The fate of porcine sperm CRISP2 from the perinuclear theca before and after in vitro fertilization

In a previous study, we reported that porcine sperm cysteine-rich secretory protein 2 (CRISP2) is localized in the post-acrosomal sheath-perinuclear theca (PT) as reduction-sensitive oligomers. In the current study, the decondensation and removal of CRISP2 was investigated during in vitro sperm capacitation, after both the induction of the acrosome reaction and in vitro fertilization. Confocal immunofluorescent imaging revealed that additional CRISP2 fluorescence appeared on the apical ridge and on the equatorial segment (EqS) of the sperm head following capacitation, likely due to cholesterol removal. After an ionophore A23187-induced acrosome reaction, CRISP2 immunofluorescence disappeared from the apical ridge and the EqS area partly not only owing to the removal of the acrosomal shroud vesicles, but to its presence in a subdomain of EqS. The fate of sperm head CRISP2 was further examined post-fertilization. In vitro matured porcine oocytes were co-incubated with boar sperm cells for 6-8 h and the zygotes were processed for CRISP2 immunofluorescent staining. Notably, decondensation of CRISP2, and thus of the sperm PT, occurred while the sperm nucleus was still fully condensed. CRISP2 was no longer detectable in fertilized oocytes in which sperm nuclear decondensation and paternal pronucleus formation were apparent. This rapid dispersal of CRISP2 in the PT is likely regulated by redox reactions for which its cysteine-rich domain is sensitive. Reduction of disulfide bridges within CRISP2 oligomers may be instrumental for PT dispersal and elimination

Quantitative Imaging of Protein-Protein Interactions by Multiphoton Fluorescence Lifetime Imaging Microscopy using a Streak camera

Fluorescence Lifetime Imaging Microscopy (FLIM) using multiphoton excitation techniques is now finding an important place in quantitative imaging of protein-protein interactions and intracellular physiology. We review here the recent developments in multiphoton FLIM methods and also present a description of a novel multiphoton FLIM system using a streak camera that was developed in our laboratory. We provide an example of a typical application of the system in which we measure the fluorescence resonance energy transfer between a donor/acceptor pair of fluorescent proteins within a cellular specimen.Comment: Overview of FLIM techniques, StreakFLIM instrument, FRET application

Weyl-Underhill-Emmrich quantization and the Stratonovich-Weyl quantizer

Weyl-Underhill-Emmrich (WUE) quantization and its generalization are considered. It is shown that an axiomatic definition of the Stratonovich-Weyl (SW) quantizer leads to severe difficulties. Quantization on the cylinder within the WUE formalism is discussed.Comment: 15+1 pages, no figure

The rigged Hilbert space approach to the Lippmann-Schwinger equation. Part II: The analytic continuation of the Lippmann-Schwinger bras and kets

The analytic continuation of the Lippmann-Schwinger bras and kets is obtained and characterized. It is shown that the natural mathematical setting for the analytic continuation of the solutions of the Lippmann-Schwinger equation is the rigged Hilbert space rather than just the Hilbert space. It is also argued that this analytic continuation entails the imposition of a time asymmetric boundary condition upon the group time evolution, resulting into a semigroup time evolution. Physically, the semigroup time evolution is simply a (retarded or advanced) propagator.Comment: 32 pages, 3 figure

The Weyl bundle as a differentiable manifold

Construction of an infinite dimensional differentiable manifold ${\mathbb R}^{\infty}$ not modelled on any Banach space is proposed. Definition, metric and differential structures of a Weyl algebra and a Weyl algebra bundle are presented. Continuity of the $\circ$-product in the Tichonov topology is proved. Construction of the $*$-product of the Fedosov type in terms of theory of connection in a fibre bundle is explained.Comment: 31 pages; revised version - some typoes have been eliminated, notation has been simplifie

Microscopic and flow cytometric semen assessment of Dutch AI-bucks : effect of semen processing procedures and their correlation to fertility

