Talk:Paper - The relations of endogenous and exogenous factors in bone and tooth development (1937)

From Embryology



By HANS GRUNEBERG Department of Zoology, University College, London


‘Tue grey-lethal mutation in the mouse (Griineberg, 1935, 1936 a, b) is characterized by a complex of symptoms which is unparalleled in human pathology. The fur lacks the yellow pigment. No tooth ever erupts. Without artificial feeding death invariably occurs between the 22nd and the 30th day of life. Liquid food prolongs the life for a fortnight or so, but does not materially change the condition. Death is thus not due to mechanical troubles in feeding, but its actual cause remains unknown.

The main feature of the skeleton is the entire lack of all secondary absorption processes. Thus bone substance once laid down in development is preserved in its entirety. The geometrical proportions of bones can only be. maintained during growth by secondary remodelling processes by which some of the bone temporarily laid down is eventually removed. This has been thoroughly worked out in detail by means of madder feeding in the pig by Brash (1984). If, however, these secondary modelling processes fail to act, very characteristic anomalies in bone shape are bound to develop. The greylethal bones correspond excellently to these expectations. There is, therefore, no doubt that the grey-lethal complex of symptoms is in fact largely due to a failure of the secondary resorption processes on the bones. In addition there is incomplete calcification, both in bone and dentine. This leads to failure of eruption of the teeth as has been described in detail elsewhere.

In this paper a more thorough description of the crown shape of the greylethal molars will be given. The anomalies to be described are constantly found. Generally, the grey-lethals are remarkably constant in the abnormal conditions they show: the differences between individuals other than those due to age are practically negligible. It will be discussed below whether and to what extent the anomalies in the shape of teeth can be explained in terms of the peculiarities already well established in the grey-lethal mouse. Finally, the bearing of these facts on the general conception of the action of exogenous stimuli and the response of: the bones will be considered. Factors in Bone and Tooth Development 237


For comparison, developmental stages of normal teeth must be used in which the shape of the cusps has not yet been altered by wearing off. The illustrations (Text-figs. 1-4) show camera lucida drawings of a 14 days old grey-lethal mouse and a normal litter-mate. The roots of first molars in the normal are still incomplete, while the uncalcified vestiges of roots in the greylethal, dried up and shrivelled and largely lost during the preparation, are here of no concern.

As already mentioned in the first paper (1985), the lower first molar is strongly compressed laterally in the grey-lethal. At the same time it is noticeably longer than the corresponding tooth of the normal. The median and lateral surfaces are considerably flattened, while those of the normal are bulging and bulbous. This applies particularly to the fissures between the

Text-fig. 1. Left lower first molar of grey-lethal (c, d) and normal litter-mate (a, b), 14 days old. a and c, median view; 6 and d, lateral view.

cusps, which are shallow grooves or impressions only, while they are deep and wide in the normal. The cusps themselves are a little more erect and project slightly more over the tooth neck. The flattened surfaces are not smooth and regular, but show minute and irregular depressions, the majority of which are directed parallel to the axis of the mandible. All these anomalies are more strongly developed on the lateral surface which as a whole is considerably lower. In lateral view a good deal of the inside cavity of the tooth is therefore visible (Text-figs. 1-2).

In the upper first molars, the most conspicuous anomaly is that all the cusps are erected, though to a varying extent. In a normal first upper molar, all the cusps have their axes practically parallel to each other, including angles of between 40 and 45° with the plane of the neck of the tooth. In the greylethal tooth the axes of the cusps have an inclination of between 55 and 75°. This difference is most clearly seen in lateral view, since-the lateral surface is 238 Hans Griineberg

comparatively flat while the median surface in both types bulges towards the sagittal plane (Text-fig. 8). Comparison of such teeth strongly suggests that


b Text-fig. 2. Top view of same teeth as in Text-fig. 1. a, normal; b, grey-lethal. The arrows indicate the approximate planes of the sections on Pl. I carrying corresponding numbers.

