bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effect of albumin on fertilization of mouse ova in vitro
Authors: Quinn P. , Stanger J. D. , Whittingham D. G. 
Published: February 2005

AbstractSerum albumin is an obligatory component of the incubation medium for the fertilization of mouse ova. Normal, untreated bovine serum albumin supports high rates of fertilization of cumulus‐free ova both with and without their zonae pellucidae. Heat‐treated or trichloroacetic acid‐extracted bovine serum albumin is unable to support the fertilization of a majority of zona‐intact ova but fertilization of zona‐free ova is unimpaired. Spermatozoa incubated in medium containing heated bovine serum albumin fertilize zona‐intact ova when 2 mM caffeine is present but the progress of sperm head decondensation is delayed when compared to normal controls. Trichloroacetic acid extracted BSA preferentially and irreversibly inhibits zona penetration by spermatozoa, but this effect is not mediated by an inhibition of spermatozoal motility or zona‐binding ability. This effect occurs after only a 10‐min preincubation of the spermatozoa in the extracted BSA or when the medium contains only a 10% (v/v) proportion of this albumin. It is estimated that mouse spermatozoa under the conditions used take 2 hr to penetrate the zonae pellucidae of 50% of ova and effect fertilization.
Acrosome reaction in sperm of the toad, bufo bufo japonicus
Authors: Yoshizaki Norio , Katagiri Chiaki 
Published: February 2005

AbstractThe acrosome in the sperm of the toad, Bufo bufo japonicus, consists of a membrane‐limited acrosomal cap and a fibrous perforatorium. When sperm are incubated with the oviducal pars recta extract (PRE) for 30–60 min, the outer acrosomal membrane fuses with the overlying plasma membrane at several points with concomitant loss of the contents of the acrosomal cap. The inner acrosomal membrane thus exposed fuses with the plasma membrane at the caudal end of the acrosomal region. This PRE‐induced acrosome reaction is completely inhibited by soybean trypsin inhibitor. Sperm found in the innermost jelly layer of inseminated eggs possess an intact acrosome, but those either passing through the vitelline coat or localizing in the perivitelline space are acrosome‐reacted in the same manner as when treated with PRE. These observations, combined with recent evidence showing involvement of the pars recta substance in fertilization, indicate that the acrosome reaction occurring in a fertilizing sperm at or near the surface of the vitelline coat is a response to a substance that is derived from the pars recta and deposited in the vitelline coat.
The effect of catecholamines and their antagonists on the fertilization of cumulus‐free mouse ova in vitro at a suboptimal spermatozoal density
Authors: Stanger J. D. 
Published: February 2005

AbstractA suboptimal sperm concentration was used to assess the capacity of catecholamines to stimulate the fertilization of cumulus free F1,(C57BL × CBA) mouse ova in vitro. At a concentration of 50 μM, (L) epinephrine significantly increased the proportion of ova fertilized at 2 × l05 spermatozoa/ml. However, when (D, L) propranolol at an equimolar concentration was tested for inhibition of the (L) epinephrine effect, fertilization was inhibited in both the test and control dishes. At l0μM, propanolol by itself or in the presence of 50μM (L) epinephrine significantly increased the number of ova fertilized at 2 × l05 sperm/ml. Norepinephrine (50 μM) and phentolamine (50 μM), either alone or together, were also slightly stimulatory. Some data are presented to suggest that propranolol may act in a nonadrenergic manner to precipitate the acrosome reaction and that the stimulatory effect is maximised when it is added to spermatozoa at the same time as ova addition. It was suggested that propranolol may act to trigger calcium influx by a nonspecific alteration in membrane function for example in (Ca + Mg) ATPase activity. It was concluded that spermatozoa at suboptimal densities are capable of achieving fertilization and that sperm concentration dependency in fertilization in vitro may be a reflection of the proportion of spermatozoa achieving capacitation.
Reduced survival in utero from transferred mouse blastocysts compared with morulae
Authors: Hoppe Peter C. , Coman Dale Rex 
Published: February 2005

