Biomedical subjects

Kogiku Shiba

Publications and source records attributed to Kogiku Shiba.

5 recordsLinked to original sources

PubMed2006

Na+/Ca2+ exchanger modulates the flagellar wave pattern for the regulation of motility activation and chemotaxis in the ascidian spermatozoa.

Ion channels and ion exchangers are known to be important participants in various aspects of sperm physiology, e.g. motility activation, chemotaxis, the maintenance of motility and the acrosome reaction in the sperm. We report here on a role of the K+ -independent Na+/Ca2+ exchanger (NCX) on ascidian sperm. Reverse-transcriptase PCR reveals that the NCX is expressed in the testis while immunoblotting and immunolocalization demonstrate that the NCX exists on the sperm in the ascidian Ciona savignyi and C. intestinalis. A potent blocker of the NCX, KB-R7943 was found to block sperm-activating and -attracting factor (SAAF)-induced motility activation, sperm motility and sperm chemotaxis. We further analyzed the effects of this blocker on motility parameters such as the flagellar waveform, curvature, beat frequency, amplitude and wavelength of the sperm flagella. Inhibition of the NCX caused two distinct effects: a low concentration of KB-R7943 induced symmetric bending, whereas a high concentration of KB-R7943 resulted in asymmetric flagellar bending. These findings suggest that the NCX plays important roles in the regulation of SAAF-induced sperm chemotaxis, motility activation and motility maintenance in the ascidian. This study provides new information toward an understanding of Ca2+ transport systems in sperm motility and chemotaxis.

Animals↗

PubMed2006

Peptide-induced hyperactivation-like vigorous flagellar movement in starfish sperm.

Asterosap, a sperm-activating peptide (SAP) from the starfish egg jelly coat, is diffusible and controls a cGMP-signalling pathway in starfish sperm in the same manner as resact, a potent chemoattracting SAP in sea urchins. This fact suggests that asterosap may serve as a chemoattractant like resact at concentrations with appropriate gradients. Since asterosap is one of three egg jelly components, which in concert induce the acrosome reaction, it is still worthwhile to evaluate how asterosap modulates sperm motility prior to this reaction. We analysed the flagellar movement of sperm of the starfish Aphelasterias japonica in artificial seawater (ASW) containing the asterosap isoform P15 at 1 micromol l(-1). We found that sperm swim straighter with more symmetrical flagellar movement in P15 than in ASW, but without any significant difference in the flagellar beat frequency and the swimming velocity. The flagellar movement is, however, dramatically different between sperm firmly attached to the solid surface by the head in P15 and those attached in ASW: in P15 the flagellum bends to a greater extent, with higher curvature and with higher shear angle up to a right angle to the flagellar wave axis, and beats at an increased frequency. The vigorous flagellar movement of sperm, which can be activated when sperm are placed in high-load circumstances just as entering into a jelly layer, may increase propulsive forces and hydrodynamic resistances, allowing sperm to undergo the acrosome reaction as effectively as possible.

Acrosome Reaction↗

PubMed2006

Stroboscopic illumination using light-emitting diodes reduces phototoxicity in fluorescence cell imaging.

Excited fluorophores produce reactive oxygen species that are toxic toward many live cells (phototoxicity) and accelerate bleaching of the fluorophores during the course of extended or repeated measurements (photobleaching). We recently developed an illumination system for fluorescence microscopy using a high power light-emitting diode (LED), which can emit short pulses of light (0.5-2 ms) to excite fluorophores. This system minimizes illumination time, thus reducing phototoxicity and photobleaching artifacts. To demonstrate the usefulness of the new system, we compared images of human sperm loaded with various fluorescent indicators and excited with either a conventional mercury lamp as a continuous excitation light source or the LED as a source of pulsed illumination. We found that sperm motility decreased rapidly and photobleaching was relatively rapid under continuous illumination, whereas under pulsed LED illumination, motility was maintained and photobleaching was much reduced. Therefore, fluorescence microscopy using LED-based pulsed illumination offers significant advantages for long-term live cell imaging, reducing the degree of phototoxicity, and extending the effective lifetime of fluorophores.

Calcium↗

PubMed2005

Sperm-activating peptide induces asymmetric flagellar bending in sea urchin sperm.

Speract, a sperm-activating peptide (SAP) from sea urchin eggs, induces various sperm responses including a transient increase in the intracellular Ca2+ concentration. However, it has not been clarified how speract modulates sperm motility and whether it functions as a chemoattractant. To confirm the effect of speract on sperm motility, we observed the flagellar bending response to speract in sperm of Hemicentrotus pulcherrimus, in experiments using caged speract and a lighting system for a microscope newly developed with a power LED. We found that speract induces increases in curvature of swimming paths and changes flagellar bending shape to asymmetric. These facts show that speract directly regulates flagellar motility, and suggest that speract-induced increases in intracellular Ca2+ concentration play an actual role in regulation of the flagellar movement.

Animals↗

PubMed2004

A sea urchin egg jelly peptide induces a cGMP-mediated decrease in sperm intracellular Ca(2+) before its increase.

Speract, a sperm-activating peptide (SAP) from sea urchin eggs, increases the intracellular concentration of Ca(2+) ([Ca(2+)]i) and modulates sperm motility. We measured the initial sperm response to speract using its caged analog and observed, for the first time, a small but significant decrease in sperm [Ca(2+)]i before the increase. Both directions of the [Ca(2+)]i change were completely blocked in high K(+) seawater. Using membrane-permeant caged cyclic nucleotides (cNMP), only cGMP induced the decrease in [Ca(2+)]i although both cGMP and cAMP increased the [Ca(2+)]i. The decrease in the [Ca(2+)]i induced by cGMP was more notable following a second photolytic event, once [Ca(2+)]i had been elevated by an initial flash. This pattern of [Ca(2+)]i change was confirmed in individual sperm. These results together with pharmacological evidence suggest that the initial [Ca(2+)]i decrease is due to a Na(+)/Ca(2+) exchanger activity, stimulated by hyperpolarization mediated by K(+) efflux through cGMP-regulated K(+) channels.

1-Methyl-3-isobutylxanthine↗