Context
1 class session in Introductory Biology for majors Taught in a large lecture hall Pre-existing information (from previous lectures)
Cell parts (including cytoskeleton) Membrane structure Exocytosis
Explain steps in endocytosis using the concepts of molecular recognition and sorting Explain the function of endocytosis in cell metabolism Predict how perturbations of endocytic processes affects cells Explain how the endocytic process may be hijacked by pathogens Analyze and interpret experimental data Connect graphical data with visual representations
Assessments
Identify and order the steps of endocytosis Use concepts of molecular recognition and sorting to
explain specificity in the process Interpret data about how a perturbation affects endocytosis and predict consequences for cells Given information about how a drug affects endocytosis, analyze experimental data to determine how a pathogen enters a cell Predict how drug treatment might prevent a pathogen from reaching the cytosol
Diversity
New material is presented in multiple ways including extensive use of animations.
Homework assignment
Salmonella ?
Think-Pair-Share: Endocytosis
A) THINK INDIVIDUALLY (4 minutes) Read the terms on next slide Choose endocytosis terms Draw a labeled diagram using those terms B) PAIR (2 minutes) Discuss your diagram with your neighbor Modify your diagram as needed C) SHARE (2 minutes) We will use one stellar example in lecture Modify your diagram as we discuss
Lecture starts
Mini-lecture + student input to cover the concepts on the next few slides and videos Remind students to add to their diagram: Phagocytosis, pinocytosis differences and similarities
Types of Endocytosis
1. Multiple pathways of endocytosis 2. Molecular recognition between cell (receptor) and cargo (internalized object)
Receptor-mediated Endocytosis
Animated Video: Key Points 1. Recognition of SPECIFIC cargo 2. Multiple steps + compartments 3. Consequences for cargo
http://www.youtube.com/watch?v=lNKKg6hIBcM
Lecture continues
Cover steps clathrin acidification-2 steps Concepts: Molecular recognition Sorting cargo-receptor vesicle-vesicle
Why do we need this?
Molecular Biology of the Cell (2002) Alberts et al.
Phagocytosis
Animated Video: Key Points 1. Similarities to receptor-mediated endocytosis Molecular recognition Sorting 2. Differences No clathrin http://www.youtube.com/watch?v=7VQU28itVVw
Animation: McGraw-Hill
Phagocytosis
Molecular recognition between cell and bacterium
Lysosomes fuse with phagosome: Sorting: vesicles specific places Molecular recognition: lysosomes phagosomes What would happen if lysosomes fused with any (all) cellular membranes?
C
nucleus
nucleus
C
nucleus
nucleus
?
? Drug C
% of cells infected
Test cells received Drug C + virus. Untreated cells received only virus. Control cells received no virus. 3 viruses: Ebola, VSV and HIV. Infection is when virus reached cytosol.
Ebola
% of cells infected
Test cells received Drug C + virus. Untreated cells received only virus. Control cells received no virus. 3 viruses: Ebola, VSV and HIV. Infection is when virus reached cytosol. Clicker Q1 (1 min): Which virus(es) enter the cell by clathrin-mediated endocytosis?
Ebola
% of cells infected
Test cells received Drug C + virus. Untreated cells received only virus. Control cells received no virus. 3 viruses: Ebola, VSV and HIV. Infection is when virus reached cytosol. Clicker Q2 (1 min): Which virus(es) require receptors to enter the cell? A. Ebola only B. VSV only C. HIV only D. Not enough information
Ebola
C V
Bafilomycin
?
? Drug C
Homework
This experiment is like the one in class, except cells were treated with Bafilomycin, which inhibits endosome acidification. Q1: What is the most likely method by which VSVg enters the cell? A. Clathrin-mediated endocytosis B. phagocytosis C. pinocytosis D. not enough information
% of cells infected
Q3: Based on experiments with Chlorpromazine (discussed in class) and Bafilomycin, what method do you think HIV uses to enter the cell? Q4: What would happen to a normal cell treated with Bafilomycin, i.e. what are the side effects of this drug?