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04 Laboratory Manual Sbc3043 Semester 1, 2018 2019 A181
04 Laboratory Manual Sbc3043 Semester 1, 2018 2019 A181
04 Laboratory Manual Sbc3043 Semester 1, 2018 2019 A181
SBC3043 DEVELOPMENTALBIOLOGY
SEMESTER
S 1,S
EMESTER2, SESSION 2018/2019
ESSION2017/2018
DEPARTMENTOFBIOLOGY
FACULTYOFSCIENCE&MATHEMATICS
UNIVERSITI PENDIDIKANSULTANIDRIS
0
CONTENTS
Laboratory Syllabus
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SBC 3043 Developmental Biology
Introduction
In the lab portion of Developmental Biology students will learn and practice various
experimental approaches for investigating early embryonic development using different
model organisms. The lab course will emphasize experimental design, methodology, and
writing scientific reports. By the end of the term, Developmental Biology students will be
able to plan and execute experiments with living organisms, gather and interpret the
results, and present the outcome of these experiments in clear and concise scientific
reports. Exercises conducted during Developmental Biology Lab will relate to material
covered during lectures and material presented in lab. Lab grades will be based on
production of reports and the lab notebook.
Student Responsibilities.
You are responsible for leaving your work area clean and for rinsing out your own
glassware and dissecting equipment. You are responsible for maintaining and returning
your microscopes and dissecting equipment in good working condition. Shared equipment
(microscopes, dissecting kits) should always be left in perfect condition, so that the next
person can use it without having to clean it or to fix it. If you have questions about how to
use a piece of equipment, do not hesitate to ask the tutor or lab assistants. If you find
problems with tools or equipment, or if supplies or equipment are damaged in the course of
use, please report the problem to the instructor.
Lab report
Reflection
This reflection is a part of component relate to laboratory practical work. At the end of the
laboratory session, students will write a reflection. It should be submitted /attached with the
laboratory report.
1. Students’ will work individually to prepare a reflection. Those who are not participating
will not be given any mark.
2. Please refer to Laboratory Reflection Questions provided.
Lab Reflection Questions
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1. Lab Exercise: __________________________
8. Explain what student have learned and got something from doing this experiment?
Scoring Rubric
Details will be explained by the lecturer/tutor and laboratory during the course.
Late submissions your written reports will result in a reduction in marks available for the
course work.
Lab report will be given 10% and reflective writing is 5%.
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A description of observations are
reported
Data interpreted
5. Discussion Supported by evidence
Section is well thought out and
organized
6. Mechanics Grammatical, spelling or punctuation
errors.
At least 2 was cited in report Poor Good
7. References Proper format (sources in italic or (1) (2)
underlined)
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learning experience and content courses, life experience future
content courses, life experience and /or future goals
experience and /or future goals
goals.
Not submitted
Submitted on time (2 Submitted, but delay
Submission reflection writing -
mark) (1 mark)
(0 mark)
Total Mark= 15
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Laboratory 1 : Reproduction of Flowering Plants
Learning Objectives:
Flowers. The basic structural unit of sexual reproduction in angiosperms is the flower.
Although flowers come in a spectacular array of sizes, shapes, and colors, they are, in
essence, believed to have developed from shoots that have become modified for
reproductive purposes.
Flowers typically have fours kinds of foliar appendages usually arranged in a series of
whorls: sepals, petals, stamens, and carpels, arranged in this order from the bottom to the
top of the floral axis. Although the typical flower displays all of these types of appendages,
sometimes the parts are modified, reduced in size, fused with other parts, or they fail to
develop or abort. These developmental changes contribute to the complexity of flowers.
The two outermost sets, sepals, and petals, are sterile. That is, they do not produce
spores.
Sepals are typically green, and petals are often brightly colored. However, both sepals and
petals can be green or showy. Collectively, the sepals are known as the calyx and the
petals as a unit, the corolla. Together the calyx and corolla comprise the perianth. The term
“perianth” is especially useful in flowers which do not have a distinct whorl of petals and
sepals i.e. only one set of appendage is present instead of the usual two.
