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Biomedical subjects

R Yanagimachi

Publications and source records attributed to R Yanagimachi.

At least 181 records · Page 10Linked to original sources

Temperature dependence of sperm-egg fusion and post-fusion events in hamster fertilization.

The effects of temperature (4--37 degrees C) on sperm-egg fusion and the post-fusion events were studied. At 4--10 degrees C, acrosome-reacted spermatozoa bound to egg plasma membranes, but could not fuse with them. At 25 degrees C or above, both binding and fusion took place. The post-fusion events could occur over a broad temperature range (4--37 degrees C) but the events progressed faster with increasing temperature. An abnormal development of egg pronucleus, possibly due to an incomplete functioning of the meiotic spindle mechanism, was observed in eggs inseminated at 37 degrees C and cultured at 25 degrees C.

Animals↗

Detrimental effect of visible light on meiosis of mammalian eggs in vitro.

Short wavelength visible light (less than 470-480 nm) emmitted from ordinary light sources is detrimental to unfertilized hamster eggs in that prolonged exposure to the light disturbs the completion of normal meiosis after the eggs are penetrated by spermatozoa. The fluorescent light commonly used in modern laboratories is more harmful than the light from incandescent lamps. In experiments involving the handling of eggs in vitro, minimal exposure to the light or the use of appropriate filters (e.g., red cellophane sheets) is recommended.

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Movement characteristics and power output of guinea-pig and hamster spermatozoa in relation to activation.

Spermatozoa were collected from the cauda epididymidis of golden hamsters and guinea-pigs, and the acrosome reaction was induced in vitro. Movement characteristics of the spermatozoa were assessed with high-speed cinemicrography. Before the initiation of the acrosome reaction (preactivated spermatozoa), sperm movement in both species was characterized by progressive swimming by regular flagellar waves of moderate amplitude and relative high frequency. After the acrosome reaction (activated spermatozoa), sperm movement in both species was not progressive, and was characterized by whiplash-like flagellar undulations of significantly (P less than 0.05) higher amplitude and lower frequency. Calculation of the hydrodynamic power output by a new theory indicated that no significant change occurred after activation.

Acrosome↗

Inhibition of hamster sperm acrosome reaction and fertilization by oligomycin, antimycin A, and rotenone.

Effects of respiratory inhibitors (oligomycin, antimycin A and rotenone) on hamster sperm acrosome reaction and fertilization were studied. Hamster spermatozoa were incubated in a mixture of a modified Tyrode's solution and heat-treated human serum in the presence and absence of inhibitors. Oligomycin (2.4 x 10(-6) M), antimycin A (2.5 x 10(-6) M) and rotenone (2.5 x 10(-6) M) all reduced the incidence of the sperm acrosome reaction and fertilization without markedly affecting sperm motility. Antimycin A was the most effective in reducing the incidence of acrosome reaction. A reduction in the rate of fertilization was found in the presence of all of these respiratory inhibitors. The reduction in the incidence of acrosome reaction and fertilization by respiratory inhibitors implies an intimate relationship between high energy production (via respiration and oxidative phosphorylation) and capacitation and the acrosome reaction of spermatozoa. The necessity of oxidative metabolism for efficient capacitation and acrosome reaction of spermatozoa is suggested.

Acrosome↗

Activation of hamster eggs by pricking.

When mature unfertilized eggs of the golden hamster were picked with a glass needle, nearly 80% of them were activated. The diameter of needle was of critical importance for the successful induction of activation. It appeared that extracellular calcium was not necessary for the establishment (reception) of stimulation by pricking but was necessary for later phases of activation. The possible mechanisms by which pricking with a needle triggers the activation of the egg were compared with the mechanisms by which divalent ionophore induces activation.

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Membrane particle changes attending the acrosome reaction in guinea pig spermatozoa.

To examine the freeze-fracture appearance of membrane alterations accompanying the preparation of sperm membranes for fusions-the first preparatory stage occurring before physiological release of the acrosomal content, the second afterward-we induced the acrosome reaction in capacitated guinea pig spermatozoa by adding calcium to the mixture. The most common features observed before fusion of the acrosomal and plasma membranes were the deletion of fibrillar intramembranous particles from the E-fracture faces of both membranes, and the clearance of globular particles from the P face of the plasma membrane-events taking place near the terminus of the equatorial segment. Large particles, >12nm, remained not far from the cleared E-face patches. The P face of the outer acrosomal membrane is virtually clear from the outset. In addition, when fusion was completed, occasional double lines of large particles transiently embossed the P face of the plasma membrane (postacrosomal) side of the fusion zone. Behind the line of fusion, another series of particle-cleared foci emerged. We interpreted these postfusion membrane clearances as a second adaptation for sperm-egg interaction. Induction of the acrosome reaction in media containing phosphatidylcholine liposomes resulted in their apparent attachment, incorporation, or exchange in both the originally and secondarily cleared regions. Our observations support the concepts that membranes become receptive to union at particle- deficient interfaces, and that the physiologically created barren areas in freeze-fracture replicas may herald incipient membrane fusion.

