To the unaided eye the main difference between the newly formed zygotes of different species of animals is that of bulk, and this is due to the amount of food-yolk held in suspension in the protoplasm. The ovum of the fowl is 30 mm. in diameter, that of the frog 1·75 mm., while the ova of the rabbit and Amphioxus have a diameter of ·l mm. The food-yolk is deposited in the ovum as a result of the vital activity of its protoplasm, while the ovum is still a part of the ovary of the parent. It is an inert substance which is used as food later on by the developing embryo, and it acts as a dilutant of the living matter of the ovum. It has a profound influence on the subsequent developmental process. The newly formed zygotes of different species of animals have undoubtedly, as staved above, a certain family resemblance to one another; but however great this superficial resemblance may be, the differences must be most profound, and this fact becomes at once obvious when the properties of these remarkable masses of matter are closely investigated.
As in the case of so many other forms of matter, the more important properties of the zygote do not become apparent until it is submitted to the action of external forces. These forces constitute the external conditions of Causes of development. existence, and the properties which are called forth by their action are called the acquired characters of the organism. The investigation of these properties, particularly of those which are called forth in the early stages of the process, constitutes the science of Embryology. With regard to the manifestation of these properties, certain points must be clearly understood at the outset:—(1) If the zygote is withheld from the appropriate external influences, e.g. if a plant-seed be kept in a box free from moisture or at a low temperature, no properties are evolved, and the zygote remains apparently unchanged; (2) the acquisition of the properties which constitutes the growth and development of the organism proceeds in a perfectly definite sequence, which, so far as is known, cannot be altered; (3) just as the features of the growing organism change under the continued action of the external conditions, so the external conditions themselves must change as the organism is progressively evolved. With regard to this last change, it may be said generally that it is usually, if not always, effected by the organism itself, making use of the properties which it has acquired at earlier stages of its growth, and acting in response to the external conditions. There is, to use a phrase of Mr Herbert Spencer, a continuous adjustment between the external and internal relations. For every organism a certain succession of conditions is necessary if the complete and normal evolution of properties is to take place. Within certain limits, these conditions may vary without interfering with the normal evolution of the properties, though such variations are generally responded to by slight but unimportant variation of the properties (variation of acquired characters). But if the variation of the conditions is too great, the evolved properties become abnormal, and are of such a nature as to preclude the normal evolution of the organism; in other words, the action of the conditions upon the organism is injurious, causing abortions and, ultimately, death. For many organisms the conditions of existence are well known for all stages of life, and can be easily imitated, so that they can be reared artificially and kept alive and made to breed in confinement—e.g. the common fowl. But in a large number of cases it is not possible, through ignorance of the proper conditions, or on account of the difficulty of imitating them, to make the organism evolve all its properties. For instance, there are many marine larvae which have never been reared beyond a certain point, and there are some organisms which, even when nearly full-grown—a stage of life at which it is generally most easy to ascertain and imitate the natural conditions—will not live, or at any rate will not breed, in captivity. Of late years some naturalists have largely occupied themselves with experimental observation of the effects on certain organisms of marked and definite changes of the conditions, and the name of Developmental Mechanics (or Physiology of Development) has been applied to this branch of study (see below).
