Mitophagy in Huntington's disease.

Huntington's disease (HD), as well as Parkinson's disease and Alzheimer's disease, belong to a group of neurodegenerative diseases characterized by common features, such as the progressive loss of neurons and the presence of pathogenic forms of misfolded protein aggregates. A quality control system such as autophagy is crucial for the clearance of protein aggregates and dysfunctional organelles and thus essential for the maintenance of neuronal homeostasis. The constant high energy demand of neuronal tissue links neurodegeneration to mitochondria. Inefficient removal of damaged mitochondria is thought to contribute to the pathogenesis of neurodegenerative diseases such as HD. In addition, direct involvement of the huntingtin protein in the autophagic machinery has been described. In this review, we focus on mitophagy, a selective form of autophagy responsible for mitochondrial turnover. We also discuss the relevance of pharmacological regulation of mitophagy in the future therapeutic approach to neurodegenerations, including HD.

Association of the transcription profile of bovine oocytes and embryos with developmental potential.

Although improvements in culture system have enhanced in vitro embryo production, success rates are still not adequate. The reasons for developmental arrest of a part of in vitro produced embryos are unknown, but are connected in part with low cytoplasmic competence of oocytes. The immaturity of cytoplasm can negatively influence fertilization efficiency and subsequent progression through embryonic genome activation (EGA), which are necessary steps in further pre-implantation development. A large number of studies have compared mRNA abundance among oocytes with different developmental competence with the aim to find markers of the normal embryo development. The amount of mitochondrial DNA (mtDNA) and mRNA for mitochondrial transcriptional factors directing oxidative phosphorylation belongs to such promising markers. Nevertheless, recently published studies revealed that the mammalian embryo is able to compensate for a reduced level of mtDNA in oocyte during subsequent pre-implantation development. The search for other molecular markers is in progress. Characterization of oocyte and embryonic mRNA expression patterns during the pre-implantation period, and their relationship to the successful in vitro and in vivo development will be essential for defining the optimized culture conditions or the nuclear transfer protocols. Microarrays technology enables us to reveal the differentially expressed genes during EGA, and to compare the expression profile of in vivo and in vitro produced embryos. Recent evidence indicates that the depletion of the pool of stored maternal mRNAs is critical for subsequent embryo development. All these experiments gradually offer a list of possible candidates for quality and developmental competence markers for mammalian oocytes and pre-implantation embryos.

Transcriptomic analysis of in vivo and in vitro produced bovine embryos revealed a developmental change in cullin 1 expression during maternal-to-embryonic transition.

Pre-implantation embryos derived by in vitro fertilization differ in their developmental potential from embryos obtained in vivo. In order to characterize changes in gene expression profiles caused by in vitro culture environment, we employed microarray constructed from bovine oocyte and preimplantation embryo-specific cDNAs (BlueChip, Université Laval, Québec). The analysis revealed changes in the level of 134 transcripts between in vitro derived (cultured in COOK BVC/BVB media) and in vivo derived 4-cell stage embryos and 97 transcripts were differentially expressed between 8-cell stage in vitro and in vivo embryos. The expression profiles of 7 selected transcripts (BUB3, CUL1, FBL, NOLC1, PCAF, GABPA and CNOT4) were studied in detail. We have identified a switch from Cullin 1-like transcript variant 1 to Cullin 1 transcript variant 3 (UniGene IDs BT.36789 and BT.6490, respectively) expressions around the time of bovine major gene activation (8-cell stage). New fibrillarin protein was detected by immunofluorescence already in early 8-cell stage and this detection correlated with increased level of fibrillarin mRNA. The qRT-PCR analysis revealed significant differences in the level of BUB3, NOLC1, PCAF, GABPA and CNOT4 gene transcripts between in vivo derived (IVD) and in vitro produced (IVP) embryos in late 8-cell stage. The combination of these genes represents a suitable tool for addressing questions concerning normal IVD embryo development and can be potentially useful as a marker of embryo quality in future attempts to optimize in vitro culture conditions.

Gene expression during minor genome activation in preimplantation bovine development.

The main goal of this study was to identify mRNA transcripts whose content increases during bovine minor embryonic genome activation. We compared the gene expression profile of the bovine 4-cell-stage embryo and MII oocyte using the technique of suppression subtractive hybridization. Differentially expressed amplicons were subcloned, and 60 of them were sequenced. The resulting DNA sequences were compared with GenBank databases using BLAST search. The expression of five differentially expressed genes with an apparent function in cell cycle progression, chromatin remodeling, and splicing or translation initiation was further characterized by a real-time RT-PCR. Centromere protein F, 350/400ka (CENPF), and splicing factor arginine/serine-rich 3 (SRFS3) show an increase in mRNA content during the 2- to 4-cell and late 8-cell stages. For the high mobility group nucleosomal binding domain 2 (HMGN2), the level of mRNA increases in 2- to 4-cell and morula embryos. The transcription of splicing factor SRFS3 is alpha-amanitin sensitive both during 4-cell and late 8-cell stages. The transcription of CENPF and HMGN2 is alpha-amanitin sensitive only at late 8-cell stage and morula, respectively. SRFS3 represents the first described gene with an important function in preimplantation development, which is also expressed during bovine minor genome activation, and it is alpha-amanitin sensitive during this period. All described genes can play an important role in the preimplantation development of bovine embryos.

[Various indices of microclimate in the newly built and re-adapted stables for race horses]. / Nekteré ukazatele mikroklimatu v nových a adaptovaných stájích sportovních koní

A study was performed to examine microclimate in 14 stables belonging to 10 horsemen's teams and clubs; five of these houses were new-built. In five race-horse stables housing 16 horses each, on an average, where the optimum air temperature ranged from 10 degrees C to 12 degrees C, measurements and examinations were performed in the winter period and the following results were obtained: space per 1 horse housed 42.9 plus or minus 8.7 m-3, relative air humidity 74.3 plus or minus 3.8%, CO2 concentration 0.175 plus or minus 0.027%, NH3 concentration 0.00135 plus or minus 0.00044%. A large majority of horse stables under our conditions lack suitable ventilating equipment for winter and for cold periods. Together with the present-day building technology and with excessive space of box-type houses, this implies that microclimate conditions are unsuitable and harmful to health; in particular, this is true of cold and wet conditions. In the existing stables this problem can be solved by additional heating, preferably with the hot-air system. It is necessary that horse stables should have good thermal-insulation characteristics, with plastered brick walls 45 cm in thickness and with thermally insulated loft. Floors must be solid, hard, and plane. Modern building technology and new materials must secure all the required parameters, with due respect to all factors constituting microclimate and to purposeful layout of race horse stables. It appears desirable to issue a state standard for the construction of horse stables.