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BIOLOGY 453 - COMPARATIVE VERT.

ANATOMY
WEEK 1, LAB 2: Embryology of Frog & Chick

Learning Goals

1. Know both the traditional taxonomy & major clades to which the frog & chick embryos belong.
a. Know at least 1 shared derived trait that can “define” each clade.
2. Be able to identify the following in a frog & chick embryo:
a. Relative amount of yolk in eggs: microlecithal, mesolecithal, macrolecithal
b. Cleavage patterns: holoblastic, meroblastic & discoidal.
c. Identify these developmental stages & associated structures in embryonic frog development:
i. Fertilized egg or zygote: animal & vegetal pole
ii. Blastula (blastocoel, micromeres & macromeres)
iii. Gastrula (blastocoel, archenteron, notochord, yolk plug, blastopore, anterior vs posterior regions)
iv. Neurula (neural plate & neural folds, anterior vs posterior regions)
d. In the chick find these homologous structures to the blastopore: Hensen’s node & primitive streak.
e. Primary germ layers (ectoderm, mesoderm or endoderm) & examples of adult tissues each may form.
i. Identify the 3 major types of mesoderm: epimere (somites), mesomere (intermediate) & hypomere
(lateral plate mesoderm). Give an example of an adult tissue that is derived from each type.
f. Identify these structures or organs in both frog & chick embryos, as noted: eye, lens, otic capsule, notochord,
somites, neural tube or neural folds, brain, pharyngeal arches and heart.
g. Identify these additional organs in the tadpole (they remain small or difficult to see in early chick stages): liver,
kidney, and gut or intestine.
3. Compare the lamprey larva & amphioxus specimens to the frog & chick embryos. What traits do the frog & chick
embryos share with the lamprey larva & Amphioxus?

Specimens to Examine

Misc. 35 mm slides Microscope slide: Whole mount Microscope slide: Sagittal Sec.
Frog embryo X X X
Chick embryo X X X

Traditional Taxonomy & Major Clades

Traditional Taxonomy Major Clades


Subphylum Vertebrata (Craniata) Clade Vertebrata: fishes, amphibians, “reptiles”, birds & mammals
Class Lissamphibia (frogs, salamanders...) Shared derived trait: vertebral column surrounds spinal cord
Class Aves (birds)
& more!! Clade Tetrapoda: amphibian, lizard, snake, alligator, bird, mammal
Shared derived traits: digits on limbs, loss of internal gills...

Clade Amniota: lizards, snakes, alligators, birds & mammals


Shared derived traits: amniotic egg with new extra-embryonic
membranes amnion, chorion & allantois
Additional Sources of Information

Topic Egg type Cleavage Coelom formation Mesoderm diff. Neurulation


Kardong (text) pg. 159-160 162-165 165, 168-170 170-175 167, 170-174

GENERAL
Cell & Developmental Biology Online! 2009. Univ. of Guelph, Zoology Dept.
http://www.uoguelph.ca/zoology/devobio/dbindex.htm
Mallery, C. 2009. Animal Development. Biol. 150. Univ. of Miami.
http://www.bio.miami.edu/~cmallery/150/devel/animal_development.htm
Nouvelles Technologies Educatives del l'universite de Lyon1. 2000. Atlas d'embryologie descriptive des vertébrés.
http://nte-serveur.univ-lyon1.fr/nte/embryon/tp2000/correction.htm
PBS. 2008. Morphing Embryos. Odyssey of Life. (Quicktime videos of embryos)
http://www.pbs.org/wgbh/nova/odyssey/clips/
Slonczewski, J. 2009. Chapter 14. Gastrulation & Neurulation. Biology 114, Biology Dept., Kenyon College
http://biology.kenyon.edu/courses/biol114/Chap14/Chapter_14.html
Univ. of Calgary. 2000. The Virtual Embryo.
http://people.ucalgary.ca/~browder/virtualembryo/db_tutorial.html
Wasserman, B. Movies/Pics. Bill Wasserman’s Developmental Biology Page, Loyola Univ., Chicago.
http://www.luc.edu/faculty/wwasser/dev/devm.htm