In normal fertilization, as a rule, only one spermatozoon fuses with the ovum. It has been observed in some eggs that a membrane, formed round the ovum immediately after the entrance of the spermatozoon, prevents the entrance of others. If Gametogeny. than one spermatozoon enters, a corresponding number of male pronuclei are formed, and the subsequent development, if it takes place at all, is abnormal and soon ceases. An egg by ill-treatment (influence of chloroform, carbonic acid, &c.) can be made to take more than one spermatozoon. In some animals it appears that several spermatozoa may normally enter the ovum (some Arthropoda, Selachians, Amphibians and Mammals), but of these only one forms a male pronucleus (see below), the rest being absorbed. Gametogeny is the name applied to the formation of the gametes, i.e. of the ova and spermatozoa. The cells of the reproductive glands are the germ cells (oögonia, spermatogonia). They undergo division and give rise to the progametes, which in the case of the female are sometimes called oöcytes, in the case of the male spermatocytes. The oöcytes are more familiarly called the ovarian ova. The nucleus of the oöcyte is called the germinal vesicle. The oöcyte (progamete) gives rise by division to the ovum or true gamete, the nucleus of which is called the female pronucleus. As a general rule the oöcyte divides unequally twice, giving rise to two small cells called polar bodies, and to the ovum. The first formed polar body frequently divides when the oöcyte undergoes its second and final division, so that there are three polar bodies as well as the ovum resulting from the division of the oöcyte or progamete. Sometimes the ovum arises from the oöcyte by one division only, and there is only one polar body (e.g. mouse, Sobotta, Arch. f. mikr. Anat., 1895, p. 15). The polar bodies are oval, but as a rule they are so small as to be incapable of fertilization. They may therefore be regarded as abortive ova. In one case, however (see Francotte, Bull. Acad. Belg. (3), xxxiii., 1897, p. 278), the first formed polar body is nearly as large as the ovum, and is sometimes fertilized and develops. The spermatogonia are the cells of the testis; these produce by division the spermatocytes (progametes), which divide and give rise to the spermatids. In most cases which have been investigated the divisions by which the spermatids arise from the spermatocytes are two in number, so that each spermatocyte gives origin to four spermatids. Each spermatid becomes a functional spermatozoon or male gamete. The gametogeny of the male therefore closely resembles that of the female, differing from it only in the fact that all the four products of the progamete become functional gametes, whereas in the female only one, the ovum, becomes functional, the other three (polar bodies) being abortive. In the spermatogenesis of the bee, however, the spermatocyte only divides once, giving rise to a small polar-body-like structure and one spermatid (Meves, Anat. Anzeiger, 24, 1904, pp. 29-32). The nucleus of the male gamete is not called the male pronucleus, as would be expected, that term being reserved for the second nucleus which appears in the ovum after fertilization. As this is in all probability derived entirely from the nucleus of the spermatozoon, we should be almost justified in calling the nucleus of the spermatozoon the male pronucleus. In most forms in which the formation of the gametes from the progamete has been accurately followed, and in which the progamete of both sexes divides twice in forming the gametes, the division of the nucleus presents certain peculiarities. In the first place, between the first division and the second it does not enter into the resting state, but immediately proceeds to the second division. In the second place, the number of chromosomes which appear in the final divisions of the progametes and assist in constituting the nuclei of the gametes is half the number which go to constitute the new nuclei in the ordinary nuclear divisions of the animal. The number of chromosomes of the nucleus of the gamete is therefore reduced, and the divisions by which the gametes arise from the progametes are called reducing (maiotic) divisions. It is not certain, however, that this phenomenon is of universal occurrence, or has the significance which is ordinarily attributed to it. In the parthenogenetic ova of certain insects, e.g. Rhodites rosae (Henking), Nematus lacteus (Doncaster, Quart. Journal Mic. Science, 49, 1906, pp. 561-589), reduction does not occur, though two polar bodies are formed.
As soon as the spermatozoon has conjugated with the ovum, a second nucleus appears in the ovum. This is undoubtedly derived from the spermatozoon, possibly from its nucleus only, and is called the male pronucleus. It possesses in the Fertilization. adjacent protoplasm a well-marked centrosome. The general rule appears to be that the female pronucleus is without a centrosome, and that no centrosome appears in the female in the divisions by which the gamete arises from the progamete. If this is true, the centrosome of the zygote nucleus must be entirely derived from that of the male pronucleus. This accounts for the fact, which has been often observed, that the female pronucleus is not surrounded by protoplasmic radiations, whereas such radiations are present round the male pronucleus in its approach to the female. In the mouse the subsequent events are as follow:—Both pronuclei assume the resting form, the chromatin being distributed over the nuclear network, and the nuclei come to lie side by side in the centre of the egg. A long loop of chromatin then appears in each nucleus and divides up into twelve pieces, the chromosomes. The centrosome now divides, the membranes of both nuclei disappear, and a spindle is formed. The twenty-four chromosomes arrange themselves at the centre of this spindle and split longitudinally, so that forty-eight chromosomes are formed. Twenty-four of these, twelve male and twelve female, as it is supposed, travel to each pole of the spindle and assist in giving rise to the two nuclei. At the next nuclear division twenty-four chromosomes appear in each nucleus, each of which divides longitudinally; and so in all subsequent divisions. The fusion of the two pronuclei is sometimes effected in a manner slightly different from that described for the mouse. In Echinus, for instance, the two pronuclei fuse, and the spindle and chromosomes are formed from the zygote nucleus, whereas in the mouse the two pronuclei retain their distinctness during the formation of the chromosomes. There appears, however, to be some variation in this respect: cases have been observed in the mouse in which fusion of the pronuclei occurs before the separation of the chromosomes.
