The Excitement of
Developmental Biology
Assoc. Prof. Loida R. Medina, PhD, RMT
Department of Biological Sciences
UST – College of Science
Introduction
Historical Background
¤ Aristotle (384-2322 B.C.)
¤ Galen (ca. 130-200 A.D.) structure of relatively advanced
fetuses
¤ Regnier de Graaf (1672) - described the ovarian follicles
¤ Jon Hamm and Anton van Leeuwenhoek (1677) – first saw
the human sperm
¤ Charles Bonnet (1745) – discovered that the eggs of some
insects can develop parthenogeneticaly
Historical Background
¤ Lazzaro Spallanzani (1729-1799) – both male and female sex
products are necessary for the initiation of development
¤ C a s p a r F r i e d r i c h Wo l f f ( 1 7 3 3 - 1 7 9 4 ) – e m b r y o n i c
development occurs through progressive remodeling and
growth
¤ Karl Ernst von Baer (1828) – basic features appear early in
development than do special features; existence of germ
layers in embryo.
¤ Matthias Schleiden and Theodore Schwann (1839) –
foundation of modern embryology was laid down and the
science began.
Developmental biology
¤ Science of how biological form changes
in time.
¤ Ontogeny – individual’s entire life span
¤ O c c u r s m o s t l y i n e m b r y o , d u r i n g
regeneration, metamorphosis, and cell
differentiation.
¤ Fusion of experimental embryology,
developmental genetics, and molecular
biology.
Experimental Embryology
¤ Started at the beginning of the 20th century and
demonstrated the existence of embryonic
induction.
¤ We must not hide from ourselves the fact that the
causal investigation of organisms is one of the most
difficult, if not the most difficult, problem which the
human intellect has attempted to solve … since
every new cause ascertained only gives rise to
fresh questions regarding the cause of this cause.
Developmental Genetics
¤ Flowered in the late 1970s when
mass genetic screens were
carried out on Drosophila.
¤ Resulted in the identification of
a high proportion of the genes
that control development
Molecular Biology
¤ Started with the discovery
of the 3D structure of DNA
in 1953.
¤ Key innovations include
molecular cloning,
n u c l e i c a c i d
hybridization, and
methods for DNA
sequencing.
Activity
Write two (2) possible impacts of
developmental biology in the following key
areas.
1. Health and disease
2. Human genetic manipulation
3. Pharmacological industry
Methods in the Study of
Embryology
Methods Used in the Study of
Embryonic Development
1. Direct Observation
2. Examination of Fixed Material
3. Histochemical Methods
4. Autoradiography
5. Tracing Methods
6. Immunological Methods
Methods Used in the Study of
Embryonic Development
7. Microsurgical techniques
8. Culture Techniques
9. Biochemical and Molecular Techniques
10. Irradiation Techniques
11. Inhibitory Agents and Teratogens
Direct Observation
¤ Microcinematography: technique provides a moving picture
of developing embryo. Vital dyes are used to trace cell
movements
¤ Above is a sample images from time-lapse
microcinematography of cardiac neural crest cell migration in
vitro
Examination of Fixed Material
(Fluorescence Microscopy)
¤ Fixation
¤ preservation of a
structure by treating
the tissue with
chemicals that
preserve structures
without causing
distortion
Fluorescence micrograph showing
the chromosomes from developing
sperm cells.
The blue stain is outlining the
nucleus of the cell and the red
strands represent the chromosomes
within those nuclei
Examination of Fixed Material
(Scanning Electron Microscope)
¤ T h i s t e c h n i q u e
produces a three-
dimensional view of
the surfaces of a
part or the entire
embryo
Scanning electron
micrograph (SEM) of a
round-headed sperm cell
Histochemical Methods
¤ Method of localizing
specific chemical
substances or sites of
chemical activity on
morphological
structures
¤ Typically the tissue or
embryo is rapidly frozen
in liquid Nitrogen
¤ T h e t i s s u e i s t h e n
sectioned with a low-
t e m p e r a t u r e
Histochemical study in microtome called
cancer cryostat.
