. \ [ab Embryonic Development ous In developmental biology, embryonic development, also known as embryogenesis, is the developmental stage of ananimal embryo. Embryonic development starts with the ferilization of an egg cell (ovum) by a sperm cell, (Spermatozoon). Once fertilized, the ‘ovum becomes a single diploid cell known as a zygote. The zygote undergoes mitotic divisions with no significant growth (a process known as cleavage) and cellular differentiation, leading to development of a multicellular embryo after passing through an organizational checkpoint during mid-embryogen: ‘The main stages of animal embryonic development are as follows: + The zygote undergoes a series of cell divisions (called cleavage) to forma structure called a morula, The morula develops into a structure called a blastula through a process called blastulation, ‘The blastula develops into a structure called a gastrula through a process called gastrulation, The gastrula then undergoes further development, including the formation of organs (organogenesis), Fertilized Dyes: The fertilized eggs become gradually more and more transparent and the vitelline membrane separates from egg proper and develops a space known as perivitelline space, which is filled with Mui, Formation of Blastodise: Just after fertilization, the eytoplasm which is present on the periphery, starts flowing towards the area where the sperm has probably entered the egg. The accumu pass through the equator and at the animal n of eytoplasm is due to the contraction wave which is set in vegetal pole, pole. The polarity af this stage is set up. ‘The completion of one contraction cycle takes about 2 minutes. About swenty ofthese eycles follows one another, each cycle adding more and more eytoplasm at the animal pole and soon forms « cap-like structure, the blastodermie cap or blastodise,‘The blastodise i aa he blastodise in teleost is dise shape. Most of the eggs have two principal regions in common, acentre, which is relatively stable to cent ization and an endoplasm which is displaceable containing yolk and other inclusion. Fig.213eh; Diagrammatic reprenntatin of deve ston of develpienal sage. (0) Teritgd es Pera ace ey Teatauge ceed nge st etsap eed nk {Beaty morte soe (Late rovle tage B BlooeB,Bastomere: CYL, eyteplasmic layer, VM. viteline membrane, Y, yelk Source, Verna (1970) " Cleavage: ‘The further development in major teloosts is almost identical, followed by the process of cleavage. The teleost egg has blastoderm inthe form of blastodis, the cleavage ismeroblastic, i.e, limited to blastodise, the entie zygote i not divided. ‘The segmentation starts from 1 to 13/4 hours after fertilization, The factors which bring the cleavage are many but the major changes are the orientation of nuclear spindle and viscosity Visible. They are parallel and on either side ofthe sevond cleavage plane and right angles tothe first and third, In this way 16 cells are formed,32-Cell Stage: The 6-cell stage undergoes further division, but fom now, the cleavage furrows are horizontal as will as vertical. The four central cells are divided by a horizontal division into 8 cells which are arranged in two layers of four cells each. With the exception of the four coiner cells in which the division is more or less diagonal, the rest of the cells are divided by vertical division. These are either parallel to the first or to the second cleavage furrow. In this way the 32-cell stage is formed. Early Morul: At theend of cleavage, a ball of cells the morula is formed. The total area occupied by the cells of the early morula is more or less the same as that of the original blastodermic disc, The superficial view of egg showing cleavage and formation of morula is given in diagrams. 086 666 88 6 FIE-2141 Superficial view of the eatly developmental stages. (A-K) Acipencer. (AE Crenllbrus pavo. Source : Gresse (1957) " , Late Morula; The cells of morula divide further and becomes smaller insize, Ina side view, this stage appears as a mass of cells with prominent hemispherical projection and the convex base which rests inhollow concavity of the yolk. A large number of oil granules pass out from the cell mass into the yolk, where they combine to form bigger globules. The cells of morula loose and they become separated from one another under slight pressure. A syncytial layer is formed between yolk and the convex base of the cell mass. This syneytium is called periblast. Cleavages result in the formation of two kinds of cells, blastoderm or periblast. The blastoderm cells are distinct and produce the embryo. The periblast or trophoblast cells lie between the yolk and cells of blastoderm and cover the entire yolk mass, having originated from the most marginal and outlying blastomeres. This syncytial layer helps in the mobilization of yolk reserves. ‘The nuclei arise at the edge of the blastoderm from division of marginal cell nuclei, each resultant nuclei are drawn into the divided yolk protoplasm or the periblast. The periblastis syncytial i.e, multinucleated cytoplasm. These nuclei resemble the nuclei of the blastomeres and since spindle, asters and chromosomes were