Embryology   Biology 441   Spring 2013   Albert Harris


More on Driesch, Roux, and regulative development



Hans Driesch (1890s), doing research at marine laboratories in Trieste and Naples, (originally using early embryos of starfish), discovered:

"Embryonic Regulation"

But please don't be misled by the word "regulation",
which can seem to imply some exterior control telling somebody what to do.

This kind of regulation is internal adjustment, to fix damage or overcome abnormality or effects of some external disturbance.

Embryos of many kinds of animals are not regulative.
They are said to have "mosaic development"

More regulative development    <------------->    More mosaic development:

Mammals    Echinoderms    Amphibians         Sea Squirts    Flies    Snails     Nematodes

      If species use ooplasmic segregation
      their embryos development will be mosaic.

      Very consistent cleavage patterns,
      and consistent cell lineages
      occur in species with mosaic development.

But there isn't any exact quantitative measure of how regulative or how mosaic embryonic development is in any given kind of animal.
(Because the mechanistic causes are not yet understood well enough.)

Many people think the key difference is just how soon in development cells become irreversibly committed to differentiate into specific cell types, and to form particular parts of the body.
The sooner cells become irreversibly committed,
The more mosaic the development of those kind of embryos will be.

Other people think that some fundamental differences in mechanisms are the cause of development being more regulative or more mosaic.

(for example if differentiation were controlled by special proteins or other molecules being put into particular cells at the time of cleavage, that would cause development to be more mosaic.)

Regulative development would be impossible without cell-cell communication. (I guess?)

The theory called "Positional Information" claims that regulative development proves that cell differentiation is controlled by diffusion gradients of "morphogens", the local concentrations of which control which differentiated cell type each embryonic cell will differentiate into.

Driesch, himself, concluded from regulation that the mechanisms that control embryonic development must be in some sense supernatural.

He used the word "entelechy" to mean the controlling consciousness.

(My PhD. Major Professor, J. P. Trinkaus used to joke that embryos would need at least TWO entelechies, one for each separated cell.)

If you are willing to include cellular slime mold amoebae, and their formation of stalked "fruiting bodies" as an example of something like embryos, then their development is the most regulative of all, because you can cut a Dictyostelium "slug" into 10 or 100 pieces, and the cells of each fragment will reorganize to form a normally-proportioned slug, and then a scale model stalk and spore mass, one one-hundredth the size the original "slug" would have formed.

(Please understand that these "slugs" are masses of a kind of amoebae, & only happen to resemble real slugs, which are a kind of snail.)

About ten years before Driesch's discovery of embryonic regulation, another great German embryologist named Wilhelm Roux tested the effect of 'killing' one of the first two cells of amphibian embryos at the two-cell stage. (He also tried killing two of the first 4 cells, and different combinations of 4 of the first 8 cells.)

(He 'killed' these cells by poking them with red-hot needles! And if you read Roux's actual paper, you find out that the cells Were not quite dead, and sometimes started dividing and developing later.)

Roux's results were very different from Driesch's.
His unkilled cells developed into half-embryos, in the sense that only half the organs and structures formed.

Roux's famous "hot needle" experiment has been successfully repeated by others,
Including in some other kinds of animals (including sea squirts)
And had led everyone to believe that embryonic development works by each dividing cell being given different information (perhaps different genes) at the time of each mitotic division.

Driesch's discovery of "Embryonic Regulation" conclusively disproved Roux's theory. But nobody knows (and nobody even ASKS!) why "killed" damaged cells prevent embryonic regulation.
No theory explaining Driesch's discovery can be true unless it also can explain Roux's observations.

Page 135 or Wolpert and Tickle (starting at the third line from the top of the page:

"We can now understand the results of Roux's classic experiment (see Fig. 1.8) in which he destroyed one cell of a frog embryo at the two cell stage and got a half-embryo rather than a half-sized whole embryo. The crucial feature of his experiment, it turns out, was that the killed cell remained attached, but the embryo did not 'know' it was dead. The remaining living cell could still develop a functional dorsal dorsal organizer but it developed as if the other half of the embryo was still there. If the killed cell had been separated from the living blastomere, the embryo could have regulated and developed into a small-sized whole embryo"


An English translation of Roux' paper has been published in a paper-back book. If we can find it, I will bring it to class. It contains original drawings. It also described variations of the experiment in which 2 of the first four cells were 'killed" by poking with hot needles, and other variations in which 4 of the first 8 cells were "killed". These variations included cases in which the entire anterior half was killed, leaving the posterior half alive. He also describes and includes drawings of what happens when the 4 (out of 8) posterior cells were killed, leaving the anterior cells alive to develop as a half-embryo. Variations were also done in which the dorsal half was killed, leaving the ventral half alive - and the exact reverse experiment.