Acrosome↗

Lectin-binding sites on the plasma membranes of rabbit spermatozoa. Changes in surface receptors during epididymal Maturation and after ejaculation.

MODIFICATIONS IN RABBIT SPERM PLASMA MEMBRANES DURING EPIDIDYMAL PASSAGE AND AFTER EJACULATION WERE INVESTIGATED BY USED OF THREE LECTINS: concanavalin A (Con A); Ricinus communis I (RCA(I)); and wheat germ agglutinin (WGA). During sperm passage from caput to cauda epididymis, agglutination by WGA drastically decreased, and agglutination by RCA(I) slightly decreased, although agglutination by Con A remained approximately unchanged. After ejaculation, spermatozoa were agglutinated to a similar degree or slightly less by Con A, WGA, and RCA(I), compared to cauda epididymal spermatozoa. Ultrastructural examination of sperm lectin-binding sites with ferritin- lectin conjugates revealed differences in the densities of lectin receptors in various sperm regions, and changes in the same regions during epididymal passage and after ejaculation. Ferritin-RCA(I) showed abrupt changes in lectin site densities between acrosomal and postacrosomal regions of sperm heads. The relative amounts of ferritin-RCA(I) bound to heads of caput epididymal or ejaculated spermatozoa. Tail regions were labeled by ferritin RCA(I) almost equally on caput and cauda epididymal spermatozoa, but the middle-piece region of ejaculated spermatozoa was slightly more densely labeled than the principal-piece region, and these two regions on ejaculated spermatozoa were labeled less than on caput and cuada epididymal spermatozoa. Ferritin-WGA densely labeled the acrosomal region of caput epididymal spermatozoa, although labeling of cauda epidiymal spermatozoa was relatively sparse except in the apical area of the acrosomal region. Ejaculated spermatozoa bound only a few molecules of ferritin-WGA, even at the highest conjugate concentrations used. Caput epididymal, but not cauda epididymal or ejaculated spermatozoa, bound ferritin-WGA in the tail regions. Dramatic differences in labeling densities during epididymal passage and after ejaculation were not found with ferritin-Con A.

Animals↗

Maturation and sperm penetration of canine ovarian oocytes in vitro.

Canine ovarian oocytes were cultured in a medium consisting of TC medium 199, fetal calf serum and antibiotics. Ninety-nine percent of the apparently healthy oocytes were in the germinal vesicle (dictyate) stage when recovered from the ovaries; 25% of them reached metaphase I or II by 72 hours of culture. Washed ejaculated spermatozoa were added to BWW medium containing oocytes which had either been removed directly from the follicles or which had been cultured for 24--72 hours. The earliest acrosome reaction and zona penetration by spermatozoa were seen at seven hours after insemination. Seventy-four percent of the oocytes examined between 11 and 24 hours after insemination showed evidence of zona penetration by spermatozoa. Neither the condition of the oocyte vitellus nor the stage of nuclear maturation influenced the incidence of zona penetration. Decondensing sperm nuclei were found in the vitellus of 27% of the oocytes which had not been cultured and in the vitellus of 20% of those which had been cultured for 24--72 hours and were in various stages of maturation. These results indicate that (1) canine ovarian oocytes can be matured in vitro, (2) the spermatozoa require capacitation which takes approximately seven hours in vitro and (3) maturation of the oocytes is not required for sperm passage through the zona pellucida or entry into the vitellus nor for sperm nuclear decondensation.

Animals↗

Immunological block to mammalian fertilization: survival and organ distribution of immunoglobulin which inhibits fertilization in vivo.

Fertilization of golden hamster eggs was blocked both in vitro and in vivo by antibodies produced in rabbits against specific hamster ovarian antigens (HOA). Antibodies against HOA bound to surfaces of the hamster egg zona pellucida and prevented spermatozoa from attaching to the zona and entering eggs in vitro. Fertilization in animals could be blocked for four estrous cycles by a single intraperitoneal injection of anti-HOA immunoglobulin, but not by control immunoglobulin. The in vivo fate of anti-HOA immunoglobulin was analyzed by simultaneous injection of 125I-anti-HOA IgG and control 131I-IgG. Both anti-HOA IgG and control IgG appeared in a variety of organs (lung, kidney, spleen, liver, and uterus) shortly after injection, but disappeared rapidly with no detectable differences in organ half lives. However, in the ovary anti-HOA IgG (but not control IgG) persisted at high levels during the period of infertility. Quantitative precipitin analysis of organ homogenates indicated that a high percentage of anti-HOA IgG in the ovary (but not in the other organs tested) was immunologically indistinguishable from IgG indicating lack of radiolabel metabolism and reincorporation. The results are discussed in terms of the development of a specific immunological block to fertility.

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