Cut-the-head-off-the-frog Lab

MATERIALS NEEDED:

• Frogs (Rana pipiens).

• Plastic containers to recapture the frogs.
• Several aquaria or other suitable containers with enough water to allow
the frogs to swim.
• Wood platforms to float within the containers.
• Paper towels to cover frogs when not being observed.
• 15 % acetic acid solution.

During this exercise you will determine the nervous system location of various
reflexes in a frog. In brief, the behavior of a normal frog will be observed and then
portions of the nervous system will be sequentially destroyed. If a behavior is still
observed in animals with a lesioned system, then the portion that was destroyed
does not have the nervous pathways for the reflex.
PROCEDURE:
1. Work in groups of three or four and read through the entire procedure before
beginning. Obtain a living frog and capture container from your instructor. Collect
several paper towels and wet them. Use the paper towels to cover the animal
when it is not being observed. Always wet your hands before handling the animal
so as not to cause it undue discomfort. When carrying your frog, make sure that
its legs are held in an extended position. If you allow it to assume a flexed leg
position, it can push against the palm of your hand and will get away. Should the
frog escape, do not stop the frog with your foot. Instead, confine it in the capture
container provided by your instructor.
2. Frog Reflex Procedure. The frog will serve as it's own control in this
experiment. Begin by observing the behaviors (listed below; step 5) in a living
frog. When recording the behaviors you may find it helpful to make a few simple
sketches. Move slowly and speak quietly when near the animal. Fast movements
or loud noises will startle the frog, and you will spend much of your time retrieving
escaped animals. In addition, a frightened animal is not as likely to perform all of
the behaviors.3. After the frog's normal behavior has been recorded, take the
animal to your instructor, and he or she will remove the cerebrum from the animal
by cutting directly behind the eyes (Figure 1). If the procedure is quickly done,
the animal will experience little or no discomfort, much as when you get a really
bad burn or cut (it takes a few moments to experience the pain). Once the
cerebral hemispheres are removed, the frog is dead and is termed a decerebrate
frog. Although you may find it unsettling to watch a headless frog move about,
keep in mind that the animal is dead and therefore feels no pain. In order to feel
 OLFACTORY LOBE

CEREBRUM

OPTIC LOBE

FIRST CUT
(DECEREBRATE FROG)
REMOVAL OF MEDULLA
(DEMEDULATED)
MEDULLA
REMOVAL OF SPINAL
CORD (SPINAL FROG)

SPINAL CORD

Figure 1. Procedure to create a decerebrate frog.

pain, it must perceive the pain. Centers for perception of pain are located in the
cerebrum, which has been removed. Cover the frog with moist towels and allow it
to recover for a few minutes and then record the behavior of the decerebrate
frog. Expect to see some decrease in the spontaneous movements of the animal
(you may have to try for a response several times or prod it). When you have
finished recording the behavior of the decerebrate frog, return it to your instructor
for destruction of the medulla. The medulla is destroyed by inserting a dissecting
needle into the opening created by the cut and then moving it back and forth.
Care must be taken that the spinal cord is not damaged. These animals are
demedullated frogs and are, of course, still dead. Again, cover the animal with
moist towels, wait a few minutes, and re-record the behaviors. Finally, your
instructor will destroy the spinal cord by inserting and twisting a dissecting needle
down the hole previously occupied by the medulla. The procedure is finished
when the hind legs straighten out completely and then go limp. Return to your
station, cover the frog for several minutes, and rerecord the behaviors. Dispose
of the frog as directed by your instructor.
4. As portions of the central nervous system are removed, behaviors that require
those structures will drop out. Thus, if a behavior depends on the presence of the
medulla, it will be seen in the normal and decerebrate frog, but will disappear
when the medulla is removed. If a behavior is not immediately seen, retest the
animal before continuing with the rest of the lesion experiment. Use your data to
complete the Results Section table.
5. Behaviors to Record. Cover the animal with moist towels and allow it to
adjust for a few minutes. Always moisten your hands before handling the animal.