Text-fig. 3. Left upper first molar of grey-lethal (c, d) and normal litter-mate (a, 6), 14 days old. a and c, lateral view; b and d, median view.

Text-fig. 4. Top view of same teeth as in Text-fig. 3. a, normal; 6, grey-lethal. The arrows indicate the approximate planes of the sections on Pl. I carrying corresponding numbers.

pressure from the front has acted on the grey-lethal tooth so as strongly to erect the cusps.

Another striking difference is found in the outline of the crown when seen in top view (Text-fig. 4). Whereas the normal tooth bulges broadly towards Factors in Bone and Tooth Development 239

the middle line of the body, this protrusion is very much less pronounced in the grey-lethal. Still more pronounced is the lack of curvature in the region of the lateral front cusp which has a very irregular shape, is wrinkled and merges irregularly into its surroundings. The cingulum is completely suppressed. Evidently this is the most abnormal region. Again, as in the lower first molars of the grey-lethal, the surface is not smooth and regular, but shows small depressions, chiefly on the lateral surface and the front parts. The irregular surface of the lateral front cusp has already been mentioned.

All these anomalies of shape suggest that the grey-lethal teeth have grown under pressure. What has happened seems to be this. At the time when the then uncalcified tooth germ gets enclosed into the socket, it has not yet reached its definite size. Now, when it grows, in a normal animal the surrounding bone responds by absorption to fit the requirements of the growing germ. Under normal conditions, therefore, shape and size of the socket is always in harmony with the dimensions of the tooth. In other words, the growth tendencies of the tooth determine size and shape of the socket.

This situation seems to be reversed in the grey-lethal. Here the bone of the socket does not yield to the growth pressure of the tooth germ. Accordingly the tooth germ has to adapt itself to the space available. It does not respond by stopping growth altogether. The growing germ, chiefly the enamel organ, is therefore bound to get deformed. In the lower first molars, the socket does not allow for a transverse expansion, but there is no obstacle to a longitudinal growth. The tooth, therefore, gets compressed in bucco-lingual direction, but compensates by longitudinal elongation. In the upper first molar, too, there are obstacles to a transverse expansion. In the present case, however, the tooth cannot compensate for this by longitudinal evasion. Under such circumstances, the only possible answer can be erection of the cusps. When calcification starts, the cusps are petrified in these positions.

The upper second molars of the grey-lethal show no anomalies of crown shape whatever. The lower second molars and the wisdom teeth are very nearly normal, the differences being almost negligible. In particular, there are no irregularities on the surface. That would suggest that their growth tendencies were practically exhausted at the time they were enclosed in the socket. Their development was therefore not hampered by too narrow walls of unyielding bone.


If this interpretation of the facts is correct, it should be demonstrable histologically. Suitable stages of development are those in which the tooth has already formed some enamel and dentine, but in which the tooth of the normal has not yet started erupting. For the first molars this stage is found at about the 9th or 10th day after birth. Longitudinal and transverse sections were therefore cut serially through grey-lethals and normal litter-mates of 240 Hans Griineberg

that age. The results are shown on Pl. I. Fig. 1 represents a longitudinal section through all the upper molars of a normal. As is clearly seen, the first molar is everywhere clear of bone, there being an interval of normal width all round it. Fig. 2, a corresponding section through a grey-lethal jaw, exhibits a totally different situation. A big mass of cancellous bone presses hard against the tooth on its front part, i.e. at the expected place. The erection of the cusps which results from this is clearly seen, and it is further apparent that the effect is strongest in the neighbourhood of bone pressure. Single spicules of the spongy bone mass project towards the tooth, leaving here impressions on the surface mentioned previously in this paper. Furthermore, it is seen that the pressure, under which the tooth grew, could not be passed on to the second molar since a bony interdental septum closes the socket from behind. This offers the explanation why that tooth is not at all affected by the pressure under which its neighbour developed. In fact it develops under perfectly normal space conditions, as might have been predicted from its normal shape.

