Involvement of L (-) rhamnose in sea urchin gastrulation: a live embryo assay

126 Pages·2012·7.22 MB·English

California State University, Northridge

INVOLVEMENT OF L (-) RHAMNOSE IN SEA URCHIN GASTRULATION: A
LIVE EMBRYO ASSAY

A thesis in partial fulfillment of the requirements
For the degree of Master of Science
in Biology

By

Tiffany N. Smith

December 2012
The thesis of Tiffany Smith is approved:

__________________________________________________________
Randy W. Cohen, Ph.D. Date

__________________________________________________________
Stan T. Metzenberg, Ph.D. Date

__________________________________________________________
Steven B. Oppenheimer, Ph.D., Chair Date

California State University, Northridge
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Acknowledgements

Foremost, I would like to express my sincere gratitude to my mentor and advisor, Steven
B. Oppenheimer, PhD for his guidance, support, and advisement throughout my graduate
curriculum. He has truly been inspirational to me. His expertise and knowledge has
influenced me to pursue a career in cell and molecular biology research. I really
appreciate him for giving me the opportunity to join his lab to gain experience and
expertise in laboratory skill and technique.

Besides my advisor, I would also like to thank Randy W. Cohen PhD and Stan T.
Metzenberg PhD for their insightful comments and for taking time to serve on my
committee. In addition, I would like to thank Aida Metzenberg PhD for serving as my
external thesis evaluator and for contributing to my thesis edits as well.

I would like to thank my lab partner, Kathy D. Fernando, who has helped me throughout
the rigorous curriculum and has accompanied me in long nights of experimentation.
Thank you for teaming up with me and being supportive by working around my work
schedule. I would, also, like to thank Christine Spence and Erika Lorenzo who have
helped me with data entry and with the tedious task of counting embryos.

My sincere thanks go to my parents who have provided love, support, and encouragement
throughout the program. I would like to particularly thank my mom, Julia Smith who
helped me with data formatting and my dad, Harold Smith for dropping off the numerous
copies of my thesis to the biology office when I was unable to.

I really appreciate all of you for your contributions in helping me complete my thesis.

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Table of Contents

Signature page ii
Acknowledgements iii
List of Figures v
List of Tables xi
Abstract viii
Introduction 1
Materials & Method 11
Results 17
Discussion 23
Conclusion 33
References 34
Appendix 38

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List of Figures
Figure 1: Mechanism of Gastrulation 38
Figure 2: Mechanism of Exogastrulation 39
Figure 3: Carbohyrdate Mediated/Inhibition Cellular Adhesion 40
Figure 4: α-Cyclodextrin 24 Hour Treatments: Overview with Percentages 41
Figure 5: α-Cyclodextrin 24 Hours: Overview with Standard Error 43
Figure 6A: Controls α-Cyclodextrin 24 hours 45
Figure 6B: 0.03M α-Cyclodextrin 24 hours 45
Figure 6C: 0.015M α-Cyclodextrin 24 hours 45
Figure 6D: 0.003M α-Cyclodextrin 24 hours 46
Figure 6E: 0.0015M α-Cyclodextrin 24 hours 46
Figure 6F: 0.0009 M α-Cyclodextrin 24 hours 46
Figure 7: α-Cyclodextrin 30 Hour Treatments: Overview with Percentages 47
Figure 8: α-Cyclodextrin 24 Hour Treatments: Overview with Standard Error 49
Figure 9A: Controls α-Cyclodextrin 30 hours 51
Figure 9B: 0.03M α-Cyclodextrin 30 hours 51
Figure 9C: 0.015M α-Cyclodextrin 30 hours 51
Figure 9D: 0.003M α-Cyclodextrin 30 hours 52
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Figure 9E: 0.0015M α-Cyclodextrin 30 hours 52
Figure 9F: 0.0009 M α-Cyclodextrin 30 hours 52
Figure 10: Melibiose 24 Hour Treatments: Overview with Percentages 53
Figure 11: Melibiose 24 Hour Treatments: Overview with Standard Error 55
Figure 12A: Controls Melibiose 24 hours 57
Figure 12B: 0.03M Melibiose 24 hours 57
Figure 12C: 0.015M Melibiose 24 hours 57
Figure 12D: 0.003M Melibiose 24 hours 58
Figure 12E: 0.0015M Melibiose 24 hours 58
Figure 12F: 0.0009 M Melibiose 24 hours 58
Figure 13: Melibiose 30 Hour Treatments: Overview with Percentages 59
Figure 14: Melibiose 30 Hour Treatments: Overview with Standard Error 61
Figure 15A: Controls Melibiose 30 hours 63
Figure 15B: 0.03M Melibiose 30 hours 63
Figure 15C: 0.015M Melibiose 30 hours 63
Figure 15D: 0.003M Melibiose 30 hours 64
Figure 15E: 0.0015M Melibiose 30 hours 64
Figure 15F: 0.0009 M Melibiose 30 hours 64
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Figure 16: L (-) Rhamnose 24 Hour Treatments: Overview with Percentages 65

