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CSB346H1 Lecture Notes - Lecture 4: Immunohistochemistry, Pipette, Patch Clamp

103 views9 pages
coraleel39
10 Apr 2013
School
UTSG
Department
Cell and Systems Biology
Course
CSB346H1
Professor
John Peever

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Document Summary

Csb346 lecture 4 respiratory rhythm generation (january 22, 2013) We are going to focus on the neurons in the brainstem, how do they function and how do we understand how breathing is generated by those neurons. We will go back to the idea that the neurons are there. The neurons in the pre-b tc give you the regular rhythm, so just simple respiration. All the other neurons in the vrg, drg, and prg are talking to each other and creating the shape of breathing, not so much when we breathe, but how we breathe. The main controller of breathing is in the brainstem. All the neurons that create breathing (the rhythm and the shape) are the components of the rrg. The neurons become active at different parts of the respiratory cycle. The muscles contract and you get thoracic expansion so air can come in. The main respiratory muscle is the diaphragm, which is controlled by the phrenic nerve.

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Related Questions

Background

Almost all animal and plant cells require oxygen and give off carbon dioxide. This is called gas exchange and it occurs by passive diffusion. Some animals do all or most of their gas exchange across their skin (for example amphibians) but they have very low metabolisms and use very little oxygen and get rid of very little carbon dioxide.

On the other hand humans (and other mammals) have very high metabolic requirements and we can’t rely on gas exchange over the limited 1.5 to 2.0 square meters of our skin to get enough oxygen in and carbon dioxide out fast enough. Our gas exchange takes place across a very thin membrane in the Alveoli of the lung and the gas exchange surface in humans is estimated to be between 50 and 100 square meters, or between 25 and 67 times our skin surface area.

Our lungs provide a huge surface for gas exchange but they are a dead-end system. Once the oxygen in the lungs is used up it needs to be replenished and carbon dioxide builds up in the air spaces in the lungs and needs to get out. This is where the diaphragm and a few of the skeletal muscles become important. In its relaxed state the diaphragm is at its longest and the space in the thoracic cavity (and the lungs) is small. When the diaphragm contracts it shortens, which increases the volume of the thoracic cavity (and the lungs). The increase in size means the pressure is lower in the lungs than outside so air comes into the lungs (inhalation). As the diaphragm relaxes, the lungs decrease in size enough that the pressure inside the lungs is higher than the external air pressure so air comes out (exhalation).

Introduction

In this lab you will be building a model of a lung and experimenting with different ways to breath.

Supplies:


In the Lab Kit

One round 9" balloon (one lung, we don’t need two for this)
medium-sized plastic bag large enough to fit over bottom end of bottle (diaphragm)
medium-size, thin rubber band

You supply

plastic 2-liter soda bottle with bottom cut off (thoracic cavity)

Tape (optional)

Directions:

1. Insert the balloon through the open bottom of a modified 2-liter soda bottle and up over the neck of the bottle. Pull the end of the balloon over the edges of the neck of the bottle. The balloon is held open by the neck of the bottle.

2. Place the plastic bag over the bottom (open) end of the 2-liter bottle and use the rubber band to hold it in place. This will serve as the diaphragm. To inflate the lung, blow into the balloon just before you fasten the rubber band. You can also wrap tape over the edges of the plastic bag to minimize air leaks

3. Grasp the bottom of the plastic bag and pull it down and push it up. Watch as the "lungs" (the balloon) expand and contract as you do this. The balloon has to change size because the air space between the balloon and bottle must remain constant. (Well, it would if the seal around the plastic bag and bottle were perfect).

You may block the "trachea" (the neck of the bottle) so that the air supply is cut off and watch as nothing happens when the "diaphragm" is pushed in and out.

This model isn’t perfect. None of the joints are air tight as they would be in a real thoracic cavity. Pull the “diaphragm down and watch the lungs expand . If you hold it in place, the lungs will collapse again over time. This is because of leaks and is not how real lungs work. Pushing the diaphragm in and pulling it out over and over does give you an idea of how lungs work. As the diaphragm tightens, contracts (pull down on the plastic bag) the lungs fill with air. As the diaphragm relaxes (push up on the plastic bag) air is expelled from the lungs .

There are several different ways we breath, i.e. expand our thoracic cavity so that our lungs can expand. You have just made a demonstration model of the diaphragm at work. You can also use your rib cage to expand and contract so that you can inhale and exhale. There are muscles between and around your ribcage which you can use to change the size of your thoracic cavity.

This is approximately what your model of a lung will look like.

Answer the questions on the Report Page

Report Page Name

1. Put a hand on your lower ribcage and try moving your ribs out and in. Does air also move in and out?

2. Someone with an upper back or neck injury may not have the options of using diaphragm or ribs to breath. They can learn to use the clavicle and the muscles attached to expand their thoracic cavity a small amount and breathe independently for short periods of time. Put your hand on you clavicle and try to raise and lower just that bone without using your diaphragm or your ribs. It isn’t easy but you should be able to get a little air into and out of your lungs this way. How effective do you think this way of breathing is?

3. When you take a really deep breath you use all these muscle groups to help. Take a really deep breath and notice that your ribs expand, your lower abdomen comes out (a sign that the diaphragm has contracted) and your shoulders lift a little (the upper chest and clavicle are working). Now take a relaxed breath. Only your abdomen should go in and out. The movement of your abdomen is a result of the contracting diaphragm pushing on the contents of your “gut” which forces your abdomen to push out.

