CHAPTER I

INTRODUCTION

A. Background
One feature that signifies that it is sentient life is able to reproduce. in animals,
fertilization does is external fertilization internal fertilization. Fertilization is external
fertilization is done outside the body. Whereas internal fertilization is fertilization
outside the body. In animals that have two living like amphibians, eggs are done in
water. There are lenders on egg-slime to protect the eggs of amphibians are animals
because of the pressure that does fertilization externally.
Organ are complex structures composed types of tissues. In the vertebrate eye, for
example, light is transmitted through the transparent corneal tissue and focused by
the lens tissue, eventually impinging on the tissue of the neural retina. The precise
arrangement of tissues in this organ cannot be disturbed without impairing its
function. Such coordination in the construction of organs is accomplished by one
group of cells changing the behavior of an adjacent set of cells, thereby causing them
to change their shape, mitotic rate, or fate. This kind of interaction at close range
between two or more cells or tissues of different history and properties is called
proximate interaction or induction.
Ovulation could be induced at any season by injection more efficiently than by
implantation of the anterior pituitary. Ovulation is not an all or none reaction but
depends upon the amount of anterior pituitary hormone available. Male and female
pituitaries are not equally potent, the female anterior pituitary being approximately
twice as polent as the corresponding male gland in respect to inducing ovulation or
amplexus. There is no hormone sex specifically of the hormone, mammalian extracts
have on the whole prove to be rather ineffective in inducing ovulation in Rana is
unsuccessfully attempted when the male is alone sexually stimulated.
B. Purpose
1. Obtain eggs and frog at the time of fertilization process desired in great
numbers.
2. Study the development of the embryo to form the Zygote starts frog larvae.
 3. Learn the type and fertilization at frog
4. Study the formation of future organs of frog originating from each layer
embryo.
C. Benefit
From this experiment, we can get some benefit :
1. The college student can know about the process of fertilization on frogs.
2. The college student can know about development of the embryo begins to
form a Zygote larvae.
3. The college student can know the type and pattern of cleavage of the frog.
4. The college student can know the way to establishment of the organs of frog
originating from each layer embryo.
 CHAPTER II
BASIC THEORY