observed, it is concluded that they divide mitotically. According to Beer's law, percent transmission of a nucleus would be inversely proportional to the number of absorbing molecules in that nucleus, and the relationship would be logarithmic rather than linear. Blastula: ‘The cleavages or segmentations results inthe formation of two kinds of cells, the ‘blastoderm’ and ‘periblast’. The embryo is formed by the blastoderm, while the perblast or trophoblast eels, which lie between yolk and cells of blastoderm, which is syncytial in nature helps in mobilization of yolk reserves. There are substantial cohesive forces between developing blastomeres and the surrounding periblast which are important in the subsequent morphogenetic movement, It is suggested tht periblast acts as an intermediary between two ‘non-vettable’ components—blastoderm and yolk, When the blastoderms diameter is 4/5 of the egg diameter, itis converted into blastula.oo Dp Fig. 2130-51 Diagrammatic representation of developmental stages. (3) Blastuls sage (2) Gastuliton age gem ring descend to the queef the gp 6 The quater lof the egg surface covered. (showing aril yelk put. () Showing clnure of biastopere, side view. (D Same, dara view, (g) Formation ef somites, BUC batocoe EM ery; EMS. erbryonie Wied GR, germ ng, NK neural eel OF opilobe OPV opi vei somite, iY, os YP, yo ug The hemispherical mass of cells of morula project from the yolk. The cells then flattened and extend outwards. The periphery of the blastodise lines with the periphery of the yolk. The marginal or peripheral cells remain in close contact with the periblast. Whereas the central cells of the floor of the blastodise are raised, ‘As these cells are raised, a space is developed. This space is called as segmentation cavity or blastocoel (Fig. 21.5a). Soon blastocoel becomes well developed. The blastula formation starts 15 hours after fertilization in stickleback while it takes 8 hours after fertilization in Cyprinus carpio. ‘Atthe end of segmentation, the blastodise becomes radially symmetcical, The radial syrnmetty changes to bilateral symmetry because the flattening of cell mass is expressed in one sector and thus this region becomes thicker.The thicker sector is very important because itis embryonic material and future emtyyo develops from it, and its median plane becomes the median plane of the embryo. At this stage the anterior and posterior sides of future embryo are also fixed. The distal part of the thicker Sector is the prospective posterior end of the embryo and its central part corresponds tothe Prospective anterior end of the embryo. Gastrula: The appearance of distinct primitive streak on the embryonic shields the beginning of gastrula. ‘The gastrulation generally ends withthe closure ofblatopore. According to Riley (1974), this distinction is arbitrary. Both epiboly and emboly actively take part inthe formation of gastrula Invagination or Emboly: takes place at about 21-26 hours after fertilization, the cells of the thicker sector invaginate at the limit of cytoplasm and yolk. This marks the beginning of gastrulation. The invagination which originally starts at one point, then extends laterally around the edge of the blastoderm and soon spread to the periphery of the entire blastodise. The invaginated layer does not extend over the floor of the sub-germinal cavity but is confined to the edges of the blastoderm thus forming a prominent ring, known as ‘germ ring’. The only Part which shows further invagination is the region of the germ ring which is formed by the thick sector of blastoderm disc. « Fig, 21444 + Gana, (0) Acipener, (0) Amis cle, () Pach, (4) Archenteron (Source: Cs 1958)As soon as the germ ring becomes established, it moves towards the yolk of the egg, The width Nn, it remains constant but it increases in its circumference. Regarding further invagin: advances more rapidly at one place than round the rest of the periphery of the blastoderm making itin triangular shape. The apex is pointing towards the animal pole of the egg. The embryo loses its triangular shape and becomes clongated. If blastoderm is seen from above, the posterior pole is roughly triangular which is thicker than the adjacent area. This makes the embryonic shield more prominent. The embryonic shield has been differentiated as embryonic and extra embryonic area, ‘The embryonic shield is made up of an epiblast of polygonic cells covered by epidermic stratum and a complex lower layer which is known as entochordamesoblast. This entochordamesoblast is the analogue of mesoderm and endoderm. The thickened median portion will become the prechordal plate and chorda, while somewhat loosely arranged cells will form entoderm. In the extra embryonic region, an elongated sub-germinal cavity bounded laterally by germ ring extends between the periblast and epiblast. The presumptive mesoderm in the meantime has coverage towards the dorsolateral edges of the blastodise where it involutes, passing to the inside between the entoderm and ectoderm, The ‘mesoderm becomes arranged on either side of the median notochordal material in developing embryo. The notochord, prechordal plate and