Wolpert's explanation seems to me to imply that Roux only tried the version of the experiment in which either the right half or the left half was killed. They seem not to realize what happens if you destroy the top half, bottom half, anterior half, or posterior half.

But I am not sure, and would like to hear students' opinions on this question.

A variation on the experiment that Roux did not try (and that a UNC honors student and I did not succeed in doing) is the take embryos at the 1 cell stage (or 2 or 4 or 8) and poke ALL the cells in one embryo, and push this embryo up against the side of an undamaged embryo. Our hypothesis was that the undamaged embryo would develop into a half-embryo; in other words, that the closeness of the embryo in which all the cells had been damaged would somehow suppress differentiation of half of the structures of the undamaged embryo next to it.

The student and I had difficulty getting frogs to lay eggs at convenient times (all ours laid eggs between 3 and 5 AM). Also, Xenopus embryos are fragile, and killing one cell out of 2 or 4 resulted in yolk leaking out through the hole poked in the surface, and all the embryos lysed within the next 5 of 10 hours. To repeat and extend Roux' classic experiments, you need to have lab-cultured amphibians (or echinoderms?) that will lay a few eggs at a time over a long time period, and whose early embryos won't just die a few hours after you poke a few of their cells with a hot needle. If you had such an organism (and don't forget that they need to have very regulative development! So Nematodes can't very well be used!) then you could do many simple but important experiments, the results of which would go into the textbooks immediately.

This is a good example of quite simple experiments being impractical, for lack of the right properties in eggs of any suitable organism.


Regulation is one of several embryological phenomena in which the same end results are reached by any of two or more series of intermediates.

Another example is "secondary neurulation".
(The neural tube can be formed either by epithelial folding, or formed by cavitation of a solid mass of cells)

The most extreme example is sorting out of separated embryonic cells to form functional, or sometimes even normal anatomy.
(Cell sorting will be discussed in a later lecture).

My own theory has been that all such regulation phenomena Should be regarded as logically equivalent to homeostasis
(as in physiological homeostatic control of body temperature) with the difference that geometric properties are being regulated instead of simple quantitative variables.)

Homeostatic control depends on stable balances of opposing forces.

Many people think that stable balances between opposing forces is the same thing as minimization of thermodynamic free energy or minimization of potential energy.

This is a common mistake.

Some forces are "conservative" in the sense that they do not continue to expend energy in a situation of static counter-balance.

Other forces are "non-conservative" in the sense that the use up energy even when not producing any change, and exactly counter-balanced by some other force.

Not understanding this leads people to make two further mistakes:

    1) They think that stable counter-balances are the same thing as minimization of free energy.

    2) They think that only conservative forces can cause stable counter-balance.

For example, these fallacies are accepted as true by Scott Gilbert's (excellent) textbook that I have used many times in this course.
(I am sort of a fanatic on this subject, because I think these fallacies hold back embryology, including accurate understanding of cancer.)

Stable counterbalances of both non-conservative forces (like contraction of acto-myosin) and conservative forces can explain shape and size control of many structures and organs.

A big problem is how such mechanisms adjust sizes of parts in proportion to sizes of wholes, which is closely related to adjusting sizes (and relative strengths of forces) in perpendicular directions.
(which means keeping the same shape at different sizes)

This sometimes occurs in non-living mechanical patterns, from which we can learn important lessons.

When a separated embryonic cell rounds up, few people have trouble this rounding is directly caused by mechanical forces. But the later adjustment of fates of cells is not thought of as having anything to do with mechanical forces.

Notice, however, that in Roux's experiment the continued presence of the "killed" cells prevents the remaining undamaged parts of the embryo from rounding up.
So maybe anything that prevents reshaping would block regulation?

There are lots of important phenomena left for you to discover.
(But I warn you that frogs tend to lay eggs in the middle of the night on weekends, so it's not so easy to get their eggs at the 2 or 4 cell stages!
So, it's not so easy to repeat variations of Roux's hot needle experiment.)

One way to avoid having to explain embryonic regulation is to concentrate research on the kind of animals whose development is more mosaic than any others'! (Specifically, nematodes)

Possible topics for discussion:

Do Nematodes not have entelechies?

What if Driesch had done his experiments on embryos of a species that has mosaic development?

(Roux' false conclusions about unequal distribution of genetic material would have seemed to be confirmed?)