Figs. 3 and 4, Pl. I, represent transverse sections through the front part of the first upper molar, in the region of the deformed lateral front cusp. Here again, the reason for the deformity in the grey-lethal is clearly a mass of cancellous bone which in the normal course of events is removed by resorption. Figs. 5 and 6 show transverse sections through the tooth farther back. Here there is hardly any lateral pressure on the grey-lethal tooth, while a spicule hindering its median expansion is obvious.

Figs. 7 and 8 represent transverse sections through the lower first molar. The compression of the grey-lethal tooth acts mainly from the lateral side as seen by the position of projecting spongiosa. This situation corresponds exactly with Brash’s conclusions about the growth of the mandible.

The same type of causation is found throughout. Where there is a deformity in any of the teeth, its cause, in the form of persistent spicules, is readily found. Histological evidence, therefore, leaves no doubt of the correctness of the interpretation that the tooth shape anomalies are due to lack of bone absorption.


A few words may be added about the upper incisors. In a normal mouse, this tooth has an outline which is almost exactly a segment of a circle. This applies already to the original tip, which is soon worn off by use. The axis of the tooth lies exactly in one plane. The corresponding tooth of the grey-lethal has an outline like an eagle’s beak. The tip is strongly turned downwards so that the dorsal surface has two different curvatures meeting each other in an obtuse and rounded angle. It seems that the extreme tip immediately after the first dentine and enamel is laid down is subjected to some pressure from behind. This pressure is caused by the fact that the growing back end of the tooth finds resistance against unyielding bone. That results in an angle being Factors in Bone and Tooth Development 241

formed between the tip which is already calcified or at least stiffened by dentine and enamel, and the rest of the tooth which is still flexible. Actually, this pressure does not act exactly along the axis of the tooth. In consequence the tooth is forced into a very shallow spiral, the tip pointing somewhat towards the middle plane of the body whereas the back opening faces laterally.

The position of the bending on the tooth allows for an estimate of the time at which it occurs during development. If calcification of this tooth follows the same course as in the rat, this should have happened during the first day after birth (Addison & Appleton, 1915).

A very similar situation to that described here for the upper incisor sometimes occurs in human teeth due to an external trauma. If the calcified parts of a developing tooth are forcibly dislocated as compared with the uncalcified parts, but both continue their development, a bend with a distinct angle between the two parts may result. This condition, chiefly found in the most exposed front teeth, is called “‘dilaceration” in Dental Surgery (Bennett, 1931).

When comparing the width of the dentine and of the pulp cavity of upper incisors in older animals, one is struck by the fact that the grey-lethal has, at least in the front parts of the tooth, a considerably thicker layer of dentine, but a much narrower pulp cavity. This, however, is not yet so in young animals of about 10 days of age which do not exhibit any appreciable difference. The reason for this is very simple. In a normal tooth which is worn off by use at a regular rate, the single odontoblast has only a limited period during which it may lay down dentine. This period lasts from the time it starts functioning at the back end of the tooth until it approaches the tip where its activities are automatically stopped by the formation of secondary dentine by which the pulp cavity is obliterated. In the grey-lethal, however, the tooth is not cut and therefore not worn off. No secondary dentine is therefore formed near the tooth tip. The odontoblasts are therefore able to go on laying down dentine as long as the animal lives and their physiological condition allows. This results in a very thick layer of dentine, but a very narrow pulp cavity.


The histological investigation leaves no doubt that the explanation deduced from the shape of the grey-lethal molars is essentially correct. We have, therefore, a complete reversal of normal development. Whereas in the normal mouse or any other mammal size and shape of socket is determined by the growth of the tooth, the size and shape of the tooth in the grey-lethal is limited and influenced by that of the socket at the time it becomes enclosed in it. Teeth which have still to grow after that time are deformed, while teeth whose growth was completed then develop normally.