Parthenogenesis, or development of the female gamete without fertilization, is known to occur in many groups of the animal kingdom. Attempts have been made to connect this phenomenon with peculiarities in the gametogeny. For Parthenogenesis. instance, it has been said that parthenogenetic ova form only one polar body. But, as we have seen, this is sometimes the case in eggs which are fertilized, and parthenogenetic ova are known which form two polar bodies, e.g. ova of the honey-bee which produce drones (Morph. Jahrb. xv., 1889, p. 85). ova of Rotifera which produce males (Zool. Anzeiger, xx., 1897, p. 455), ova of some saw-flies and gall flies which produce females (L. Doncaster, Quart. Journ. Mic. Sc., 49, 1906, pp. 561-589). Again it has been asserted that in parthenogenetic eggs the polar bodies are not extruded from the ovum; in such cases, though the nucleus divides, those of its products which would in other cases be extruded in polar bodies remain in the protoplasm of the ovum. But this is not a universal rule, for in some cases of parthenogenesis polar bodies are extruded in the usual way (Aphis, some Lepidoptera), and in some fertilized eggs the polar bodies are retained in the ovum.
It is quite probable that parthenogenesis is more common than has been supposed, and it appears that there is some evidence to show that ova, which in normal conditions are incapable of developing without fertilization, may yet develop if subjected to an altered environment. For instance, it has been asserted that the addition of a certain quantity of chloride of magnesium and other substances to sea-water will cause the unfertilized ova of certain marine animals (Arbacia, Chaetopterus) to develop (J. Loeb, American Journal of Physiology, ix., 1901, p. 423); and according to M.Y. Delage (Comptes rendus, 135, 1902. Nos. 15 and 16) such development may occur after the formation of polar bodies, the chromosomes undergoing reduction and the full number being regained in the segmenting stage. These experiments, if authenticated, suggest that ova have the power of development, but are not able to exercise it in their normal surroundings. There is reason to believe that the same assertion may be made of spermatozoa. Phenomena of the nature of parthenogenesis have never been observed in the male gamete, but it has been suggested by A. Giard (Cinquantenaire de la Soc. de Biol., 1900) that the phenomenon of the so-called fertilization of an enucleated ovum which has been described by T. Boveri and Delage in various eggs, and which results in development up to the larval form (merogony), is in reality a case in which the male gamete, unable to undergo development in ordinary circumstances on account of its small size and specialization of structure has obtained a nutritive environment which enables it to display its latent power of development. Moreover, A.M. Giard suggests that in some cases of apparently normal fertilization one of the pronuclei may degenerate, the resultant embryo being the product of one pronucleus only. In this way he explains certain cases of hybridization in which the paternal (rarely the maternal) type is exclusively reproduced. For instance, in the batrachiate Amphibia, Héron Royer succeeded in 1883 in rearing, out of a vast number of attempts, a few hybrids between a female Pelobates fuscus and a male Rana fusca; the product was a Rana fusca. He also crossed a female Bufo vulgaris with a male Bufo calamita; in the few cases which reached maturity the product was obviously a Bufo calamita. Finally, H.E. Ziegler (Arch. f. Ent.-Mech., 1898, p. 249) divided the just-fertilized ovum of a sea-urchin in such a way that each half had one pronucleus; the half with the male pronucleus segmented and formed a blastula, the other degenerated. It is said that in a few species of animals males do not occur, and that parthenogenesis is the sole means of reproduction (a species of Ostracoda among Crustacea; species of Tenthredinidae, Cynipidae and Coccidae among Insecta); this is the thelytoky of K.T.E. von Siebold. The number of species in which males are unknown is constantly decreasing, and it is quite possible that the phenomenon does not exist. Parthenogenesis, however, is undoubtedly of frequent occurrence, and is of four kinds, namely, (1) that in which males alone are produced, e.g. honey-bees (arrhenotoky); (2) that in which females only are produced (thelytoky), as in some saw-flies; (3) that in which both sexes are produced (deuterotoky), as in some saw-flies; (4) that in which there is an alternation of sexual and parthenogenetic generations, as in Aphidae, many Cynipidae, &c. It would appear that “parthenogenesis does not favour the production of one sex more than another, but it is clear that it decidedly favours the production of a brood that is entirely of one sex, but which sex that is differs according to circumstances” (D. Sharp, Cambridge Natural History, “Insects,” pt. i. p. 498). In some Insecta and Crustacea exceptional parthenogenesis occurs: a certain proportion of the eggs laid are capable of undergoing either the whole or a part of development parthenogenetically, e.g. Bombyx mori, &c. (A. Brauer, Arch. f. mikr. Anat., 1893; consult also E. Maupas on parthenogenesis of Rotifera, Comp. rend., 1889-1891, and R. Lauterborn, Biol. Centralblatt, xviii., 1898, p. 173).
