Dictyostelium discoideum is initiated by the fusion of cells that are of opposite mating types (e.g. NC4- and HM1-type cells). Cells grown in light on agar plates are not capable of sexual cell fusion, but become capable when cultured in the dark in a liquid medium. Cells in the incapable state are called fusion-incompetent cells, and cells in the latter state, fusion-competent cells. To gain some understanding of the mechanism of cell fusion, cell ghosts prepared by freeze-thawing intact cells were incubated with intact cells. The cell ghosts killed the intact cells by directly fusing with them, the extent of fusion depending on the particular strains employed and the fusion-competency of the intact cells and of the cells from which the cell ghosts had been prepared. A detailed examination revealed that fusion-competent NC4 cells were always more easily killed by cell ghosts than fusion-incompetent NC4 cells. It also became apparent that cell ghosts prepared from fusion-competent NC4 cells killed all cell types far more efficiently than did those prepared from fusion-incompetent NC4 cells. However, fusion-competent and fusion-incompetent HM1 cells were equally sensitive to cell ghosts, and cell ghosts prepared from fusion-competent HM1 cells had the same ability to kill as those prepared from fusion-incompetent HM1 cells. From these findings, it thus appears that opposite mating-type cells have distinct membrane properties related to sexual cell fusion.     

Ca2+-dependent and phorbol ester activating phosphorylation of a 36K-dalton protein of rat basophilic leukemia cell membranes and immunoprecipitation of the phosphorylated protein with IgE-anti IgE system

Endogeneous phosphorylation of rat basophilic leukemia cell membranes was investigated. EGTA specifically inhibited the phosphorylation of a protein having an approximate molecular weight of 36,000 dalton (36K-Da protein). Phosphorylation of this protein was enhanced by phorbol-12-myristate-13-acetate in the presence of phosphatidylserine. The phosphorylated 36K-Da protein was specifically immunoprecipitated with IgE and anti IgE antibody. These results suggest that the phosphorylated 36K-Da protein is the beta-chain of the receptor for IgE and that protein kinase C is involved in the phosphorylation mechanism.     

Sex differences in song perception in Bengalese finches measured by the cardiac response

Birdsong may be perceived and processed differently by males and females because the production and function of this behaviour are often sexually dimorphic. However, examination of this hypothesis has been difficult, since different behavioural measures have been used to describe the perceptual process for each sex. We analysed changes in heart rate as an index of song perception in Bengalese finches, Lonchura striata var. domestica. In this species, only males sing, and song is used exclusively for mate attraction. This species is not territorial and songs are never used in aggressive contexts. In both sexes, repeated presentation of a song resulted in a waning of the heart rate response. Presentation of heterospecific songs did not increase the heart rate. When a novel conspecific song was presented, the heart rate increased only in females with each presentation of the stimulus, not in males. These results correspond to the sex differences in song usage in this species. Copyright 2003 The Association for the Study of Animal Behaviour. Published by Elsevier Science Ltd. All rights reserved.      

中国东北地区兴安落叶松林树干呼吸的研究

树干呼吸是森林碳平衡估计中的一个重要项目同时还能够显示树木的活力.对于如何准确估计森林树干呼吸释放CO2总量还存在争论.在本项研究中,2001～2002连续两年在一个33年生的兴安落叶松(Larix gmelini Rupr.)人工林内对树干呼吸进行了测定,同时测定了不同高度树干呼吸、呼吸的日变化、同龄落叶松林内不同个体的树干呼吸以及相关生长状态因子、水分因子和温度因子.结果显示:1)树干上部的呼吸速率在不同季节都高于下部呼吸速率,树干温度的差异能够一定程度上解释这种差异;2)树干呼吸有午间降低的现象,上午的测定结果树干温度与树干呼吸速率紧密相关,而下午则温度依赖性很小,土壤、空气、小枝木质部水势、叶片蒸腾速率和气孔导度都显示下午植物水分亏缺下午较上午严重,呼吸的这种上下午温度相关性的差异可能受这种水分亏缺的影响;3)在同龄林内,树木个体生长状态包括平均生长速率和树冠投影面积与树干呼吸速率有显著相关关系,而树干温度与之相关性很小.幂指数模型和S曲线模型能够产生较好的拟合效果;4)树干呼吸季节变化明显,7月份出现最大值,但同一月份的年间差异较大.自然指数模型能够较好地拟合温度与树干呼吸的季节变化规律.Q10值在2.22(2001年)和3.53(2002年)之间,与以往研究的结果相当.从以上结果可以看出,通过单一的Q10值估计森林树干呼吸总量会产生偏差,要想得到准确的估计,至少应该考虑生长状态的差异和水分状态的差异.     