Actually, a similar situation was described in the grey-lethal previously but not fully understood then. The lower incisors of these animals, instead of growing in the normal way backwards till they reach the basis of the condyloid

Anatomy LxXxI 16 242 Hans Griineberg

process, find a way out of the mandible through the mental foramen. This phenomenon is essentially the same as the much less obvious anomalies described in this paper. Here again, the bone does not yield to the pressure of the backward-growing end of the tooth.

This situation is of some importance for the understanding of the bone anomaly of the grey-lethal. As mentioned above, there is no secondary bone absorption. Now it was doubtful whether this was due to a lack of stimulus for absorption or to some other reason. What the actual stimulus is which causes bone absorption, for example, at the surface of a long bone, is unknown. It was therefore doubtful whether the unknown stimulus was absent for one reason or another, or whether the stimulus acted normally, but the bone could not respond to it. Now the situation is clear. For, in the case of a socket, the stimulus for absorption is known to be the growing tooth germ. If that germ grows vigorously, but the socket does not respond to its proper stimulus, it is evident that the lack of secondary bone absorption is due to the inability of the bone to respond, not to the absence of the stimulus.

This conclusion is of some importance for the general conception of the relations of external and internal factors in bone development. It is well known that bone responds to pressure by absorption. That has been worked out for the relation of tooth and socket in great detail by Gottlieb & Orban (1931). From work of this kind it was concluded that there is a direct response of bone to pressure, a conclusion which was fully justified by the facts then known. In the light of the facts described in this paper, this relationship turns out to be more complex than then assumed. If the bone is to respond to pressure by absorption, it must be able to do so. The ability of bone to react, however, is evidently rooted in hereditary factors. If these factors act normally, as is the case in all known animals with the exception of the greylethal mouse, the relationship between pressure and absorption appears to be direct. If, however, this hereditary mechanism is upset, as in the grey-lethal, the stimulus by itself is unable to induce absorption in the bone. The old scheme was:

STIMULUS -—-> RESPONSE (Pressure) (Absorption)

Now we have to fit in a new link:


This link is disturbed in the grey-lethal. No response, therefore, can take place. The physiological mechanism of the hereditary basis is unknown as yet and is the subject of further studies.

In the light of these facts, certain suggestions previously made by the author need revision. It was shown that in the grey-lethal the infra-orbital foramen is considerably narrower than in a normal mouse. Incidentally, the Factors in Bone and Tooth Development 243

anterior part of the Masseter which originates in that neighbourhood is very poorly developed. As a tentative suggestion the explanation was put forward that this is due to the comparative inactivity of the.muscle. Since the greylethal does not cut its teeth it does not nibble, and consequently uses its masticatory muscles to a much lesser extent than a normal mouse. The explanation given then amounted to the assumption that in that case the narrowness of the foramen was due to the lack of a proper stimulus for absorption. It seems now that that assumption is untenable. The causal relationship is probably reversed. Not: the hole is small since there is only a weakly working muscle which does not stimulate enlargement. But: the muscle cannot grow larger since the foramen is narrow. In this connexion it is irrelevant that the muscle is little used anyhow.

As has been shown in this paper, all the anomalies in tooth shape in the grey-lethal are secondary consequences of the lack of bone absorption, and they all develop after birth. In the hierarchy of causes, therefore, the bone anomaly is nearer the original gene action than are the tooth shape anomalies dependent on it.


In normal development, size and shape of socket are determined by the growth tendencies of the tooth germ. In the grey-lethal mouse the growth tendencies of the tooth are limited by the width and size the socket had when the tooth became enclosed in it. This leads to the conclusion that bone reacts by absorption to pressure only if the hereditary basis for doing so is undisturbed. The relation between stimulus and response is therefore not direct, as assumed hitherto, but subject to conditions based in the hereditary constitution of an animal.

It is shown that all the anomalies of tooth shape in the grey-lethal are caused by the lack of secondary bone absorption which is therefore a step nearer to the original gene action.