Next along the floral axis, inside of the perianth, are the stamens. Stamens are considered
to be microsporophyll. A stamen typically consists of a four-chambered anther, in which
pollen is produced, and a supporting stalk called the filament. Each anther chamber is
equivalent to a microsporangium.
The zygote undergoes mitosis and cytokinesis to reproduce a two-cells embryo. Numerous
subsequent divisions produce an increasingly large and complex embryo.
Materials:
Fresh flowers
Forceps
Razor blade
Dissecting microscope
Mature pollen of Impateins ; prepared slide
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Experimental procedure:
i) Female structure
ii) Male structure
iii) Other structures
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Laboratory Exercise 2: Reproduction of Flowering Plants:
Microsporogenesis / Microgametophyte
Overview
The pollen, or male gametophyte (gamete-producing plant) and the ovule, or female
gametophyte, represents the sexual generation in the life-cycle of the flowering plant. The
pollen may be transferred to the stigma of the same flower or to the stigma of another
flower of the same species. In the former case the process is called self-pollination, in the
latter case, cross-pollination.
On the stigma, the pollen germinates, and produces the male gametes, which are
transported to the ovule in a pollen tube. One of these male gametes fuses with the
female gamete within the ovule. This fusion, called fertilization, marks the beginning of the
development of the embryo plant.
The tissues of the ovule surrounding the embryo form the seed, while the tissues of the
ovary, sometimes with some of the surrounding structures, form the fruit. The fruit is the
ripened ovary (ovaries) with the enclosed seeds, if any, and in some cases may include
other parts of the parent flower. The characteristics of the fruit are thus closely related to
the characteristic of the flower (or flowers), and especially the ovary (or ovaries) from which
it was produced.
Objectives
Materials:
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1. Examine a prepared slide of a lily anther cross section under microscope. First, look
at the lower power. Identify the pollen sacs (microsporangia) where microspores
arise.
2. Use high power to examine the cells in the interior of the pollen sacs. The cells may
be mature pollen grains or microspores that are in stage of pollen development. The
walls of the anther split open the pollen is fully developed.
3. Identify and draw the structure of microspore, and stages of pollen development and
mature pollen grains. Include your drawing sketches of microspores and mature
pollen grains and pollen tubes.
Instruction:
iii) Observe and sketch your field of view at time = 0, after 30 to 40 minutes, after one
hour and after 24 hours
Procedure:
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Laboratory Exercise 3: Reproduction of Plants:
Megasporogenesis / Megagametophyte
Materials:
1. Examine a prepared slide of lily ovary cross section using lower power.
Identify three chamber within ovary contain ovules. You may be able to see the
embryo sac (female gametophyte) as an oval arrangement of larger cells in the
center of the ovule.
3. Draw the lily ovary in cross section. Include in your drawing of a mature
embryo sac and indicate which nuclei are involved in double fertilization.
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Observation: Embryo Development
2. The slide shows globular stage; spherical mass of cells, heart-shaped stage, and
torpedo shaped stage.
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Laboratory Exercise 4 & 5: Mammalian Reproductive System and
Development
Objectives
1) To study the major structures of the male and female reproductive system using
both models and diagrams
2) To understand the major functions of major male and female reproductive system
structures.
3) To examine basic histology of select male and female reproductive structures
4) To study spermatogenesis and understand how sperm are produced the male
5) To study oogenesis and understand events leading to ovulation and production
of an ovum
Preparation
Materials:
Procedure:
Testes; scrotal sac, epididymis, vas deferens, seminal vesicle, Prostate gland
Bulbourethral, and gland penis
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B. Female reproductive structures
1. Using your textbook and the available models, identify the following reproductive
structures vulva vagina cervix uterus, ovary fallopian tubes broad ligament urethra
D. Human Sperm
Focus the specimen using the low power objective. Switch to high power and increase the
amount of light by opening the diaphragm. Sketch the appearance of human sperm under
high power magnification.
