Effect of Ca2+ channel antagonists on the acrosome reaction of guinea pig and golden hamster spermatozoa

The effects of Ca²⁺ channel antagonists on the motility and acrosome reaction of guinea pig spermatozoa were examined by incubating the spermatozoa continuously in Ca²⁺-containing capacitating media with 10^?6 M to 10^?4 M antagonist. Antagonists tested were four voltage-gated Ca²⁺ channel antagonists (verapamil, nifedipine, nimodipine, and FR–34235) and two ligand-gated channel antagonists (NaNO₂ and Na-nitroprusside). None of these antagonists could block the acrosome reaction. Instead, three antagonists (verapamil, nimodipine, and FR-34235, each at 10^?4 M) accelerated the onset of the acrosome reaction with a subsequent decrease in sperm motility. Nifedipine and Na-nitroprusside at the same concentration caused a complete loss of sperm motility by 4 hr of incubation with no substantial effect on the rate of acrosome reaction. The detrimental effect of antagonists on the motility of spermatozoa appears to be due to a direct, Ca²⁺-independent, membrane-perturbing action of the reagents. The acrosome reaction was not inhibited when guinea pig spermatozoa were precapacitated in Ca²⁺-free medium (with a low concentration of lysolecithin) in the continuous presence of antagonists. An acceleration of the onset of the acrosome reaction by verapamil (10^?4 M) was also demonstrated in the golden hamster. These results may be interpreted as indicating that the entry of extracellular Ca²⁺ into spermatozoa, which triggers the acrosome reaction of guinea pig and hamster spermatozoa, is not mediated by Ca²⁺ channels. This is in marked contrast with the case reported in invertebrate spermatozoa. Possible mechanisms by which some of the antagonists stimulate the acrosome reaction and affect the motility of mammalian spermatozoa are discussed.     

A minimum mechanism for Na+−Ca++ exchange: Net and unidirectional Ca++ fluxes as functions of ion composition and membrane potential

Summary Both simultaneous and consecutive mechanisms for Na⁺–Ca⁺⁺ exchange are formulated and the associated systems of steady-state equations are solved numerically, and the net and unidirectional Ca⁺⁺ fluxes computed for a variety of ionic and electrical boundary conditions. A simultaneous mechanism is shown to be consistent with a broad range of experimental data from the squid giant axon, cardiac muscle and isolated sarcolemmal vesicles. In this mechanism, random binding of three Na⁺ ions and one Ca⁺⁺ on apposing sides of a membrane are required before a conformational change can occur, translocating the binding sites to the opposite sides of the membranes. A similar (return) translocation step is also permitted if all the sites are empty. None of the other states of binding can undergo such translocating conformational changes. The resulting reaction scheme has 22 reaction steps involving 16 ion-binding intermediates. The voltage dependence of the equilibrium constant for the overall reaction, required by the 31 Na⁺Ca⁺⁺ stoichiometry was obtained by multiplying and dividing, respectively, the forward and reverse rate constants of one of the translocational steps by exp(–FV/2RT). With reasonable values for the membrane density of the enzyme (120 sites m²) and an upper limit for the rate constants of both translocational steps of 10⁵·sec^–1, satisfactory behavior was obtainable with identical binding constants for Ca⁺⁺ on the two sides of the membrane (10⁶ m ^–1), similar symmetry also being assumed for the Na⁺ binding constant (12 to 60m ^–1). Introduction of order into the ion-binding process eliminates behavior that is consistent with experimental findings.     

Effects of short-term treatment with calcium on the parathyroid gland of the rat,under particular consideration of the alteration of storage granules

Summary Short-term effects of CaCl₂-treatment on parathyroid cells of the rat, especially on their storage granules, were studied at the ultrastructural level. After an injection of 4% CaCl₂, serum calcium levels (SCL) rapidly increased from 9.1 mg/dl (controls) to a maximum of 14.9 mg/dl at 20 min. At 5 min after the injection, the number of type-I storage granules (large core) [NSG-I] and that of type-II storage granules (small core) [NSG-II] remained unchanged, in spite of elevated SCL (12.4 mg/dl). As soon as SCL rose to 13.2 mg/dl at 7.5 min, NSG-I gradually decreased to a minimum at 30 min; in contrast, NSG-II gradually increased to a maximum at 30 min. Vacuolar bodies also increased together with the augmentation of type-II storage granules. The average diameter of the core of the storage granules decreased significantly after the injection. Protein A-gold method for immunocytochemistry showed that the cores of these granules contain parathormone. Acid-phosphatase activity was occasionally found in storage granules of both types, especially in those of type II. It is concluded (i) that type-I storage granules may be transformed into vacuolar bodies via type-II granules as a result of hydrolysis, and (ii) that these processes may be accelerated during hypercalcemia.     