The heart develops from lateral plate mesoderm, at its extreme lateral edge where the left and right lateral plates come together. If a barrier is put between them, so that they cannot touch, then each develops into an entire heart. Two hearts! Another example of embryonic regulation.

Other examples are that branched legs will form if you split limb buds, that double eyes will develop if you split the optic cup, and that single cyclops eyes will develop (only 1) if you put the optic cups too close together.











Stimulation of embryonic cell differentiation by cell-cell signaling is called "embryonic induction".

Hans Spemann won a Nobel prize for his graduate student's (Hilde Pröscholdt ) discovery that skin ectoderm can be stimulated by notochord cells to differentiate into a second neural tube. (See photographs on p. 102 and 103 of the textbook)

The experiment was to cut a piece of future notochord out of one salamander embryo and insert it into the blastocoel of another gastrula-stage salamander. Different colored species of salamanders were used in order to distinguish which cells formed what organs.

Somatic endoderm of the "host" (the embryo into which the 'graft' is made) forms a second neural tube.
(The first semester I was a graduate student, we did this as a lab experiment & it really works well!)

Another graduate student of Spemann at the same time (Hans Holtfreter) observed cases of exogastrulation in which the mesoderm bulges outward, instead of invaginating, one of the results of which is that no neural tube forms.

The conclusion is that in normal development the notochord sends some signal to the ectoderm closest to it, and this signal causes that ectoderm to form a neural tube (& brain etc.)

Much inconclusive research was done trying to discover what molecule is the signal. A major problem was that salami, etc. could also induce neural tubes!

Dozens of other examples of induction have been discovered.

    1) The eye induces the skin to differentiate into lens. (which was discovered before Spemann)

    2) The lens induces the skin to re-differentiate into cornea.

    3) The inner and outer layers of teeth both induce each other to differentiate.

    3.5 ) It is reported that bird mouth tissue can be induced to form teeth by mouse mouth cells.

    4) Endodermal cells can be induced to form either salivary glands, lungs or pancreas, by contact with mesenchymal cells from those.

    5) Grafting Hensen's node to abnormal locations in chicken embryos can induce formation of a second neural tube and entire new embryos. It's what forms the notochord after all.

This last fact was discovered in the late 1930s by C. H. Waddington, who later designed anti-submarine strategies for the Royal Navy in WWII, and is given credit as a major inventor of "operations" research.

He smoked a pipe, and at seminars would sit in front and blow smoke rings at the speaker. I was among those subjected to this when I was a graduate student.

UNC Biology Department's own Bob Goldstein was the first to prove an example of embryonic induction in the development of nematode worms (C. elegans), which made him world famous. See http://www.bio.unc.edu/faculty/goldstein/lab/Nature.pdf

-------------------------------------------------------------------------------------------------------------------------------- a) If you separate the first two cells of ... ? (What kinds of animal?) What happens?

b) Who was Hans Driesch and about when did he make a revolutionary discovery?

c) What is an entelechy? Do they really exist? *Would a negative feedback control be one?

d) Does embryonic regulation mean external control of embryos? What does it mean?

e) Whose embryos are more regulative that any other group of animals?

f) If embryonic development in a certain kind of animals is very non-regulative, then it is ...?

g) What is a chimera? Where does this word come from?

h) What are some differences between chimeras and genetic hybrids?

*i) If you mated two chimeric animals, would the offspring be hybrids? (Sometimes? Depending on?)

*j) Can you devise an experiment, using a chimera, to test whether or not sekeletal muscle cells become multinuclear by fusion of separate cells versus by mitosis without cell division?

*k) Suppose that cells of the each species tended only to fuse with cells of the same species; why could that confuse your experiment (in the sense of giving you a mistaken result)?

l) Can embryonic induction change the fates of cells that have been grafted to abnormal locations? What about if they are grafted long after being formed by cleavage?

*m) If there weren't so many invaginations and folding of embryonic cells from place to place, then would induction be such a good means of pattern formation?

n) Do you think that birds still have the genes for making teeth. What about the genes for whatever receptors of inductive signals that induce tooth formation?

o) By what embryological experiment might you prove that snakes still have the genes for making legs?
(On analogy to the experiments in which bird stomodeum epithelium was induced to form teeth, which they lost in evolution over 50 million years ago, when snakes were first evolving.)

*p) If Spemann hadn't yet imagined the possibility of induction at the time of the discovery. then what would you guess they were trying to test?

q) How might you test for cases of induction by inserting thin pieces of plastic of mica into early embryos?



contact the webmaster

back to index page