scholarly journals An adaptable chromosome preparation methodology for use in invertebrate research organisms

2017 ◽  
Author(s):  
Longhua Guo ◽  
Alice Accorsi ◽  
Shuonan He ◽  
Carlos Guerrero-Hernández ◽  
Shamilene Sivagnanam ◽  
...  
Keyword(s):  
Cultured Cells ◽  
Chromosome Preparation ◽  
Pomacea Canaliculata ◽  
Apple Snail ◽  
Metaphase Chromosomes ◽  
Chromosome Architecture ◽  
Complete Life Cycle ◽  
In The Wild ◽  
New Biology

AbstractBackgroundThe ability to efficiently visualize and manipulate chromosomes is fundamental to understand the genetic architecture of organisms. This can be challenging for many invertebrates because conventional chromosome preparation protocols deal primarily hypotonic conditions tailored for use with vertebrates and rely extensively on cultured cells which are rarely available for invertebrates. Hence, a simple and inexpensive chromosome preparation protocol, adaptable to multiple invertebrate model species is needed.ResultsWe optimized a chromosome preparation protocol and applied it to several planarian species (Phylum: Platyhelminthes), the freshwater apple snailPomacea canaliculata(Phylum: Mollusca), and the starlet sea anemoneNematostella vectensis(Phylum: Cnidaria). We showed that both mitotically active adult tissues and embryos can be used as sources of metaphase chromosomes, expanding the potential use of this technique to invertebrates lacking cell lines and/or with limited access to the complete life cycle. Simple hypotonic treatment with DI water was sufficient for karyotyping. The karyotypes we obtained allowed the identification of differences in ploidy and chromosome architecture among otherwise morphologically indistinguishable organisms, as in the case of a mixed population of planarians collected in the wild. Furthermore, we showed that in all tested organisms representing three different phyla, this protocol can be effectively coupled with downstream applications, such as chromosome fluorescentin situhybridization.ConclusionThe simple and inexpensive chromosome preparation protocol reported here can be readily adapted to new invertebrate research organisms in order to expand and accelerate the discovery of new biology in understudied branches of the tree of life.

scholarly journals The Immune Response of the Invasive Golden Apple Snail to a Nematode-Based Molluscicide Involves Different Organs

Biology ◽  
2020 ◽  
Vol 9 (11) ◽  
pp. 371
Author(s):  
Alice Montanari ◽  
Giulia Bergamini ◽  
Agnese Ferrari ◽  
Anita Ferri ◽  
Milena Nasi ◽  
...  
Keyword(s):  
Invasive Species ◽  
Immune Response ◽  
Well Being ◽  
Pomacea Canaliculata ◽  
Apple Snail ◽  
Organ Specific ◽  
Golden Apple Snail ◽  
The Central Nervous System ◽  
Basic And Applied Research

The spreading of alien and invasive species poses new challenges for the ecosystem services, the sustainable production of food, and human well-being. Unveiling and targeting the immune system of invasive species can prove helpful for basic and applied research. Here, we present evidence that a nematode (Phasmarhabditis hermaphrodita)-based molluscicide exerts dose-dependent lethal effects on the golden apple snail, Pomacea canaliculata. When used at 1.7 g/L, this biopesticide kills about 30% of snails within one week and promotes a change in the expression of Pc-bpi, an orthologue of mammalian bactericidal/permeability increasing protein (BPI). Changes in Pc-bpi expression, as monitored by quantitative PCR (qPCR), occurred in two immune-related organs, namely the anterior kidney and the gills, after exposure at 18 and 25 °C, respectively. Histological analyses revealed the presence of the nematode in the snail anterior kidney and the gills at both 18 and 25 °C. The mantle and the central nervous system had a stable Pc-bpi expression and seemed not affected by the nematodes. Fluorescence in situ hybridization (FISH) experiments demonstrated the expression of Pc-bpi in circulating hemocytes, nurturing the possibility that increased Pc-bpi expression in the anterior kidney and gills may be due to the hemocytes patrolling the organs. While suggesting that P. hermaphrodita-based biopesticides enable the sustainable control of P. canaliculata spread, our experiments also unveiled an organ-specific and temperature-dependent response in the snails exposed to the nematodes. Overall, our data indicate that, after exposure to a pathogen, the snail P. canaliculata can mount a complex, multi-organ innate immune response.

