Investigating The Investigating The Effects Of Effects Of Caffeine Caffeine On The 6-Day Chicken On The 6-Day Chicken

Heart RateHeart Rate

By: Keila Ortiz ,Kaltrina Sylejmani By: Keila Ortiz ,Kaltrina Sylejmani and Dr. Jacqueline McLaughlinand Dr. Jacqueline McLaughlin

The Pennsylvania State The Pennsylvania State University, Berks/Lehigh Valley University, Berks/Lehigh Valley College, Fogelsville, PA 18051College, Fogelsville, PA 18051

Physiology of the Chicken Physiology of the Chicken HeartHeart

The chicken heart has 4 The chicken heart has 4 chamberschambers

- a right atrium and - a right atrium and ventricleventricle

- left atrium and ventricle- left atrium and ventricle

Its body temperature is 41-Its body temperature is 41-45 45 ˚C˚C

Pulse Rate of an adult Pulse Rate of an adult chicken is as high as 400 chicken is as high as 400 beats/min. beats/min.

Caffeine BackgroundCaffeine Background

Caffeine is a Caffeine is a trimethylaxanthine and trimethylaxanthine and directly affects the directly affects the contractility of the heart and contractility of the heart and blood vessels.blood vessels.

Stimulates the central Stimulates the central nervous system.nervous system.

Stimulant that enhances and Stimulant that enhances and prolongs thermogenesis (heat prolongs thermogenesis (heat creation) .creation) .

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www.seeleywis.com/mooselips.htmwww.seeleywis.com/mooselips.htm

Caffeine FormulaCaffeine Formula

www. chem.purdue.edu/gchelp/molecules/caff.html

Caffeine MoleculeCaffeine MoleculeIt belongs to a group of chemicals called It belongs to a group of chemicals called xanthinesxanthines

It resembles the molecule adenosineIt resembles the molecule adenosine

It is necessary for the construction and It is necessary for the construction and maintenance of DNA.maintenance of DNA.

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Calcium Blockers and the Calcium Blockers and the Electrophysiology of the HeartElectrophysiology of the Heart

Calcium channel blockers are used to regulate heart beat Calcium channel blockers are used to regulate heart beat and lower the blood pressure. and lower the blood pressure. Calcium blockers are used in the treatment of heart failure Calcium blockers are used in the treatment of heart failure because they reduce peripheral resistance and expand because they reduce peripheral resistance and expand arteries. arteries. The main kinds of calcium blockers are as following: The main kinds of calcium blockers are as following: diltiazem, nicardipine, nifedipine, verapamil, amlodipine diltiazem, nicardipine, nifedipine, verapamil, amlodipine besylate. besylate. They are used to treat chest pain or to manage hypertension.They are used to treat chest pain or to manage hypertension.Caffeine has shown to increase the intracellular calcium by promoting its release from the endoplasmic reticulum and by facilitating its influx through “Slow Channels” in the cell membrane.

Effects of CaffeineEffects of Caffeine

Affects the neurotransmission Affects the neurotransmission in the central and peripheral in the central and peripheral nervous system causing cardiac nervous system causing cardiac arrhythmias arrhythmias

Can increase the heartbeatCan increase the heartbeat

Increases the risk of Increases the risk of spontaneous abortion and spontaneous abortion and retardation retardation

It also affects the change in the It also affects the change in the electrophysiology of the human electrophysiology of the human heartheart

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Advantages of CaffeineAdvantages of CaffeineMakes people more Makes people more alertalert

Less drowsyLess drowsy

Improves coordinationImproves coordination

Helps relive headachesHelps relive headaches

Caffeine consumption Caffeine consumption helps the memory and helps the memory and reduces the incidence reduces the incidence of Parkinson'sof Parkinson'sdiseasedisease

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PurposePurpose

Different caffeine concentrations, Different caffeine concentrations, 0.002%,0.02%, and 0.2%, were added to the 0.002%,0.02%, and 0.2%, were added to the in vitro chicken heart. It was hypothesized in vitro chicken heart. It was hypothesized that the chicken heart rate (BPM) would that the chicken heart rate (BPM) would respond in such a way that various respond in such a way that various arrhythmias would be noted, specially arrhythmias would be noted, specially sporatic episodes of tachycardia and sporatic episodes of tachycardia and bradycardia due to increasing concentrations bradycardia due to increasing concentrations of this drugof this drug. .