AbstractStudies on the survival of mouse embryos revealed that fewer offspring were produced when blastocysts, rather than morulae, were transferred to foster mothers. Approximately 8–10 h after fertilization F1 hybrid eggs (C57BL/6J × LT/Sv) were collected and cultured to morulae (day 4) or blastocysts (day 5 ) before transfer into uteri of day 3 foster mothers. A few recipients were killed on day 8 of gestation and deciduae were examined histologically. Embryos developing from transferred morulae were found to lie deep within the deciduae and were surrounded by numerous, large blood islands. Conversely, embryos developing from transferred blastocysts implanted more distally to the maternal blood vessels with only a few blood islands surrounding the embryos. These observations, suggesting abnormal implantation with insufficient embyro nourishment, were confirmed when uteri of foster mothers were examined on day 19 of gestation. Although the proportion of implantations from transferred morulae or blastocysts was similar (42 % and 47%, respectively), significantly more of the implantations were resorbed after transfer of blastocysts (78%) as compared with morulae (15%). These results demonstrate that transfer of day 5 cultured blastocysts into uteri of foster mothers increases embryonic mortality as a consequence of improper implantation.
Selective binding of specific rat epididymal secretory proteins to spermatozoa and erythrocytes
Authors: Brooks D. E. 
Published: February 2005

AbstractTissue pieces from the caput epididymidis of the rat were incubated in vitro with (35S) methionine to produce radioactive secretory proteins. The radioactive secretory proteins so formed were tested for their ability to bind to washed rat spermatozoa collected from the rete testis and cauda epididymidis, and to rat erythrocytes. The sperm and erythrocytes bound approximately 5% of the total radioactive protein. Binding was protein‐specific in that only selected proteins became associated with the cells. Binding was not cell‐specific, however, since testicular spermatozoa, caudal spermatozoa, and erythrocytes all bound the same proteins to a similar degree.
Parameters influencing ovum pickup by oviductal fimbria in the golden hamster
Authors: Mahi‐Brown Cherrie A. , Yanagimachi Ryuzo 
Published: February 2005

AbstractThe process whereby hamster oviductal fimbria transport ova to the ampulla was investigated in vitro and in vivo with the intent of determining whether cumulus‐free ova were transported and resolving some of the parameters involved in cumulus‐isolated from ovaries could not be picked up until 16–17 h before it would be ovulated. This was apparently related to the time at which the cumulus matrix begins to expand. The fimbria were not species‐specific, as they readily picked up rat, mouse, and rabbit ovulated cumul, but they were partially tissue‐specific as they would only pick up tissues such as the vitreous humor and loose connective tissue, which contain considerable extracellular glycosaminoglycan. Interaction of cumulus and fimbria could be prevented b treating either structure with polycatinic macromolecules such as poly‐l‐lysine, cationic ferritin, and protamine. Treatment of fimbria with proteolytic enzymes or hyaluronidase did not prevent pickup, but neuraminidase did. The only “artificial cumulus” the fimbria would pick up was chicken egg‐white. Cumulus‐free ova were not picked up in vitro, but part of them placed into the ovarian bursa were transported to the ampulla in vivo. Coating the ova with chicken egg‐white allowed their pickup in vitro, but had little effect on the success of transport of ova transplanted into the ovarian bursa in vivo. The transport of ova from the ovarian bursa to the oviductal ampulla is apparently a rather complex process involving interaction of some component of the cilia surfaces with the cumulus as well as other factors not necessarily related to this interaction.
Immunocytochemical localization of protamine in the spermatids of the ram
Authors: Courtens J. L. , Delaleu B. , Dubois M. P. , Lanneau M. , Loir M. , Rozinek J. 
Published: February 2005

AbstractPure ram protamine isolated from epididymal spermatozoa was used to raise antisera in castrated rabbits. The antibodies were visualized in the electron microscope using the method of Moriarty and Halmi [1972] with either peroxidase or coupling with colloidal gold. The gold gave better contrast but lower afinity than the peroxidase method. With the use of fixation according to Thiery and Rambourg [1976] and thick sections treated with hydrogen peroxide, it was possible to detect the protamine in the cytoplasm near the flagellum and the chromatoid body of step 12 spermatids. It was concluded that protamine enters the nucleus and concentrates at that step in the ram.
The localization of Ca2+‐ATPase and Ca2+ binding proteins in the flagellum of guinea pig sperm
Authors: Gordon Mildred , Morris Eugene G. , Young Ronald J. 
Published: February 2005