CHICK ONLY
Burke, AC. 2009. Body wall formation in the chick embryo. Learning Objects Team, Wesleyan University.
http://learningobjects.wesleyan.edu/musc_dev/
Cebra-Thomas, J. 2003. Chick Embryo Staging. Developmental Biology, Swarthmore Univ.
http://www.swarthmore.edu/NatSci/sgilber1/DB_lab/Chick/chick_stage.html
DevBio.com 2009. Coelom Formation (animation). Developmental Biology Online. Companion to Developmental Biology,
8th Edition by Scott F. Gilbert, Sinauer Associates. http://8e.devbio.com/article.php?ch=15&id=138
Gilbert, SF. 2000. Early Development in Birds. Cleavage in Bird Eggs. Developmental Biology, 6 th Ed.
http://www.ncbi.nlm.nih.gov/bookshelf/br.fcgi?book=dbio&part=A2581
Hill, M. 2008. Embryology: Chicken Development Stages. Univ. of New South Whales Embryology.
http://embryology.med.unsw.edu.au/OtherEmb/chick2.htm
Muneoka, K. 2006. Online Developmental Atlas. CELL 413 - Embryology Lab., Dept. of Cell & Mol. Biol., Tulane Univ.
http://www.tulane.edu/~embryo/labsyllabus.htm
Shaw, M. 2005. Gametogenesis & Development. Univ. of Manitoba - series of labeled chick slides with descriptions
http://www.umanitoba.ca/faculties/science/biological_sciences/lab14/biolab14_4.html
Temkin, M. 2001. Atlas of Developmental Stages, Developmental Biology - 31 St. Lawrence Univ.
http://it.stlawu.edu/~mtem/devbiol/db99atlas.htm
Univ. of Illinois Urbana-Champaign. 1998. Chickscope (Go into a stage to find great microscopic views!)
http://chickscope.beckman.uiuc.edu/explore/embryology/

FROG ONLY
Cebra-Thomas, J. 2003. Xenopus Embryo Staging. Developmental Biology, Swarthmore Univ.
http://www.swarthmore.edu/NatSci/sgilber1/DB_lab/Frog/frog_staging.html
Frontiers in Bioscience. 2007. Germ Layer Derivatives. Atlas of Xenopus Embryology
http://www.bioscience.org/atlases/fert/htm/develhum/5germper.htm
Kimball, J. 2009. Frog Embryology. Kimball’s Biology Pages.
http://users.rcn.com/jkimball.ma.ultranet/BiologyPages/F/FrogEmbryology.html
Munson, D. 2009. Histology. (frog images) MCB 116: Experimental Embryology. Harvard Univ.
http://www.courses.fas.harvard.edu/~mcb116/topics/histology.html
Sievert, L. 2004. Frog Development Models. Vertebrate Structure and Development, ZO 515 - 516. Emporia State Univ.
http://academic.emporia.edu/sievertl/verstruc/frmodel.htm
Univ. of Wisconsin. 2002. Amphibian Embryology Tutorial! (QuickTime movie & more)
http://worms.zoology.wisc.edu/frogs/welcome.html
Embryology Basics
EGG TYPES
Eggs vary in size primarily because of differences in the amount of yolk, but eggs of similar size may also vary in the proportionate
amount of yolk. Eggs are described in the table.
Microlecithal [lecith = yolk] Mesolecithal Macrolecithal
small amount of yolk moderate amount of yolk large amount of yolk
In fact, eggs within some taxa such as amphibians and fishes show a wide range of size & yolk proportion. Technically, they fall
into a continuum of variation rather than absolute categories. Most vertebrate eggs have an unequal distribution of yolk in the egg.
These eggs have separate regions with non-yolky material at one end of the egg & yolk concentrated at the opposite side. This
type of egg is called telolecithal [telo = end] with separate animal & "vegetal" or yolk poles.

CLEAVAGE PATTERNS
Cleavage (mitotic cell division) through yolk is slower than cleavage through the animal pole. As a result, cells may get smaller in
the animal pole region than in the vegetal pole. If cleavage occurs through both the animal & vegetal poles it is called holoblastic
[holo = whole; blast = bud] cleavage. It will be equal & holoblastic in microlecithal eggs but there is usually unequal, holoblastic
cleavage in mesolecithal eggs with more yolk. These eggs produce smaller animal pole cells & larger, yolk filled cells.

If the vegetal pole has a very large mass of yolk, i.e. its a macrolecithal egg, the egg doesn't cleave completely. If only the animal
pole undergoes cleavage this is called meroblastic [mero = a part] cleavage. A specialized form of meroblastic cleavage is called
discoidal cleavage [disc = round plate]: the animal pole cells are extremely small & the yolk amount is huge.

GERM LAYER INTRODUCTION


Ectoderm forms the epidermis of the skin, the lens of the eye & neural ectoderm forms brain & spinal cord.
Mesoderm divides into 4 major regions that form specific tissues of the body:
Chordamesoderm - forms the notochord
Epimere (Paraxial mesoderm) - forms segmented blocks called somites
Mesomere (Intermediate mesoderm) - forms the kidneys & gonads
Hypomere (Lateral plate mesoderm) - forms smooth muscle, heart, coelomic cavity
Endoderm forms the epithelial lining of the gut or archenteron, epithelial lining of the lungs & the yolk, if present.

The mesodermal somites form in pairs along either side of the notochord in young embryos. Each somite divides into 3
subdivisions that form some of the segmented structures of the body & part of the skin:
Sclerotome - vertebrae Dermatome - dermis of skin Myotome - skeletal or somatic muscle
Myotomes develop into a series of V-shaped, segmentally arranged muscles or myomeres that are visible in the tail of the frog
embryo. Myomeres form segmentally arranged muscles in all vertebrates although the pattern may be modified. Myosepta are
connective tissue layers that separate the myomeres.

Germ layer
development in a
transversely
sectioned amniote.
Class Lissamphibia - Frog Embryos
Identify the listed structures for each slide. More features may be present; you are responsible for only those listed.