DNA sequence mapping in interphase and metaphase chromosomes by fluorescence in situ hybridization

1992 ◽  
Vol 50 (1) ◽  
pp. 496-497
Author(s):  
Barbara Trask ◽  
Susan Allen ◽  
Anne Bergmann ◽  
Mari Christensen ◽  
Anne Fertitta ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Dna Sequence ◽  
Dna Sequences ◽  
Dual Band ◽  
Nick Translation ◽  
Metaphase Chromosomes ◽  
Band Pass ◽  
Texas Red ◽  
Fluorescent Spot

Using fluorescence in situ hybridization (FISH), the positions of DNA sequences can be discretely marked with a fluorescent spot. The efficiency of marking DNA sequences of the size cloned in cosmids is 90-95%, and the fluorescent spots produced after FISH are ≈0.3 μm in diameter. Sites of two sequences can be distinguished using two-color FISH. Different reporter molecules, such as biotin or digoxigenin, are incorporated into DNA sequence probes by nick translation. These reporter molecules are labeled after hybridization with different fluorochromes, e.g., FITC and Texas Red. The development of dual band pass filters (Chromatechnology) allows these fluorochromes to be photographed simultaneously without registration shift.

THE ESSENTIAL CONTRIBUTION OF CAPTIVE SUMATRAN ELEPHANT IN ELEPHANT TRAINING CENTER, WAY KAMBAS NATIONAL PARK FOR WILDLIFE GENETICS CONSERVATION

2020 ◽  
Vol 6 (1) ◽  
Author(s):  
Elly Lestari Rustiati ◽  
Priyambodo Priyambodo ◽  
Yanti Yulianti ◽  
Eko Agus Srihanto ◽  
Dian Neli Pratiwi ◽  
...  
Keyword(s):  
National Park ◽  
In Situ Conservation ◽  
Training Center ◽  
Conservation Effort ◽  
In The Wild ◽  
Wildlife Genetics ◽  
Way Kambas National Park ◽  
Human Conflict ◽  
Essential Contribution

Way Kambas National Park (WKNP) is home of five protected big mammals including sumatran elephants.  It shares its border with 22 of 37 villages surrounding the national park.  Understanding their existence in the wild is a priority, and  wildlife genetics is a crucially needed. Besides poaching and habitat fragmentation, wildlife-human conflict is one big issue.  Elephant Training Center (ETC) in WKNP is built for semi in-situ conservation effort on captive sumatran elephants that mainly have conflict histories with local people.  Participative observation and bio-molecular analysis were conducted to learn the importance of captive Sumatran elephant for conservation effort.  Through captive sumatran elephants, database and applicable methods are expected to be developed supporting the conservation of their population in the wild.  Participative observation and molecular identification was carried on captive sumatran elephants in ETC, WKNP under multiple year Terapan grant of Ministry of Research and Technology Higher Education, Indonesia. Gene sequence and cytological analyses showed that the captive sumatran elephants are closely related and tend to be domesticated.  Translocation among ETC to avoid inbreeding, and maintaining the captive sumatran elephant as natural as possible are highly recommended. Developing genetic database can be a reference for both captive and wild sumatran elephants.

scholarly journals Growth, lifetime, directional movement and myosin-dependent motility of mutant keratin granules in cultured cells

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
S. M. Lehmann ◽  
R. E. Leube ◽  
R. Windoffer
Keyword(s):  
Digital Image Analysis ◽  
Cultured Cells ◽  
Epidermolysis Bullosa Simplex ◽  
Continuous Growth ◽  
Relative Contribution ◽  
Complete Life Cycle ◽  
Directional Movement ◽  
Multiple Characteristics ◽  
Cell Centre

