Drosophila and Agriculture Pest insects Core

Coordinator: Prof. Giuseppe Saccone

email:giuseppe.saccone@unina.it

The Department of Biology at the University of Naples Federico II operates an Insect Molecular Genetic Facility (IMGF) that houses state of the art equipment for developing transgenic organisms and applying gene editing approaches through the use of microinjection technology. A dedicated insect genetics team provides access to functional genomics technologies, including genetic modification to the soma and germ-line of insects, screening services for genetically modified insects, genetically modified insect line development, mapping, and validation. IMGF offers access to researchers and students to learn how to use the model insects Drosophila melanogaster and Ceratitis capitata (Medfly), a major agricultural pest, to investigate biological problems ranging from the genetics of development to gene regulation and biocontrol of invasive pest species.  IMGF offers its know-how and expertise to meet the demands of scientists interested in using the evolutionary approach for their functional studies of genes and proteins. Drosophila is used worldwide as a model organism for basic research and for human disease modeling, the dissection of cellular morphogenesis, behavioral studies, and aging. The Medfly is an emerging model for developing novel eco-friendly biocontrol strategies targeting many other related agricultural pest species.

The IMGF Facility allows students to learn how to apply the cutting-edge CRISPR/CAS9 technology. This short-term lab experience will help them understand and investigate the link between gene and phenotype in a pluricellular complex organism. The student will be able to induce and observe mutant phenotypes within 1-3 weeks from the embryos microinjection in non-vertebrate animals, hence more accessible also from a regulatory standpoint.

The equipment includes:

• An insect rearing facility for containment of the insects, having a room with controlled temperature and day/night cycle, insect traps/attractors, separated from the external environment by three other rooms and four doors/barriers.
• Laser-based needle puller for quartz needles (useful also for injections of mosquitoes eggs)
• Micromanipulator and microinjector
• Light and fluorescent microscopes
• Cameras to capture color images

The IMGF Facility allows researchers to perform the following experimental procedures:

• Gain- (by transient embryonic-larval expression or transgenesis) and Loss-of-function (by RNA interference, CRISPR/Cas9) of genes
• Generation of transgenic insect lines
• Generation of Knock-in and Knock-out insect transgenic lines.
• Morphological inspection and molecular studies (embryos and tissues in situ hybridizations, protein immunolocalization)
• Pesticides Assays
• Behavioral Assays

ACKNOWLEDGEMENTS

All the users and collaborators of the IMGF Facility are obligated to acknowledge it in publications as it follows: “The authors acknowledge the Drosophila and Agriculture Pest insects Core at the Department of Biology of University of Naples Federico II for their scientific support”.

Medaka Fish Core

Coordinator: Prof. Ivan Conte

email: ivan.conte@unina.it

The Department of Biology at University of Naples Federico II offers researchers and Ph.D students access to its Medaka Fish Core which provide resources for research projects and  teaching classes for Ph.D and ungraduated students using Medaka fish as model system. There is a specific room in Animal House with controlled temperature and photoperiod, and one microinjection room for teaching and research.

Medaka fish develop externally and both embryos and adults are optically transparent, allowing direct observation of pathological tissue alterations. Medaka fish breed regularly, produce large clutches of offspring and can be easily handled and maintained in the laboratory allowing for small or large-scale genetic and molecular screens. Transgenic expression of exogenous genes and experimental knockout of endogenous genes can be carried out using relatively straightforward techniques such as targeted mutagenesis using CRISPR/Cas9 system.

The Medaka Core allows researchers to take advantage of the unique features of the Medaka embryos and provides useful tools to test hypotheses concerning genetic disorders, aging, cancer, sex determination, toxicology, oxidative stress, behavior, epigenetics, and evolution by using the following experimental procedures:

• Gain- and Loss-of-function of genes

• Generation of transgenic fish lines

• Generation of Knock-in and, Knock-out Medaka lines.

