The Old Martyr of Science:

The Frog in Experimental Physiology

FREDERIC L. HOLMES

History of Medicine
Yale University School of Medicine
333 Cedar Street
New Haven, Connecticut 06510-8015

During a lecture at the International Physiological Congress in

1929, August Krogh stated: "For a large number of problems there
will be some animal of choice, or a few such animals on which it
can be most conveniently studied." After recounting the example
of a certain kind of tortoise whose unusual lung structure made it
exceptionally suited for respiration experiments conducted by his
teacher, Christian Bohr, Krogh commented: "We used to say as a
laboratory joke that this animal had been created expressly for
the study of respiration physiology. I have no doubt that there is
quite a number of animals which are similarly 'created' for special
laboratory purposes, but I am afraid that most of them are unknown
to the men for whom they are 'created' and we must apply to the
zoologists to find them." Krogh's assertion, scarcely elaborated
beyond these brief passages, made so lasting an impression on a
young biochemist named Hans Krebs, who attended the lecture, that
Krebs wrote a paper forty-fiv e years later on what he named "The
August Krogh Principle. ''I
When he heard Krogh's lecture, Krebs was already aware that
several of the scientific successes of his own mentor, Otto Warburg,
had depended on Warburg's choice of biological organisms
peculiarly well adapted to the particular type of experiments he
aimed to perform: the sea urchin egg for the increase of energy
metabolism after fertilization, because of its high proportion of
active cell constituents to yolk; suspensions of a unicellular green
alga for photosynthesis, because it was readily available and could
be pipetted; and transplantable experimental rat tumors to measure

1. August Krogh, "The Progress of Physiology," Amer. J. Physiol., 90 (1929),

247; Hans A. Krebs, "The August Krogh Principle: 'For Many Problems There
Is an Animal on Which It Can Be Most Conveniently Studied,'" J. Exp. Zool.,
194 (1975), 221.

Journal of the History of Biology, vol. 26, no. 2 (Summer 1993), pp. 311-328,
9 1993 Kluwer Academic Publishers. Printed in the Netherlands.
312 FREDERIC L. HOLMES

the metabolism of cancer cells, because they contained only actively

growing cells. 2
Kreb's own achievements benefited also from such especially
convenient "materials" - most prominently, from the use of pigeon
breast muscle tissue to study oxidative metabolism. The excep-
tionally high rate of metabolism maintained by this tissue after it
was minced and placed in suspension in a manometer flask made
measurements of its metabolic products and exchanges larger and
more reliable than anything that could be obtained at the time
from any other easily accessible tissue. The peculiar properties of
pigeon breast muscle were instrumental to the discovery of the
Krebs cycle in 1937. 3 In the paper he wrote in 1975, Krebs recalled
these examples from his own experience and enumerated other
cases from diverse biological fields. Then he made a few gener-
alizations. "It is an important point," he wrote, "that a relatively
minor modification of a standard situation may present great advan-
tages in studying a phenomenon without affecting basic principles."
Among the advantageous modifications he mentioned were unusu-
ally large size, the magnitude of the process under study, and a
convenient anatomical arrangement. The "general lesson" he drew
from the Krogh Principle was simple and pragmatic: It is impor-
tant to "look out for a good experimental material when trying to
tackle a specific biological problem. ''4
Krogh and Krebs were each protesting, in part, the tendency
of physiologists and biochemists to use the same organisms over
and over, without searching through the rich diversity of the animal
or plant world for just that organism which might be best suited
to their particular experimental needs. In many subfields of the
experimental life sciences, one or a few organisms have, in fact,
become the dominant subjects of investigation. Have they done
so because they proved "most convenient" for the special require-
ments of a problem central to the definition of such fields, or
perhaps because their characteristics made them convenient for a
larger range of problems than other organisms? Or is such domi-
nance often attributable more to the inertia that leads investigators
to continue using organisms with which they are familiar rather than
to explore less-known ones? Undoubtedly the historical patterns
differ from example to example, and we should examine partic-
ular cases in detail before erecting broad generations.