AbstractFerritin‐labeled antibody to calmodulin was localized in flagella of guinea pig sperm. Ferritin granules were present on the surface of the medial portion of the coarse fibers, and in the matrix immediately surrounding this segment of the fibers. The results demonstrate that calmodulin is confined to regions close to the tubulin‐dynein systems os the axoneme. A calcium‐dependent ATPase was localized in the same region by trapping the reaction product with cadmium. The presence together of calmodulin and the Ca2+‐ATPase close to the axonemal complex suggests that they function to control the concentration of Ca2+, which in turn may regulate the action of the dynein ATPase, and sperm motility. The remainder of the coarse fiber surface also contains specialized material as evidenced by intense staining with ethanolic phosphotungstic acid. These data suggest that the role of the coarse fibers is complex and may not be solely structural.
Nuclear maturation of follicle‐enclosed rat oocytes during inhibition of steriodogenesis
Authors: Billig Håkan , Hillensjö Torbjörn , Tsafriri Alexander , Mangusson Claes , Brodie Angela 
Published: February 2005

AbstractThis study examined the possible role of steriods in meiotic maturation of preovulatory oocytes. Follicles were isolated from PMSG‐treated immature rats and incubated with or withour LH in the presence of one of four inhibitors of steroidogenesis. The inhibitors employed had different sites of attack in the steriodogenic pathway and were aminoglutethimide, cyanoketone, SU 10603 (17β‐hydroxylase inhibitor), and 4‐OH‐androstenedione (aromatase inhibitor). As predicted, the inhibitors drastically altered the pattern of steroid production. None of the inhibitors, however, changed the proportion of oocytes resuming or completing meiosis in response to LH, and there was also no effect of the inhibitors on the oocytes in the absence of LH. It was concluded that steriods are not required for preovulatory nuclear maturation of oocytes in the rat.
Gametes and fertilization in the Chinese hamster
Authors: Yanagimachi R. , Kamiguchi Y. , Sugawara S. , Mikamo K. 
Published: February 2005

AbstractFreshly ovulated eggs are each surrounded by a compact cumulus oophorus. The overall diameter of the normal egg (including the zona pellucida) is about 100 μm. Cumulus cells, particularly those near the egg, are arranged redially in a viscous noncellular matrix. The spermatozoon is about 250 μm in length. The head a large acrosome, changes in which can be readily examined with the light (phase‐ contrast) microsope. When exposed to physiological salt solutions, testicular spermatozoa either were motionless or flexed the posterior half of their tails slowly. Spermatozoa from the caput epididymis were highly motile, flexing the entire tail. A few of them moved progressively. Mature spermatozoa from the vas deferens were highly motile and moved either straightforward or in a circle. They vibrated their tails stiffly without flexing them.In normally mated females, fertilization began sometime between 2 and 3 h after ovulation and was completed within the next 4 to 5 h. Spermatozoa swimming in the ampullary fluid or within the cumulus oophorus about the time of fertilization flexed the anterior half (which roughly corresponds to the midpieac region) of their tails. This peculiar movement may be homologous to the so‐called “hyperactivation” of spermatozoa as reported in several other mammalian species. Actively motile spermatozoa within the cumulus or no the zona pellucida had either modified (“collapsed”) or no acrosomal caps. The sperm head usually passed verticually or nearly through the zona, but the path was oblique in some instances. In 54% of the recently fertilized eggs examined, the entire length of the sperm tail was within the perivitelline space; in the other 46% of the eggs varying lenghts of the tail remined the perivitelline space, the tails were extruded from the vitellus of many eggs even before the eggs began their first cleavage.When unfertilized eggs in the cumulus oophorus were inseminated with vas deferens spermatozoa in a modified Tyrode's solution (m‐TALP), about 80% of them were ferrtilized by 4–6 h after insemination. The vast majority were monospermic. When eggs were freed from the cumulus prior to insemination, none were fertilized, suggesting that the cumulus cells or their matrix assisted capacitation and/or the acrosome reaction of the spermatozoa under the in vitro conditions employed. No eggs were fertilized by the testicular or caput epididymal spermatozoa regardless of the presence or absence of cumulus oophorus around the eggs at the time of insemination.
p15, A nuclear‐associated protein of human sperm: Identification of four variants and their occurrence in normal and abnormal seminal cells
Authors: Rodman Tody C. , Pruslin Fred H. , Chiorazzi Nicholas , Michelis Mary Ann , Winston Ronald 
Published: February 2005