This study was done to determine the effects of processing techniques on the quality of semen from Dutch AI-bucks with the view on improving pregnancy rates after artificial insemination (AI) with liquid or frozen–thawed semen. Motility of spermatozoa was estimated under a microscope whereas the percentage live spermatozoa and the percentage live spermatozoa with intact acrosomes were determined by means of flow cytometry. Aspects of semen processing that were investigated are storage temperature of liquid semen (i), the effect of glycerol on liquid-stored semen (ii), removal of seminal plasma (iii) and type of extender (iv). The correlation between semen quality and fertility rates in inseminated does was also investigated. The percentage motile spermatozoa in semen stored in liquid form for 72 h progressively declined over time, irrespective of whether storage occurred at 4 or 18 °C. The percentage motile spermatozoa in semen stored at 18 °C was similar to that in semen stored at 4 °C if stored for 24 h but lower if stored for 48 h. Goats differ in the sensitivity of their spermatozoa to the deleterious effects of glycerol. Neither the removal of seminal plasma nor the type of extender had any effect on semen quality before freezing but semen frozen in a Tris–citric acid-glucose (TCG) buffer with egg yolk without removal of the seminal plasma had better quality after thawing than semen frozen in another diluent or after removal of seminal plasma. Remarkably no significant correlation between fertility and membrane integrity of spermatozoa could be found. Thus, although integrity assays for spermatozoa are useful to asses resistance to semen handling, the validity of these assays for predicting fertility is questioned.The authors acknowledge the contributions of the Dutch Goat AI Co-operation (GKN) by granting us access to their breeding bucks and thereby facilitating this research. This study was performed as part of the EU-supported projects: CRAFT FA-S2-9207 to strengthen the European goat industry by developing a new reproduction strategy, including the distribution of kidding during the year and AITECH for dissemination of Animal Insemination Technology for goats in Europe (Contract No. QLK5-CT-2002-30252)

The rigged Hilbert space approach to the Lippmann-Schwinger equation. Part I

We exemplify the way the rigged Hilbert space deals with the Lippmann-Schwinger equation by way of the spherical shell potential. We explicitly construct the Lippmann-Schwinger bras and kets along with their energy representation, their time evolution and the rigged Hilbert spaces to which they belong. It will be concluded that the natural setting for the solutions of the Lippmann-Schwinger equation--and therefore for scattering theory--is the rigged Hilbert space rather than just the Hilbert space.Comment: 34 pages, 1 figur

Deformation Quantization of Bosonic Strings

Deformation quantization of bosonic strings is considered. We show that the light-cone gauge is the most convenient classical description to perform the quantization of bosonic strings in the deformation quantization formalism. Similar to the field theory case, the oscillator variables greatly facilitates the analysis. The mass spectrum, propagators and the Virasoro algebra are finally described within this deformation quantization scheme.Comment: 33+1 pages, harvmac file, no figure

On two superintegrable nonlinear oscillators in N dimensions

We consider the classical superintegrable Hamiltonian system given by $H=T+U={p^2}/{2(1+\lambda q^2)}+{{\omega}^2 q^2}/{2(1+\lambda q^2)}$, where U is known to be the "intrinsic" oscillator potential on the Darboux spaces of nonconstant curvature determined by the kinetic energy term T and parametrized by {\lambda}. We show that H is Stackel equivalent to the free Euclidean motion, a fact that directly provides a curved Fradkin tensor of constants of motion for H. Furthermore, we analyze in terms of {\lambda} the three different underlying manifolds whose geodesic motion is provided by T. As a consequence, we find that H comprises three different nonlinear physical models that, by constructing their radial effective potentials, are shown to be two different nonlinear oscillators and an infinite barrier potential. The quantization of these two oscillators and its connection with spherical confinement models is briefly discussed.Comment: 11 pages; based on the contribution to the Manolo Gadella Fest-60 years-in-pucelandia, "Recent advances in time-asymmetric quantum mechanics, quantization and related topics" hold in Valladolid (Spain), 14-16th july 201