ABCG2 is a High-Capacity Urate Transporter and its Genetic Impairment Increases Serum Uric Acid Levels in Humans

The ATP-binding cassette, subfamily G, member 2 (ABCG2/BCRP) gene encodes a well-known transporter, which exports various substrates including nucleotide analogs such as 3′-azido-3′-deoxythymidine (AZT). ABCG2 is also located in a gout-susceptibility locus (MIM 138900) on chromosome 4q, and has recently been identified by genome-wide association studies to relate to serum uric acid (SUA) and gout. Becuase urate is structurally similar to nucleotide analogs, we hypothesized that ABCG2 might be a urate exporter. To demonstrate our hypothesis, transport assays were performed with membrane vesicles prepared from ABCG2-overexpressing cells. Transport of estrone-3-sulfate (ES), a typical substrate of ABCG2, is inhibited by urate as well as AZT and ES. ATP-dependent transport of urate was then detected in ABCG2-expressing vesicles but not in control vesicles. Kinetic analysis revealed that ABCG2 is a high-capacity urate transporter that maintained its function even under high-urate concentration. The calculated parameters of ABCG2-mediated transport of urate were a Km of 8.24 ± 1.44 mM and a Vmax of 6.96 ± 0.89 nmol/min per mg of protein. Moreover, the quantitative trait locus (QTL) analysis performed in 739 Japanese individuals revealed that a dysfunctional variant of ABCG2 increased SUA as the number of minor alleles of the variant increased (p = 6.60 × 10^?5). Because ABCG2 is expressed on the apical membrane in several tissues, including kidney, intestine, and liver, these findings indicate that ABCG2, a high-capacity urate exporter, has a physiological role of urate homeostasis in the human body through both renal and extrarenal urate excretion.     

A novel mutation of ALK2, L196P, found in the most benign case of fibrodysplasia ossificans progressiva activates BMP-specific intracellular signaling equivalent to a typical mutation, R206H

Fibrodysplasia ossificans progressiva (FOP) is a rare autosomal dominant congenital disorder characterized by progressive heterotopic ossification in muscle tissues. Constitutively activated mutants of a bone morphogenetic protein (BMP) receptor, ALK2, have been identified in patients with FOP. Recently, a novel ALK2 mutation, L196P, was found in the most benign case of FOP reported thus far. In the present study, we examined the biological activities of ALK2(L196P) in vitro. Over-expression of ALK2(L196P) induced BMP-specific activities, including the suppression of myogenesis, the induction of alkaline phosphatase activity, increased BMP-specific luciferase reporter activity, and increased phosphorylation of Smad1/5 but not Erk1/2 or p38. The activities of ALK2(L196P) were higher than those of ALK2(G356D), another mutant ALK2 allele found in patients with FOP and were equivalent to those of ALK2(R206H), a typical mutation found in patients with FOP. ALK2(L196P) was equally or more resistant to inhibitors in comparison to ALK2(R206H). These findings suggest that ALK2(L196P) is an activated BMP receptor equivalent to ALK2(R206H) and that ALK2(L196P) activity may be suppressed in vivo by a novel molecular mechanism in patients with this mutation.     

Rate-dependent shortening of action potential duration increases ventricular vulnerability in failing rabbit heart

Congestive heart failure (CHF) predisposes to ventricular fibrillation (VF) in association with electrical remodeling of the ventricle. However, much remains unknown about the rate-dependent electrophysiological properties in a failing heart. Action potential properties in the left ventricular subepicardial muscles during dynamic pacing were examined with optical mapping in pacing-induced CHF (n=18) and control (n=17) rabbit hearts perfused in vitro. Action potential durations (APDs) in CHF were significantly longer than those observed for controls at basic cycle lengths (BCLs)>1,000 ms but significantly shorter at BCLs<400 ms. Spatial APD dispersions were significantly increased in CHF versus control (by 17-81%), and conduction velocity was significantly decreased in CHF (by 6-20%). In both groups, high-frequency stimulation (BCLs<150 ms) always caused spatial APD alternans; spatially concordant alternans and spatially discordant alternans (SDA) were induced at 60% and 40% in control, respectively, whereas 18% and 82% in CHF. SDA in CHF caused wavebreaks followed by reentrant excitations, giving rise to VF. Incidence of ventricular tachycardia/VFs elicited by high-frequency dynamic pacing (BCLs<150 ms) was significantly higher in CHF versus control (93% vs. 20%). In CHF, left ventricular subepicardial muscles show significant APD shortenings at short BCLs favoring reentry formations following wavebreaks in association with SDA. High-frequency excitation itself may increase the vulnerability to VF in CHF.