• Morphological inspection and molecular studies (RNA ISH and immunofluorescence)

• Toxicology Assays

• Live-imaging Assays

• Behavioral Assays

The use of Medaka Fish at University of Naples is normed in accordance with national and European regulations in force (Legislative Decree 26/2014, Directive 2010/63/EU), to guarantee the best housing conditions and the use of Medaka model system and promote good scientific practice as well.

Access to the Medaka Facility is allowed only for personnel included in approved projects by the OPBA (Animal Welfare Body) and by the Italian Ministry of Health.

ACKNOWLEDGEMENTS

All the users and collaborators of the Medaka Fish Core are obligated to acknowledge the Core in publications: “The authors acknowledge the Medaka Fish Core at Department of Biology of University of Naples Federico II for their scientific support.”

Mosquito Genetics and Control Core

Coordinator: Prof. Marco Salvemini

email:marco.salvemini@unina.it

The Department of Biology at the University of Naples Federico II operates a Mosquito Genetics and Control Facility (MGCF) that houses state-of-the-art equipment for the study of molecular genetics and for field research activities of insect vector species, with a special focus on the Asian tiger mosquito Aedes albopictus and the sand fly species Phlebotomus perniciosus. The Asian tiger mosquito, Aedes albopictus is a major arboviruses vector and represents an outstanding emerging model organism for the study of developmental genetics, of the host-parasite interactions and the evolution of adaptation mechanisms. It is considered one of the most invasive mosquito species worldwide. It is native of the East Asia area, and, in the last 30 years, it has rapidly spread across the world, in every continent except Antarctica.

A dedicated mosquito team provides access to functional genomics technologies, including in silico gene identification and validation and in vivo functional assays. MGCF offers access to researchers, students,and individuals to learn how to use the model insect Aedes albopictusto investigate biological problems ranging from the genetics of sex determination, larval vision system and lifespan as well as to apply state-of-the-art field techniques for the field monitoring and eco-friendly control. MGCF offers its know-how and expertise to meet the demands of scientists, private companies or public administrations interested in using the study of genes and proteins of the Asian tiger mosquito or to develop ecofriendly field control programs. In addition, the MGCF facility makes its expertise available to companies and public administrations to plan and implement community-engagement and Citizen science campaigns to raise awareness and involvement of local communities and to increase the probability of success of insect vector control programs.

The MGCF allows researchers to perform the following experimental procedures:

• In silico gene study
• In vivo gene silencing by dsRNA microinjections
• In vivo gene silencing by dsRNA oral delivery
• Toxicology assays on mosquito larvae
• Biological assays for mosquito repellents
• Behavioral assays at larval and adult stage
• Field monitoring of mosquito and sand fly species by state-of-the-art trapping systems

The MGCF equipment include:

• Servers for bioinformatics analysis of insect vector genomes.
• A rearing facility for containment of the insects, with a double-door system and with controlled temperature, humidity, and day/night cycle.
• Hemotek PS6 blood feeding system for hematophagous insects.
• Micromanipulation and microinjection devices.• Light and fluorescence microscopes.
• HD cameras for insect video recording.
• Equipment for field analysis of natural populations of Asian tiger mosquitoes and sand flies (ovitraps, gravitraps, CDC Light traps, BG-Sentinel-2 traps, manual electric aspirators for analysis byHuman Landing Catches).
• Ten stereomicroscopy stations for didactic activity.

ACKNOWLEDGEMENTS

All the users and collaborators of the MGCF are obligated to acknowledge it in publications as it follows: “The authors acknowledge the Mosquito Genetics and Control Facility at the Department of Biology of University of Naples Federico II for their scientific support.”

