bluuebunny/crossref_metadata_2025_split huggingface dataset:
Prediction of human sperm penetrating ability using computerized motion parameters
Authors: Fetterolf P. M. , Rogers B. J. 
Published: February 2005

AbstractThe sperm penetration assay (SPA) is used to assess male fertilizing potential but it is tedious and costly. Computer analysis could replace the need for the SPA in some cases, if computerized sperm motility parameters are highly predictive of SPA performance. The objective of this study was to determine whether computerized motility parameters from fresh semen samples could be used to predict SPA performance. Computer automated semen analysis (CASA; CellSoft, Cryo Resources) was used to quantitate sperm concentration (CONC), percent motility (MOT), curvilinear velocity (VEL), linearity of swimming trajectory (LIN), mean amplitude of lateral head displacement (ALH), and beat/cross frequency (B/CF). The SPA was performed using either Biggers, Whitten, and Whittingham's medium (BWW) or TEST‐yolk buffer (TYB). Patients were divided into three groups depending on SPA performance: group 1, BWW‐treated, 0% versus > 0% penetration; group 2, TYB‐treated, 0% versus > 0% penetration; and group 3, TYB‐treated, < 20% versus ≥ 20% penetration. SPA performance was highly correlated with CONC, MOT, VEL, and B/CF. CONC, MOT, VEL, and B/CF were significantly higher for patients who penetrated in the SPA than for those who failed to penetrate. Discriminant function analysis (DFA) successfully classified 76% of all patients treated with TYB (group 2) who penetrated and 86% of nonpenetrators based on their computerized motility parameters. For group 2 DFA predicted that 93 men would penetrate in the SPA with TYB. Of these, 90 (96%) successfully penetrated at least one egg. It is concluded that DFA of computerized motility parameters can accurately predict whether a patient will penetrate (> 0%) in the TYB‐treated SPA.
Immunoreactive inhibin in follicular fluid is related to meiotic stage of the oocyte during final maturation of the porcine follicle
Authors: Miller K. F. , Xie S. , Pope W. F. 
Published: February 2005

AbstractThe concentration and content of inhibin was determined in individual porcine follicles from gilts ovariectomized at various times after the onset of estrus. In one experiment, gilts (n = 5) were ovariectomized at 0, 10, or 20 hr after the onset of estrus and the follicular fluids from all large follicles individually aspirated. In a second experiment, gilts (n = 6) were ovariectomized at 21, 24, 27, 30, or 34 hr after the onset of estrus; follicular fluids were aspirated; and each oocyte was stained and evaluated for cytogenetic stage of meiotic maturation. Inhibin was determined in diluted follicular fluids with a radioimmunoassay based on a synthetic peptide replica of part of the α subunit of porcine inhibin. Inhibin values were expressed in terms of thousands of units (kU) of a World Health Organization inhibin standard (86/690). Concentration of inhibin did not vary among hours (overall mean 248 kU/ml). Total follicular content of inhibin also was not different among hours (overall mean 57 kU/follicle). When follicles were classified on the basis of the maturation of the oocyte, significant differences were found. Concentration of inhibin in follicles with a germinal vesicle‐stage oocyte was 138 kU/ml, whereas follicles with more mature oocytes had concentrations of between 204 and 254 kU/ml. Follicular content of inhibin showed a similar pattern with 34.9 kU/follicle at germinal vesicle stage, increasing to 42.5–56.1 kU/follicle at later stages. Quantities of inhibin were also negatively skewed and were positively correlated to follicular content of estradiol and dermatan sulfate.
Identification and characterization of the major components of the Oncorhynchus mykiss Egg Chorion
Authors: Brivio Maurizio F. , Bassi Rosaria , Cotelli Franco 
Published: February 2005

AbstractThe extracellular coat surrounding the fish egg, commonly called the chorion, is a primary envelope that confers biochemical and morphological identity typical of the species. Purified chorions can be easily isolated from either oocytes or ovulated eggs. The aim of this work was to analyze the macromolecular composition of the various chorion components in Oncorhynchus mykiss (Salmonids). SDS‐PAGE analysis of purified chorion showed a reproducible pattern of four major components (129, 62, 54, and 47 kD), representing about 80% of total chorion proteins. The 129 and 47 kD polypeptides were periodic‐acid Schiff (PAS) and concanavalin A positive. After chemical and enzymatic deglycosylation treatments only the 129 and 47 kD components proved to be glycosylated and to belong to the “asparagine‐linked” glycoprotein family. Furthermore, peptide mapping performed on isolated polypeptides showed comigrating fragments on SDS‐PAGE. These results suggest that the four main chorion polypeptides might share common structural features.
Fertilizing competency of multiple ovulated eggs in the domestic fowl (Gallus domesticus)
Authors: Nakanishi A. , Miyake M. , Utsumi K. , Iritani A. 
Published: February 2005

