Characteristics of the inward-rectifying potassium current in mouse ventricular myocytes and its relation to early after-depolarization

The properties of the inward-rectifying potassium current (IK1) were studied in the single myocytes isolated from adult mouse ventricles by the whole-cell patch-damp technique for the first time. Most of the properties of IK1 including channel conductances, activation, inactivation, rectification and external K sensitivity in mouse ventricular myocyte were similar to those in other species, but the current-voltage (1-V) curve of mouse ventricular myocyte showed no negative slope, i.e the slope in the range of membrane potential 50 mV positive to the reversal potential (VRev) was virtually flat and remained at a low current level ((59±39) pA). Under the superfusion of Tyrode's solution with 3mmol/L K and 3mmol/L Cs , IK1 in the above region nearly decreased to zero, and then the early after-depolarization (EAD) occurred. The results suggest that this distinctive characteristic of IK1 in mouse ventricular myocyte may relate to the high susceptibility to EA0 in mouse myocardium. The inhibition of IK1 se     

The development of female gametophyte and antipodal embryo formation in Sedum fabaria

In Sedum fabaria, the ovule is anantropus, bitegmic and crassinucellate. The development of the nucellus conforms to the Sedum type. The development of the embryo sac is of the Allium type. The antipodal cells in unfertilized embryo sac occasionally divide and one of them forms four-celled structures resembling embryos and remaining once elongate in the form of haustoria. The entry of the pollen tube is porogamous. After division the primary endosperm nucleus forms two cells: the apical one develops into cellular endosperm according to the Acre type and the basal one acts as the endosperm haustorium of the Sempervivum type. The embryogeny corresponds to the Caryophyllad type. This revised version was published online in July 2006 with corrections to the Cover Date.     

Activation of inwardly rectifying potassium channels by muscarinic receptor-linked G protein in isolated human ventricular myocytes

Muscarinic receptor-linked G protein, G _i , can directely activate the specific K⁺ channel (I _K(ACh)) in the atrium and in pacemaker tissues in the heart. Coupling of G _i to the K⁺ channel in the ventricle has not been well defined. G protein regulation of K⁺ channels in isolated human ventricular myocytes was examined using the patch-clamp technique. Bath application of 1 μm acetylcholine (ACh) reversibly shortened the action potential duration to 74.4 ± 12.1% of control (at 90% repolarization, mean ±sd, n= 8) and increased the whole-cell membrane current conductance without prior β-adrenergic stimulation in human ventricular myocytes. The ACh effect was reversed by atropine (1 μm). In excised inside-out patch configurations, application of GTPγS (100 μm) to the bath solution (internal surface) caused activation of I _K(ACh) and/or the background inwardly-rectifying K⁺ channel (I _K1) in ventricular cell membranes. I _K(ACh) exhibited rapid gating behavior with a slope conductance of 44 ± 2 pS (n= 25) and a mean open lifetime of 1.8 ± 0.3 msec (n= 21). Single channel activity of GTPγS-activated I _K1 demonstrated long-lasting bursts with a slope conductance of 30 ± 2 pS (n= 16) and a mean open lifetime of 36.4 ± 4.1 msec (n= 12). Unlike I _K(ACh), G protein-activated I _K1 did not require GTP to maintain channel activity, suggesting that these two channels may be controlled by G proteins with different underlying mechanisms. The concentration of GTP at half-maximal channel activation was 0.22 μm in I _K(ACh) and 1.2 μm in I _K1. Myocytes pretreated with pertussis toxin (PTX) prevented GTP from activating these channels, indicating that muscarinic receptor-linked PTX-sensitive G protein, G _i , is essential for activation of both channels. G protein-activated channel characteristics from patients with terminal heart failure did not differ from those without heart failure or guinea pig. These results suggest that ACh can shorten the action potential by activating I _K(ACh) and I _K1 via muscarinic receptor-linked G _i proteins in human ventricular myocytes. Received: 23 September 1996/Revised: 18 December 1996     

Transfer of macromolecules into living adult cardiomyocytes by microinjection

Among techniques commonly used to deliver bioactive molecules into living cells, microinjection is a very efficient method. Microinjection has been used extensively for gene transfer into different cell types. We applied the microinjection technique to the adult rat ventricular cardiac muscle cells (AVC) in primary culture and optimized microinjection parameters and the appropriate cell culture conditions. We also optimized the use of particular agents (i.e. 2,3-butanedione monoxime, verapamil) for the prevention of the cell damage caused by the micropuncture. We obtained the expression of a CMV--galactosidase reporter gene in up to 20% of the injected cells with efficient maintenance of long term cell viability. Under our experimental conditions direct microinjection is a very advantageous technique to transfer macromolecules into living adult cardiac muscle cells and a powerful system to study and manipulate the biochemistry and molecular biology of the cardiac myocyte.     

Estimation of larval dry weight of Chaoborus americanus

A method for estimating larval dry weight as a function of tracheal front air sac length and instar was developed for second, third, and fourth instar Chaoborus americanus using linear regression analysis. The following equation: where W = larval dry weight (μg), L = front air sac length (mm), a = the y-intercept, and b = the slope provided accurate estimates of larval dry weight for three C. americanus populations. No detectable change in size occurred in the air sacs of larvae preserved for one year in 5% formalin. This indicates that air sacs can be used to accurately estimate larval dry weight for preserved as well as living specimens.     

