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**Biological Principles Exam Overview**

Prepare for the upcoming exam covering topics on fluorescence microscopy, gastrulation, gene therapy, ECM components, and cell junctions. Be familiar with lectures by Stratman, Mecham, and Miner.

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**Biological Principles Exam Overview**

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  1. Exam 1 Review Session Allen Yen and McKenna Wilhelm 9/18/19

  2. Exam format Questions will be short-answer style (concise responses are encouraged), matching, fill-in-the-blank, and some that require drawing/diagraming.  The exam will be Friday (9/20) from 8:30-10:30am in Holden (here).  Makeup exam will be Monday (9/23) from 2-4pm in FLTC 204.  Lecturers/TAs will not be available for questions during the exam. Stratman and Mecham materials will be weighted more than Miner.

  3. Fitzpatrick's Lectures - Basics of light and fluorescence microscopy - Immunostaining

  4. Fluorescent probes and microscopy Absorbance: wavelength of light that excites photon Emission: wavelength of light emitted by photon returning to ground state Stoke's shift: difference in wavelength between the maximums of absorbance and emission spectra

  5. Brightfield vs fluorescence microscopy Key differences: - excitation light - use of dichroic mirror - sample prep

  6. Immunofluorescence staining

  7. Stratman's Lectures L1: gastrulation and germ layers; gradients and tissue patterning; transcriptional regulation and TFs L2: cell junctions; Cerebral cavernous malformations L3: how does CRISPR/Cas9 work and gene therapy applications;

  8. Gastrulation and germ layers

  9. Gastrulation and germ layers

  10. Gradients and zebrafish embryo patterning

  11. Vessel Formation and ECM Expression in the Vessel Wall

  12. Chromatin state affects TF binding

  13. TFs as activators and repressors

  14. How do enhancers affect gene expression?

  15. Stratman's Lectures L1: gastrulation and germ layers; gradients and tissue patterning; transcriptional regulation and TFs L2: cell junctions; Cerebral cavernous malformations L3: how does CRISPR/Cas9 work and gene therapy applications;

  16. Main types of cell junctions Be able to name a function of each junction

  17. Adherens               vs.            Desmosomes

  18. Gap junctions couple electrical signals What is the function of adherens junctions? Where would this be particularly important?

  19. Cerebral cavernous malformations (CCMs)

  20. CRISPR/Cas9 system

  21. Mecham's Lectures ECM and Tissue integrity Content: -Different types of ECM proteins and structural components  -Diseases related to ECM proteins -Understand how post translational modifications relate to ECM components -Proteoglycans structure and function ECM and Mechanical Signals Content: -Know how Cre-loxP systems work:   1) to inactivate/activate genes  2) to lineage trace cells in tissues  3) as conditional and inducible systems 4) with FLP-FRT system

  22. Properties of ECM

  23. Structural Organization of Collagens Mutations in glycine lead to multiple diseases due to H-bond disruption

  24. Regulation of Growth Factor Signaling What disease did we talk about in class relating to mutation in Fibrillin? Large latent complex small latent complex Latency Associated Peptide (LAP)

  25. Post Translational Modifications How does this relate to Scurvy?

  26. Elastin and Related Diseases Inherited- supravalvular aortic stenosis and Williams syndrome Acquired- emphysema and aneurysms

  27. Proteoglycan vs. glycoprotein Glycoprotein: -oligosaccharide chains covalently attached to polypeptide side chain -short or branched chains may or may not be charged - connected through O or N-linked glycosylation  -found on cell surface -play a role in cell-cell recognition Proteoglycan: -glycosaminoglycan (GAG) chains attached to a polypeptide side chain -GAGs are long, linear, and negatively charged due to sulfates -found in connective tissues -provide structural support and mechanical strength to ECM Both: -comprised of protein and glycan 

  28. Cre-Lox • -Arrow means promoter present • -Poly A sequence signals a stop Gene X

  29. Miner's Lectures  Content: • Basement membrane components, function, biological processes involved  • Laminin structure, function • Collagen IV network, mechanism 

  30. Agrin: a heparan sulfate proteoglycan

  31. Agrin in neuromuscular junctions (NMJs)

  32. Integins, dystroglycan, basement membrane, and actin

  33. Anoikis and cancer

  34. ECM dysfunction in disease Basic pathophysiology and the therapeutic strategy (if discussed) of: Alport syndrome Epidermolysis bullosa Muscular dystrophy

  35. Good luck!

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