Mucuna pruriens (Velvet bean) Rescues Motor, Olfactory, Mitochondrial and Synaptic Impairment in PINK1(B9) Drosophila melanogaster Genetic Model of Parkinson's Disease

The fruit fly Drosophila melanogaster (Dm) mutant for PTEN-induced putative kinase 1 (PINK1B9) gene is a powerful tool to investigate physiopathology of Parkinson's disease (PD). Using PINK1B9 mutant Dm we sought to explore the effects of Mucuna pruriens methanolic extract (Mpe), a L-Dopa-containing herbal remedy of PD. The effects of Mpe on PINK1B9 mutants, supplied with standard diet to larvae and adults, were assayed on 3–6 (I), 10–15 (II) and 20–25 (III) days old flies. Mpe 0.1% significantly extended lifespan of PINK1B9 and fully rescued olfactory response to 1-hexanol and improved climbing behavior of PINK1B9 of all ages; in contrast, L-Dopa (0.01%, percentage at which it is present in Mpe 0.1%) ameliorated climbing of only PINK1B9 flies of age step II. Transmission electron microscopy analysis of antennal lobes and thoracic ganglia of PINK1B9 revealed that Mpe restored to wild type (WT) levels both T-bars and damaged mitochondria. Western blot analysis of whole brain showed that Mpe, but not L-Dopa on its own, restored bruchpilot (BRP) and tyrosine hydroxylase (TH) expression to age-matched WT control levels. These results highlight multiple sites of action of Mpe, suggesting that its effects cannot only depend upon its L-Dopa content and support the clinical observation of Mpe as an effective medication with intrinsic ability of delaying the onset of chronic L-Dopa-induced long-term motor complications. Overall, this study strengthens the relevance of using PINK1B9 Dm as a translational model to study the properties of Mucuna pruriens for PD treatment

The efficiency of in-situ hybridization on human chromosomes with alphoid DNAs is enhanced by previous digestion with AluI and TaqI

Centromeric alphoid DNAs of human chromosomes 6, 9, 16 and Y were employed to obtain information on the molecular mechanism(s) determining cytological effects produced by digestion in situ with AluI and TaqI restriction enzymes, possibly related to the structure of the above-cited areas. The following cytological and biochemical experiments were carried out using the above-mentioned alphoid sequences as probes: (1) standard in-situ hybridization and in-situ hybridization after chromosome cleavage with AluI/TaqI, and (2) filter hybridization on the DNA fractions obtained from the material solubilized and that retained on the slides after digestion in situ with AluI/TaqI. Biochemical data show that cleavage of alphoid DNAs is not prevented by the peculiar organization of centromeric heterochromatin, but such cleavage is not necessarily followed by complete DNA solubilization. The analysis of alphoid sequence cleavage in naked genomic DNA as well as during digestion of fixed chromosomes shows that (1) AluI cuts more efficiently than TaqI, (2) DNA fragments as large as 3-5 kb can be solubilized, and (3) DNA fragments of the same size are found in both fractions of DNA, i.e. that retained on the chromosomes as well as that solubilized from chromosomes. Cytological data show that previous chromosome digestion, mostly with TaqI, increases the hybridization signal area, suggesting that this fact might be due to (1) chromatin reorganization produced by enzyme attack and/or (2) the presence of alphoid DNAs which might be restricted not only to the kinetochore area but also to para/peri-centromeric heterochromatin. Lastly, centromere DNA solubilization as a consequence of restriction enzyme cleavage seems to vary from chromosome to chromosome, thus suggesting that centromeric regions do not represent a homogeneous class of constitutive heterochromatin

Cardiac responses to sucrose reception and intake in the adult blowfly Protophormia terraenovae

