Analyse comparative de la séquence sexuelle chez les Ipopodes Oniscoidea de la série ligienne en relation avec le développement de la volvation chez les Armadillidiidae et les Armadillidae

A comparative study of the sexual sequence is carried out in four Oniscoidea of the ligiid series Porcellio laevis, Metoponorthus sexfasciatus (Porcellionidae), Armadillidium vulgare (Armadillidiidae) and Armadillo officinalis (Armadillidae). The differences between the four species chiefly appear in the precopulatory manoeuvres. The integration of coiling or uncoiling reactions in the sexual sequence of coiling forms is studied in two genera: Armadillidium and Armadillo. Results of the behavioral study provide information about the evolution of the group.     

Evidence for pseudopregnancy and induced ovulation in captive wolverines (Gulo gulo)

Progesterone levels and vaginal smears were monitored to detect estrus and formation of corpora lutea during the first year of a 4-year study of reproduction in captive wolverines. No evidence of spontaneous ovulation was detected during the first year, and most females did not attain complete vaginal cornification. Follicle stimulating hormone was used in subsequent years to induce estrus in several females, and human chorionic gonadotropin (hCG) was used to induce ovulation. Females treated with hCG were artificially inseminated with fresh wolverine semen. Prolonged elevation of serum progesterone above 1 ng/ml was only observed in females that received hCG. The profiles and duration of the progesterone secretory pattern of these females closely resembled that of other mustelids that exhibit a prolonged delay of implantation. Progesterone remained below 1 ng/ml throughout the year in all females that did not receive hCG. No kits were produced. The data suggest that ovulation in this species is normally induced by coitus, and that pseudopregnancy can occur, lasting as long as pregnancy. © 1993 Wiley-Liss, Inc.     

Fungal artificial chromosomes for mining of the fungal secondary metabolome

Background

With thousands of fungal genomes being sequenced, each genome containing up to 70 secondary metabolite (SM) clusters 30–80 kb in size, breakthrough techniques are needed to characterize this SM wealth.

Results

Here we describe a novel system-level methodology for unbiased cloning of intact large SM clusters from a single fungal genome for one-step transformation and expression in a model host. All 56 intact SM clusters from Aspergillus terreus were individually captured in self-replicating fungal artificial chromosomes (FACs) containing both the E. coli F replicon and an Aspergillus autonomously replicating sequence (AMA1). Candidate FACs were successfully shuttled between E. coli and the heterologous expression host A. nidulans. As proof-of-concept, an A. nidulans FAC strain was characterized in a novel liquid chromatography-high resolution mass spectrometry (LC-HRMS) and data analysis pipeline, leading to the discovery of the A. terreus astechrome biosynthetic machinery.

Conclusion

The method we present can be used to capture the entire set of intact SM gene clusters and/or pathways from fungal species for heterologous expression in A. nidulans and natural product discovery.

Electronic supplementary material

The online version of this article (doi:10.1186/s12864-015-1561-x) contains supplementary material, which is available to authorized users.     

Comparative Evaluation of Methods for Estimating Retinal Ganglion Cell Loss in Retinal Sections and Wholemounts

To investigate the reliability of different methods of quantifying retinal ganglion cells (RGCs) in rat retinal sections and wholemounts from eyes with either intact optic nerves or those axotomised after optic nerve crush (ONC). Adult rats received a unilateral ONC and after 21 days the numbers of Brn3a⁺, βIII-tubulin⁺ and Islet-1⁺ RGCs were quantified in either retinal radial sections or wholemounts in which FluoroGold (FG) was injected 48 h before harvesting. Phenotypic antibody markers were used to distinguish RGCs from astrocytes, macrophages/microglia and amacrine cells. In wholemounted retinae, counts of FG⁺ and Brn3a⁺ RGCs were of similar magnitude in eyes with intact optic nerves and were similarly reduced after ONC. Larger differences in RGC number were detected between intact and ONC groups when images were taken closer to the optic nerve head. In radial sections, Brn3a did not stain astrocytes, macrophages/microglia or amacrine cells, whereas βIII-tubulin and Islet-1 did localize to amacrine cells as well as RGCs. The numbers of βIII-tubulin⁺ RGCs was greater than Brn3a⁺ RGCs, both in retinae from eyes with intact optic nerves and eyes 21 days after ONC. Islet-1 staining also overestimated the number of RGCs compared to Brn3a, but only after ONC. Estimates of RGC loss were similar in Brn3a-stained radial retinal sections compared to both Brn3a-stained wholemounts and retinal wholemounts in which RGCs were backfilled with FG, with sections having the added advantage of reducing experimental animal usage.