Do you breath in or out when your diaphragm contracts?\

4. If a hole were poked in your thoracic wall (the wall of the pop bottle – the hole should be at least the diameter of a pencil)

a. what would happen when you tried to breath? (This is a pneumothorax).

b. Is the space between the balloon and plastic bottle still required to remain constant?

5. Would it be possible to live with a paralyzed diaphragm? Explain your answer.

As you may recall in the work of Triplett (1898), the presence of others in a bicycle race was found to enhance individual performance. This is called social facilitation (Williams, Harkins, & Karau, 2007). At times, however, the presence of others can detract from individual performance, which is termed social inhibition (Crisp & Turner, 2007). Similarly, social loafing can occur when an individual’s performance within a group decreases due to his or her reliance on others in the group (Crisp & Turner, 2007).

For this Discussion, review the media program, Week 5: The Virtual Office. Focus on the characters in the media and determine which characters displayed social facilitation, social inhibition, and social loafing.

With these thoughts in mind:

Post by Day 3 a description of the characters in the media program demonstrating social loafing, social inhibition, and social facilitation. Explain how you identified them. Then, select one of the characters and, using the current literature, explain two ways you might mitigate their behavior.

Media Program

(Onscreen Text: The employees at the United Cardboard Company often work individually on projects, but as with most environments, they are occasionally asked to participate in group projects. One such group is about to meet right now to plan a presentation for the Board of Directors. Observe the conversation and reactions of various group members to determine which group members exhibit signs of the following: Social Loafing, Social Inhibition, and Social Facilitation.)

(Interior of an office. Ken stands at the front of the conference room.)

KEN: OK, I guess we might as well all get started. As we all know, the big man has requested the five of us to get together and write and conceptualize a presentation to the board on our new packaging system. Maybe we can start by going around and throwing out some initial ideas.

KATHY: Sounds good to me. I think a PowerPoint presentation would do the job well.

GAVIN: I agree. We could follow it up with a physical demonstration of how the packaging looks and folds-- maybe a video of the larger pieces that won't fit into the boardroom.

KEN: Sounds perfect to me. Amanda, Brian, what do you guys think?

BRIAN: Yeah, what you guys said sounds fine.

(Amanda thinks, “I don’t know what to say!”)

KEN: Amanda?

AMANDA: I don't know. I didn't really get much chance to think about this. We only found out we needed to do this together three days ago.

KEN: OK, no worries. Let's focus on the PowerPoint idea. I've been doing a lot of them lately, so I'm happy to take the lead on the design aspect. What do you guys think we should all incorporate into the slides?

GAVIN: I think we should start with where we got the idea for this new packaging -- the company that designed it and the different sizes that it comes in.

KATHY: We should follow that with a graph comparing the old prices to the new ones to illustrate how much we'll be saving over time. Amanda, didn't you have most of that information collected at some point?

(Amanda thinks, “Oh no! What if I deleted that?)

AMANDA: I don't know, maybe, it's been so long. I didn't think I'd need it again for a group project like this.

(Gavin thinks, “I wonder what’s gotten into her. She’s not usually this tense.”)

KEN: That's OK, Amanda, I'm sure we can easily dig it up again. Brian, do you have any other ideas for the PowerPoint slides?

BRIAN: Nah, I think what you guys mentioned sounds right.

(Kathy thinks, “Wow. Way to contribute Brian.”)

KEN: OK then, Kathy and Gavin why don't you send me all of the info and I'll write it up and make the slides. Amanda, don't worry about the cost saving document, I'm sure I can dig it up from my old emails and put it into that slide too.

AMANDA: Are you sure? I'm happy to look. I mean I'm sure I have it somewhere. (Kathy thinks, “She really needs to get it together. Why is she so nervous?”) At least I think I have it somewhere. You won't tell the Board of Directors if I lost it, will you?

KEN: Of course not, we're in it together. So as I was saying, I'll write up and prepare all the PowerPoint slides. Then we can start our presentation with reading and explaining each slide. Then, what?

KATHY: Well, and then I think we should show them the packaging live, and the quick deconstructing techniques.

KEN: Great idea. You and Amanda can demonstrate the folding, while I narrate what you're doing.

(Amanda thinks, “Is he doing too much of the work? And what if I drop everything while he’s talking?”)

GAVIN: And after you all demonstrate the smaller packaging, we could show a quick video of how the really big ones function down at the warehouse. Brian, would you be able to go film that quickly one of these days?

BRIAN: I don't really have any experience with video cameras or anything like that, so...

(Gavin thinks, “I should have seen that one coming.”)

KEN: That's OK. My wife actually just got me this great little camera for our anniversary. It takes great videos. So I can just run down to the warehouse, get some footage, then put it together on this new editing software I just downloaded. Maybe add a little music. It's going to be really slick!

KATHY: Are you sure, I wouldn't mind doing that too.

KEN: Oh no, it's fine. I love this kind of stuff; just leave it to me. But I suppose that's enough planning. Now it's time to get to work and make the presentation a knock out. Go team!

(Everyone except Ken thinks, “Team?”) ©2012 Laureate Education, Inc.