2. Krebs, "Krogh Principle," p. 221.

3. Ibid., p. 222; H. A. Krebs and W. A. Johnson, "The Role of Citric Acid
in Intermediate Metabolism in Animal Tissues," Enzymologia, 4 (1937), 148-156.
4. Krebs, "Krogh Principle," pp. 222-226.
The Frog in Experimental Physiology 313

We may note also that the validity of the strategy of relying

on an organism with special properties when studying a general
biological phenomenon rests on the assumption expressed in
Krebs's statement that the features unique to the organism in
question do not alter "basic principles." Throughout the history
of biological investigation, however, the question of how far one
can extend conclusions drawn from particular organisms to other
organisms (often most crucially to humans) has remained prob-
lematic.
The fundamental idea that a problem examined in a specific
animal can be used to draw conclusions applicable to other animals
- as well as the idea that there are limits on the degree to which
it can be done - can be traced back to Aristotle. Aristotle's view
that all animals have some essential parts in common; that the
majority have additional parts in common, but that other parts
may be identical or different; and that the parts that are common
may differ in character or position and in "excess or defect," was
grounded in his rich knowledge of the comparative anatomy of
the major classes of animals. His own efforts to draw general
inferences from the examination of specific animals illustrated both
the benefits and the risks that investigators ever since have
encountered in following his powerful precedent. 5
Galen displayed a well-developed sense for the appropriateness
of certain animals for particular purposes. For detailed dissections
intended to elucidate human anatomy, which he could not directly
examine because of the prohibitions against human dissection, he
chose the Barbary ape - that is, of all the animals available to
him, the one whose anatomy most closely resembled human
anatomy. For some of his most sophisticated vivisection experi-
ments, however, he used pigs, because they more closely resembled
humans in the functional arrangements most pertinent to his
investigation. 6
As is well known, William Harvey made creative use of a wide
diversity of animals, each especially convenient for studying certain
aspects of the heartbeat. They ranged from humans (for the non-
invasive ligature experiments) to dogs, pigs, frogs, snakes, eels,
fish, and shrimp. Underlying his resort to all these animals was

5. Aristotle, Historia animalium, trans. A. L. Peck (Cambridge, Mass.: Harvard

University Press, 1965), pp. 3-31.
6. Galen, On Anatomical Procedures, trans. Charles Singer (London: Oxford
University Press, 1956), pp. 2-3; Galen, On Anatomical Procedures: The Later
Books, trans. W. L. H. Duckworth (Cambridge: Cambridge University Press, 1962),
pp. 88-99.
314 FREDERIC L. HOLMES

Harvey's conviction that, despite the enormous differences in the

size, configuration, and rate of activity of their hearts, the funda-
mental nature of the heartbeat was the same in all animals that
possessed one. 7

THE RISE OF THE FROG

Of all the animals that have proved particularly convenient for

the study of special problems, few have served so prominently
for so long as has the familiar frog. In the seventeenth century,
as Harvey's discovery of the circulation stimulated other investi-
gators to reexamine other animal functions, they quickly learned
that frogs lent themselves very well to the resolution of some
questions that were more difficult to answer with warm-blooded
animals. Frogs continued to be especially useful to eighteenth
century experimenters. By the nineteenth century they were
experimented on so frequently that, when Hermann Helmholtz
turned to them in 1845 for his first experiments on muscle action,
he referred to these creatures as "the old martyrs of science. ''8 In
the rest of this paper I intend to illustrate, through several examples
of landmark experiments performed on frogs, some of the prop-
erties that made them so popular with and so important to
physiologists.
In September 1660, Marcello Malpighi began to investigate
sheep lungs to clarify the relations between their arteries, veins, and
trachea. After inflating and injecting the lungs, he observed them
with a microscope. He found that they did not consist of a "fleshy"
porous parenchyma, as he had thought, but were composed "solely
of membranous, air-filled vesicles." He published a letter on this
discovery in January 1661. At that point he believed that the arteries
and veins opened into these vesicles. Afterward, however, he began
to examine frog lungs under his microscope; these were simpler
and more transparent than the sheep lungs, and could be kept
inflated while exposed to view. Only then did he find the "fan-
tastic red network" connecting arteries and veins in a closed system,
making the observation that is regarded as the historical comple-
tion of Harvey's discovery of the circulation of the blood. 9

7. William Harvey, Anatomical Studies on the Motion of the Heart and Blood,
trans. Chauneey D. Leake (Springfield, Ili.: Charles C. Thomas, 1928), pp. 28,
42-46.
8. Hermann Helmholtz, "Ueber Stoffverbrauch bei der Muskelaction," Arch.
Anat., 1845, p. 74.
9. Marcel Malpighi, Discours anatomiques sur la structure des visc~res, 2nd
ed. (Paris: L. d'Houry, 1687), p. 334; Howard B. Adelmann, Marcello Malpighi
The Frog in Experimental Physiology 315

