Welcome to SPEP 2009!

1. Welcome toSPEP 2009! The University of Cincinnati Heather Hale Elise Demitrack

2. Getting to know you… • Name • School • Field of interest

3. Key Terms • Physiology • Study of Organ function • Regulation/Interaction of organ systems • Homeostasis • Body performs functions to maintain constituents of extracellular fluid • Ability to maintain constant internal environment • “Steady-state” (equal mixing)

4. Units • Mass % (g%) = gram amount per 100 mL (1dL) • Equivalent (Eq) = gram amount of one mole of a substance divided by it’s valence • i.e. 1Eq of Ca2+ is 40 gm / 2 • Osmoles = # particles released into solution when solute is dissolved in H2O • Osmolar = 1 Osmole/mole dissolved in 1L H2O • Osmolal = 1 Osmole/mole dissolved in 1kg H2O • One gram-molecular weight of any substance represents and consists of 6 x 1023 molecules

5. Physiology I Membrane PhysiologyHeather HaleJune 23, 2009

6. Head group Hydrocarbon tail The Plasma Membrane • Lipid bilayer • Key Constituents: • Phospholipids • Amphipathic (polar head & non-polar tails) • Fluidity • Cholesterol • Rigidity • Glycoproteins (protein + carbohydrate) • “Float” throughout bilayer • Forms receptor substances (glycocalyx) http://www.cytochemistry.net/Cell-biology/membrane_intro.htm

7. Crucial for creating an electrochemical gradient! Membrane Permeability • Selective! • Simple Diffusion • Rate depends on molecule’s: • Lipid solubility • Size • Charge • Assisted by: • Ion channels • Transporters http://en.wikipedia.org/wiki/Semipermeable_membrane

8. Selective Permeability

9. Plasma Membrane Proteins • Integral • Permanently associated with membrane • Transmembrane: spans entire bilayer • Peripheral • Associate with bilayer or another protein • Temporarily attached www.ultranet.com/~jkimball/BiologyPages/C/CellMembranes.html

10. Summary: Plasma Membrane

11. Membrane Proteins: Channels • Spans bilayer to form “pore” • Moves substances across bilayer • Gating: • Selectivity! • Ligand-gated • Mechanically gated • Voltage-gated Passive transport!

12. “Carrier” proteins Transport specific substance across bilayer (selective!) Channel changes shape/orientation Membrane proteins: Transporters Active transport! http://phy.asu.edu/phy598-bio/D5%20Notes%2006.htm

13. Types of Transporter Proteins Symport video http://www.biologie.uni-hamburg.de/b-online/library/biology107/bi107vc/fa99/terry/images/SymporA.gif http://library.thinkquest.org/C004535/cell_membranes.html

14. Membrane Proteins: Enzymes • Protein active site (intracellular or extracellular) catalyzes reactions Catalyze reactions inside/outside cell Associated with membrane to increase efficiency http://www.biochem.arizona.edu/classes/bioc462/462b/Miesfeld/Photosynthesis.html

15. Examples of Membrane Proteins • Channels: Ca2+ and Na2+ channels • Transporters: • Proteins transporters • Glucose transporters • Enzymes: Mitochondrial membrane proteins

16. Movement Across Plasma Membrane • Transport of material across bilayer • Can be direct (non-facilitated) • Some requires proteins: • Diffusion (facilitated) • Active transport (energy!)

17. No proteins required! Passive Movement • No cell energy required! • Simple diffusion • [high] [low] • Based on molecule‘s properties • Gases, nutrients, ions • Limited by diffusion rate of molecule! http://www.indiana.edu/~phys215/lecture/lecnotes/diff.html

18. H2O moves toward compartment with high [solute] Passive Movement of Water Movement of H2O across cell Moves from [H2O]high [H2O]low H2O keeps osmotic pressure equal across membrane http://www.indiana.edu/~phys215/lecture/lecnotes/diff.html

19. Solutions & Osmotic Pressure • Solutions • Isotonic: equal [solute] inside/outside • Hypotonic: low [solute]; H2O moves out • Hypertonic: high [solute]; H2O moves in • Osmotic Pressure • Required to stop osmotic H2O movement • Determined by # particles/unit volume • Osmole = # particles in 1 gram (MW) of un-dissociated solute

20. Pressure & Water Movement H2O moves from Phigh to Plow • Influenced by two forces • Hydrostatic pressure: caused by gravity on a column of fluid • Hydraulic pressure: caused by action of a pump (active!) • Osmotic pressure = only pressure to initiate water flow in/out of cell

21. Osmotic Pressure ECF and ICF have [osmotic] = 300 mOs/L • Calculated osmotic pressure (π = CsRT) • Cs = osmolar concentration • R = universal gas constant • T = absolute temperature • (RT = 22.4 ATM/osmole at 37˚C) • Effective osmotic pressure: • Depends on permeability of membrane to specific solute

22. Osmotic Pressure • Fig A: semipermeable membrane • Solute cannot pass • Pos will equal the Phydrostatic as water flow into tube • Fig B: solute-permeable membrane • Solute equilibrates • Effective Pos of solution is zero

23. Passive Movement: Facilitated • Examples: • Glucose transport • K+, Na+, Cl- transport Requires membrane proteins! Forms water-filled pore Solutes move down [conc] gradient (high  low)

24. Active Movement: 1˚ Transport • Requires cellular energy! • Transporters bind ATP • Hydrolysis of ATP to ADP + Pi • Drives transport of solute against concentration gradient! • Examples: • Na+/K+ ATPase pump • Ca2+ ATPase