Cardiac activity of intact Protophormia terraenovae [Phormia terraenovae] flies and its taste- and feeding-induced variations were analysed by electrocardiogram recording either simultaneously with nervous spike discharge from stimulated labellar taste sensilla, or during feeding. Results indicate that the cardiac response to taste stimulation consists of the inhibition of fast beating activity and the premature setting in of tachycardic slow beating activity. Although the response was evoked by taste stimulation with solutions of sucrose or NaCl, the former were far more effective than the latter. During feeding, variations in cardiac activity consisted of a sharp reduction in the duration of periods of fast and slow activity, that continued for 20 minutes after the end of feeding. Variations in haemolymphatic circulatory dynamics which, in all likelihood, are consequent to taste- and feeding-induced cardiac changes may influence haemolymphatic volume and pressure in extracardiac compartments. The hypothesis is advanced that the biological meaning of variations in cardiac activity and thus in haemolymphatic pressure, is that of creating conditions favourable to the behaviour the insect has to manifest, in particular as concerns the ingestion of food

Characterization of olfactory sensilla of the olive fly: Behavioral and electrophysiological response to volatile organic compounds from the host plant and bacterial filtrate

The responses of olive fly (Bactrocera oleae) antennal and palpal olfactory receptors to odors emitted by Pseudomonas putida bacterial filtrate and to volatiles from a host plant were evaluated using electrophysiological and behavioral bioassays. Morphological identification of olfactory receptors was also performed. The third antennal segment (flagellum) bears four types of multiporous sensilla: trichoid, short basiconica, clavate and grooved. Maxillary palps have mechanosensory bristles and multiporous basiconica sensilla. In wind-tunnel bioassays, olive fly responses to volatiles emitted by bacterial filtrate were higher than those to culture medium. Bacterial filtrate was more attractive than ammonium carbonate or a mixture of ethyl acetate and acetic acid in ethanol. GC–MS of bacterial filtrate identified some of the chemicals produced by bacterial activity, including methyl thiolacetate, ammonia, 2-pentanone, 2- heptanone, ethyl tiglate and methyl thiocyanate. Electrophysiological investigations proved that antennal sensilla are responsive to bacterial filtrate odor, methyl thiolacetate, olive leaves and olives, as well as to a-pinene, while acetic acid elicited an inhibitory response. Electropalpgrams recorded a specific response to bacterial filtrate by mated males and females, as well as a dose-dependent response relationship to methyl thiolacetate by mated females. The identification of new active volatile compounds in the semiochemical system of the olive fly is promising for the development of innovative control strategies in areawide management

Description of antennal structures of the parasitoid Mallophora ruficauda (Diptera: Asilidae) and its relationship with resources searching behaviour

The robber fly Mallophora ruficauda is a parasitoid of white grubs (Coleoptera: Scarabaeidae) inhabiting in Pampas region of Argentina. Females locate host’s habitat and lay eggs away from the host in tall grasses. After hatching, larvae fall to the ground and actively seek hosts. Previous works suggested that female would detect the presence of host’s chemical cues, but sensory organs involved in olfaction are still unknown. However, few studies have looked at dipteran parasitoids sensilla, and no study has been undertaken in Asilidae species. The aim of this work was to determine the presence, density, distribution and morphology of chemosensilla in M. ruficauda antennae using optic and scanning microscope techniques. We found that antennae have 4 segments: scape, pedicel, postpedicel and style. We identified basiconic and trichoid sensilla, small and long bristles, and sensory pits. Basiconic sensilla are multiporous and are widely spread between the small bristles through the postpedicel. Trichoid sensilla are grouped in 6–8 units on latero-ventral margin of postpedicel, have mobile base, striated wall and an apical porous. Small bristles are present in the pedicel and postpedicel, and long bristles are found in groups on scape and pedicel. Three different types of sensory pits were observed, with basiconic sensilla, distributed along external and internal lateral side of the postpedicel. Considering the morphological characteristics of the antennae, and based on the olfaction biomechanics, the structure and distribution of these cuticular structures of the parasitoid antennae would contribute to the odour detection mechanism in adults of M. ruficauda.Fil: Groba, Hernán Federico. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; ArgentinaFil: Lauria de Cidre, Lilia Susana. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Biodiversidad y Biología Experimental. Laboratorio de Histología Animal; ArgentinaFil: Castelo, Marcela Karina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Ecología, Genética y Evolución de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Ecología, Genética y Evolución de Buenos Aires; Argentin