The salient feature of the frog in this famous case was that it
could be treated as a simplified, more accessible version of a
mammal. The anatomical feature that could, under the conditions
in which Malpighi worked, be seen only in the frog, could never-
theless be considered common also to warm-blooded animals,
including humans. Frogs had simpler lungs than mammals, but since
they looked similar, were similarly placed, and were similarly
connected to blood vessels and trachea, there was no reason to doubt
that their basic structure and functions corresponded to those of
the lungs in "higher" animals.
In the mid-seventeenth century several people, including Ren6
Descartes and William Croone, proposed mechanical explanations
of muscular motion. These speculations invoked variants on the
theme that a subtle fluid flowing through the nerve into a muscle,
or a fermentable fluid dilating explosively within it, inflated the
muscle, causing it to expand in width and shorten in length. A
necessary corollary of such views was that the volume of the muscle
increases during its contraction. The Dutch naturalist Jan
Swammerdam considered the "real causes of muscular action" to
be shrouded "in the thickest clouds of obscurity." In an effort to
dissipate some of these mists, Swammerdam "made many experi-
ments, at different times, on the motion of the muscles." He
performed them "chiefly on the frog; for the nerves are very
conspicuous in these animals, and may be easily discovered and
laid bare"; moreover, the "motions of the muscles are not so
considerable in animals which have warm blood, or rather they
do not last so long." Swammerdam conducted his most decisive
experiments on "the largest m u s c l e s . . , separated from the thigh
of a frog, and together with its adherent nerve, prepared in such
a manner as to remain unhurt." The very fact that he could stim-
ulate the muscle to move by irritating a short segment of nerve
attached to it but isolated from the rest of the frog was sufficient
to satisfy him that he had disproved the idea that "any matter of
s e n s i b l e . . , bulk flows through the nerves into the muscles. ''1~
Swammerdam carried the investigation much further. In an extra-
ordinarily elegant experiment, he enclosed a frog thigh muscle with
its adherent nerve, to which he attached a fine silver wire, within

and the Evolution of Embryology (Ithaca, N.Y: Cornell University Press, 1966),
I, 172-198.
10. L.G. Wilson, "William Croone's Theory of Muscular Contraction," Not.
Rec. Roy. Soc. London, 16 (1961), 158-178; Jan Swammerdam, The Book of Nature,
or the History oflnsects, trans. Thomas F. Lloyd (London: C. G. Seyffert, 1757),
pt. 2, pp. 122-124.
316 FREDERIC L. HOLMES

a tube whose upper end was extended into a fine capillary; within
the capillary was a drop of water (see Fig. 1). By pulling on the
wire, which extended from the airtight interior to the exterior of the
tube, he irritated the nerve, inducing the muscle to contract. The
drop of water did not rise in the capillary, but actually sank
somewhat, indicating that the muscle - far from expanding -
occupied "a smaller space" when contracted than when relaxed.
This experiment was, Swammerdam remarked, "very difficultly
sensible, and requires so many conditions to be exactly performed,
that it must be tedious to make it." Nevertheless it was so beauti-

Figure 1. Drawing of the experimental arrangement for Jan Swammderdam's

experiment on volume change in muscle contraction, from The Book of Nature,
trans. T. F. Lloyd (London: C. G. Seyffert, 1757), plate XLIX.
The Frog in Experimental Physiology 317

fully conceived as a crucial test of contemporary theories of

muscular contraction, that those to whom Swammerdam reported
the result, in particular Nicolas Steno, were henceforth constrained
to formulate theories of muscular contraction compatible with the
condition that muscles do not expand in the p r o c e s s , u
The success of Swammerdam's experiment depended critically
on properties peculiar to frogs. The convenient size of their thigh
muscles, the fact that the muscle could be dissected out together
with its nerve, and the considerable time for which this isolated
preparation retains its ability to contract vigorously when its nerve
is stimulated, enabled Swammerdam to perform a "delicate and
useful experiment" that would have been far harder to do on any
other available organism. Despite these particularities, he had no
doubt that the result was generalizable. The section of his Book
of Nature in which this investigation is discussed is headed
"Experiments on the particular motion of the muscles in the frog,
which may be also, in general, applied to all the motions of the
muscles in men and brutes. ''12 To put the matter in the language
of Krebs's "August Krogh Principle," the "great advantages" offered
by the convenient size of the frog muscle and by the magnitude and
tenacity of the contractions it could produce in isolation, did not
affect the "basic principles" of muscle contraction.
In the mid-eighteenth century, Albrecht von Hailer and Robert
Whytt engaged in a critical, prolonged, and ultimately unresolved
debate about irritability, sensibility, and the question of whether the
sentient soul is confined to the brain or extends throughout the body.
Each side adduced evidence drawn from a wide range of experi-
mental and pathological observations. During the course of their
exchanges Whytt developed the novel and fundamental idea that
there exist in the animal body coordinated "involuntary" motions.
The most compelling evidence that he brought forth in support of
this view was an experiment on a frog: "the b o d y . . , after being
divided from its head preserves the power of motion for above
an hour; when its hind feet or toes are cut, or otherwise hurt, the
muscles of its thighs, legs, and trunk are strongly contracted, by
which it raises its body from the table, and sometimes moves from
one place to another. ''13 Whytt called these movements "sympathetic
motions," because he took the organs to be excited into their
coordinated actions by a particular sympathy for one another that

