Control of ovulation with a GnRH agonist after superstimulation of follicular growth in buffalo: fertilization and embryo recovery

The potential to use a GnRH agonist bioimplant and injection of exogenous LH to control the time of ovulation in a multiple ovulation and embryo transfer (MOET) protocol was examined in buffalo. Mixed-parity buffalo (Bubalus bubalis; 4-15-year-old; 529 13 kg LW) were randomly assigned to one of five groups (n = 6): Group 1, conventional MOET protocol; Group 2, conventional MOET with 12 It delay in injection of PGF(2alpha); Group 3, implanted with GnRH agonist to block the pre-ovulatory surge release of LH; Group 4, implanted with GnRH agonist and injected with exogenous LH (Lutropin(R), 25 mg) 24 h after 4 days of superstimulation with FSH; Group 5, implanted with GnRH agonist and injected with LH 36 h after superstimulation with FSH. Ovarian follicular growth in all buffaloes was stimulated by treatment with FSH (Folltropin-V(R), 200 mg) administered over 4 days, and was monitored by ovarian ultrasonography. At the time of estrus, the number of follicles greater than or equal to8 mm. was greater (P < 0.05) for buffaloes in Group 2 (12.8) than for buffaloes in Groups 1 (8.5), 3 (7.3), 4 (6.1) and 5 (6.8), which did not differ. All buffaloes were mated by AI after spontaneous (Groups 1-3) or induced (Groups 4 and 5) ovulation. The respective number of buffalo that ovulated, number of corpora lutea, ovulation rate (%), and embryos + oocytes recovered were: Group 1 (2, 1.8 +/- 1.6, 18.0 +/- 13.6, 0.2 +/- 0.2); Group 2 (4, 6.1 +/- 2.9, 40.5 +/- 17.5, 3.7 +/- 2.1); Group 3 (0, 0, 0, 0); Group 4 (6, 4.3 +/- 1.2, 69.3 +/- 14.2, 2.0 +/- 0.9); and Group 5 (1, 2.5 +/- 2.5, 15.5 +/- 15.5, 2.1 +/- 2.1). All buffaloes in Group 4 ovulated after injection of LH and had a relatively high ovulation rate (69%) and embryo recovery (46%). It has been shown that the GnRH agonist-LH protocol can be used to improve the efficiency of MOET in buffalo. (C) 2002 Elsevier Science Inc. All rights reserved

Caratteristiche di fermentazione in vitro di alcuni foraggi tropicali.

Roma (Italy

Architecture of tree species of different strata developing in environments with the same light intensity in a semideciduous forest in southern Brazil

We aimed to answer the following questions related to the architecture of individuals 0.5-3.0 m in height belonging to understory or canopy/emergent layer tree species: "Is there a difference between individuals belonging to different strata developing in environments with the same light intensity, in terms of their architecture?"; and "Given the same light intensity, do understory species exhibit less crown plasticity than do canopy/emergent layer species?" Thirteen architectural variables were evaluated in 80 individuals per species. We found that understory species showed greater increases in stem thickness and leaf number, as well as wider, deeper crowns, longer branches, greater self-shading and less crown plasticity. Stems and crowns were more slender in the canopy species than in the understory species. These differences might be due to the trade-off between vertical and lateral growth. Our results indicate that, regardless of the group to which they belong, species are best able to take advantage of light conditions in the understory of the forest. However, because they demand more light, canopy species showed a growth form that resulted in an architecture that is likely to enable better light capture in the understory

Cell Fate Analysis of Embryonic Ventral Mesencephalic Grafts in the 6-OHDA Model of Parkinson's Disease

<div><p>Evidence from carefully conducted open label clinical trials suggested that therapeutic benefit can be achieved by grafting fetal dopaminergic (DAergic) neurons derived from ventral mesencephalon (VM) into the denervated striatum of Parkinson's disease (PD) patients. However, two double-blind trials generated negative results reporting deleterious side effects such as prominent dyskinesias. Heterogeneous composition of VM grafts is likely to account for suboptimal clinical efficacy.</p> <p>We consider that gene expression patterns of the VM tissue needs to be better understood by comparing the genetic signature of the surviving and functioning grafts with the cell suspensions used for transplantation. In addition, it is crucial to assess whether the grafted cells exhibit the DAergic phenotype of adult substantia nigra pars compacta (SNpc). To investigate this further, we used a GFP reporter mouse as source of VM tissue that enabled the detection and dissection of the grafts 6 weeks post implantation. A comparative gene expression analysis of the VM cell suspension and grafts revealed that VM grafts continue to differentiate post-implantation. In addition, implanted grafts showed a mature SNpc-like molecular DAergic phenotype with similar expression levels of TH, Vmat2 and Dat. However, by comparing gene expression of the adult SNpc with dissected grafts we detected a higher expression of progenitor markers in the grafts. Finally, when compared to the VM cell suspension, post-grafting there was a higher expression of markers inherent to glia and other neuronal populations.</p> <p>In summary, our data highlight the dynamic development of distinctive DAergic and non-DAergic gene expression markers associated with the maturation of VM grafts in vivo. The molecular signature of VM grafts and its functional relevance should be further explored in future studies aimed at the optimization of DAergic cell therapy approaches in PD.</p> </div

List of primers′ sequences used for qRT-PCR.

<p>List of primers′ sequences used for qRT-PCR.</p

Increased TH-positive cells post-grafting. (A) Smear preparation of VM cell suspension.

<p>(<b>A</b>) TH staining (A1). DAPI staining (A2). Merged pictures (A3). GFP staining of grafts (<b>B1</b>), TH staining of grafts (<b>B2</b>). Higher magnification of the marked area showing TH cells extending axons which re-innervate the surrounding striatum (<b>B3</b>). (<b>C</b>) <b>Percentage of TH+ cells pre- and post-grafting.</b> Data are presented as mean ± SEM, n = 6. ** p<0.01 by two-tailed student's t-test. SEM, standard error of the mean.</p