Gleichgewichtsbewegungen von Blättern als Reaktion auf asymmetrische Belastung — Isoklinotropismus nach Härdtl (1927) — und ihre Beziehung zum Gravitropismus

1. Die anscheinend in Vergessenheit geratenen älteren Angaben von HÄRDTL (1927 und spärer) und PRINGSHEIM (1931), nach denen einseitig belastete Blätter Gleichgewichtsbewegungen ausführen können (Isoklinotropismus nach HÄRDTL), wurden an mehreren Arten (Chelidonium majus, Aegopodium podagra ria, Ranunculus repens, Sambucus nigra, Coleus blumei und Hibiscus rosa sinensis) bestätigt,— Die einseitige Belastung erfolgte durch Einstecken von Nadeln in die eine Hälfte der Blattspreite bzw. in eine Blattfieder. 2. Bei Blättern, die in Spreite oder Stiel epinastisch gekrtimmt sind (die überwiegende Mehrzahl) oder auch durch die Belastung abwärts gebogen sind, kommt es dabei zu zwei verschiedenen Reaktionen: a) zu einer sofort eintretenden rein physikalischen reversiblen Reaktion, da die einseitige Last eine Torsion der Spreite bewirkt, die die Spreitenspitze nach der der Last gegenüberliegenden Seite [führt, und b) zu einer spärer eintretenden physiologischen Reaktion durch eine Wachstumskrümmung nach derselben Seite, - Beide Komponenten der Bewegung ftihren zu einer Verringerung oder einem Ausgleich des Ungleichgewichtes. 3. Die physiologische Reaktion war meist nach einem oder einigen Tagen erkennbar, bei Chelidonium mitunter schon nach einigen Stunden, bei Hibiscus gelegentlich erst nach etwa einer Woche. Mitunter blieb die Reaktion auch aus. Bei den gefiederten Blattern erfolgte sie in der Rhachis, ober- und unterhalb der belasteten Fieder, mitunter auch im Blattstiel, bei Coleus im Spreitengrund und im Blattstiel, bei Hibiscus im oberen Gelenkpolster (“Sekundargelenk”) des Blattstiels. Auch ältere Blätter reagierten oft noch überraschend gut. Vielfach führte die physiologische Reaktion zu einer vollständigen Ausbalancierung der einseitig belasteten Blattspreite. Auf nachtragliche Entfernung der eingesteckten Nadeln gingen auch die Krtimmungen in 2–3 Tagen wieder weitgehend zurück. 4. Als Ursache ftir die Gleichgewichtsbewegungen der Blätter kommen zwei verschiedene Mechanismen in Betracht: a) Infolge der durch die einseitige Belastung hervorgerufenen Schräglage der Blattspreite sammelt sich das Auxin auf der tiefer liegenden Flanke von Mittelnerv bzw. Rhachis und Blattstiel an, was zu einem stärkeren Wachstum dieser Seite und einer Gleichgewichtskrümmung führen muß. Nach dieser Auffassung ordnet sich der Isoklinotropismus dem Gravitropismus ein. b) Die nach der Belastung sofort eintretende auf der Torsionsspannung beruhende rein physikalische und zunächst reversible Gleichgewichtsreaktion des Blattes wird nach einigen Stunden oder Tagen teilweise irreversibel. (Für die spannungsfreien Abschnitte der Rhachis eines gefiederten Blattes oberhalb der Belastung kann diese Erklärung natürlich nicht gelten). Vermutlich sind beide Mechanismen, vor allern wohl der erstgenannte, bei den einzelnen Arten in verschiedenem Maße, als Ursache der Gleichgewichtsbewegungen wirksam. 5. Der biologische Sinn der Ausbalancierung eines (größeren) Blattes (PRINGSHELM 1931, HÄRDTL 1927, 1937 a) liegt darin, daß a) ein ausbalanciertes Blatt den geringsten Aufwand an mechanischen Elementen erfordert, und b) Photo- und Gravitropismus nur ein ausbalanciertes Blatt ohne Komplikationen in die angestrebte Lage fuhren konnen.     