11. Swammerdam, Book of Nature, p. 127.

12. Ibid., p. 122.
13. Robert Whytt, Physiological Essays, 3rd ed. (Edinburgh: I. Balfour, 1766),
pp. 158-159.
318 FREDERIC L. HOLMES

they have through their nerve connections. If the spinal marrow

is destroyed by a hot wire, all sympathetic motions are eliminated
because these communications through the nerves are cut off;
afterward one can only evoke convulsions of individual muscles by
irritating nerves attached directly to them.
In Whytt's experiment the critical property of the frog was the
tenacious capacity of its body to function in a coordinated way after
a severe mutilation. The importance of the decapitation was that,
because sensation and will were considered properties of the brain,
the motions in question could be neither voluntary nor a response
to a sensation that had reached the brain. Snakes and tortoises could
similarly withstand decapitation, and Whytt in fact performed such
experiments also on these animals. Then and later, however, for
reasons that seem almost self-evident, frogs became the favorite
subjects for this brutal treatment. After Marshall Hall extended
Whytt's idea of "involuntary movements" to the broader and more
sharply defined concept of "reflex movements" in the 1830s, many
frogs lost their heads for such experiments. TM Among the most
important were those of Alfred Volkmann, who in 1838 examined
systematically the effects of longitudinal and transverse cuts in
the spinal cord on the reflex movements of the limbs of previ-
ously decapitated frogs. 15
The most famous frog experiment in the history of science was
the discovery by Luigi Galvani in 1791 that when an arc composed
of two metals touches a nerve, or one end touches a nerve and
the other end touches the muscle to which the nerve is attached, the
muscle contracts. 16 That Galvani happened to discover the phe-
nomenon with a pair of frog legs is one of the classic illustrations
of chance in science. It was not by chance, however, that others
who entered this new field to confirm, vary, and otherwise explore
this phenomenon also used frogs most often as their experimental
animals. Maria Trumpler has recently analyzed the work of more
than a dozen people who performed such experiments in the first
decade after Galvani's discovery. They modified the experimental
procedures in many ways, but all of them performed the experi-
ments mainly on the thigh muscles of frogs. When they sometimes

14. Marshall Hall, "On the Reflex Function of the Medulla Oblongata and
Medulla Spinalis," Phil. Trans. Roy. Soc., 1833, pp. 635-665.
15. A. W. Volkmann, "Ueber Reflexbewegungen," Arch. Anat., 1838, pp.
15-43.
16. Marcello Pera, The Ambiguous Frog: The Galvani-Volta Controversy on
Animal Electricity, trans. Jonathan Mandelbaum (Princeton: Princeton University
Press, 1992), pp. 69-90; J. L. Heilbron, "The Contributions of Bologna to
Galvanism," Hist. Stud. Phys. BioL Sci., 22 (1991), 57-85.
The Frog in Experimental Physiology 319

did test whether the phenomenon was generalizable to other

animals, particularly to mammals, they found the experiments much
more difficult to carry out. Not only were frogs the easiest to handle
- whether intact, flayed, or dissected into various-sized pieces of
nerve and muscle - but also, as Trumpler notes, the phenomenon
lasted for up to 30 hours in an isolated frog leg, whereas in cats
or dogs it disappeared within 15 minutes. 17
Frogs continued to be the preferred subject for experiments on
animal electricity through the first half of the nineteenth century.
Carlo Matteucci's "rheoscope" - an isolated frog nerve-muscle
preparation - was viewed as the most sensitive indicator of galvanic
currents, even after the "multiplicator," an instrument based on
the discovery of electromagnetism, became common. When Emil
Du Bois-Reymond took up his landmark experiments on animal
electricity during the 1840s, the frog was so well established as
the subject for such studies that he apparently never seriously con-
sidered alternatives. Du Bois-Reymond performed all of his delicate
measurements on the currents and potentials within nerves and
muscles, and the changes brought about by stimulating them, on
frogs and parts of frogs (Fig. 2). By then the current detectable
between the surface and a cut section of a muscle was known simply
as the "Froschstrom. ''18
From the fact that more than half a century of investigation of
animal electricity was dominated by experiments on frogs, can
we infer that, under the conditions of the time, this was the optimal
organism for the job? Or had the use of frogs merely become so
habitual that experimenters lacked the imagination to seek out other
animals that might sometimes have better served their purposes?
Did the accumulating experience with frogs make it increasingly
advantageous to continue experimenting with them, rather than to
start over with another organism? Did the combination of their
innate characteristics, their familiarity, and their easy availability
make them unmatchable? Were there any peculiar characteristics of
frogs that shaped the long-term direction of these investigations
in ways that might have been different if some other organism
had achieved preference? It will require fine-structured studies
of this experimental tradition, which have so far not been done,
to reveal whether any opportunities were missed, or whether