Synthesis and accumulation of ribosomes in individual cells of the female gametophyte of maize during its development

Summary The pattern of synthesis of ribosomes during three stages of development of the female gametophyte of maize has been studied by in situ hybridization using a ribosomal RNA probe. Changes in volume of individual cells of the embryo sac during its maturation have been determined by confocal microscopy. These data have permitted us to calculate the relative numbers of ribosomes in the cells of the embryo sac at different stages of their maturation. The egg apparatus and the central cell at all stages of development contain several fold greater numbers of ribosomes than are present in the antipodal cells or cells of the surrounding nucellus. The accumulation of ribosomes during embryo sac maturation appears to proceed at a constant and high rate, with the rate being highest in the developing central cell.     

Plasma protein and apolipoprotein synthesis by human yolk sac carcinoma cells in vitro

Summary Three human yolk sac carcinoma cell lines were characterized for the expression of several markers. Each of the cell lines expressed alpha-fetoprotein, without detectable levels of chorionic gonadotropin, and the level of alpha-fetoprotein expression increased dramatically when the cultures were held without passage for extended periods. The secretion of a number of plasma proteins was documented by metabolic labeling, immunoprecipitation, and gel analysis. The major plasma proteins detected were alpha-1-antitrypsin, alpha-fetoprotein, transthyretin,β-2 microglobulin, and plasminogen, with lower levels of transferrin and complement C4 released. Apolipoproteins B, E, and A1 were secreted in high levels as well and were found in the form of lipoprotein particles. Time course experiments on the synthesis of apolipoproteins E and A1 indicated that, as with alpha-fetoprotein, the level of synthesis increased substantially when the cultures were held without passage. The results indicate that these yolk sac carcinoma cells display a protein expression profile similar to that observed for the human yolk sac, and the possibility that the cells may have the potential to differentiate is discussed. This investigation was supported in part by Program Project HL 28972 from the National Heart, Lung and Blood Institute, Bethesda, MD.     

Patterns of embryo-sac organization,synergid degeneration and cotyledon orientation in Linum usitatissimum L.

The embryo sac of Linum usitatissimum consists, as in most angiosperms, of the egg, two synergids, central cell and antipodals. In Linum the embryo sac is strongly polarized in the longitudinal axis, but rotationally random relative to the vascular bundles of the ovule and ovary. Synergids designated right or left based on their orientation relative to the egg can be distinguished unambiguously if viewed from one pole of the embryo sac — in this case, the chalaza was used as the reference point. The synergid most likely to degenerate is the left (60%), proximal (52.3%) or septal-facing (52.7%) synergid. The volume and the surface area of the filiform apparatus of the left synergid is significantly smaller than that of the right synergid. Synergid-degeneration patterns varied between individual plants, indicating genetic control; however, the preference for the proximal and septal-facing synergid, although weak, indicates the possibility of some physiological influence. The cotyledons appear to assume bilateral symmetry with respect to the ovule only once endosperm digestion has begun. As the cotyledons grow, the embryo rotates to occupy the widest part of the embryo sac, thus imposing bilateral symmetry between the embryo and seed; prior to that time, the early and heart-shaped embryos are randomly oriented.This research was supported by U.S. Department of Agriculture grant 88-37261-3761. We thank Dr. Roger Meicenheimer, Department of Botany, Miami University, Oxford, USA, for providing seeds and Dr. Stephen Young, Department of Psychology, University of California, San Diego, USA, for providing the program     

急性心肌梗塞静脉溶栓后冠脉再通对左心室功能的影响

为进一步探讨静脉溶栓后冠脉再通对左室功能的有益影响,回顾性分析了88例静脉溶栓急性心肌梗塞(AMI)病人的二维超声心动图和部分病人的冠脉造影资料。根据溶栓再通标准分为再通组和未通组。在AMI发病后平均24.8±11.6天和22.9±24.4天分别进行了心超和冠造检查,观察室壁运动情况并测量左室射血分数(LVEF)。结果显示:未通组LVEF明显低于再通组,而室壁运动异常积分却显著高于再通组。左室扩大和室壁瘤的发生上,两组未发现有统计学差异。以上结果提示:AMI静脉溶栓使冠脉再通对左室功能和室壁运动障碍的改善起到有益的作用。     

Development of pressure overload induced cardiac hypertrophy is unaffected by long-term treatment with losartan

Left ventricular hypertrophy with adequate wall thickness, preserved adult phenotype and extracellular matrix may be useful in the prevention of heart failure. Because activation of subtype 1 of angiotensin II (AT₁) receptors is thought to be involved in the hypertrophic response of cardiomyocytes, we tested the potential of systemic AT₁ blockade to modify the development of left ventricular hypertrophy due to pressure overload.Sham-operated rats and rats with ascending aorta constriction were treated with losartan (30 mg/kg/day) for 8 weeks. Left ventricular geometry, dynamics of isovolumic contractions, hydroxyproline concentration as well as myosin isozymes (marker of fetal phenotype) were assessed. Rats with aortic constriction exhibited a marked increase in left ventricular weight and the diastolic pressure-volume relationship was shifted to smaller volumes. An enlarged ventricular pressure-volume area and increased (p < 0.05) peak values of +dP/dtmax and -dP/dtmax demonstrated an enhanced overall ventricular performance. Signs of congestive heart failure were not apparent. In contrast, parameters of myocardial fimction (normalized length-stress area, +d/dtmax and -d/dtmax) were depressed (p < 0.05), indicating an impaired myocardial contractility. The hydroxyproline concentration remained unaltered. However, the proportion of -myosin heavy chains (NMC) was increased (p < 0.05). Administration of losartan decreased (p < 0.05) blood pressure and body weight in sham operated and pressure overloaded rats. By contrast, neither the concentric left ventricular hypertrophy or depressed myocardial function nor the increased -MHC expression were significantly altered. Thus, activation of AT₁ receptors appears not to be involved in the initial expression of the fetal phenotype of pressure overloaded heart which may be responsible for the progressive functional deterioration of the hypertrophied ventricle.