AbstractA basic protein of apparent molecular weight 15,000 d (p15) has been identified as a tissue‐specific species‐unique component of spermatogenic cells of human semen. Cytoimmunochemical study with a monoclonal antibody indicates that p15 resides in a perinuclear space in morphologically normal spermatozoa and differs in distribution and stat in abnormal seminal cell and nonnucleated bodeis. Biochemical analysis indicateds that p15 occurs as four variiants, differentially migratory on acetic acid/urea gel and differnetially extractable by NaCl in reducing solution. By correlation of the cytologic and biochemical data, we propose that variant 1 is the unmodified form of p15; in the normal progression of spermiogenesis p15 is modified to variants 2 and 3 and in the absence of the normal progression is unmodified or aberrantly modified to variant 4. The association of molecular abnormality in p15 with morphologically abnormal sperm suggests that p15 may play a role in sperm‐head shaping.
Surface and internal antigens of rat spermatozoa distinguished using monoclonal antibodies
Authors: Jones Roy , Brown Colin R. , Cran David G. , Gaunt Stephen J. 
Published: February 2005

AbstractFluorescent antibody labeling techniques are frequently used to investigate the topography of antigens on spermatozoa. It is generally assumed that these procedures detect molecules only on the sperm surface but we now show that this assumption is not always valid. Using monoclonal antibodies that recognize either surface or internal antigens we demonstrate how spurious conclusions can be made, and we suggest simple procedures for assigning the position of an antigen to the cell surface or to an intracellular organelle. Antibodies against plasma membrane antigens should stain 100% of normal intact spermatozoa, but this proportion should be greatly reduced if the spermatozoa have previously been demembranated. If ≪ 100% of spermatozoa are stained but the proportion increases following permeabilization, then the possibility should be considered that the antigens are intracellular. We conclude that assignment of an antigen to a regional domain on the sperm surface using fluorescent antibody techniques should be validated by a demonstration that the antigen is actually located on the cell surface.
The status of acrosomal caps of hamster spermatozoa immediately before fertilization in vivo
Authors: Yanagimachi Ryuzo , Phillips David M. 
Published: February 2005

AbstractAdult female golden hamsters were induced to superovulate. When they were mated several hours prior to ovulation or artificially inseminated about the time of ovulation, nearly 100% of their eggs were subsequently fertilized monospermically. During the progression of fertilization when the eggs were still surrounded by compact cumulus oophorus, the contents of the ampullary region of the oviducts were collected and spermatozoa moving in the ampullary fluid, within the cumulus and on/in the zonae pellucidae of unfertilized eggs, were examined by light and electron microscopy to evaluate the status of their acrosomal caps.Most spermatozoa swimming in the ampullary fluid had apparently intact acrosomal caps, while the vast majority moving within the cumulus had distinctly modified acrosomal caps. Most spermatozoa that had passed through the cumulus and reached the zona surfaces had remnants of their acrosomal caps (“acrosomal ghosts”). When the ghosts were present around the sperm heads on the zona, the heads pivoted about a point roughly corresponding to the places where the ghosts were located. The ghosts seemed to firmly attach to the zona surfaces, then were split open by the sperm heads and left behind as the sperm heads advanced into the zona. A few spermatozoa on the zona surfaces had no acrosomal ghosts (at least not detectable by light microscopy). In this case, the sperm head pivoted about either the inner acrosomal membrane or the equatorial segment of the acrosome. In no instance were spermatozoa with intact acrosomal caps found on zona surfaces.We infer from these observations that most spermatozoa in vivo initiate their acrosome reactions while they are advancing through the cumulus. When they arrive at the zona surfaces, acrosomal ghosts are generally present on the sperm heads. These ghosts appear to hold sperm heads to zona surfaces as well as to restrict the direction of advancement of sperm head through the zona. In a minority of cases, ghostless spermatozoa reach the zona surfaces. As these spermatozoa appear to be able to penetrate the zona successfully, structures other than the acrosomal ghost (ie, the inner acrosomal membrane and the plasma membrane over the equatorial segment of the acrosome) may also attach to zona surfaces before spermatozoa penetrate into the zona.
Developmental ability of precociously superovulated golden hamster eggs
Authors: Yanagimachi R. 
Published: February 2005