Materials and MethodsMaterials and Methods Prepared stock solutions of caffeine: Prepared stock solutions of caffeine: “ “Window” an eggWindow” an egg

-Modified from Cruz, 1993:-Modified from Cruz, 1993:

a.a. Pick up an egg from the incubator. Pick up an egg from the incubator. b.b. Place scotch tape on top of the egg . Place scotch tape on top of the egg .

c. With the tip of the scissors, puncture the side of c. With the tip of the scissors, puncture the side of the egg the egg d. Cut out the part of the egg covered with scotch d. Cut out the part of the egg covered with scotch tape tape and use forceps to remove the top and use forceps to remove the top shell of the egg.shell of the egg.

e. e. IMMEDIATELY determine the in vivo HR, IMMEDIATELY determine the in vivo HR, five times and five times and record it. record it. 6-day chicken heart

Materials and Methods Materials and Methods Continued……Continued……

““Explanting” the Explanting” the embryo embryo

-Modified from Cruz, 1993:-Modified from Cruz, 1993:a.a. GGently position a ently position a filter filter paper doughnut on paper doughnut on top of top of the vitelline the vitelline envelope.envelope.b.b. Fill a small dish with Fill a small dish with saline and place it saline and place it on the on the stage of your stage of your dissecting dissecting microscope. microscope. c.c. Quickly transfer the Quickly transfer the

explant to the small explant to the small dish dish with warmed saline. with warmed saline. f. Record the in vitro HR f. Record the in vitro HR five times.five times. www.lv.psu.edu/jxm57

Materials and Methods Materials and Methods Continued………Continued………

Application of CaffeineApplication of Caffeine

a.a. Using a plastic pipette Using a plastic pipette apply 0.002% of caffeine apply 0.002% of caffeine

in in the in vitro embryo and let the in vitro embryo and let it it sit for 30 seconds. After sit for 30 seconds. After

sitting for 30 seconds sitting for 30 seconds record its HR five times.record its HR five times.

b. Using a plastic pipette at all b. Using a plastic pipette at all times, apply 0.02% of times, apply 0.02% of caffeine to the in vitro caffeine to the in vitro embryo, let it sit for 30 embryo, let it sit for 30 seconds and record its seconds and record its HR five times. HR five times.

c. Repeat same strategies as c. Repeat same strategies as

in step a and b but this time in step a and b but this time adding 0.2% of caffeine. adding 0.2% of caffeine.

6-day chicken embryo

ControlControl

The The in vivoin vivo and and in vitroin vitro HR of the 6-day HR of the 6-day Chicken embryo Chicken embryo

without exposure to without exposure to caffeine.caffeine.

6-day Chicken Embryo

Data InterpretationData Interpretation

A stereoscopic microscope was used to A stereoscopic microscope was used to observe both in vivo and observe both in vivo and in vitro in vitro 6-day 6-day chicken HR’s.chicken HR’s.

Histograms were constructed to plot data. Histograms were constructed to plot data. When adding different concentrations of When adding different concentrations of caffeine cardiac arrhythmias were noted such caffeine cardiac arrhythmias were noted such as bradycardia and tachycardia. as bradycardia and tachycardia.