Octopus Core

Coordinator: Prof. Anna Di Cosimo

email: dicosmo@unina.it

Octopuses have been creatures seen for a long time as curious and explorative animals. They are Cephalopods, smart active predators with a unique camouflage system and   advanced cognitive abilities. Cephalopods are mollusks but unlike general mollusks like snails or mussels they have very complex sensory systems and are highly mobile. The cephalopods brains are larger and more complex than those of any invertebrate.  During evolution, cephalopods had to compete with fish and developed from well-armed slow animals to unarmed fast and active predators. To make this possible they had to develop multiple sensory organs and evolve the molluscan ganglion ring design of the central nervous system ‘cephalized’. The neural architecture became increasingly complex, organized, and subdivided into lobes .The laboratory uses multiple different research methods from classic and advanced morphology, molecular biology, neurobiology and ethology to investigate the unique properties of cephalopods.

The Octopus Core allows researchers to take advantage of the unique features of the octopus  and provides useful tools to test hypotheses by using the following experimental procedures:

• Standard histology
• Modern imaging techniques
• Molecular Biology Methods
• Genetic Methods
• Neurobiology
• Behavioral Experiments

ACKNOWLEDGEMENTS

All the users and collaborators of the Octopus Core are obligated to acknowledge the Core in publications: “The authors acknowledge the Octopus Core at Department of Biology of University of Naples Federico II for their scientific support.”

Photosynthetic model organisms Core

Coordinator: Dr. Simone Landi

email: simone.landi@unina.it

The Department of Biology at University of Naples Federico II offers researchers and Ph.D students access to its Photosynthetic model organisms Core which provide resources for research projects and  teaching classes for Ph.D and ungraduated students using different vegetal model system including plants and algae. There are specific spaces including greenhouse and growth chambers with controlled temperature and photoperiod for teaching and research.

Photosynthetic model organisms core offers plant growth and develop in-vitro and in-soil conditions. Different plants model organisms (and genotypes of these) are available and regularly growth in the greenhouse of the Department of Biology. Different aspects of photosynthetic metabolism and physiology can be monitored including response to abiotic stresses, production of metabolites, variations in primary and secondary metabolism. Furthermore, assistance and skills can be put to employ bioinformatic, phylogenetic and NGS analyses.

The Photosynthetic model organisms Core allows researchers to take advantage of the unique features of the vegetal organisms and provides useful tools to test hypotheses concerning physiology metabolism, and molecular aspects by using the following experimental procedures:

• Analysis of protein metabolism
• Enzymatic assays
• Expression analysis
• RNA sequencing
• Immunoblotting

ACKNOWLEDGEMENTS

All the users and collaborators of the Photosynthetic model organisms core are obligated to acknowledge the facility in publications: “The authors acknowledge the Photosynthetic model organisms Core at Department of Biology of University of Naples Federico II for their scientific support.”

Plant Science Core

Coordinator: Prof. Carmen Arena

email: carmen.arena@unina.it

The Department of Biology at University of Naples Federico II offers internal and external researchers and Ph.D students access to its Plant Science Core support for ecological, investigations concerning different model plant organisms. The ecological aspects of photosynthesis related to structural and functional adaptations of plants in natural and polluted environments can be assessed, as well as the strategies of plants and macro algae in response to mono and multiple abiotic stresses.

The Plant Core has the following infrastructures:

Instruments

• Pulse Amplitude Fluorometers for conventional fluorescence emission analyses and imaging fluorometry
• Instruments for measurements of gas exchanges (CO2 e O2)
• Growth chambers equipped for environmental variables and light quality control

ACTIVITIES

• Study of photosynthesis of higher plants and macroalgae in vivo
• Performance indicators of the photosynthetic apparatus
• Assessment of environmental stress and recovery through ecological, functional and structural indicators
• Diagnostics of the functionality of photosystems

ACKNOWLEDGEMENTS

All the users and collaborators of the Plant Science Core are obligated to acknowledge the facility in publications: “The authors acknowledge the Plant Science Core at Department of Biology of University of Naples Federico II for their scientific support.”