Ovulation is a process of cell terlepasnya eggs (ova) from the ovarian follicle
rupture as a result of which it has entered. The mechanism of the occurrence of
ovulation is affected by hormonal, neural, and periodicity of light. Ovulation on the
frog occurs after the first polar body of oocyte release, wall theka externa and follicle
cells of the follicle rupture. Follicle growth due to the influence of the hormones FSH
(Follicle Stimulating Hormone) produced by anterior hypofise gland, then a follicle
cells able to produce the hormones estrogen and progesterone. These two hormones
in small amounts of giving impetus to the anterior hipofisa gland to produce
hormones LH (Luteinizing Hormone). LH hormones play an important role in the
onset of ovulation is bluffing. Hypofise frog glands taken through hyposectomy is
the source of the hormone FSH and LH can be used to induce ovulation. The
potential of the gland will decline if the are in room temperature for a few hours
(Adnan, Penuntun Praktikum Perkembangan Hewan, 2016).
Fertilization on various types of animals can be distinguished based on the
venue, i.e., fertilization in external and internal fertilization. Fertilization is external
in fertilization happens outside its mother's body. For example various types of fish,
some types of frogs, and so on. Fertilization is internal fertilization that takes place in
the body of its mother. The fertilization in animal usually take place internally
produce eggs that mature in a limited amount in one time cycle of reproduction and
usually only 1-15 fruit. Fertilization has several functions, such as the transmission
of paternal and maternal genes to the , stimulates the egg to grow further, resulting in
the occurrence of paternal genetic properties of melting and maternal conditions
diploid a particular species of its kind (syngami), and determination of gender
genetically (Adnan, Arifin, & Suryani. 2016).
Frog egg consists of two poles that is pigmented black and anima pole
vegetative not pigmented. Characteristics of eggs that have been fertilization is the
existence of grey areas the opposite shift kea rah entry of sperm. A Zygote is formed,
entered stage 1 Division. The fertilization is holoblastic type, while the fertilization
thorough from pole to pole the anima vegetative. Division 1 with merodional the
direction of the fertilization right on the line of middle gray Crescent, producing two
blastomeres. Division 2 meridional direction but 90 degrees toward the field of
Division 1, yielding 4 blastomeres. Division 3 horizontal, perpendicular to the plane
of Division 1 and 2 produce 8 blastomeres are not as great, i.e. 4 micromer and 4
macromer. Starting from the Division 3 and beyond will produce large blastomeres is
not the same. Division 4, meridional simultaneously, formed 16 blastomeres.
Division 5, horizontally above and below the plane of Division 3, yielding 32
blastomeres. After fertilization, the embryo enters a blastula. At the blastula stage
there is a cavity called the blastocoel. Blastula stage, the embryo once it entered the
stage of grastula. Gastrula is formed of a series of process of cell movement, with the
final results in the form of a layer of ectoderm embrional, namely, mesoderm, and
endoderm. After grastulasi, the next stage is neurula. At this stage the physical form
and changes transpired between layer induces embrional each other. The formation
of organs occurs after the formation of the neurulation (Adnan, Penuntun Praktikum
Perkembangan Hewan, 2016).
The pattern of cleavage in amphibians is radial cleavage holoblastic, the
division in which the blastomeres on the polar part of the anima eggs located just
above the blastomeres on the vegetative parts, so that the pattern is radial
blastomeres symmetrical. Blastula has 3 different areas, i.e., areas around the pole
anima that includes cells that form the roof of the blastocoel, the cells of the
ectoderm layer would constitute, and these cells are small and called micromer that
contains a lot of a grain of pigments : Polar area surrounding vegetative cells include
large yolk (macromer) which is the future endoderm cells and contains many details
of the yolk; sub Equatorial regions and in the form of a ring of marginal cells, covers
the gray (gray crescent), and this area is normally will form the mesoderm cells. The
formation of layers of institutions on a frog through 3 morfogenic movement, namely
the epiboly (widespread mikromer of anima to pole vegetative), invagination (stand
out cells in the area he was gray and formed blastoporus), and involution (Shaper
primitive area along epiboly).involution (the form of primitive region with epiboly).
 The number of eggs per egg clutch was recorded. Embryos were analyzed
from cleavage until tadpole hatching, and were staged according to the E. machalilla
table of stages (del Pino et al. 2004). Embryos were cultured in humid chambers at
room temperature (18–23 °C). Cleavage of D. auratus was not observed. The H.
vertebralis blastula consisted of small, pigmented cells in the animal hemisphere;
whereas, cells of the vegetal hemisphere were larger. The blastocoel developed
during cleavage, and was large in blastula and gastrula stage embryos. The blastocoel
roof, of the two species, was several cell diameters in thickness at stage 10 , and it
was reduced to one cell thickness in the late-gastrula stage of H. vertebrali. Similarly
the thickness of the blastocoel roof was reduced to one or two cells in thickness in
the early gastrula of D. auratus. Thickness of the blastocoel roof in the late-gastrula
was not documented for this frog species. In E. machalilla, expansion of the
blastocoel was accompanied by reduction in its thickness until it was a monolayer of
cells in the late-gastrula (Hervas, Torres, Larrea, & del Pino, 2015).
R. muscosa appears to require a hibernation period for successful
reproduction. The effects of hibernation were manifested in vocal advertisement
signaling, amplexus, female receptivity, and oviposition. Each of these behaviors
plays an important role, leading to successful reproduction in R. muscosa, and by
taking a behavioral approach, we were able to observe how hibernation catalyzed this
repertoire of interacting behaviors that are critical for successful reproduction. The
increased frequency of calling observed in hibernated. Male R. muscosa no doubt
served to help establish amplexus, which stimulates female egg release and
fertilization. Virtually all (88% of observations) hibernated males established
amplexus with females, while almost none (2% of observations) of the non-
hibernated males did so. Females can influence the outcome of amplexus by
signaling a lack of receptivity, and non-hibernated females were more likely to emit
release calls during amplexus compared to hibernated females. Maintaining
amplexus is costly for females because they cannot eat, have restricted mobility and
can drown if they are in amplexus simultaneously with multiple competing males
(Santana, Swaisgood, Lemm, Fisher, & Clark, 2015).
 Spawning period is the time needed for the brood fish after injected with frog
(F. cancrivora) pituitary gland to spawn. The observation time started from the time
of injection up to the moment when ovulation happened and fish ready to spawn.
Observation was conducted to each treatment to investigate the impact of pituitary
gland injection on spawning period of the brood fishes. T3 treatment yielded the
fastest spawning period with the average time was 12.03 ± 0.44 hours. It is indicated
that the use of frog (F. cancrivora) pituitary gland with a dosage of 0.5 ml/kg per
brood fish gave the best impact on acceleration of spawning period. The
mathematical correlation between the spawning period of common carp (C. carpio)
after the injection of pituitary gland with the pituitary gland dosage in a polynomial
equation as: y = 8.5085x - 9.3166x + 15.391 with R = 0.6257 and r = 0.791. Where,
correlation coefficient (r) stated the significant effect between independent variable
and dependent variable which meant that the injection of pituitary gland of frog (F.
cancrivora) significantly affected the spawning period of common carp (C. carpio by
70.10%. Theoretically, deformation of follicle layer and the egg movement into the
ovary tube are the signals which indicate that the brood fish ready to spawn. The
research shows that different treatment level of will also affect the spawning period
of brood fish at different time. Higher dosage injected of frog (F. cancrivora)
pituitary gland will cause longer spawning period. While lower dosage level will also
cause longer spawning period. It occurs because lower dosage does not affect
oocyte’s growth while higher dosage of frog (F. cancrivora) pituitary gland would
retard the growth process of oocytes as the reaction of gonadotropin works
(Rumondang, Risjani, & Fadjar, 2015).
 CHAPTER III
PRACTICAL METHOD

