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  • 1
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Gamete Research 18 (1987), S. 319-332 
    ISSN: 0148-7280
    Keywords: sperm membrane ; stallion spermatozoa ; spermatic maturation ; ultrastructural cytochemistry ; cellular microelectrophoresis ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The structure, relative density, and distribution of anionic sites on the surface of epididymal and ejaculated spermatozoa were studied using polycationic ferritin (CF), colloidal iron hydroxide (CIH), various enzymatic treatments, methylation, and de-acetylation. Macro-molecules containing sugar residues, probably sialic acid, are part of the sperm membrane and show a characteristic distribution and density that is dependent of the sperm region and of its origin. Unlike the spermatozoa of other eutheria examined, the exposure of the stallion spermatozoa to neuraminidase treatment did not produce significant changes in the density of the negative charge of the sperm surface. The ability of purified neuraminidase to act only after saponification suggests that sialic acid may be present in the acetylated form. When CIH was used it is seen that the density of the negative charge is rather uniform within a particular segment of the spermatozoa and abruptly changes at the junction of morphologically distinct segments (Between the acrosomal and post acrosomal region of the sperm head and between the post acrosomal region and middle piece of the flagellum). The acrosome presented more negative groups dissociated at pH 1.8 than the postacrosomal region. A greater concentration of anionic sites over the flagellum was also observed when CIH and CF were used. This assymetry probably represents different domains that may be related to specific functions.The cytochemical observations and the cellular electrophoretic mobility measurements did not show striking differences on the negative charge of sperm obtained from different regions of epididymis and ejaculates in contrast to previous results in other species. The spermatozoa collected from caput epididymidis bind CIH but not all population present equal response. In corpus and cauda region of epididymis the population displaying the capacity to bind CIH or CF significantly over the head and tail surface was the majority.This study corroborates that the distribution and density of terminal oligosaccharide residues on the sperm plasma membrane has species specific characteristics. The surface charge of the spermatozoa obtained either during the breeding or nonbreeding season, determined by measurements of cellular electrophoretic mobility and by the binding pattern of CIH and CF, does not show significant differences.
    Additional Material: 9 Ill.
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 35 (1993), S. 176-180 
    ISSN: 1040-452X
    Keywords: Cholesterol diet ; Polyunsaturated diet ; Fish oil ; Acrosome reaction ; Hypercholesterolemia ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In this study we examined the effect of cholesterol (Diet 2), cholesterol and fish oil (FO) polyunsaturated acid (Diet 3), and polyunsaturated acid (Diet 4) enriched diets upon the acrosome reaction (AR) of New Zealand White rabbit spermatozoa. Male rabbits fed with cholesterol alone or with FO increased their cholesterol and LDL-cholesterol serum levels after 15 days of diet. Ten semen samples were obtained after 2 months of diet. Our results suggest that hypercholesterolemia and hypertriglyceridemia in male rabbits could produce a decreased capacity of sperm AR after 4 h (0%, 0%, and 60% lower than the control), 6 h (0%, 68%, and 44%), or 8 h (58%, 52%, and 32%) of incubation in capacitating medium. Another set of experiments were made with lysophosphatidylcholine (LPC), 80 μg/ml, and the same pattern of AR was seen. Nevertheless, the high cholesterol and total lipids (TL) levels in serum did not affect the cholesterol levels in seminal plasma (SP) but affect the SP total lipids. The diminished capacity of rabbit sperm to undergo the AR was not reverted by in vitro incubation with the Shinitsky medium for cholesterol depletion (MDC). These results indirectly suggest that the cholesterol/phospholipid ratio in hypercholesterolemic sperm is similar to that of controls and are in agreement with preliminary studies made in our laboratory that evidenced the same cholesterol/ phospholipid ratio in rabbit sperm from hypercholesterolemic animals than from controls. These findings and the higher sterol-filipin complex in the acrosomal membrane of hypercholesterolemic animals compared to controls reported by Diaz-Fontdevila et al. (1992) could suggest that in this study different sperm membrane lipid domains induced by the hypercholesterolemia are the cause of the AR kinetics modification. © 1993 Wiley-Liss, Inc.
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  • 3
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Journal of Morphology 174 (1982), S. 169-184 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The gross morphology, histology, and ultrastructure of Liolaemus gravenhorsti gonads prior to and after differntiation are described. Special emphasis has been given to characterization and changes of the germ cell line throughout intrauterine development and 3 days postpartum.During the pregonadal stage, the primordial germ cell migrates toward gonadal rudiments by way of the mesenchyme. These cells can easily be identified by their great size, voluminous and lobulated nucleus, great quantities of yolk platelets, microtubules, and numerous lipid inclusions. In the undifferentiated gonad, the germ cells (type 1 gonocytes) have an ovoid or spherical shape and autodigestion of yolk platelets, great development of Golgi complex, and mitochondrial aggregation, though fewer liposomes, pseudopodes, and microtubules were noted. Concomitantly with the beginning of mitosis, a third type of germ cell appears, the type 2 gonocytes, which are smaller, with poorly defined membranous systems in various degrees of involution. The seminiferous cords are organized when somatic cells of the medullar portion of the gonad surround type 1 gonocytes. Germinal cells are centrally localized in the cords. Near birth many gonocytes migrate toward the basal lamina of cords and differentiate into spherical prespermatogonia, with few organoids. Sertoli cells eparate them from the basal lamina. In advanced pregnancy, Leyding cells become numerous with morphology typical of androgen-producing cells.