1. Using the scanning power of the microscope, identify the growing follicles.
2. Identify the corpus luteum. What is the function of this structure?
3. Sketch the field of view showing the maturing follicle.
4. Locate a follicle that is close to the surface of the ovary, getting ready to be ovulated.
5. Slide/ Model of Ovary:
a) Examine a mature follicle and locate the mature ovum within the follicle. With the
help of Microscopic ovary anatomy (from references book/internet exploring, etc),
examine a prepared slide of an ovary. Under low power you will see a large number
of small, primary follicles near the outer edge. A primary follicle contains a primary
oocyte.
b) Find the secondary follicle (use high power). Note the secondary oocyte (egg)
surrounded by numerous cells; the Graafian follicle with contain a mature secondary
oocyte. How many secondary follicle and Graafian follicle can you find on your slide?
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G. Model of mammalian ovary
1. Using your textbook, identify the following structures in the model of a mammalian ovary
maturing follicles antrum ovarian artery
mature follicle ovarian vein corpus luteurn
2. Sketch the model and label the structures identified above.
3. State the function of the structures identified above.
4. Following fertilization, how many chromosomes does the zygote, the first cell
of the new individual, have?
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Laboratory Exercise 5 & 6: Embryological development
Overview
All sexually reproducing, multicellular organisms begin life as single-celled zygotes, and
progress through stages of growth, differentiation, and morphogenesis. Growth is the
irreversible increase in size of an organism. Differentiation is the structural and functional
specialization of groups of cells. Morphogenesis is the development of pattern, shape, and
form. Embryology is the study of these processes in the early development of an organism.
Objectives
Introduction
Early stages of embryology are similar in advanced invertebrates and vertebrates, and
include formation of body cavities and multiple cell layers. During these stages the
development of embryos must overcome fundamental problems. For example,
development always requires energy (i.e., food), protection of the embryo, and a waste
disposal system. However, the specific morphologies and structures involved in
development vary significantly among groups of animals. In other words, different
organisms have different strategies to cope with the same problems.
During this lab exercise you will compare features and developmental strategies of the
three earliest stages of development of the sea-star, frog, and chick.
2. Cleavage - Zygote divides into a larger and larger number of smaller and smaller cells
that eventually form a hollow sphere of many cells. Cleavage ends when
groups of cells begin to differentiate.
3. Gastrula - Cells of the sphere formed during cleavage continue to divide and move
inward to form three germ layers.
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Materials
Laboratory Procedure
1. Before coming to lab, go to the following websites, print the appropriate pictures
from each, label them, and tape them into your notebook with citations.
Frog development
http://www.cascadia.ctc.edu/FacultyWeb/instructors/ccollin/frog_development.htm
Chick development:
http://web.grcc.edu/biosci/pictdata/104contents.htm
http://www.uoguelph.ca/zoology/devobio/splab4/
3. Sketch a sea star egg, two-celled blastula, four-celled blastula, and a gastrula.
4. Examine prepared slides of a frog egg and zygote. Sketch and label each.
5. Examine slides of early and late cleavage and a slide of a frog blastula cross section.
6. Observe the demonstration specimen of a living 33- and 72-hour chick embryo.
7. Examine 24-, 33-, and 72-hour chick embryos, and follow the formation of the brain,
heart, and eye.
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Laboratory Review
1. Explain the process of gastrulation in each of the organisms observed. What are the
major differences between them?
4. Why do frog eggs have proportionately less yolk than bird eggs?
6. At what point in its development does a human embryo implant in the uterus?
10. What structures do each of the 3 germ layers develop into in an adult?
14. Sketch a frog zygote and label the animal and vegetal poles, and the gray crescent.
Label where yolk is found, and explain how these divisions will affect the axes of
development of the adult.
16. At what stage in the development does the embryo change shape? How does this
change occur?
17. As the embryo becomes a tadpole, the tail becomes a prominent structure. How does
the adult frog lose its tail?
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