17. Maria J. Trumpler, "Questioning Nature: Experimental Investigations of

Animal Electricity in Germany, 1791-1810," Ph.D. diss., Yale University, 1992,
pp. 65-98.
18. Emil Du Bois-Reymond, Untersuchungen iiber thierische Elektricitiit,
vol. I (Berlin: G. Reimer, 1848).
320 FREDERIC L. HOLMES

Figure 2. Illustrations for experiments of Emil. Du Bois-Reymondusing frogs

and parts of frog muscles,from Untersuchungen fiber thierische Elektricitdt (Berlin:
G. Reimer, 1848), vol. I, plate IV.

obstacles that might sooner have been overcome persisted, because

experimenters did not regularly employ a wider range of animals
in their work on animal electricity. The extended study of this work
on which Maria Trumpler has embarked may eventually answer
some of these questions.
I turn next to a case in which frogs were not the primary exper-
imental subject, but in which their special characteristics helped
clinch a discovery made initially on mammals, the validity of which
had remained in doubt. In 1822 Francois Magendie made the
momentous observation that "the anterior and posterior roots of
the nerves that arise in the spinal cord have different functions, that
the posterior appear more particularly intended for sensibility,
whereas the anterior appear more particularly connected with
movement." Magendie arrived at this cautious statement of the
distinction between the anterior and posterior nerve roots by per-
forming delicate surgical operations on six-week-old puppies.
Severing the anterior roots without disturbing the posterior roots
The Frog in Experimental Physiology 321

proved particularly difficult, He succeeded by maneuvering a thin,

curved cataract knife carefully past the posterior roots while
managing to avoid cutting the closely associated blood vessels.
Magendie marshalled his considerable surgical dexterity in mini-
mizing the trauma inflicted beyond that necessary to eliminate
selectively the respective connections of the posterior and anterior
roots with the central nervous system, and thus he was able to
observe a functional differentiation that eventually became funda-
mental to the whole of modern neuroscience. 19
In its own time, however, Magendie's discovery did not go
uncontested - even if we ignore the acrimonious priority dispute
provoked by Charles Bell's spurious, but widely credited, claim
to have discovered the phenomenon before Magendie. Not only did
Magendie qualify the distinction with the phrase "more particu-
larly," but he could not invariably replicate his own results. Other
experimenters obtained conflicting results, and controversy sur-
rounded the discovery for more than a decade. 2~
The young Johannes Mtiller, concerned about the "completely
doubtful and uncertain results" of Magendie's initial experiments,
attempted in Berlin to confirm them in 1824, using rabbits. Mialler
ran into insuperable difficulties. Opening the spinal cord of these
animals subjected them to such extreme pain that he thought it
impossible to tell whether they were sensible to a stimulation of the
spinal nerve roots. Several years later, while still skeptical that
Magendie - or anyone else who had in the meantime attempted
the experiments - had overcome these obstacles, Mtiller "came upon
the happy thought to apply frogs to the controversial experiments,
animals whose life is so tenacious that they long survive the opening
of the spinal cord, whose nerves remain sensible for a long time,
and in which the thick roots of the nerves to the lower extremi-
ties run separated for a long distance within the vertebral canal
before they join together. ''21 Here both the general hardiness with
which frogs can withstand severe operations, and a particular
anatomical arrangement, made them for Mtiller the most convenient
animal available for studying the functions of the spinal nerve roots.
He experienced immediate success: "These experiments are so
easy," he reported, "that everyone can henceforth persuade himself

19. Francois Magendie, "Exptriences sur les fonctions des racines des nerfs
rachidiens," J. Physiol. Exp. Pathol., 2 (1822), 276-279.
20. Paul F. Cranefield, The Way In and the Way Out: Franfois Magendie,
Charles Bell and the Roots of the Spinal Nerves (Mount Kisco, N.Y.: Futura, 1974).
21. Johannes Miiller, "Best~itigung des Bell'schen Lehrsatzes, dass die dop-
pelten Wurzeln der Rtickenmarksnerven verschiedene Function haben," Froriep's
Not. Gebiete Nat. Heilk., 30 (1831), 114-115.
322 FREDERIC L. HOLMES