Xenopus laevis Core

Coordinator: Prof. Rosa Carotenuto

email: rosa.carotenuto@unina.it

The Department of Biology at University of Naples Federico II offers researchers and PhD students access to its Xenopus laevis Core which provide resources for research projects and teaching classes for PhD and ungraduated students using Xenopus laevis embryos as model system. There is a specific room in Animal House with controlled temperature and photoperiod, and one fertilization room for teaching and research.

Xenopus laevis embryos develop externally and are optically transparent, allowing direct observation of developmental alterations. Xenopus laevis breed regularly, produce large clutches of offspring and can be easily handled and maintained in the laboratory allowing for small or large-scale morphological and molecular screens.

The Xenopus laevis Core allows researchers to take advantage of the unique features of the Xenopus embryos and provides useful tools to test hypotheses concerning toxicology, teratogenesis (chemicals, drugs, environmental pollutants) oxidative stress, behavior, epigenetics, and morphology by using the following experimental procedures:

• Gain- and Loss-of-function of genes (with microinjection)
• Knock-out. with Morpholino
• Morphological ultrastructural inspection (histology, immunofluorescence on section and whole mount, TEM)

Molecular studies (RNA ISH and, PCR, Real time PCR)

Biochemical studies (WB, IHC)

• Toxicology Assays
• Live-imaging Assays
• Behavioral Assays

Teratogenic Assay (FETAX Assay)

The use of Xenopus laevis at University of Naples is normed in accordance with national and European regulations in force (Legislative Decree 26/2014, Directive 2010/63/EU), to guarantee the best housing conditions and the use of Xenopus model system and promote good scientific practice as well.

Access to the Xenopus model is allowed only for personnel included in approved projects by the OPBA (Animal Welfare Body) and by the Italian Ministry of Health.

ACKNOWLEDGEMENTS

All the users and collaborators of the Xenopus laevis model are obligated to acknowledge the Core in publications: “The authors acknowledge the Xenopus laevis Core at Department of Biology of University of Naples Federico II for their scientific support.”

Zebrafish Core

Coordinator: Prof. Ida Ferrandino

email: ida.ferrandino@unina.it

Zebrafish Core is a part of the Animal House of the Biology Department and is equipped for the stabling and use of Danio rerio fish (Zebrafish) to support the scientific activity of university researchers, allowing them to carry out research “in vivo” in that model animal under the legislation in force.

Zebrafish is an important model organism in the study of vertebrate biology and more recently in eco-toxicological research. Traditionally used by developmental biologists, zebrafish are also useful for modeling a wide variety of human diseases and for screening small molecules for toxicity or pharmacological function. It is an ideal model organism in molecular biology, and biogenetics research fields, such as drug screening, disease research, and aquatic ecotoxicology research. The main advantage of zebrafish as a model organism is that it is easy to rear, handle, observe, and analyzeduring its three stages of development (embryo, juvenile, and adult).

The goal of this facility is to enable access to the zebrafish model system for local research groups.

Zebrafish Core allows researchers to take advantage of the unique features of this organism model through various experimental methods such as the preparation, treatment, and imaging of embryos, larvae, and adults to test:

• Toxicology Assays
• Live-imaging Assays
• Behavioural Assays
• Teratogenicity
• Genotoxicity
• Cito-histochemical and Ultrastructural analysis
• Oxidative stress
• Gene expression analysis

The use of Zebrafish at the University of Naples is normed following National and European regulations in force (Legislative Decree 26/2014, Directive 2010/63/EU) concerning the protection of animals used for scientific or educational purposes.

Accede to the Zebrafish Facility only for employees included in approved projects by the OPBA (Animal Welfare Body) and by the Italian Ministry of Health.

ACKNOWLEDGEMENTS

All the collaborators of the Zebrafish Facility are obligated to acknowledge the facility in publications: “The authors acknowledge the Zebrafish Core at Department of Biology of University of Naples Federico II for their scientific support.”