Embryo’s In Vitro HREmbryo’s In Vitro HR

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Embryo’s In Vivo HREmbryo’s In Vivo HR

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0.002% Caffeine 0.002% Caffeine ConcentrationConcentration

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0.02% Caffeine 0.02% Caffeine ConcentrationConcentration

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0.2% Caffeine 0.2% Caffeine ConcentrationConcentration

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Averages of In Vivo and In vitro Averages of In Vivo and In vitro HRHR

Average of In Vivo and In Vitro Heart Rates

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In Vivo

In Vitro

Averages of Different Caffeine Averages of Different Caffeine ConcentrationsConcentrations

Averages of 0.002%, 0.02% and 0.2% Caffeine concentration

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0.002%

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ConclusionConclusion

Since the in vitro chicken heart Since the in vitro chicken heart experienced more arrhythmias as the experienced more arrhythmias as the concentrations of caffeine increased, concentrations of caffeine increased,

the proposed hypothesis was then the proposed hypothesis was then supported. supported.

Future ExperimentsFuture Experiments

For a future experiment, it For a future experiment, it would be interesting to take a would be interesting to take a 1- day egg, carefully remove a 1- day egg, carefully remove a piece of shell, then using a piece of shell, then using a syringe apply half a mL of syringe apply half a mL of caffeine (0.2%, 0.02%, caffeine (0.2%, 0.02%, 0.002%) to the chorionic 0.002%) to the chorionic membrane. After 21 days membrane. After 21 days wherein the chicken is fully wherein the chicken is fully developed, see if the developed, see if the concentrations applied caused concentrations applied caused any abnormalities in the any abnormalities in the developed chicken heart.developed chicken heart.

Any Questions…………?Any Questions…………?

ReferencesReferences1. 1. Morgan, J.G. and Brown Carter, M.E., Morgan, J.G. and Brown Carter, M.E., Investigating BiologyInvestigating Biology, ,

Fourth Edition. Fourth Edition. San Francisco: Benjamin/Cummings Publishing Co., Inc. San Francisco: Benjamin/Cummings Publishing Co., Inc. 2002. 2002.

2. Skinner. B, Braun. N, 2. Skinner. B, Braun. N, Caffeine and chick heartsCaffeine and chick hearts. 2 Aug. 2001. . 2 Aug. 2001. 12 Feb. 2004 <http://www.swartmore.edu>. 12 Feb. 2004 <http://www.swartmore.edu>.

3. 3. Caffeine. Caffeine. 5 Mar. 2000. 12 Feb. 2004 5 Mar. 2000. 12 Feb. 2004 <http://www.geocities.com>.<http://www.geocities.com>.

4. 4. Cruz, Y. P. 1993. Laboratory exercises in developmental Cruz, Y. P. 1993. Laboratory exercises in developmental biology. Academic Press, San Diego, California, 241 pages. biology. Academic Press, San Diego, California, 241 pages. [ISBN 0-12-198390-0] [Book][ISBN 0-12-198390-0] [Book]

5. Groski, Dawn , and Janet Sinn-Hanlon. 5. Groski, Dawn , and Janet Sinn-Hanlon. ChickscopeChickscope. 22 Mar. . 22 Mar. 2004 http://www.chickscope.beckman.uiuc.edu>. 2004 http://www.chickscope.beckman.uiuc.edu>.

6. 6. Null M.S, Gary, and Bulton Sanford Ph.D. Null M.S, Gary, and Bulton Sanford Ph.D. Caffeine: Psychological Effects, Caffeine: Psychological Effects, Use & AbuseUse & Abuse. 3 Nov. 1981. Orthomolecular Psychiatry. 15 Mar. 2003. 3 Nov. 1981. Orthomolecular Psychiatry. 15 Mar. 2003

<http://www.garynull.com/Documents/CaffeineEffects.htm >. <http://www.garynull.com/Documents/CaffeineEffects.htm >. 7. McCain, Dr. Elizabeth E., and Dr. Jacqueline Mclaughlin. 7. McCain, Dr. Elizabeth E., and Dr. Jacqueline Mclaughlin. Developmental andDevelopmental and Physiological Aspects of the Chicken Embryonic HeartPhysiological Aspects of the Chicken Embryonic Heart . 18 Mar. 1999. Copyright. 15 . 18 Mar. 1999. Copyright. 15 Mar.Mar. 2004 <http://www.lv.psu.edu/jxm57>. 2004 <http://www.lv.psu.edu/jxm57>.