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NROC34 Lec 2.docx

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Karen Williams

NROC34 Lec 2 - Difference between review and primary articles: o Primary  have a hypothesis and experiment to find out about it o Review  any different primary articles put together - Blog: o How does the lecture relate to the material  Can all be asked about in the exam o 10 posts  2 for each lecture o Takes information from the blog journal and puts on the exams o Worth 2%  post to discussion board = 1%, posts to blog journal = 1% - Video assignment: all done in class o NO LECTURE CAST ON THESE DAYS o Watch an animal behave and describe their behaviour o JAN 25 o 10 minutes to write description and hand in - Critical review: o Review of a single, primary article o Everyone is doing the same review of the same article - Poster: o First part of critical review reading a paper, review, primary, and making comments about them - Midterm: March 8 - Midterm and final  short answer and multiple choice questions  Some questions directly from lecture notes, class discussions and blog posts FRUIT FLY FORAGING - Able to find fruit, know when you’re drinking herbal tea and seem to find their way to your picnic (back track) Candidate gene approach - Used to find info about honey bee foraging - Identifying and categorizing behaviour of interest, nominating a candidate gene and doing literature or data-based searches o Looking at primary articles looking for foraging - Nominated foraging gene which In drosophila melanogaster, has been described for larvae (sitters) vs long distances (rovers) o Similar to nurse and foraging bees How to discover the genetic basis of larval foraging behaviour? - First step in very analysis: describe the behaviour. What do you see? What is the animal doing? - Picture: larva just sitting there o If foraging, we know the larva lives in their food, and emerge from their pupil as adults o If it is foraging, does it have to look far for food? No, is sitting in it o This larva can get food into itself by:  Putting the food in its mouth  Need to look for mouth end  Mouth is on the end where the food are going in o Eyes are deep inside the larva – cannot see o When it gets to a large enough size to pupate, out of the larva will come a single fly o If is a forager, need to know behaviour and something about life cycle  Larva (egg)  then hatches into 1 , 2 and 3 instar larva  pupates into pupae  closes into adult in 3-4 days  Genetic information stored on flybase Foraging is a food-related behaviour - How to look at larval foraging in drosophila - LIVE IN THEIR FOOD - Have to be able to analyze behaviour and quantify distance the larva moves when looking for food o Food-related: different from when there is no food o Food-related = yeast based, no-food = agar o Can make a food and no food situation by putting food patches in the mitts of agar - On agar, larvae move similar distances. On yeast, some larvae move long and some move short - The distance on agar is the same but changes in the presence of food therefore is FOOD- RELATED - If larva can smell yeast paced patch in agar, the larvae will move between patches as they search for food o Rovers do this a bit more than sitters Foraging trails - Can plot in terms of cm - See some move only short distances, some long distances but some fall in the middle - SITTER o Need to find some mean to consider it a sitter o Important in psych  don’t have those that can’t learn; those that can, have distribution - Look at peaks to distinguish rovers and sitters - Cross sitter males, rover females and look at their progeny to see if there is a pattern from one generation to the next o Manipulate offspring - Define mean foraging trail length to distinguish between the 2 Generations - F1 = findt filial generation - F2 = 2 filial generation How to discover the genetic basis of a behaviour? - Look at f1 and f2 - Found no difference when female was sitter or rover o Therefore, the gene is NOT sex-linked o Is an autosomal gene A simplified model of the factors contributing to the expression of a behavioural phenotype - For behavioural phenotype, have influence of hereditary and environment - Generally, we are always looking at gene AND environment 1 - Partition heredity and environment - Can look at chromosomal and non-chromosomal o Chromosomal = nuclear material from parents o Non-chromosomal = maternal effect - Epigenetic signalling on behaviour  non-chromosomal component of contribution o Very big right now o Transient maternal factors - Looking mainly at autosomal 3.19 Genetic differences cause behavioural differences in fruit fly larvae - Rover is dominant - Easiest explanation: simple autosomal Mendelian gene o Pattern of 3:1 Drosophila chromosomes nd - Gene was on the 2 chromosome on the left arm Foraging: cyclic GMP dependent protein kinase - Tells us a whole variety of things could also be involved Expressing for changes foraging behaviour and enzyme activity - Foraging DNA (cDNA) was put into sitter host o Made transgenic animal - See that transgenics were much more rover in behaviour - Sitter host have short path lengths, transgenics and rovers have ~9 - PKG enzyme activity also changes: o Low in sitters o Increased in rovers Adult flies foraging - Is foraging the same in flies as larva and adult? - Dev’l differences in flies: o Larvae live in their food o Adults go to a droplet or banana peel, etc. when it is gone, they look for another source - Vince Dethier o Described behaviour of flies o Pattern of behaviour:  Find a drop of sucrose, have immediate search response, then look around a larger area, and keep looking further and further  While the fly was eating, if there was a small noise, the fly would go away - Rovers make a larger trail around for sucrose than do sitters - Define foraging behaviour of adults, after this, ask if the transformed flies do the same thing - Made genetic mutants: would they respond the same? o Sitter mutants with rover-like background,  Just changing foraging changes the behaviour and PKG activity Foraging: cyclic GMP dependent protein kinase - Where is PKG highest? - PKG enzyme activity in sitter and sitter mutants, see lower activity in the heads than rovers - But in the body, the enzyme activity of rovers and sitters was around the same - PKG enzyme activity is lower in rovers than in sitters Sucrose response and foraging - Where does PKG act on? o Head o Antennae - Immobilize flies in pipet tip, allowed one leg to stick out and measured mouth response to sucrose - If detecting sucrose concentration, they move their proboscis - How often does fly stick proboscis out? Antibody to FOR in the adult brain - Expressed all over the head, some places might receive neurons from the tarsi - Can look at parts of brain, amounts of enzyme activity and look at behaviour Sucrose response and foraging - Behavioural measure Targeting foraging expression to the nervous system - Can target expression of gene to particular areas from immunoreactivity in the head - Can we change the behaviour if we make the fly rover in certain areas of the brain? - One fly has promoter region, one has recognition sequence + target gene lead to expression of that bit of DNA in a particular pattern in the fly - ELAV expresses entire nervous system Sucrose response and foraging - Do we see a difference in behaviour? - Overexpress foraging in the nervous system, see an increase in sucrose responsiveness o Make fly a rover all over the nervous system o Sticks out proboscis more in response to sucrose - Regardless of where they are placed
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