AbstractSuperovulated hamster eggs obtained by injections of PMSG on day 1 (the day of ovulation) and hCG on day 3 (2 days before expected day of next ovulation) have been used widely for assessing the fertilizing capacity of spermatozoa of the homologous as well as heterologous species. The developmental ability of these eggs was assessed by transferring them to host females (recipients) that had ovulated and mated naturally. The proportion of transplanted eggs developing into normal fetuses and young was the same regardless of whether the eggs had been collected from superovulated or naturally ovulating donors. The young originating from superovulated eggs were normal in that they grew at a normal rate and produced normal progeny. It may be concluded that the precociously superovulated eggs were as normal as the control (naturally ovulated) eggs. Thus, the use of precociously superovulated hamster eggs in evaluating the fertilizability of spermatozoa is justified.
Inhibition of mouse embryonic development by calmodulin antagonists
Authors: Lenz R. W. , Hart R. , Ax R. L. , Cormier M. J. 
Published: February 2005

AbstractTwo‐cell mouse embryos were incubated in the presence of calmodulin inhibitors to determine their effect on embryonic development to the blastocyst stage. Calmodulin, a Ca2+‐dependent regulatory protein, has been localized in the cytoplasm and has been implicated in regulation of many cellular events, such as mitosis. Several concentrations of either commercial or synthesized calmodulin inhibitors were tested. Several phenothiazine sulfoxide derivatives were more effective than the three naphthalene sulonic acid derivatives tested; 2‐chloro‐10‐aminopropyl phenothiazine and 10‐aminopropyl phenothiazine were the most potent phenothiazines to inhibit embryonic development at the two‐cell stage. The interesting aspect of this study is that phenothiazine sulfoxide derivatives are not potent inhibitors of calmodulin, however, they were successful in inhibiting embryonic development. Potent inhibitors of calmodulin apparently did not penetrate the embryonic membranes because they had no effect.
Spermatozoa of chromosomally heterozygous mice and their fate in male and female genital tracts
Authors: Redi C. A. , Garagna S. , Pellicciari C. , Romanini M. G. Manfredi , Capanna E. , Winking H. , Gropp A. 
Published: February 2005

AbstractThe fate of morphologically normal but chromosomally abnormal spermatozoa derived from mice with variable degrees and complexity of Robertsonian heterozygosity was studied at different sites along the male and female genital tract by Feulgen‐DNA measurements. In addition, the percentage frequencies of morphologically abnormal spermatozoa in transit along the male and female genital tracts were studied.It was found that during transit from the epididymis to the vas deferens the distribution of the Feulgen‐DNA contents of morphologically normal spermatozoa changed: Spermatozoa with chromatin with the extremely low or high Feulgen staining intensity disappeared. The percentages of morphologically abnormal sperm cells did not change at these levels. In the female genital tracts, the distribution of Feulgen‐DNA content of morphologically normal spermatozoa did not show significant changes. This indicates that spermatozoa are able to reach the fallopian tube in spite of gross genome unbalance. There is evidence that unbalanced spermatozoa take part in the fertilization process, producing abnormal zygotes subject to postzygotic loss. Conversely morphologically abnormal spermatozoa were preferentially lost before they reached the fallopian tube, suggesting they had been eliminated prezygotically.
Reproduction in mice: Behavioral changes induced in female mice by mating
Authors: Chaykin Sterling 
Published: February 2005

AbstractMated C57BL/6J females experienced in food‐motivated maze running were found to make more choices and choose more correctly than virgins. The differences correlated with the receipt of spermatozoa and prior pseudopregnancy. Pregnancy, active pseudopregnancy, and male‐female social interactions did not appear to be required determinants. The biologic ramifications of mating in mice may extend beyond the fusion of sperm and egg and the induction of hormonal changes that regulate the earliest stages of pregnancy.