AbstractFertilizing competency of multiple ovulated eggs in the domestic fowl was examined by fertilization in vitro and early development in culture. Normal laying hens (White Leghorn) were treated with 75 IU of PMSG for 7 days followed by injection of anterior pituitary extracts from chickens (CAPE). Ovulation began to occur 7.5 h after injection of CAPE. These hens ovulated 1–7 ova but some premature ovulation of GV stage ova were observed. In vitro fertilization of the multiple ovulated ova was examined by inseminating 106–107 sperm onto the germinal disks in m‐Ringer's solution. The gamete or zygote nuclei were detected by DNA specific fluorescence using DAPI (4′,6′‐diamidino‐2‐phenylindole) in the histological section prepared from the germinal disk. Process of fertilization was examined in the eggs incubated for 4 h after insemination in DMEM+ liquid albumen at 41°C under the atmosphere of 5% CO2 in air. Fertilization rate of the total multiple ovulated eggs was 55% (11/20), in which 90% (9/10) and 10% (1/10) in the eggs recovered 7.5–8.5 h and 9.0–9.5 h after CAPE injection were obtained, respectively. Normal pronuclei were formed in five eggs of those recovered 7.5–8.5 h after CAPE injection. Early development after fertilization in vitro was also examined by incubation for 12 h in DMEM + liquid albumen at 41°C under the atmosphere of 5% CO2 in air. Although development in vitro was delayed compared to that in utero condition, normal development was observed in naturally and multiple ovulated eggs. The developmental rate of the multiple ovulated eggs was 67% (20/30) in total, in which 94% (15/16) and 5% (5/14) were obtained in the eggs recovered 7.5–8.5 h and 9.0–9.5 h after CAPE injection, respectively. Both fertilization and developmental rate of the eggs recovered 9.0–9.5 h after CAPE injection were significantly lower (P<0.01) than those recovered 7.5–8.5 h after CAPE injection or naturally ovulated. These results suggested that multiple ovulated fowl eggs recovered 7.5–8.5 h after CAPE injection had normal fertilizing and developmental ability in vitro.
In vitro phosphorylation of sea urchin sperm adenylate cyclase by cyclic adenosine monophosphate‐dependent protein kinase
Authors: Bookbinder Louis H. , Moy Gary W. , Vacquier Victor D. 
Published: February 2005

AbstractAddition of [γ‐32P]ATP to a 2% Brij‐78 40,000g supernatant of sea urchin sperm results in the cAMP‐dependent phosphorylation of eight to ten proteins. One phosphoprotein of Mr 190 kD is sperm adenylate cyclase (AC). An antiserum to the AC immunoprecipitates the Mr 190 kD protein. Peptide maps of immunoprecipitates show that the AC is the only phosphoprotein present in the Mr 200 kD range. With respect to the in vitro phosphorylation of AC, the endogenous kinase has a Km for ATP of 5.2 μM and is maximally stimulated by 4–8 μM cAMP. The protein kinase inhibitors H8 (9 μM) and PKI (30 U/ml) inhibit the phosphorylation of the AC. The catalytic subunit of bovine cAMP‐dependent protein kinase phosphorylates the AC on the same peptides as the endogenous protein kinase. Cyanogen bromide generated peptide maps of the phosphorylated AC show a minimum of five sites of phosphorylation. No change in the Km or Vmax of the sperm AC resulted from the additional phosphorylation by bovine kinase. Calcium ions at submicromolar concentrations completely block the in vitro phosphorylation of the AC, suggesting the presence in the preparation of a Ca2+‐activated protein phosphatase. To our knowledge, this is the first report of the phosphorylation of an AC by cAMP‐dependent protein kinase.
Presence and localization of the 60 KD calicin in human spermatozoa presenting postacrosomal sheath defects: Preliminary results
Authors: Courtot Anne‐Marie 
Published: February 2005