The question of the determination of sex may be alluded to here. Is sex determined at the act of conjugation of the two gametes? Is it, in other words, an unalterable property of the zygote, a genetic character? Or does it depend Determination of sex. upon the conditions to which the zygote is subjected in its development? In other words, is it an acquired character? It is impossible in the present state of knowledge to answer these questions satisfactorily, but the balance of evidence appears to favour the view that sex is an unalterable, inborn character. Thus those twins which are believed to come from a split zygote are always of the same sex, members of the same litter which have been submitted to exactly similar conditions are of different sexes, and all attempts to determine the sex of offspring in the higher animals by treatment have failed. On the other hand, the male bee is a portion of a female zygote—the queen-bee. The same remark applies to the male Rotifer, in which the zygote always gives rise to a female, from which the male arises parthenogenetically, but in these cases it does not appear that the production of males is in any way affected by external conditions (see R.C. Punnett, Proc. Royal Soc., 78 B, 1906, p. 223). It is said that in human societies the number of males born increases after wars and famines, but this, if true, is probably due to an affection of the gametes and not of the young zygote. For a review of the whole subject see L. Cuénot, Bull. sci. France et Belgique, xxxii., 1899, pp. 462-535.
The first change the zygote undergoes in all animals is what is generally called the segmentation or cleavage of the ovum. This consists essentially of the division of the nucleus into a number of nuclei, around which the protoplasm sooner or later Cleavage. becomes arranged in the manner ordinarily spoken of as cellular. This division of the nucleus is effected by the process called binary fission; that is to say, it first divides into two, then each of these divides simultaneously again into two, giving four nuclei; each of these after a pause again simultaneously divides into two. So the process continues for some time until the ovum becomes possessed of a large number of nuclei, all of which have proceeded from the original nucleus by a series of binary fissions. This division of the nucleus, which constitutes the essential part of the cleavage of the ovum, continues through the whole of life, but it is only in the earliest period that it is distinguished by a distinct name and used to characterize a stage of development. The nuclear division of cleavage is usually at first a rhythmical process; all the nuclei divide simultaneously, and periods of nuclear activity alternate with periods of rest. Nuclear divisions may be said to be of three kinds, according to the accompanying changes in the surrounding protoplasm: (1) accompanied by no visible change, e.g. the multinucleated Protozoon Actinosphaerium; (2) accompanied by a rearrangement of the protoplasm around each nucleus, but not by its division into two separate masses, e.g. the division which results in the formation of a colony of Protozoa; (3) accompanied by the division of the protoplasm into two parts, so that two distinct cells result, e.g. the divisions by which the free wandering leucocytes are produced, the reproduction of uninuclear Protozoa, &c. In the cleavage of the ovum the first two of these methods of division are found, but probably not the third. At one time it was thought that the nuclear divisions of cleavage were always of the third kind, and the result of cleavage was supposed to be a mass of isolated cells, which became reunited in the subsequent development to give rise to the later connexion between the tissues which were known to exist. But in 1885 it was noticed that in the ovum of Peripatus capensis (A. Sedgwick, Quart. Journ. Mic. Science, xxv., 1885, p. 449) the extra-nuclear protoplasm did not divide in the cleavage of the ovum, but merely became rearranged round the increasing nuclei; the continuity of the protoplasm was not broken, but persisted into the later stages of growth, and gave rise to the tissue-connexions which undoubtedly exist in the adult. This discovery was of some importance, because it rendered intelligible the unity of the embryo so far as its developmental processes are concerned, the maintenance of this unity being somewhat surprising on the previous view. On further inquiry and examination it was found