of one of the most important truths in physiology." Mtiller could

state his results without qualifications. For example, "the stimulus
of the anterior roots through a galvanic current immediately causes
the strongest contraction. Galvanic stimulation of the posterior roots
never causes a trace of a contraction.''22
Perhaps there was some exaggeration in Mtiller's claims. By
emphasizing the uncertainty of earlier results and the absolute
nature of his own, he constructed his case for a share in the credit
for a great discovery. We might also view the situation as an
instance of Harry Collins's generalization that scientists never
replicate exactly a previously published experiment. There is no
originality in merely duplicating a result - whereas to attain the
same result, if possible with greater reliability, by some variation
in the earlier experiments, is viewed as helping to move the field
forward. 23 Nevertheless, we can say at least that Mailer showed
the frog to be ideally suited to demonstrate Magendie's discovery,
and at most that Miiller solidified a discovery that had, until then,
been incomplete.
It may also be that Mtiller and others could not reproduce
Magendie's results with warm-blooded animals because they were
not as skilled in the surgical techniques required. As John Lesch
has shown, Magendie was the most prominent representative of
the emergence in France of an experimental physiology centered
upon vivisection and dependent upon the application of surgical
experience gained by training within the reformed medical insti-
tutions of postrevolutionary Paris. 24 It is not accidental that
Magendie's student, Claude Bernard, who was at least the equal
of his mentor in operative virtuosity, pursued the problems that
had rendered Magendie's experiments on the spinal roots uncer-
tain by refining the experimental techniques on mammals, by
examining in finer detail the anatomical organization of the spinal
nerves, and by tracking down all of the experimental conditions that
could cause the experiment to succeed or to fail.
It is probably also not coincidental that Johannes Mtiller, who
was a fine comparative anatomist and zoologist, rather than a
trained surgeon, should have turned instead to an animal for which
the vivisection procedures required were simpler. What consti-

22. Ibid., pp. 115-118.

23. H. M. Collins, Changing Order: Replication and Induction in Scientific
Practice (London: Sage Productions, 1985), pp. 34-38.
24. John E. Lesch, Science and Medicine in France: The Emergence of
Experimental Physiology, 1790-1855 (Cambridge, Mass.: Harvard University Press,
1984), pp. 1-98.
The Frog in Experimental Physiology 323

tutes the best organism for a particular experiment is not entirely

an objective matter: it can depend as much on the subjective talent
and prior experience of the individual investigator as it does on
the nature of the task to be performed.
Hermann Helmholtz relied mainly on frogs for quite different
reasons, in the classic series of experiments on muscle and nerve
action that he performed between 1845 and 1851. In the first of
his papers on this subject, entitled "On the Consumption of Matter
in Muscle Action," he explained why the frog was the animal best
suited to his needs. His purpose was to prove that a measurable
transformation of matter takes place "in the muscle itself' when the
muscle is in action:

In order to bring forth noticeable changes in the chemical com-

position of muscles through their own activity, the muscles
must be continuously withdrawn from the compensatory influ-
ences of the circulation, therefore the experiments must be
performed on isolated parts or on dead animals. This require-
ment makes it necessary to turn again to the old martyrs of
science, the frogs, because in warm-blooded animals the
excitability diminishes rapidly after death, and fish muscles react
comparatively much more weakly to an intense stimulus.25

Helmholtz proceeded by isolating pairs of leg muscles from two

to four frogs, leaving half of them at rest while he stimulated the
other half with a rapid series of electrical discharges until they were
exhausted. Using the limited but refined techniques then avail-
able for extracting substances from animal tissue, he tried to
determine whether the quantities of the few identifiable substances
were increased or decreased in the contracted muscles relative to
the quantities in the uncontracted ones. He could observe no
measurable differences in any specific substances, but he was able
to establish that "in all the experiments, without exception, the
substances extractable in water were diminished in the electrified
portions of muscle; conversely the alcohol extracts were increased
with respect to the unelectrified portions. ''26
Simple though this result may appear, it was, in view of the
complexity of the problem and of the fact that Helmholtz was
making the first direct experimental attack on the fundamental
question of what chemical changes occur within muscles during
their activity, a stunningly successful outcome. "In order to test