25. Active Movement: Na+/K+ Pump video http://images.google.com/imgres?imgurl=http://student.ccbcmd.edu/~gkaiser/biotutorials/eustruct/images/sppump.gif&imgrefurl=http://student.ccbcmd.edu/~gkaiser/biotutorials/eustruct/sppump.html&h=290&w=290&sz=515&hl=en&start=1&um=1&tbnid=RE2RGHk1UT

26. Uses energy of a “driving ion” moving down [conc] gradient to move a 2nd molecule against [conc] gradient Driving ion usually Na+ using gradient created by the Na+/K+ pump Active Movement: 2˚ Transport Examples: -Na+/Ca2+ exchanger -Na+/glu transporter http://courses.cm.utexas.edu/jrobertus/ch339k/overheads-2/ch12_Na-gluc-trans

27. Active Movement: Bulk Transport • Endocytosis: transport into cell • Phagocytosis = ingest large particles • Pinocytosis = ingest small vesicles • Receptor-mediated ingestion • Exocytosis: transport out of cell http://upload.wikimedia.org/wikipedia/commons/thumb/1/1a/Endocytosis

28. Physiology I Body Fluid SpacesHeather HaleJune 24, 2008

29. Body Fluid Spaces • The human body is mostly water • Total H2O content of the human body = 45-60% of body weight • Total Body Water (% of body weight) • Males: 60% (ages 17-34) 54% (ages 50+) • Females: 55% (ages 17-34) 46% (ages 50+)

30. Major Body Fluid Spaces • Two major “compartments” • Intracellular fluid (ICF): • fluid contained within cells • Accounts for 40% of body fluid weight • Extracellular fluid (ECF): • fluid outside of the cell • Accounts for 20% of body fluid weight • Total body water = ICF + ECF

31. i.e. 70kg × 0.4 = 28 L of ICF fluid Major Body Fluid Spaces • The 60, 40, 20 rule • 60 = All fluid is 60% of total body weight • 40 = ECF is 40% of body weight • 20 = ICF is 20% of body weight

32. Major Body Fluid Spaces • ECF is subdivided even further: • Plasma space = 5% of total body weight • Interstitial fluid (ISF) = 15% of total body weight • ISF = ultrafiltrate of plasma

33. Ratio of red blood cell (RBC) volume to whole blood volume Hct is typically 40% of whole blood volume Whole blood volume = 7-9% of body weight (about 6 L) Body Fluids: Hematocrit (Hct) Hct = (vol)RBC (vol)whole blood 1 - Hct = (vol)plasma (vol)whole blood

34. Learn these values! Body Fluids: Cations/Anions • Total [Osmolar] = 280-296 mOs/L • Na+ = 13-145 mEq/L • Cl- = 100-106 mEq/L • Ca2+ = 4.3-5.3 mEq/L • Glucose = 70-110 mg% • Total protein = 6-8 g%

35. Body Fluids: Cations/Anions • ECF is high in Na+ & Cl- • ICF is high in K+

36. C1 V1 = C2 V2 V2 = (C1 V1) / C2 Body Fluid Space Measurements • To estimate the size of body fluid spaces, use a dye indicator dilution • Based on conservation of mass principle • [conc]  vol = mass • Only applicable during steady-state • No loss/gain of substance during measurement

37. Body Fluid Spaces: Fick Principle • If some solute is lost/gained: • C1 V1 = C2  V2 (+ amount gained) • orC1 V1 = C2  V2 (- amount lost) • Analysis represents Fick Principle • Commonly used to measure blood flow or cardiac output

38. 3 component system (plasma, ISF, ICF) Inject substances into plasma Assumptions Equal amnt x, y, z x, y, z not present before injection Body Fluid Space Measurements

39. Body Fluid Space Measurements • Capillaries • separate plasma/ISF • Permeable to y and z but not to x • Cell membrane • Separate ISF/ICF • Permeable to z only

40. Body Fluid Space Measurements • Volume distribution of x = plasma • Volume distribution of y = plasma + ISF • Volume distribution of x = ECF + ISF

41. Body Fluid Spaces: Markers • Plasma fluid markers (“x”) • Do not cross capillaries • Examples: • Radioiodinated serum albumin • Evan’s Blue (dye that binds albumin) • RBCs with radioactive iron or chromium

42. Body Fluid Spaces: Markers • ECF markers (“y”) • Represents plasma + ISF • Cross capillary but not cell membrane • Examples: • Isotopic Cl- or Na+ • Inulin • Mannitol

43. Body Fluid Spaces: Markers • Markers for total body water (“z”) must be permeable to both capillaries and cell membranes • Examples: • 3H water (tritiated “heavy” water) • Urea (carbon labeled, or tritiated) • Lipid soluble substances

44. Capillary Fluid Movement • Capillaries separate plasma from ISF • ECF ions move across capillaries between plasma and ISF • But, proteins are restricted to plasma • Creates osmotic pressure ~15-25 mmHg • This is the colloid osmotic pressure

45. Starling’s Law of the Capillary • Principle of pressure differences • Pressures: • Reabsorptive forces: • Capillary colloid osmotic pressure • ISF hydrostatic pressure • Filtration forces: • Capillary blood pressure • ISF colloid osmotic pressure FM = Kf [(BPcap + COPISF) - (COPcap + HPISF)]

46. Filtration Reabsorption Reabsorption Filtration Capillary Fluid Movement FM = Kf [(BPcap + COPISF) - (COPcap + HPISF)]

47. Email: heather.hale55@gmail.com Lab location: Health professionals Building Room 234 (off of Eden Ave, across from Eden Garage and MSB) Contact Info

48. Supplementary Figures

49. Supplementary Figures

50. Supplementary Figures