AbstractUsing an indirect immunofluorescence technique, we studied the anticalicin antibody distribution in four teratozoospermic human sperm samples with a high rate of spermatozoa with incomplete postacrosomal sheath structure (PAS) detected at the ultrastructural level. The calicin distribution obtained in these samples is compared with the distribution obtained in two normal human sperm samples with a high proportion of normal PAS and in 3 human sperm samples without PAS (“round‐headed syndrome”). In contrast to the homogeneous calicin distribution found in the majority of the normal group, similar to that reported by Longo et al. (J Cell Biol 105:1105–1120, 1987) and Paranko et al. (Differentiation 38:21–27, 1988), and the absence of calicin in the three non‐PAS samples, we observed new patterns of calicin distribution in the four sperm samples studied. In the light of these preliminary results, the use of anticalicin antibody as a tool for human postacrosomal region evaluation is discussed.
Localization of microfilaments and a tubulin‐like protein in crustacean (Rhynchocinetes typus) spermatozoon
Authors: Perez C. , Roco M. , Castro A. , Dupre E. , Schatten G. , Barros C. 
Published: February 2005

AbstractSperm from the decapod crustacean Rhynchocinetes typus undergo dramatic shape changes as they pass from the vas deferens to seawater and interact with the oocyte envelopes. Using FITC‐phalloidin and antitubulin antibodies, we were able to localize microfilaments and a tubulin‐like protein in R. typus spermatozoon. Microfilaments and the tubulin‐like protein were associated with the sperm rays and spines, but were absent at the spike and at its base. Folded and unfolded spermatozoa display similar fluorescence patterns. SDS‐PAGE of whole spermatozoa and electrotransfer to nitrocellulose confirmed the presence of actin and two proteins at 97 kd and 120 kd that bind to tubulin antibodies (tubulin‐like proteins). These results demonstrate the presence of actin, but not tubulin, and localize microfilaments in these sperm. It is proposed that this cytoskeletal component is active in sperm during crustacean fertilization.
Artificial activation of porcine oocytes matured in vitro
Authors: Prather R. S. , Eichen P. A. , Nicks D. K. , Peters M. S. 
Published: February 2005

AbstractThese studies were undertaken to understand the biological basis of artificially induced activation of meiotic metaphase II oocytes and to develop a source of oocytes as recipients for cloning by nuclear transfer. In vitro matured porcine oocytes were pulsed with various voltages of electricity and evalulated for pronuclear formation. The percentage of eggs that activated was significantly greater for the higher voltages. The effect on activation of the temperature of the ovaries returning from the abattoir was evaluated and it was found that oocytes derived from ovaries returning at 29°C activated at lower rates (45.5%) than those returning at 36°C (78.9%). An experiment was designed to evaluate the pH of electroporation medium (EM) and the duration of exposure to EM on activation. Oocytes were placed in EM at various pHs for 5 minutes, pulsed, and immediately removed to TL‐Hepes or allowed an additional 2 minutes in EM prior to rinsing in TL‐Hepes. The results indicate an optimum activation rate at a pH of 7.0 and allowing the additional 2 minutes in EM. Additional glucosamine (5 mM) had no affect on development of the oocyte to metaphase but reduced the percent pronuclear formation from 61% and 47%. A final experiment evaluated the developmental competence of oocytes subjected to a optimum combination of the above treatments and illustrated that a significant portion of the activated oocytes can show limited signs of cleavage. Thus in vitro matured pig oocytes can be induced to activate at high rates.
Localization of wheat germ agglutinin and antibody binding sites on the plasma membranes of sea urchin sperm heads as revealed by label‐fracture and fracture‐flip
Authors: Ru‐Long Shen , Ward Rita D. , Da Silva Pedro Pinto , Nishioka David 
Published: February 2005

AbstractFreeze‐fracture electron microscopy reveals that intramembrane particles are concentrated in a band encircling the posterior portion of the acrosome of Strongylocentrotus purpuratus sperm. Two colloidal gold labeling methods, label‐fracture and replicastaining fracture‐flip, were employed to show that the plant lectin wheat germ agglutinin, which recognizes a 210 kDa sperm surface glycoprotein, binds to this localized band of intramembrane particles. Monoclonal antibody J18/2, which also recognizes the 210 kDa surface glycoprotein, shows this localized binding in ≈20% of the sperm observed in this study. The majority of sperm displayed a uniform distribution of receptor sites for monoclonal antibody J18/2. Since wheat germ agglutinin and monoclonal antibody J18/2 are known to agglutinate Strongylocentrotus purpuratus sperm but not sperm of another sea urchin, Lytechinus pictus, similar determinations were made for the latter species. Lytechinus pictus sperm are not labeled with wheat germ agglutinin and are only sparsely labeled with monoclonal antibody J18/2. The acrosomal localizations of wheat germ agglutinin and monoclonal antibody J18/2 receptors in Strongylocentrotus purpuratus sperm are consistent with the involvement of the 210 kDa surface glycoprotein in an egg jelly‐induced sperm acrosome reaction. Low‐temperature post‐embed labeling of thin sections with wheat germ agglutinin and monoclonal antibody J18/2 show concentrations of label within the acrosomal vesicle of Strongylocentrotus purpuratus sperm, suggesting the presence of an intracellular storage site for the 210 kDa glycoprotein.
Intercellular communication in the early human embryo
Authors: Dale B. , Gualtieri R. , Talevi R. , Tosti E. , Santella L. , Elder K. 
Published: February 2005

