Frog Heart Lab

Introduction
Although the frog has a 3-chambered heart - two atria and one ventricle - its function
and control is essentially the same as that of humans. The cardiac muscle has
myogenic origin of contraction (intrinsic stimulation) but can be influenced by extrinsic
factors. The primary pacemaker region of the heart is the sinoatrial (S-A) node near
where the venae cavae enter the heart. The S-A node sets the basic rhythm of the heart
and, in the frog, the normal heart rate at room temperature is about 40 beats per
minute. Its function is modified by input from the central nervous system. The sinoatrial
node is regulated by the amount of acetylcholine (ACh) secreted by the autonomic
nervous system’s parasympathetic neurons. The parasympathetic division is referred
to, sometimes, as the “rest-and-digest” system, as opposed to the sympathetic division
which is the “fight-or -flight” system. Pain, strong emotions, the anticipation of exercise
and “fight or flight” all can increase the secretion of norepinephrine from sympathetic
neurons. Nicotine initially mimics parasympathetic actions, but eventually blocks
parasympathetic activities and has the opposite effect. Caffeine is a central nervous
system stimulant.
The ion & temperature environment can also influences heart rate. Calcium affects
two aspects of nerve-muscle physiology. Ca++ stimulates the release of ACh and
muscle contraction. Potassium helps to maintain the resting membrane potential of
neurons & muscles. High levels of K+ in the extracellular fluid can have various effects
depending on how rapid the increase in K +. Slow increases result in making cardiac
cells less excitable, while rapid increases make cardiac cells more excitable. Because
frogs are ectothermic, their heart rate reflects metabolic rate which increases with
increasing body temperature and vice versa.
The objectives of this exercise are to:
1) compare heart rate in a normal frog versus a doubly-pithed frog
2) investigate the effect of temperature on heart rate and strength of contraction,
3) investigate the effects of ions on heart rate and strength of contraction, and,
4) investigate the effect of different neurotransmitters and drugs on heart rate and
strength of contraction of the heart.

Procedure
We will be using electrocardiograms (ECG) to measure heart rate and strength of
contraction. ECGs record electrical activities of the heart as it progresses through the
cardiac cycle. Each vertical movement of the ECG is called a deflection wave and
indicates a contraction or a relaxation of different heart chambers. The frog’s ECG will
look different than the human’s because it has only three chambers. There should be 2
deflection waves, the first weaker than the second. The first represents atrial contraction
and the second ventricular contraction. We can measure heart rate by counting how
many cycles (both atrial and ventricular contractions) occur during a 15-sec recording.
We then multiply this number by 4 to get beats per minute (bpm). The strength of the
contraction can be determined by observing the amplitude of ventricular contraction
waves.
Before experiment
1) formulate hypotheses for each test as to whether you think heart rate and strength of
contraction will increase or decrease or remain stable.
2) develop a data table for recording information from ECGs.

Baseline ECG – apply leads to live frog held by a classmate – the black lead goes to
one leg and the green goes to the arm on the same side, and red to the other arm.
- record the ECG for 15-secs and calculate heart rate

Pithing & dissection procedure

1) Grab frog tightly by its trunk
2) Bend head forward and insert a sharp probe at base of skull toward the brain and
maneuver probe to macerate the brain matter.
3) Remove probe and insert probe at the same site but in the opposite direction so it goes
inside of the vertebral column where the spinal cord is. Maneuver probe to macerate
the cord.
4) Remove probe and lay frog on its back.
5) Cut skin and muscle away from chest area and then remove sternum and ribs to
expose its heart. Removal of some tissue, e.g., fat bodies, may be necessary to
make access to heart easier.
6) Clip ECG’s alligator clips to arms and leg.

Important note: Between each administration of experimental treatments, rinse

the frog’s heart with Ringer’s solution and get another baseline reading.
Effect of temperature on heart rate & contraction
1) Record 15-secs of contractions. Measure a normal heart rate & contraction.
2) Bathe the heart with warm water.
3) Measure heart rate & contraction.
4) Rinse with Ringer’s solution until the heart rate returns to normal and record ECG.
5) Bathe the heart with cold water.
6) Measure heart rate & contraction.
7) Rinse with Ringer’s until heart rate returns to normal.

Effect of ion environment on heart rate & contraction

Follow procedure as above with first 2.0% solution of CaCl 2 and then 2.0% of KCl with a
rinse of Ringer’s in between each test and a return back to normal.
Effect of drugs on heart rate & contraction
Follow procedure as above with first caffeine, then 0.2% solution of nicotine, and then
acetylcholine bromide with a rinse of Ringer’s in between each test and a return back to
normal.

Report should include tables & graphs as well as at least three references – one has to
be from a source different than your textbook or lab manual.
 Vertebrate Bone Lab

Compare among the vertebrate classes the following features of vertebrate skeletons.
How might these changes in these skeletal structures made the animal better adapted to
its environment and lifestyle? (see skeletal sections in Ex. 18- 22)

Axial Skeleton – skull, vertebral column, ribs & sternum

Specimens: shark, bony fish, frog, snake, turtle, bird, mammal
- skull – compare whether upper jaw bones are fused with cranium or not

- lower jaw – many bones, a few or one?

- homodont or heterodont teeth or no teeth

- mammals – compare dentition among mammals of different dietary habits

- vertebral column – are differences in the vertebrae in different parts of the column
(regionalization) – what are the regions in tetrapods? same length for all regions?

- vertebrae – fusion in tetrapods?

- frogs - urostyle

- birds - synsacrum

- mammals - sacrum

- ribcage
- ribs – note extent of ribs among different vertebrate classes

- shape of sternum – is it different in different groups

Appendicular skeleton – girdles & limb bones
- girdles – look at support for fins in fishes

- pectoral girdles – clavicle, coracoid & scapula – note size & position in tetrapods

- pelvic – os coxae (ilium, ischium, pubis) - note shape & position and fusion of bones
– connection w/vertebral column

- anterior/superior appendages – in tetrapods

- know bones:
- humerus
- ulna
- radius
- carpals
- metacarpals
- phalanges (phalanx, s)
- compare among tetrapods, including frogs, turtles, bats, bird, humans, cats vs. horse

- posterior/inferior appendages – in tetrapods

- know bones (sometimes these bones are fused):
- femur
- tibia
- fibula
- tarsals
- metatarsals
- phalanges
- compare among tetrapods, including frogs, turtles, bats, bird, humans, cats vs. horse
 Bird and Mammalian Taxonomy

Class Aves – 25 extant orders

- recognize representative pictures of the following taxa

Superorder Paleognathae
- ratites (unkeeled sternum) – ostriches, emus, kiwis
- tinamous

Superorder Neognathae
O. Gaviiformes
O. Anseriformes
O. Falconiformes
O. Galliformes
O. Passeriformes

Class Mammalia – 27 orders

- recognize representative pictures and skulls or limbs of the following orders

Subclass Prototheria
O. Monotremata
Subclass Theria
Infraclass Metatheria – marsupial mammals
O. Didelphimorphia
Infraclass Eutheria
O. Primates
O. Lagomorpha
O. Rodentia
O. Carnivora
O. Perissodactyla
O. Artiodactyla