一种快速、灵敏的血小板释放功能检测方法及其在抗血小板药物研究中的应用

本文报道了一种快速、灵敏的血小板释放功能检测方法:利用荧光素-荧光素酶在有ATP、Mg~(2+)、O_2存在时产生的生物发光素测定血小板ATP的释放量,以反映血小板的释放功能;研究了ADP、AA、胶原、凝血酶等四种诱导剂对血小板释放功能的作用,发现ADP的诱导释放能力较其他三者为弱;观察在不同剂量ADP和AA的诱导下,血小板聚集强度和释放能力之间的关系,研究了血小板数等因素对ATP释放功能测定的影响。应用该方法研究了Aspirin及活血化淤药物川芎嗪,毛冬青甲素对血小板释放功能的影响,发现Aspirin对AA诱导的释放反应有强烈的抑制作用。在以ADP诱导的释放反应中,川芎嗪的抑制作用较毛冬青甲素更为强烈。     

Acrosomal reaction and early events at fertilization in Marthasterias glacialis (Echinodermata: Asteroidea)

When the spermatozoon of M glacialis contacts the mature oocyte jelly it adheres to it. Following this, there is a slight tumefaction of the acrosome, which is followed by the disruption of the apical acrosomal vesicle and cytoplasmic membranes. Acrosomal vesicle contents are liberated and spread along the outer surface of the oocyte jelly. Meanwhile, the acrosomal process begins to extend, penetrates all the jelly extension, then the vitelline layer, and finally contacts the cytoplasmic egg membrane. Nevertheless, the sperm cell continues lying at the outer border of the jelly. From the beginning of the acrosome reaction the dense and finely fibrillar subacrosomal material is connected, by some expansions, to the basal acrosomal vesicle membrane. Both nuclear and mitochondrial diameters have diminished.     

A cortical granule-specific enzyme,B-1,3-glucanase,in sea urchin eggs

The ultrastructural localization of B-1,3-glucanase in three species of sea urchin eggs was determined using a monospecific antibody in an electronmicroscopic immunogold procedure. In all three species, Lytechinus variegatus, Strongylocentrotus purpuratus, and Arbacia punctulata, B-1,3-glucanase was localized specifically to the cortical granules. No other organelle within the egg contained significant label. During the fertilization reaction, B-1,3-glucanase was released from cortical granules into the perivitelline space and became associated with the hyaline layer. No significant label was found in association with the fertilization envelope.     

The Ene Reaction of Singlet Oxygen with Olefins

The reactions of singlet oxygen (¹O₂) with cis and trans butenes-1,1,1-d₃, at—80°C in Freon-11, show a product isotope effect (k_H/k_D) of 1.38 and 1.25 respectively. Isomerization of the starting materials or formation of dioxetanes were not observed during the course of the photooxygenation. Together with the isotope effects on the reactions of tetramethylethylene-d₆ isomers with singlet oxygen, these results require the reversible formation of a perepoxide or charge transfer intermediate.     

水稻品种超氧物歧化酶(SOD)活性与氧抑光合的关系

O_2抑光合程度不同的水稻品种,SOD活性存在差异。在40％O_2下,SOD活性被诱导增加水平高、延续时间长的品种,表现O_2抑光合程度小,反之则O_2抑光合程度大。在自然条件下,强光、高温都是诱导SOD活性变化的因素。选择SOD活性高、O_2抑光合程度小的种质资源可能有利于适应对光合不利的逆境条件。     

Hydrogen peroxide formation and lipid peroxidation in rat uterus—effect of hormones and vitamin E

The effect of estradiol-17β and progesterone given separately as well as in combination on the rate of hydrogen peroxide formation and lipid peroxidation in the uteri of ovariectomized rats was studied. Estradiol in 3μg dose per day per animal elicited maximum stimulatory response and progesterone (100μg), on the other hand, was without any such effect. However, progesterone given along with estradiol completely prevented the effect due to the latter. In the same way, vitamin E, a well known antioxidant was found to be extremelv effective in protecting the uterus from the highly peroxidative action of estradiol-17β.