that the ova of many other animals presented a cleavage essentially similar to that of Peripatus. Indeed, it was found that the nuclear divisions of cleavage were of the first two kinds just described. In some eggs, e.g. the Alcyonaria, the first nuclear divisions are effected on the first plan, i.e. they take place without at first producing any visible effect upon the protoplasm of the egg. But in the later stages of cleavage the protoplasm becomes arranged around each nucleus and related to it as to a centre. In the majority of eggs, however, the protoplasm, though not undergoing complete cleavage, becomes rearranged round each nucleus as these are formed. The best and clearest instance of this is afforded by many Arthropodan eggs, in which the nucleus of the just-formed zygote takes up a central position, where it undergoes its first division, subsequent divisions taking place entirely within the egg and not in any way affecting its exterior. The result is to give rise to a nucleated network or foam-work of protoplasm, ramifying through the yolk-particles and containing these in its meshes.
In other Arthropodan eggs the cleavage is on the so-called centrolecithal type, in which the dividing nuclei pass to the cortex of the ovum, and the surface of the ovum becomes indented with grooves corresponding to each nucleus. In this kind of cleavage all the so-called segments are continuous with the central undivided yolk-mass. It sometimes happens that in Arthropods the egg breaks up into masses, which cannot be said to have the value of cells, as they are frequently without nuclei. In other eggs, characterized by a considerable amount of yolk, e.g. the ova of Cephalopoda, and of the Vertebrata with much yolk, the first nucleus takes up an eccentric position in a small patch of protoplasm which is comparatively free from yolk-particles. This patch is the germinal disc, and the nuclear divisions are confined to it and to the transitional region, where it merges into the denser yolk which makes up the bulk of the egg. At the close of segmentation the germinal disc consists of a number of nuclei, each surrounded by its own mass of protoplasm, which is, however, not separated from the protoplasm round the neighbouring nuclei, as was formerly supposed, but is continuous at the points of contact. In this manner the germinal disc has become converted into the blastoderm, which consists of a small watch-glass-shaped mass of so-called cells resting on, but continuous with, the large yolk-mass. It is characteristic of this kind of ovum that there is always a row of nuclei, called the yolk-nuclei, placed in the denser yolk immediately adjacent to the blastoderm. These nuclei are continually undergoing division, one of the products of division, together with a little of the sparse yolk protoplasm, passing into the blastoderm to reinforce it (so-called formative cells). The other product of the dividing yolk-nuclei remains in the yolk, in readiness for the next division. In this manner nucleated masses of protoplasm are continually being added to the periphery of the blastoderm and assisting in its growth. But it must be borne in mind that all the nucleated masses of which the blastoderm consists are in continuity with each other and with the sparse protoplasmic reticulum of the subjacent yolk.
In the great majority of eggs, then, the nuclear division of cleavage is not accompanied by a complete division of the ovum into separate cells, but only by a rearrangement of the protoplasm, which produces, indeed, the so-called cellular arrangement, and an appearance only of separate cells. But there still remain to be mentioned those small eggs in which the amount of yolk is inconsiderable, and in which division of the nuclei does appear to be accompanied by a complete division of the surrounding protoplasm into separate unconnected cells—ova of many Annelida, Mollusca, Echinoderma, &c., and of Mammalia amongst Vertebrata. In the case of these also (G.F. Andrews, Zool. Bulletin, ii., 1898) it has been shown that the apparently separate spheres are connected by a number of fine anastomosing threads of a hyaline protoplasm, which are not easy to detect and are readily destroyed by the action of reagents. It is therefore probable that the divisions of the nuclei in cleavage are in no case accompanied by complete division of the surrounding protoplasm, and the organism in the cleavage stage is a continuous whole, as it is in all the other stages of its existence.