25. Helmholtz, "Stoffverbrauch" (above, n: 8), pp. 73-74.

26. Ibid., p, 77.
324 FREDERIC L. HOLMES

the validity of the result for other classes of animals," he repeated

the experiment on the tail of an eel and the breast muscle of a
pigeon. He found it "much more difficult to conduct" in the
warm-blooded animal. Part of the effect lost by the rapid loss of
excitability in the pigeon muscle was compensated for by "the much
stronger action of the stimulated muscle"; he did, therefore, attain
a "sensible result, although much smaller. ''27 The decisive experi-
ments were those performed on the frog muscles. They initiated
a long tradition of investigation of the chemical changes in muscle
action, in which many of the most productive experiments, until
well into the twentieth century, were conducted with frog muscles.
Helmholtz faced analogous problems in 1847, when he decided
to determine whether the rise in temperature during muscular
activity was due to "processes occurring in the muscle itself," or
to a richer flow of arterial blood. Here, too, it was necessary to
measure the process in muscles isolated from the circulation. Again
he chose frog muscles, this time both "on account of the duration
of their excitability" and because "their vanishingly small internal
heat made it easier to attain a constant temperature equilibrium."
Again he proceeded by comparing the condition of a stimulated and
an unstimulated muscle, measuring the temperature difference with
a specially designed thermocouple-multiplicator apparatus sensi-
tive enough to measure temperature changes of as little as 1/1000
degree. The result was again decisive. 28
These two examples illustrate that the criteria for selection of
the organism most convenient for a given problem can often be less
direct than Krebs's rendition of the Krogh Principle implies. Rigid
adherence to the idea that one ought to measure a process in an
organism in which its magnitude is maximized would have ruled
out frog muscles, because both their material exchanges and their
heat production were very small relative to what Helmholtz could
expect for warm-blooded muscles. To compensate for these obvious
disadvantages, Helmholtz had to improve the precision with which
he measured both processes well beyond the standards of the time.
By doing so, he could benefit from other characteristics peculiar
to frog leg muscles that were particularly advantageous within the
special context of his resourceful experimental design.
By the time Helmholtz moved on in 1848 to study the mechan-
ical characteristics of muscle action, he had the precedent of not
only his own prior work, but that of his most prominent predecessor

27. !bid., pp. 81-82.

28. H. Helmholtz, "0bet die W~rmeentwickelung bei der Muskelaction," Arch.
Anat., 1847, pp. 144-164.
The Frog in Experimental Physiology 325

in this field, Eduard Weber, to induce him to continue to work

with frog muscles. There is no evidence that Helmholtz consid-
ered any other organism during the formative stages of his
investigation. To measure the course of a muscle contraction over
the very brief time interval it occupied, he suspended the isolated
thigh muscle, attached to a section of its nerve, within an
apparatus designed so that its movement would be recorded on
the revolving drum of a kymograph (an instrument recently invented
by his friend Carl Ludwig). Here, too, Helmholtz had to accept
an obvious quantitative disadvantage of the frog muscle - that the
magnitude of its contraction was very small compared to that of the
muscles of large mammals that he might have chosen - so that
he could obtain the benefit of the capacity of a frog muscle, even
isolated and suspended in the air, to continue contracting over an
extended time period. In his early experiments the motions recorded
on the kymograph were so tiny that he had to observe them
through a microscope. Later he was able to redesign his recording
apparatus, so reducing the friction and inertia in its movements that
he could attain precise, characteristic curves even from the very
small movements of the otherwise exceptionally responsive thigh
muscles taken from the old martyrs of scienceY

THE PERSISTENT FROG

I need not further multiply examples to show how central frogs

were to the development of experimental physiology. The promi-
nence of their role was well recognized. In 1840 C. Dum6ril
published a "Historical Notice on the Discoveries Made in the
Observational Sciences by the Study of the Organization of Frogs,"
which began, "Because of their very singular organization, animals
from the order of frogs have provided for persons dedicated to
the sciences of observation extremely favorable circumstances to
interrogate nature in a great number of important researches. ''3~
In 1865 Claude Bernard declared:

29. H. Helmholtz, "Ober den zeitlichen Verlauf der Zuckung animalischer

Muskeln und die Fortpflanzungsgeschwindigkeit der Reizung in den Nerven," Arch.
Anat., 1850, pp. 276-364; idem, "Messungen tiber Fortpflanzungsgeschwindigkeit
der Reizung in den Nerven," Arch. Anat., 1851, pp. 199-216. For fuller
discussion of Helmholtz's experiments, see Kathryn M. Olesko and Frederic L.
Holmes, "Experiment, Quantification, and Discovery: Helmholtz's Early
Physiological Researches, 1843-1850," in The Borders o f Science: Essays on
Hermann yon Helmholtz, ed. David Cahan (Berkeley: University of California Press,
in press).
30. C. Dum6ril, "Notice historique sur les d6couvertes faites dans les sciences
326 FREDERIC L. HOLMES

the animals most used by physiologists are those procured most

easily, and here we must set in the front rank domestic animals
such as dogs, cats, horses, rabbits, oxen, sheep, pigs, barnyard
fowl, etc., but if we had to reckon up the services rendered to
science, frogs would deserve first place. No other animal has
been used for greater or more numerous discoveries, at all points
in science; and even today, physiology without frogs would be
impossible. If the f r o g . . , is the Job of physiology, that is to
say, the animal most maltreated by experimenters, it is certainly
the animal most closely associated with their labors and their
scientific glory. 31