AbstractIntermediate filament polypeptides (IFPs) are prominent components of cytoplasmic aggregates, which are pathognomonic for multiple diseases. Recent observations in cultured cells suggest that they are dynamic and subject to regulated turnover. The emerging concept is that multiple factors contribute to motility and turnover of IFP-containing aggregates. To understand their relative contribution, quantitative tools are needed. The current study addresses this need using epithelial cells producing mutant keratin IFPs that have been identified as the cause of the hereditary blister-forming skin disease epidermolysis bullosa simplex. Digital image analysis of individual granules allowed mapping of their complete life cycle, with information on multiple characteristics at any given time-point. The deduced signet features revealed rapid granule fusion and directed transport from the periphery towards the cell centre, and a limited, ~ 30 min lifetime with a slow, continuous growth phase followed by fast disassembly. As paradigmatic proof-of-principle, we demonstrate that inhibition of myosin II selectively reduces granule movement, linking keratin granule motility to retrograde cortical acto-myosin flow. The newly developed methods and established parameters will help in the characterization of known and the identification of novel regulators of IFP-containing aggregates.

scholarly journals Comparison of Invasive Apple Snail (Pomacea canaliculata) Behaviors in Different Water Temperature Gradients

Water ◽  
2021 ◽  
Vol 13 (9) ◽  
pp. 1149
Author(s):  
Mi-Jung Bae ◽  
Eui-Jin Kim ◽  
Young-Seuk Park
Keyword(s):  
Water Temperature ◽  
Farming Systems ◽  
Freshwater Snail ◽  
Freshwater Ecosystems ◽  
Pomacea Canaliculata ◽  
Activity Levels ◽  
Self Organizing Map ◽  
Apple Snail ◽  
Temperature Changes ◽  
Organic Farming Systems

Pomacea canaliculata (known as invasive apple snail) is a freshwater snail native to South America that was introduced into many countries (including Asia and North America) as a food source or for organic farming systems. However, it has invaded freshwater ecosystems and become a serious agricultural pest in paddy fields. Water temperature is an important factor determining behavior and successful establishment in new areas. We examined the behavioral responses of P. canaliculata with water temperature changes from 25 °C to 30 °C, 20 °C, and 15 °C by quantifying changes in nine behaviors. At the acclimated temperature (25 °C), the mobility of P. canaliculata was low during the day, but high at night. Clinging behavior increased as the water temperature decreased from 25 °C to 20 °C or 15 °C. Conversely, ventilation and food consumption increased when the water temperature increased from 25 °C to 30 °C. A self-organizing map (an unsupervised artificial neural network) was used to classify the behavioral patterns into seven clusters at different water temperatures. These results suggest that the activity levels or certain behaviors of P. canaliculata vary with the water temperature conditions. Understanding the thermal biology of P. canaliculata may be crucial for managing this invasive snail.

Inhibition of egg hatching with apple wax solvent as a novel method for controlling golden apple snail (Pomacea canaliculata)

2005 ◽  
Vol 24 (5) ◽  
pp. 483-486 ◽  
Author(s):  
Der-Chung Wu ◽  
Jih-Zu Yu ◽  
Bing-Huei Chen ◽  
Chien-Yih Lin ◽  
Wen-Hsiung Ko
Keyword(s):  
Pomacea Canaliculata ◽  
Apple Snail ◽  
Egg Hatching ◽  
Golden Apple Snail ◽  
Novel Method

Bioindication of mercury, arsenic and uranium in the apple snail Pomacea canaliculata (Caenogastropoda, Ampullariidae): Bioconcentration and depuration in tissues and symbiotic corpuscles

Chemosphere ◽  
2018 ◽  
Vol 196 ◽  
pp. 196-205 ◽  
Author(s):  
Alejandra D. Campoy-Diaz ◽  
María A. Arribére ◽  
Sergio Ribeiro Guevara ◽  
Israel A. Vega
Keyword(s):  
Pomacea Canaliculata ◽  
Apple Snail
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