AbstractA preliminary study on intercecullar communicative devices in the early human embryo has been made using dye‐coupling techniques and electron microscopy (EM). Lucifer yellow injected into single blastomeres of embryos at the 4‐cell stage up to the late morula stage did not spread to neighbouring cells, indicating that gap junctions and cytoplasmic bridges are not significant pathways for information transfer. Dye spread was first observed in the blastocyst stage, where trophectoderm cells and inner mass cells were shown to be in communication through gap junctions. Studies at the EM level confirmed this finding. Tight junctions and desmosome‐like structures, apparent from the 6‐cell stage onward, were located both peripherally and centrally and were initially nonzonular. The role of intercellular devices in the primary differentiation of the human embryo is discussed.
Reduction of the fertilizing capacity of sea urchin sperm by cannabinoids derived from marihuana. I. Inhibition of the acrosome reaction induced by egg jelly
Authors: Schuel Herbert , Berkery Diana , Schuel Regina , Chang Michael C. , Zimmerman Arthur M. , Zimmerman Selma 
Published: February 2005

AbstractΔ9‐Tetrahydrocannabinol (THC) and two other major cannabinoids derived from marihuana–cannabidiol (CBD) and cannabinol (CBN)–inhibit fertilization in the sea urchin Strongylocentrotus purpuratus by reducing the fertilizing capacity of sperm (Schuel et al., 1987). Sperm fertility depends on their motility and on their ability to undergo the acrosome reaction upon encountering the egg's jelly coat. Pretreatment of S. purpuratus sperm with THC prevents triggering of the acrosome reaction by solubilized egg jelly in a dose (0.1–100 μM) and time (0–5 min)‐dependent manner. Induction of the acrosome reaction is inhibited in 88.9±2.3% of sperm pretreated with 100 μM THC for 5 min, while motility of THC‐treated sperm is not reduced compared to solvent (vehicle) and seawater‐treated controls. The acrosome reaction is inhibited 50% by pretreatment with 6.6 μM THC for 5 min and with 100 μM THC after 20.8 sec. CBN and CBD at comparable concentrations inhibit the acrosome reaction by egg jelly in a manner similar to THC. THC does not inhibit the acrosome reaction artificially induced by ionomycin, which promotes Ca2+ influx, and nigericin, which promotes K+ efflux. THC partially inhibits (20–30%) the acrosome reaction induced by A23187, which promotes Ca2+ influx, and NH4OH, which raises the internal pH of the sperm. Addition of monensin, which promotes Na+ influx to egg jelly or to A23187, does not overcome the THC inhibition. Inhibition of the egg jelly‐induced acrosome reaction by THC produces a corresponding reduction in the fertilizing capacity of the sperm. The adverse effects of THC on the acrosome reaction and sperm fertility are reversible. These findings show that cannabinoids reduce the fertilizing capacity of sea urchin sperm by inhibiting the induction of the acrosome reaction by egg jelly. THC may affect events in the stimulus–secretion coupling mechanism before the opening of ion channels.
Reduction of the fertilizing capacity of sea urchin sperm by cannabinoids derived from marihuana. II. Ultrastructural changes associated with inhibition of the acrosome reaction
Authors: Chang Michael C. , Schuel Herbert 
Published: February 2005