    Additional Material: 30 Ill.
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  • 4
    ISSN: 0730-2312
    Keywords: chromatin ; chromatin remodeling ; fertilization ; embriogenesis ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: To obtain information on the remodeling of sperm chromatin during male pronuclei formation, we have followed the sperm specific histones (SpH) that form the nucleosomal core by Western imunoblot analysis with policlonal antibodies directed against the core SpH. The results obtained indicate that the complete set of SpH is absent from zygote chromatin at the beginning of the first S phase. The disappearance of SpH is not coincidental for the five histone classes: SpH4 and SpH3 are lost 5-15 min post insemination (p.i.), SpH2B and SpH2A disappear 20-40 min p.i., and SpH1 is progressively diminished up to 30 min p.i. This order of sperm chromatin remodeling is not affected by the inhibition of protein synthesis by emetine, indicating that the factor(s) responsible for SpH disappearance are present in unfertilized eggs. The lost SpH's are not replaced by newly synthesized CS variants, since the basic proteins synthesized de novo during male pronuclei formation are not incorporated into chromatin remaining in the cytoplasm. These newly synthesized proteins are different from the CS variants as judged by their electrophoretic migration.
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  • 5
    ISSN: 0730-2312
    Keywords: chromatin ; pronucleus ; nucleoprotein particles ; sea urchins ; zygotes ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: To determine the changes in chromatin organization during male pronucleus remodeling, we have compared the composition of nucleoprotein particles (NP-ps) resulting from digestion with endogenous nuclease (ENase) and with micrococcal nuclease (MNase). Whole nuclei were isolated from sea urchin gametes and zygotes containing a partially decondensed (15 min postinsemination, p.i.) or a fully decondensed (40 min p.i.) male pronucleus and digested with nucleases. The NP-ps generated were analyzed in agarose gels, and their histone composition was determined. Sperm core histones (SpH) and cleavage stage (CS) variants were identified by Western immunoblots revealed with specific antibodies. A single NP-ps was generated after digestion of sperm nucleus with MNase, which migrated in agarose gels between DNA fragments of 1.78-1.26 Kb. Sperm chromatin remained undigested after incubation in ENases activating buffer, indicating that these nuclei do not contain ENases. One type of NP-ps was obtained by digestion of unfertilized egg nuclei, either with ENase or MNase; the NP-ps was located in the region of the agarose gel corresponding to DNA fragments of 3.4-1.95 Kb [Imschenetzky et al. (1989): Exp Cell Res 182:436-444]. When whole nuclei from zygotes containing the female pronucleus and a partially remodeled male pronucleus were digested with ENase, a single NP-ps was generated, which migrated between DNA fragments of 2.5-1.9 Kb. This particle contained only CS histone variants. Alternatively, when these nuclei were digested with MNase, two NP-ps were generated; the slower migrating NP-ps (s) was located in the same position of the agarose gel as those resulting from ENase digestion and the faster migrating NP-ps (f) migrated between DNA fragments of 1.95-1.26 Kb. It was found that Np-ps (s) contained only CS histone variants, whereas NP-ps (f) were formed by a subset of SpH and by CS histone variants. When nuclei from zygotes containing a fully decondensed male pronucleus were digested either with ENase or MNase, a single type of NP-ps was observed, which migrated in the same position as NP-ps (s) in agarose gels. This particle contained only CS histone variants. On the basis of the histone compositions and on electrophoretic similarities, it was concluded that NP-ps (s) originated from the female pronucleus and that NP-ps (f) were generated from the partially remodeled male pronucleus. Consequently, our results indicate that at an intermediate stage of male pronucleus remodeling the chromatin is formed by NP-ps containing a subset of both SpH and of CS histone variants, whereas at final stages of male pronucleus decondensation chromatin organization is similar to that of the female pronucleus. © 1996 Wiley-Liss, Inc.
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 92 (1977), S. 43-47 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: 32P-uptake into non-histones from bone cell cultures was selectively stimulated in the presence of calcitonin. Comparison of the control and experimental radioactivity profiles of non-histones fractionated by SDS gel electrophoresis showed that, in response to calcitonin stimulation, there was a 2- to 3-fold increase in the specific activity associated with non-histone proteins in the molecular weight range of 10,000 to 45,000 daltons while that of bands between 50,000 to 200,000 decreased.
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