• Carefully remove the towels and observe the resting position of your animal.
(Careful! It may jump when the towels are removed.) How are the front and
back legs positioned? Is the body parallel with the surface of the table or at an
angle? If an experimental animal is not standing in a normal position, prod it at
the front to try and make it stand.
• Surround the animal. One member of the group should gently prod the frog into
jumping (this procedure is probably best done on the floor). Record the general
form of the jumping behavior. Does the frog pull its legs in while in the air, or
are they extended?
• Restrain the animal while one member of the group gently pulls on one of the
hind legs to straighten it. When the leg is released, is it returned to the flexed
position? This is the extension reflex.
• Turn over the animal and record the presence or absence of a "righting
response." How are the front and hind legs used when the animal rights itself?
This behavior is fairly rapid, and you'll probably have to try it several times to
see the movements of the legs and body.
• Turn your frog over and hold it upside down. Moisten your finger and gently
stroke its belly. Male frogs may clasp your finger with their forelegs. This is part
of their sexual behavior and is termed the "clasping response." Don't worry; you
won't get "frog stuff" on you. Females and males of low sexual tone may only
be quieted by the stroking (the "quieting response").
• Take a small piece of paper towel (about 5 mm X 5 mm) and soak it in 15%
acetic acid. Place the towel on the body of the frog near the rear of the animal.
Depending on the condition of the animal, the frog may attempt to remove the
towel with its hind leg. This "scratch reflex" may be seen in a different form
when the frog's head is removed. The frog may reach forward toward its
missing head with its forelegs. The animal is not searching for its head; the
central nervous system is merely showing a scratch response to irritated nerve
endings (thus, count this movement as a positive scratch reflex). If the animal
does not show a scratch response within 30 seconds, remove the towel and
wash the area with a few drops of water. The absence of a scratch reflex in a
normal frog simply indicates that the animal is too agitated to show the
behavior. In experimental frogs, the absence of a scratch reflex should be
considered as being caused by the treatment.
• Take the animal over to a swimming container and drop it in to record the
movements used in swimming. If the frog contacts the floating board, it may
attempt to climb on the board. Move the board closer to the frog if it does not
contact the board after a few minutes. For dead frogs, help them climb by
placing the forelegs on the board. Describe the movements involved both in
swimming and climbing. Turn the frog over and record the righting response in
water.
• Remove the frog from the water and watch the motion of the frog's throat.
Frogs force air into their lungs by gulping air and then swallowing it (similar to
when you swallow air to force a burp). Count the number of throat motions over
a one-minute period to determine the frog's respiration rate. Record for a
second minute, average the results, and record this as the rate if respiration.
Headless frogs will show less conspicuous throat movements and no
movement of their sides.
• Take the frog to your instructor for the next part of the experiment and begin
again with the first step if you have not finished recording with a spinal frog.
• When you have completed the spinal frog portion of the experiment, you should
open your frog and look for signs of a heartbeat, identify the sex of the animal,
and locate parasites (if any). The following illustrations will serve as an aid to
the identification of frog structure and are included so that this lab follows Dr.
Bill's Conservation of Meat Law.

LINGUAL ARTERY
INTERNAL CAROTID

EXTERNAL CAROTID
CAROTID

CAROTID BODY
CUTANEOUS

PULMONARY
SUBCLAVIAN

SYSTEMIC ARCH

LUNG LIVER

STOMACH
DORSAL
AORTA
HEPATIC ARTERY
FAT BODY
GASTRIC ARTERY
OVARIAN
ARTERY MESENTERIC ARTERY

SMALL INTESTINE
OVARY

RENAL ARTERIES
KIDNEY
ILIAC ARTERY

Frog Arteries
 INNOMINATE
LINGUAL
INTERNAL JUGULAR
MANDIBULAR

EXTERNAL SUBCLAVIAN
JUGULAR
BRACHIAL

CUTANEOUS
PULMONARY

VENA CAVA
LUNG
HEPATIC
LIVER
FAT BODY
HEPATIC PORTAL
TESTIS
SPERMATIC
RENAL VEINS

KIDNEY
SMALL INTESTINE

RENAL PORTAL VENTRAL ABDOMINAL

VEIN
PELVIC VEIN

FEMORAL VEIN

SCIATIC VEIN

Frog Veins
 TRUNCUS
ARTERIOSUS
LEFT ATRIUM

RIGHT
ATRIUM COELIACO-
MESENTERIC
LUNG ARTERY

VENTRICLE STOMACH
LIVER
PANCREAS
VENA CAVA
KIDNEY

VENTRAL SPLEEN
ABDOMINAL
VEIN SMALL
INTESTINE
BLADDER
DORSAL AORTA
COLON

LEGS (YUM!)

Frog Gutz
 SUPRASCAPULA PHALANGES PREMAXILLA
A NASAL B
METACARPALS SPHENETHMOID

CARPALS
FRONTAL-PARIETAL

RADIOULNA
PTERYGOID
HUMERUS
VERTEBRAE
FEMUR MAXILLA
SACRAL VERTEBRA

QUADRATO-
JUNGAL

PHALANGES PRO-OTIC ATLAS

ASTRAGALUS
METATARSALS EXOCCIPITAL SQUAMOSAL

TIBIOFIBULA ISCHIUM CALCANEUM

TARSALS ILIUM UROSTYLE