AbstractPretreatment of Strongylocentrotus purpuratus sperm with Δ9‐tetrahydrocannabinol (THC) prevents the triggering of the acrosome reaction by egg jelly. Examination of THC‐treated sperm by transmission electron microscopy reveals that the membrane fusion reaction between the sperm plasma membrane and the acrosomal membrane is completely blocked. Electrondense deposits are present in the subacrosomal fossa and in the centriolar fossa. The nuclear envelope is fragmented in close proximity to the electron‐dense deposits. The electron‐dense deposits are not bound by a limiting membrane, stain positively for lipid with thymol and farnesol, and disappear from THC‐treated sperm that are extracted with chloroform:methanol (2:1) after glutaraldehyde fixation. The electron‐dense deposits are lipid in nature and may be a hydrolytic product of the nuclear envelope. Electron‐dense deposits are seen in sperm after 1–10 min treatment with 5–100 μM THC. The electrondense deposits disappear after removal of THC from the sperm by washing, but the fragmented nuclear envelope in the subacrosomal fossa persists. Cannabidiol (CBD) and cannabinol (CBN) also inhibit the triggering of the acrosome reaction by egg jelly and produce ultrastructural changes in the sperm identical to those elicited by THC. Enhanced phospholipase activity stimulated by THC, CBD, and CBN may be the cause of the accumulation of lipid deposits in the sperm. Metabolites derived from this modification of membrane phospholipids may prevent triggering of the acrosome reaction by egg jelly and thereby inhibit fertilization.
Hamster oocyte penetrability during preovulatory maturation
Authors: Cuasnicu Patricia S. , Bedford J. M. 
Published: February 2005

AbstractIn vitro fertilization techniques were used to analyze the penetrability of preovulatory hamster oocytes. The zonas of granulosa cell‐free primary (GV) oocytes were penetrated in vitro in 2–3 h as readily as those of ovulated secondary oocytes (80% vs. 88%), whether inseminated separately or as mixed oocyte groups. In fact, a significantly higher (P<0.05) mean number of perivitelline spermatozoa was present in immature (3.6) compared with secondary (1.9) oocytes, primarily reflecting a lack of the zona block to polyspermy in the immature population. By contrast, when granulosa cells remained around GV oocytes, zona penetration was low and more were penetrated, with more spermatozoa incorporated into the vitellus as a function of increasing time of oocyte recovery after hCG. We conclude, contrary to previous reports, that the zona pellucida of the hamster GV oocyte is readily penetrable by spermatozoa in vitro. However, the resumption of meiosis brings an increase in the penetrability of the granulosa cell vestment as well as the capacity for cortical granule exocytosis and the ability to decondense and transform the fertilizing sperm nucleus. The fact that the zona pellucida of the immature oocyte has proved to be penetrable in vitro and/or in vivo in all the mammals studied in this respect is discussed with particular reference to the situation in man.
Ribosomal gene amplification in oocytes of the lizard Podarcis sicula
Authors: Motta Chiara M. , Andreuccetti Piero , Filosa Silvana 
Published: February 2005

AbstractIn order to provide cytological evidence of amplification, Podarcis sicula oocytes were studied by cytophotometry, thymidine incorporation and in situ DNA‐DNA hybridization.Our results show that DNA replication is completed during the preleptotene stage, the leptotene oocytes having the typical 4C nuclear DNA content. Between the zygotene and the mid‐pachytene stages further DNA synthesis occurs with consequent increase of the ribosomal nuclear DNA content.These results and the variations in nucleolar organization observed during differentiation give clear evidence of the existence of ribosomal gene amplification in Podarcis sicula oocytes.
Embryonic transcription in in vitro cultured bovine embryos
Authors: Barnes F. L. , First N. L. 
Published: February 2005

AbstractThe rate of cleavage and the onset of embryonic transcription of bovine embryos cultured in vitro (IVC) has been investigated. Embryos were derived from in vitro matured, in vitro fertilized oocytes (IVM/IVF) to improve developmental synchrony. The rate of cleavage was assessed by morphological evaluation between the one‐ and eight‐ to 16‐cell stage. The rate of cleavage was found to be equivalent to that reported for in vivo recovered embryos. To assess the onset of embryonic transcription, embryos were cultured to the eight‐ to 16‐cell stage in the presence of α‐amanitin for various periods of time followed by two‐dimensional polyacrylamide gel electrophoresis. Embryos readily cleaved to the eight‐ to 16‐cell stage in the presence of inhibitor α‐Amanitin‐sensitive protein synthesis was first detected at 36–48 h post‐insemination (hpi) and continued up to 84 hpi. We conclude that bovine embryos produced by IVM/IVF/IVC are competent to initiate embryonic transcription at 36–48 h post‐insemination and suggest that in vitro‐induced cleavage arrest is not due to failure of the embryonic genome to initiate transcription.