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.