35 mm Slides: Images from Whole views Various sectioned views


http://www.swarthmore.edu/NatSci/sgilber1/DB_lab/Frog/frog_staging.html
Fertilized egg:
Mesolecithal (moderately yolked).
Telolecithal egg with animal (pigmented) & vegetal poles

Cleavage:
Eight cell stage or slightly higher.
Holoblastic (complete cleavage)
Radial cleavage: Describe this term.
Meroblastic cleavage: means unequal sized blastomeres.
Label the micromeres: animal cells & macromeres: yolk-filled cells.

Blastula stage:
Identify the animal pole & vegetal pole.
Label the blastula cavity.

Early gastrulation:
Find the dorsal lip of the blastopore, as animal pole cells begin to
migrate over yolk & into interior space of embryo.
Cells migrating inward will form endoderm, mesoderm.
Remnant of blastocoel still apparent in Sagittal section.

Late Gastrulation:
Find the blastopore filled with the yolk plug.
Label the gut (archenteron).
Label germ layers: ectoderm, mesoderm & endoderm.

Early Neurulation:
Label the neural folds & neural groove.
In the transverse section, label notochord, somites & gut.

Late Neurulation:
Neural tube has closed.
In the transverse section find & label the 3 primary germ layers;
neural tube, notochord, archenteron & somites
Whole Mount Frog Embryo (4-8 mm) Microscope Slide Sagittal Section 10 mm Frog Embryo Microscope slide

Identify the following: eye, otic vesicle (future ear), brain, spinal Each slide has a series of sagittal sections.
cord, notochord, pharyngeal arches, heart, & myomeres. Otic Identify the following structures (they won't all be visible on any given
vesicle & pharyngeal arches may be hard to see on some slides. section or perhaps any 1 slide): eye & lens, otic vesicle, brain, spinal cord,
notochord, pharynx, heart, kidney, liver, intestine, myomeres.
B
A

http://www.uoguelph.ca/zoology/devobio/210labs/34frogwm.htm
What type of sectional view is this? Is this more or less mature What type of sectional view is this?
than the drawing in figure B? Label the brain, spinal cord, Find & label the notochord, neural tube, intestines, liver & kidney tissue.
notochord, yolk, pharynx, myotomes, anus & heart.
C D

Make your own drawings of the specimens showing para-sagittal views here:
Class Aves - Chick Embryos
Use these diagrams & illustrations to help you label the images on the following pages.
24 hr stage 33 hr hour stage

72 hour stage 96 hr stage


Identify the listed structures for each slide. More features may be present; you are responsible for only those listed.

35 mm slides - Whole View Chick Embryo


Images from http://www.swarthmore.edu/NatSci/sgilber1/DB_lab/Chick/chick_stage.html
18 hr. 21 hr.
Gastrulation Early neurulation

Find notochord, Find neural groove &


Hensen's node & the neural folds, somites,
primitive streak. primitive streak &
Hensen's node

28-33 hr. 48 hr.

Find the neural tube, Find the optic capsule,


brain, somites, blood otic vesicle/capsule,
islands in extra- heart, somites, brain &
embryonic region & neural tube.
posterior portion of
heart.

72 hr. 84 hr.

Find the eye, lens, otic Find the eye, lens, otic
capsule, brain, spinal capsule, brain, spinal
cord (neural tube), cord (neural tube),
pharyngeal grooves or pharyngeal grooves or
arches, heart, somites, arches, heart, somites &
limb buds. limb buds.

The transparent sac


near the tail is the
allantois, one of the
extra-embryonic
membranes unique to
amniotes.
Whole Mount Microscope slide 48-72 hour chick embryo
Identify the following: optic cup (eye), lens, otic capsule, brain, spinal cord, pharyngeal arches, heart, somites & limb buds.
Make your own drawing here:
48 hour embryo at left
72 hour embryo below

Sagittal Section Microscope slide 48 hour chick embryo


Each slide has a series of sagittal sections. Identify the following (not all are visible on any given section or any 1 slide): eye (lens &
eye cup shows a pigmented retina forming), otic capsule, brain, spinal cord, pharynx, heart & somites
Sagittal section of a 48 hour chick embryo. Sagittal section of a 72 hour chick embryo.

Make your own para-sagittal drawing here:


35 mm slides - Transverse Sections of Chick

#1 Gastrulation: This is a section through the primitive streak showing the formation of ectoderm, mesoderm & endoderm.

#2 Early neurulation: Label the neural plate, notochord, somites. Identify the germ layers endoderm, mesoderm & ectoderm.

#3 Late neurulation: find the neural tube, notochord, somites.

#4 Higher magnification: find somites, coelomic cavity, paired dorsal aortae, notochord & neural tube, "future" archenteron/gut
Identify the germ layers - ectoderm, mesoderm, and endoderm. Then identify the 2 divisions of the somite visible in this slide -
dermatome-myotome (darker line of tissue, dorsal to rest of somite) & sclerotome division (lateral to neural tube & notochord).