Autoradiography
¤ Autoradiography
¤ Allows localization of
a radioactive isotope
within cells or tissues
¤ T y p i c a l l y
radioactively labeled
amino acid or a
precursor of DNA or
RNA
Autoradiography of a
rat brain
Autoradiography
¤ In situ
hybridization
¤ Allows the localization
of specific types of
RNA molecules
Tracing Methods
¤ Vital dyes are used to trace cell movements
without harming the cells
¤ Changes of position of cells treated with dyes
can be followed
¤ Stains and vital dyes
-Nile Blue sulfate
-neutral red
-horseradish peroxidase (HRP)
-non-toxic fluorescent dyes
-florescein or rhodamine-
labeled dextrans
Tracing Methods
¤ Retroviruses
-contain reporter genes
(e.g. beta-galactosidase)
-have the advantage of
not being diluted with divisions
of the inflicted cells
-can only be stably
incorporated by dividing cells
Tracing Methods
¤ Adenoviruses
-agents of infecting
non-dividing cells such
as muscles and
neurons
Immunological Methods
¤ Antibodies
¤ Used as probes for the specific presence or
absence of the antigenic molecule of a cell
surface
¤ Monoclonal antibodies
¤ Designed to produces pure and specific
antibodies
Microsurgical Techniques
¤ Ablation
¤ Removal of part of
an embryo to
determine what
effect the absence
of that structure will
have on the
remainder of the
embryo
A fluorescence image of a
somite in a developing mouse
embryo
Microsurgical Techniques
¤ Transplantation and Explantation
¤ Commonly used surgical techniques
¤ Excising a small sample of embryonic tissue
and growing it an artificial environment
¤ Provides information on how the tissue can
adapt to and differentiate in a new location
¤ Self differentiation
¤ Indicates that within the transplanted cells
there is sufficient information to direct the
development of the organ
Microsurgical Techniques
¤ Autografting
¤ Embryonic tissues are transplanted to other
sites on the same embryo
¤ Heterografting
¤ Tissues or organs from a donor embryo are
grafted to hosts of a different species
¤ Xenografting
¤ Tissues or organs from a donor embryo are
grafted to hosts of a different order
Microsurgical Techniques
(Clinical application)
¤ Embryo transfer
¤ Robert Edwards and Patrick Steptoe
¤ Obtained an ovum from the woman’s
ovary
¤ Fertilized the egg in vitro
¤ Allowed the embryo to develop to the
eight-cell stage
¤ The egg was then transplanted to the
woman’s uterus
Culture Techniques
¤ Developed by Ross G. Harrison (1907)
¤ To grow components of embryos or even whole
embryos in an artificial environment
¤ The culture medium is defined chemically that
resemble the natural environment
¤ Major advantage
¤ The medium or the tissue can often be altered in
defined ways that would never be possible in vivo
¤ Disadvantage
¤ Difficult to distinguish processes that operate only in
culture condition from those that occur naturally in
embryo
Biochemical and Molecular
Techniques
¤ Spectrophotometry
¤ Analysis of enzyme activity on the metabolic
properties of embryo
¤ Chromatography and Electrophoresis
¤ First separation technique
¤ M a k e u s e o f p h y s i c a l p r o p e r t i e s o f
compounds which give migratory properties
in solutions or in electric fields
Biochemical and Molecular
Techniques
¤ Column Chromatographic Methods
¤ Develop to separate many classes of
compounds according to various physical
characteristics
¤ Hybridization Technique
¤ Involves the hybridization of simple strand of
DNA with a strand of DNA or RNA to match a
complementary sets of bases
Biochemical and Molecular
Techniques
¤ Polymerase Chain Reaction (PCR)
¤ Amplifying amounts of DNA
¤ Double stranded DNA is denatured
¤ Primers initiate the formation of new
complementary DNA strand
¤ The process is then repeated
Biochemical and Molecular
Techniques
¤ Recombinant DNA Technology
¤ Produce large quantities of a given DNA
sequence
¤ Accomplished with the help of plasmids
¤ Plasmid: Small circular molecules of DNA that replicate
independently in bacteria
¤ Both eukaryote DNA and plasmid DNA are
treated with restriction enzyme
¤ Restriction enzyme: cleave DNA strands at specific
combinations of base pairs
¤ Both types of DNA are split in the same way and some
fragments split off from the eukaryote DNA reattach to
the plasmid DNA and reform a new circle
Recombinant DNA Technology
Irradiation Techniques
¤ To inflict some form of damage on parts
of the embryo
¤ X-rays
¤ Focus on small areas of an embryo to provide
a circumscribed area of tissue injury, or to
inactivate a group of cells
¤ Laser beams
¤ Produces sharp localized lesions in embryo
Inhibitory Agents and Teratogens
¤ Chemical inhibitors that causes alteration in
normal development
¤ Attributed to a specific disturbance in
metabolic pathway
¤ Teratogens
¤ Drugs that cause abnormal development
¤ E.g., Thalidomide
Inhibitory Agents and
Teratogens
¤ Phocomelia (seal
appendage)
v In the extremities of a
patient whose mother
had taken THALIDOMIDE
during early pregnancy
Developmental Genetics and the Use
of Mutants and Genetic Markers
¤ Studies of genetic mutants that
affect specific stages of
development
¤ Lethal mutants are used in analysis of
embryonic development
¤ I d e n t i f i e s w h e r e a n d w h e n
development first goes wrong in a
mutant strain
Developmental Genetics and the Use
of Mutants and Genetic Markers
¤ T complex
¤ Series of lethal mutants in mice
¤ Located on chromosome 17
¤ Plays a role in controlling cellular interactions
involving cellular recognition in early embryo
¤ H-2 complex
¤ Found in chromosome 17
¤ Affects cellular recognition in mouse immune
system
Developmental Genetics and the Use
of Mutants and Genetic Markers
¤ Genetic Markers
¤ Tracers
¤ Identification of specific isoenzymes
by electrophoretic methods
¤ Transgenic animals
¤ Artificially manipulating the genome
Developmental Genetics and the Use
of Mutants and Genetic Markers
¤ A pair of 10-week old
mice. The one above
is a normal mouse.
The one below
carried a transferred
rate gene coding for
growth hormone