Figure 17: L (-) Rhamnose 24 Hour Treatments: Overview with Standard Error 67
Figure 18A: Controls L (-) Rhamnose 24 hours 69
Figure 18B: 0.03M L (-) Rhamnose 24 hours 69
Figure 18C: 0.015M L (-) Rhamnose 24 hours 69
Figure 18D: 0.003M L (-) Rhamnose 24 hours 70
Figure 18E: 0.0015M L (-) Rhamnose 24 hours 70
Figure 18F: 0.0009 M L (-) Rhamnose 24 hours 70
Figure 19: L (-) Rhamnose 30 Hour Treatments: Overview with Percentages 71

Figure 20: L (-) Rhamnose 30 Hour Treatments: Overview with Standard Error 73
Figure 21A: Controls L (-) Rhamnose 30 hours 75
Figure 21B: 0.03M L (-) Rhamnose 30 hours 75
Figure 21C: 0.015M L (-) Rhamnose 30 hours 75
Figure 21D: 0.003M L (-) Rhamnose 30 hours 76
Figure 21E: 0.0015M L (-) Rhamnose 30 hours 76
Figure 21F: 0.0009 M L (-) Rhamnose 30 hours 76
Figure 22: Trehalose 24 Hour Treatments: Overview with Percentages 77
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Figure 23: Trehalose 24 Hour Treatments: Overview with Standard Error 79
Figure 24A: Controls Trehalose 24 hours 81
Figure 24B: 0.03M Trehalose 24 hours 81
Figure 24C: 0.015M Trehalose 24 hours 81
Figure 24D: 0.003M Trehalose 24 hours 82
Figure 24E: 0.0015M Trehalose 24 hours 82
Figure 24F: 0.0009 M Trehalose 24 hours 82
Figure 25: Trehalose 30 Hour Treatments: Overview with Percentages 83
Figure 26: Trehalose 30 Hour Treatments: Overview with Standard Error 85
Figure 27A: Controls Trehalose 30 hours 87
Figure 27B: 0.03M Trehalose 30 hours 87
Figure 27C: 0.015M Trehalose 30 hours 87
Figure 27D: 0.003M Trehalose 30 hours 88
Figure 27E: 0.0015M Trehalose 30 hours 88
Figure 27F: 0.0009 M Trehalose 30 hours 88
Figure 28: D (+) Xylose 24 Hour Treatments: Overview with Percentages 89
Figure 29: D (+) Xylose 24 Hour Treatments: Overview with Standard Error 91
Figure 30A: Controls D (+) Xylose 24 hours 93
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Figure 30B: 0.03M D (+) Xylose 24 hours 93
Figure 30C: 0.015M D (+) Xylose 24 hours 93
Figure 30D: 0.003M D (+) Xylose 24 hours 94
Figure 30E: 0.0015M D (+) Xylose 24 hours 94
Figure 30F: 0.0009 M D (+) Xylose 24 hours 94
Figure 31: D (+) Xylose 30 Hour Treatments: Overview with Percentages 95
Figure 32: D (+) Xylose 30 Hour Treatments: Overview with Standard Error 97
Figure 33A: Controls D (+) Xylose 30 hours 99
Figure 33B: 0.03M D (+) Xylose 30 hours 99
Figure 33C: 0.015M D (+) Xylose 30 hours 99
Figure 33D: 0.003M D (+) Xylose 30 hours 100
Figure 33E: 0.0015M D (+) Xylose 30 hours 100
Figure 33F: 0.0009 M D (+) Xylose 30 hours 100
Figure 34: L (-) Xylose 24 Hour Treatments: Overview with Percentages 101
Figure 35: L (-) Xylose 24 Hour Treatments: Overview with Standard Error 103
Figure 36A: Controls L (-) Xylose 24 hours 105
Figure 36B: 0.03M L (-) Xylose 24 hours 105
Figure 36C: 0.015M L (-) Xylose 24 hours 105
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Figure 36D: 0.003M L (-) Xylose 24 hours 106
Figure 36E: 0.0015M L (-) Xylose 24 hours 106
Figure 36F: 0.0009 M L (-) Xylose 24 hours 106
Figure 37: L (-) Xylose 30 Hour Treatments: Overview with Percentages 107
Figure 38: L (-) Xylose 30 Hour Treatments: Overview with Standard Error 109
Figure 39A: Controls L (-) Xylose 30 hours 111
Figure 39B: 0.03M L (-) Xylose 30 hours 111
Figure 39C: 0.015M L (-) Xylose 30 hours 111
Figure 39D: 0.003M L (-) Xylose 30 hours 112
Figure 39E: 0.0015M L (-) Xylose 30 hours 112
Figure 39F: 0.0009 M L (-) Xylose 30 hours 112

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