mesoderm which are differentiated but continue with cntochordamesoblast, Inter ectoderm differentiates and all three germinal layers are distinguished ectoderm, mesoderm and entoderm. With the advancement of development notochord, Kuffer’s vesicle and neural plate are differentisted, Epiboly: Simultaneously with emboly, the epiboly also starts and the cells overgrow the yolk and at the Same time migrate at its periphery. The blastoderm become flattened, The flattening of blastoderm causes it to spread over yolk. Eventually the rim of blastoderm converges at or near the opposite side of the yolk and the opening closes by the contraction of rim, The rim of the blstodise corresponds to the lip of the blastopore, Later before the blastopore closes, a yolk plug isseen projecting from the blastopore,Organisation of Fish Embryo: Presumptive areas can be mapped out in the wall of the gastrula. The fate map for gastrula of Cyprinus carpio has been given by Verma (1971) Fig. 21.71 (A,8,C.D,E.F) Schematic representaton of prenumtive tenteres (Gource: Gress, 1957). Fig, 218 Faternap. 21, enter; FPDECD, epMberalectoderne MES, mesetenn¢ NLECD, neural fa ‘ectoderm; MIC, ntodhotd; FCP, peeaudal plateOrganogenesi About 27 to 50 hours after fertilization, due to further contraction of the lips, the blastopore closes Notogenesis: ‘The presumptive notochordal cells migrate inwards and roll up along the posterior edge of the blastoderm and thus form a solid string like notochord. Neurogenesis: The presumptive neural plate sinks down to the space vacated by the inwardly migrated notochordal cells. The edges of neural plates rise ‘up and fuse with each other at the middle line enclosing a cavity, ‘neurocoel”. Thus, a hollow tube-like neural tube is formed just above the notochord. The anterior part of neural tube swells to form the brain whereas the part behind it remains as such and forms the spinal cord. By the two consecutive invaginations in the brain, it is differentiated into fore, mid and hind parts. The optic lobes appear by lateral outgrowth from the forebrain. The un-segmented parts of the embryo converge towards the embryonic axis, This convergence along with the development of the central nervous system causes a thickening of the embryo proper, which now protrudes from the surface of the egg extending approximately half way round the circumference of the yolk sphere. Hatching: After the development of the various organs in the embryo, its body becomes cylindrical and bilaterally symmetrical. The connection between the body and yolk sac gradually narrows to form a stalk, The yolk sac gradually decreases in size as the embryo grows. Finally, embryo hatches into a small free-swimming larva. Larval Development: The freshly developed larva of Cyprinus carpio measures about 4.5 mm in length which is characterised by (a) the head slightly bent on the yolk, (b) mouth is open but no alimentary canal, eyes are large, pectoral fins are rudimentary and the tail is heterocereal, One day old larva increases to 5.5 mm in length. The head becomes straight than the preceding stage where the head is slightly bent, I size and the become dark black, heart enlarges alimentary canal differentiated above the yolk Mouth is bounded by the jaws but coveredby a thin membrane. Gill arches swith rudimentary gill filaments are developed which are not yet covered over by an operculum. In two days, the mouth of larva opens and becomes slit like, alimentary canal opens through with mouth. There is complete ams, Atthis tage larva starts respiration with gills and feeding W absorption of yotk at 7 mm stage larvae which is almost 4 days old. At 10 days the larva assumes the shape ofa fish with a convex dorsal profile. After two days mouth opens, caudal fin starts to move actively, and after three days the larva begins to swim. Weight increases rapidly after hatching. The weight on third day was 0. 65 mg heavier than at hatching, [this stage the larva began to take food and weight of 8.80 mg was increased and on ninth day, the unfed larvae were inactive and weight was 1.24 mg, 25% les than that on third dey. Only 1% of the unfed larvae survived to day 12. Early Zebrafish Development © mange oy Yate Yat eal oti inet (0 rate gt (a oi Vv \ sia q Vegeta pole ~ cy © oi on ir, Sie satin 1.37 Zebrafish development occurs very rapidlyFactors influencing Early Survival: Light, oxygen, temperature and feeding are some important factors which are responsible for survival during embryonic development. According to Pinus (1974), tiulka (Culpeonetta delicatula) is most abundant fish of the sea of Azon ‘with catches amounting to 40-50% of the ‘otal fish landed from this sea, He found optimum condition forthe survival or eggs of this fish, When the temperature reaches up to 15-18 &C. Abnormal development: Swarup (1958) found twin forms ifthe freshly fertilized ege of G. aculearus were subjected to heat and cold (32.5 to 37°C and 0 to 1/2°C). The malformation includes synophthalmia, ‘monophthalmia, microphthalmia and anophthalmia, Not only above-mentioned changes occur but abnormality in blastodisc also occur due to high and low temperature,