EXPERIMENTAL DEVELOPMENTAL BIOLOGY (BiSc 115)
Spring 2004
General Information:
Instructor: Dr. K.M. Brown
Office: 332 Lisner Hall (Lab--308 Bell Hall)
Phone: (202) 994-6193
Office Hours: M, W – 8-10 a.m., and by appointment
Purpose of the Course:
Morphogenetic, biochemical, and genetic mechanisms of cell differentiation in developing animal embryos will be examined through experimentation on live embryonic materials. This is a laboratory course; during lecture periods the background literature, experimental protocol, and data analysis for lab exercises will be discussed. In addition to observing embryonic development of representative animals of several animal phyla, students will also be introduced to a variety of micromanipulative, histological, immunocytochemical, cell biochemical, and molecular genetic techniques. Procedures for recording and analyzing data and for preparation of manuscripts in scientific journal format will be covered, and some original research will be performed.
Reading Materials:
Textbook: Gilbert, S.F. 2003. Developmental Biology, 7th ed., Sinauer Associates Inc.
Supplemental Readings: A reference list and journal articles will be distributed prior to each lab.
Grading Procedure:
Midterm (March 8)20%
Final20%
Lab report 120%
Lab report 225%
Notebook15%
Total100%
Week
1-3
4
5-6
7-8
March 8
9
10
Animal
sea urchin
sea urchin
sea urchin
chicken
sponge
tunicate
Experiment
Posttranslational activation of glucose-6-phosphate dehydrogenase at fertilization
Do individual blastomeres of early cleavage-stage embryos maintain the potential to form a complete organism?
Are serotonergic neurons induced during gastrulation by cells of the invaginating gut tube?
Effects of dopa synthesis inhibitors on catecholamine levels & heart formation
Midterm
Species specificity of sponge cell aggregation factor
Early specification of muscle cell fate
Techniques
in vitro fertilization, subcellular fractionation, protein assay, enzyme activity assay
in vitro fertilization, fertilization membrane removal, blastomere isolation, embryo culture
in vitro fertilization, embryo culture, lithium-induced exogastrulation, whole mount serotonin immunocytochemistry
in vitro embryo culture, whole mount preparation, high performance liquid chromatography with electrochemical detection (HPLC-EC)
sponge cell dissociation, isolation of aggregation factor, aggregation factor activity assay
in vitro fertilization, embryo culture, acetylcholinesterase histochemical assay
*Readings
Ch. 7
Ch. 3, 56-63; Ch. 8, 227-239
Ch. 8, 227-239; Ch. 13, 442-457
Ch. 11, 354-365, 381-383; Ch. 15, 491-500
Ch. 2, 44-47; Ch. 3, 70-77
Ch. 3, 57-59; Ch. 8, 246-251
Week
11
12
13
14-15
Final Exam
Animal
frog
frog
fly
sea urchin
Experiment
Disruption of dorsal-ventral axis formation by inhibition of fertilization-triggered cortical rotation; rescue by egg rotation
Growth factor-induced mesoderm induciton in prospective skin ectodermal pieces from blastula embryos
Heat-induced salivary gland gene transcriptional changes & increases in heat shock protein levels. Ecdysone-mediated eversion of isolated imaginal discs
Measurement of serotonin receptor (5-HT2B) mRNA levels in mesenchyme blastula embryos & pluteus larvae; cloning the 5HT2B cDNA
Time to be announced
Techniques
hormone-induced egg maturation, in vitro fertilization, jelly coat removal, UV disruption of microtubules, embryo culture
hormone-induced egg maturation, in vitro fertilization, jelly coat & vitelline membrane removal, animal cap isolation, tissue culture
salivary gland & imaginal disc isolation from 3rd instar larvae, chromosome squash & staining, Western blot analysis of hsp70 and hsp90 heat shock proteins
embryo culture, RT-PCR, cDNA electrophoresis, cDNA cloning, DNA sequencing (demonstration)
*Readings
Ch. 10, 305-338
Ch. 3, 65-67; Ch. 10, 305-338; Ch. 11, 363
Ch. 4, 92-93; Ch. 9, 297; Ch. 18, 583-592; Ch. 23, 776-777
Ch. 4, 93-95
*Readings are from the Gilbert text. Journal articles will also be assigned for each lab.