The author wishes to express his thanks to Prof. J. B. S. Haldane, F.R.S., and Prof. D. M. S. Watson, F.R.S., for the continued interest they took in the work. The microphotographs were taken by Mr J. R. Thomas of this department. Particular thanks are due to Dr E. W. Fish (Royal Dental Hospital, London) who very kindly put the technical experience in tooth histology of his laboratory at the author’s disposal, and to his assistant, Mr W. P. Pereira.


Appison, W. H. F. & Appteton, J. L., jr. (1915). “The structure and growth of the incisor teeth of the albino rat.” J. Morphol. vol. xxvi, pp. 43-96.

Bennett, Sir Norman (1931). The Science and Practice of Dental Surgery, 2nd ed. Oxford Medical Publications. London: Oxford Univ. Press.

16—2 244 Hans Griineberg

Brasu, J. C. (1934). “Some problems in the growth and developmental mechanics of bone.” Edinb. med. J. N.S. (tvth), vol. Xx.

Gorriies, B. & Orzan, B. (1931). Die Verdnderungen der Gewebe bei iibermassiger Beanspruchung der Zéhne. Leipzig: Georg Thieme. .

GrtneBerea, Hans (1935). “A new sublethal colour mutation in the house mouse. Soc. B, No. 809, vol. cxvi, pp. 321-42.

—— (1936 a). “Grey-lethal, a new mutation in the house mouse.” J. Heredity, vol. xxvu, pp. 105-9.

—— (1936 b). ““Komplette Retention des Gebisses, ein neuer Erbfaktor bei der Hausmaus.” Schweiz. Mschr. f. Zahnheilk. (in the press).


Proc. roy.


The technique is the same for all the preparations. Suza fixation. Haematoxylin, van Gieson; 7p; 35 times enlarged.

The animals from which Figs. 1 and 2 were made were litter-mates, 9 days old. Figs. 3, 5 and 7, and 4, 6 and 8 respectively, were taken from another pair of animals (litter-mates) of 10 days. Despite this age difference, they represent practically the same stage of tooth development (just before the eruption of the incisors), since the older animals came from a large litter which develops more slowly.

In these young stages, the enamel freshly laid down is not dissolved entirely by the process of decalcification. It is seen as a darkly staining layer in all the sections with the exception of Figs. 7 and 8, where it fades away towards the tips of the cusps. The approximate planes of the sections are indicated in Text-figs. 2 and 4.

Figs. 1 and 2. Longitudinal sections through the upper molars of normal and grey-lethal. The first molar of the normal is clear of bone all round. In the grey-lethal dense spongiosa is pressing against the front parts of the tooth, leaving marked impressions on the surface and leading to erection of the cusps. No pressure acts upon the second molar of the grey-lethal.

Figs. 3 and 4. Transverse sections through front parts of right upper first molar of normal and grey-lethal. In the grey-lethal tooth spongiosa spicules press into the tooth mainly from the lateral side where there are considerably more in the actual preparation than are brought out in the photograph. One spicule presses strongly on the median side. On the lateral side the enamel organ is complexly invaginated by spicules; such spicules are covered with a layer of adamantoblasts which have deposited enamel on the bone surface.

Figs. 5 and 6. Transverse sections through left upper first molars of normal and grey-lethal, farther backwards than the last two sections. There is little direct pressure in this region acting on the grey-lethal tooth except for one spicule pressing from the median side, strongly affecting the curvature in that neighbourhood. Note, too, the steeper shape of the middle cusp.

Figs. 7 and 8. Transverse sections through left lower first molar of normal and grey-lethal. In the abnormal tooth the resistance to growth is exclusively on the lateral side where there are many more spicules than are brought out in the photograph. They form a strong obstacle particularly for the downward growth of the lateral wall (compare Text-fig. 1). Journal of Anatomy, Vol LX XI, Part 2

GRUNEBERG—Facrors 1x Bonz anp Tooty DEVELOPMENT