A. Time and Place

Day/Date : Thursday/ November, 16th 2017
Time : 14.00-15.30
Place : Green House Biology FMIPA UNM
B. Tools and Materials
1. Tools
a. Plastic vessels
b. Surgery board
c. Scissors
d. Tweezers
e. Petri dish
f. syringes, 2 cc volume
g. Mortar and pestle
C. Materials
a. Male and female frog
b. Alcohol
c. Cotton
d. Physiology NaCl
D. Work Procedure

The frog is catch. The frog is put at

Tools and materials
surgery board.
prepared.
Take ovum and membrane Open the skin, look for Frog dissected from the bottom
tympani and throw it on the ovum on the frog, also take to the top.
petri dish which have a membrane tympani
physiology Nacl

Mashed the membrane The liquid that has been

tympani. Frog is massage so quiet
pounded and then taken by during injected.
using a syringe.

Injections of fluids that have

Frogs are sorted again so that
been obtained from the results
the injection happens
of mashing to frogs.
succeeded perfectly.
 CHAPTER IV
RESULT AND DISCUSSION

A. Result
Gambar Kegaiatan Keterangangan

1. Egg (Ovum) is a frog

that has been on
induction.
2. Rana cancarivora that
has been induced for 1
week.

2 1

B. Discussion
Based on the results of the experiments have proven that done use frog (Rana
cancarivora) which will produce a pituitary gland-injected eggs in rather a lot.
Female frogs Injection by using the hypofise glands found in the membrane tympani
frogs have somewhat yellowish orange colored glands. The glands then crushed and
then injected on the ventral abdomen of female frogs. Prior to injection, we should
sort out the part that will be injected. Based on theory, the frog that injected with
hypofise glands or pituitary with quicker reactions of formation of their eggs, this is
due to the gland hypofise gland/pituitary gland that produces a FSH which will
produce the hormones estrogen and progesterone hormone. Furthermore, the
hormone estrogen and progesterone will stimulate the pituitary gland to produce
hypofise LH. Function of LH that is stimulating the onset of ovulation or the release
of ovarian follicle-follicular towards my uterus into an ova (eggs) are prepared to
fertilized.
 CHAPTER V
CLOSING

A. Conclusion
Frogs are injected with the hypofise glands or pituitary with quicker reactions of
formation of their eggs, this is due to the gland hypofise gland/pituitary gland that
produces a FSH hormone that will result estrogen and progesterone hormone.
Furthermore, the hormone estrogen and progesterone will stimulate the pituitary
gland to produce hypofise LH. Function of LH stimulate the occurrence of ovulation
or the release of ovarian follicle-follicular towards my uterus into an ova (eggs) are
prepared to fertilized.
B. Suggestion
1. Please add lab assistant to make the experiment effective and efficiency.
2. Please wear lab uniform because it is one of rule in the laboratorium.
 BIBLIOGRAPHY

Adnan. (2016). Penuntun Praktikum Perkembangan Hewan. Makassar: Universitas

Negeri Makassar.
Adnan, Arifin, A. N., & Suryani, A. I. (2016). Perkembangan Hewan. Makassar:
Universitas Negeri Makassar.
Hervas, F., Torres, K. P., Larrea, P. M., & del Pino, E. M. (2015). Development and
gastrulation in Hyloxalus vertebralis and Dendrobates auratus (Anura:
Dendrobatidae). Amphibian & Reptile Conservation, 121-135.
Rumondang, A., Risjani, Y., & Fadjar, M. (2015). The Introduction of Pituitary
Gland Extract of Crab-eater Frog (Fejervarya Cancrivora) to Accelerate
Ovulation of Eggs and Spawning of Common Carp (Cyprinus Carpio) .
Journal of Life Science and Biomedicine , 153-158.
Santana, F. E., Swaisgood, R. R., Lemm, J. M., Fisher, R. N., & Clark, R. W. (2015).
Chilled frogs are hot: hibernation and reproduction of the Endangered
mountain yellow-legged frog Rana muscosa. ENDANGERED SPECIES
RESEARCH, 43-51.