As illustrated previously, Bernard relied less extensively on frogs

for his experiments than did some of his predecessors and con-
temporaries. Exploiting his dazzling operative prowess, he most
often chose, particularly in his early research, to perform complex
vivisection experiments with intact animals, devising delicate
procedures that succeeded best with medium-sized mammals. He
did, however, carry out his famous experiments on the physiolog-
ical action of the poison curare on frogs. The Job of physiology
contributed, therefore, also to Bernard's own scientific glory.
Frogs were, as we have seen, especially prominent in the nine-
teenth century in those types of experimentation in which an
isolated muscle connected to a segment of nerve was used to study
muscle contraction and nerve conduction; the unique combination
of the tenacity of the excitability of isolated tissues common to
cold-blooded vertebrates, with the exceptionally vigorous, rapid
response of the powerful hind legs of frogs, made them ideally
suited to such studies. They were also the most convenient
organism, however, for a broad range of problems that interested
physiologists. Whenever their simplicity, ready availability, and
capacity to survive severe injury with their remaining functions
nearly intact were pertinent, frogs rendered great service to their
tormentors.
In an era in which physiology was oriented primarily toward
the investigation of the functions of organs and systems in which
human physiology was the ultimate point of reference, the frog was
also an animal of choice because it appeared as a schematic "model"
of vertebrate anatomical and functional organization. Psycho-

d'observation par l'6tude de l'organisation des grenouilles," Ann. Sci. Nat. Zool.,
13 (1840), 65.
31. Claude Bernard, An Introduction to the Study of Experimental Medicine,
trans. Henry Copley Greene (New York: Dover, 1957), p. 115.
The Frog in Experimental Physiology 327

logically, frogs probably seemed to resemble humans more than the

fish, snakes, or tortoises that could in some cases have served
equally well for particular experimental purposes; the conformation
especially of the hind legs of frogs seems more "human" than that
of the limbs of many mammals. At a subconscious level flogs
may even have seemed to those who used them to represent
much-simplified versions of the human condition - we all know
how tempting it is in fiction to depict flogs as funny little people.
In the twentieth century, frogs have not dominated physiology
as strongly as they did during the formative stages of that science.
For many purposes small laboratory mammals, especially rats and
mice, have replaced frogs. Twentieth century advances in techniques
and in knowledge of the conditions required to sustain isolated
mammalian or avian tissues overcame the advantage that the
tenacity of surviving isolated frog tissue had once given them.
Experimenters could then exploit, in ways that were difficult in
the nineteenth century, the higher levels of general metabolism
and functional activity of warm-blooded tissues. It is symbolic of
the shift of these relative advantages and disadvantages over time,
that the pigeon breast muscle, which furnished Helmholtz much
weaker effects than frog muscle in his study of chemical change
in 1846, became by the 1930s, when isolated tissues were studied
in fluid media that duplicated the conditions of their internal envi-
ronments, an ideal material for the study of tissue metabolism.
By then the exceptionally high rate of material exchanges in pigeon
breast muscle could be fully exploited to measure chemical changes
far larger than those that take place in frog muscle.
Frogs, nevertheless, still maintain an important place in animal
experimentation. Moreover, as anyone who has taken a high-school
or college biology laboratory course knows, they have retained a
special place in science pedagogy. The properties that made the frog
so convenient for early investigators to study continue to make it
the easiest animal for fledgling experimenters to cope with in their
first efforts to acquire the requisite skills. 32 Many generations of
students have repeated the basic experiments on muscle contraction
that Helmholtz first performed with the frog leg muscle. Still more
basic to many students has been the place of the frog as the first
vertebrate they dissect in the laboratory to learn general anatom-

32. For an insightful discussion of the general historical problem of adapting

pedagogical practices to the level of students beginning to acquire scientific skills,
see Kathryn M. Olesko, "Commentary: On Institutes, Investigations, and Scientific
Training," in The Investigative Enterprise, ed. W. Coleman and F. L. Holmes
(Berkeley: University of California Press, 1988), pp. 302-304.
328 FREDERIC L. HOLMES

ical relationships. I still recall vividly my own experience in a high-

school zoology class. The internal anatomy of the previously
injected frog seemed so schematically simple and clear that its
image has forever after remained in my mind as the prototypical
structure of the vertebrate. One of the series of drawings that I
had to make at the time suggests, particularly in the pose I gave
the outlines of its limbs (Fig. 3), that I too felt that the old martyr
of science somehow mimicked our own postures.

L~- ~ H o l ~
~30
Z o o l o ~ VI
Phqlu,-~ Chordc~fo Clas~ An~ph,b,o E • Fro,:t
Dr-. ~5 Infe.~l orclar~s In pl~ce

~,dno~

Figure 3. Drawing made by F. L. Holmes for high-school zoology course, 1948.