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INFORMATION TO USERS This manuscript has been reproduced from the microfilm master. UMI films the text directly from the original or copy submitted. Thus, some thesis and dissertation copies are in typewriter face, while others may be from any type of computer printer. The quality of this reproduction is dependent upon the quality of the copy submitted. Broken or indistinct print, colored or poor quality illustrations and photographs, print bleedthrough, substandard margins, and improper alignment can adversely affect reproduction. In the unlikely event that the author did not send UMI a complete manuscript and there are missing pages, these will be noted. Also, if unauthorized copyright material had to be removed, a note will indicate the deletion. Oversize materials (e.g., maps, drawings, charts) are reproduced by sectioning the original, beginning at the upper left-hand corner and continuing from left to right in equal sections with small overlaps. Each original is also photographed in one exposure and is included in reduced form at the back of the book. Photographs included in the original manuscript have been reproduced xerographically in this copy. Higher quality 6" x 9" black and white photographic prints are available for any photographs or illustrations appearing in this copy for an additional charge. Contact UMI directly to order. University Microfilms International A Bell & Howell Information Company 300 North Zeeb Road. Ann Arbor. Ml 48106-1346 USA 313/761-4700 800/521-0600 Order Number 9130593 The impact of insect diapause on water balance physiology Yoder, Jay Alan, Ph.D. The Ohio State University, 1991 UMI 300 N. Zeeb Rd. Ann Arbor, MI 48106 THE IMPACT OF INSECT DIAPAUSE ON WATER BALANCE PHYSIOLOGY DISSERTATION Presented in Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy in the Graduate School of the Ohio State University By Jay Alan Yoder, B.A. ***** The Ohio State University 1991 Dissertation Committee: David L. Denlinger Approved by Donald E. Johnston Susan W. Fisher Brian H. Smith tyuQ — Advisor Copyright by Jay Alan Yoder 1991 In loving memory of my Grandfather, Jonas M. Yoder, and to the enduring support and strength of my family. I also dedicate this dissertation to the late Professor George W. Wharton, who inspired my interest in arthropod water balance. ACKNOWLEDGMENTS I am grateful to Dr. David L. Denlinger for his patience, guidance and support throughout my graduate work. I sincerely thank Sir Donald E. Johnston, who provided necessary guidance and fatherly advice and who is one of my greatest inspirations. A special thank you is extended to Drs Susan W. Fisher and Dana L. Wrensch for their continual emotional support and friendship, and to Dr. Brian H. Smith for serving on my dissertation committee. I thank Professor Pappachan E. Kolattukudy for allowing me to work in his laboratory and for his help with the hydrocarbon analyses. I also thank all the graduate students on whose friendship I rely heavily, in particular Ty T. Vaughn, L. Denise Boulet, Eric R. Hoffman, Thane Thay, Astri Wayadande, Brian Gilles, Robert G. and Peter W. Hancock-Kovarik, Donald S. S. Yehling, Zhi Ben Chen, various members of the OSU Zoology Department and our own departmental alumni: Drs Maria E. Casanueva and Marvin D. Sigal. I extend sincere appreciation to my fellow lab colleagues: Chen- Ping Chen, Karl H. Joplin, George D. Yocum, Mei Ling Zhang, and a special thank you to D. Courtney Smith, whose advice has strengthed my character on and off the battlefield. I owe many thanks to Dr. Karen J. Ott and Sandy Sieg, who gave me the iii incentive to begin graduate work. Mindy, your unchanging faith me is never forgotten. Lastly, I thank the Department of Entomology for supporting my graduate work. VITAE February 13, 1964, Bom - Dayton, Ohio 1982-1986_______ B.A., B.A., L.A. University of Evansville, Evansville Indiana 19 8 6 -p resen t_____ .Graduate Teaching and Research Assistant, The Ohio State University, Columbus, Ohio FIELD OF STUDY Major field: Entomology (Insect Physiology) TABLE OF CONTENTS DEDICATION_____________ :____________________________________ ii ACKNOWLEDGMENTS_______________________________________ iii VITA_________________________________________________________ v LIST OF TABLES______________________________________________ ix LIST OF FIGURES______________________________________________ x CHAPTER I. INTRODUCTION_____________________________ 1 A. Sarcophaga- A model diapause system_________ 2 B. Endocrinology of diapausing versus nondiapausing pupae of Sarcophaga__________ 3 C Cuticular lipid correlates with insect's water conservation needs____________________ 4 D. Embryonic diapause and water relation ________ 6 E Extatosoma tiaratum, a tropical walking stick possessing two stages of embryonic diapause _________________________ 8 F. The chemical and physical properties of water____________________________________ 1 0 G Water balance and basic parameters ___________ 12 H. Early water balance theory ____________________ 1 4 I. The basic water balance equation and transpiration m_____________________________ 1 5 J. Water gain by water vapor absorption and metabolic water ________________________ 1 6 K Equilibrium water mass and the energetics of mt and ms_____________________ 1 6 L. Phase barrier Ea and critical transition temperature________________________________ 1 8 M. Research objectives ___________________________ 1 9 N. References____________________________________ 2 1 II. WATER BALANCE IN FLESH FLY PUPAE AND WATER VAPOR ABSORPTION ASSOCIATED WITH DIAPAUSE 3 0 A. A bstract 3 0 B. Introduction 3 1 C Materials and Methods 3 2 D. Results 3 7 E Discussion 5 5 F. References 6 1 III. A COMPARISON OF THE WATER BALANCE CHARACTERISTICS OF TEMPERATE AND TROPICAL FLY PUPAE 6 6 A. A bstract 6 6 B. Introduction 6 7 C Materials and Methods 6 8 D. Results 7 0 E Discussion 7 7 F. References 8 1 vii IV. WATER VAPOR UPTAKE BY DIAPAUSING EGGS OF A TROPICAL WALKING STICK 8 3 A. A bstract _______________________________________ 8 3 B. Introduction ___________________________________ 8 4 C Materials and Methods_________________________ 8 5 D. Results ______________________________ „ _________ 8 9 E Discussion _____________________________________ 101 F. References____________________________________ 107 V. PUP ARIA OF DIAPAUSING FLESH FLIES ARE ENHANCED WITH ADDITIONAL HYDROCARBONS 111 A. Abstract ______________________________________ 1 11 B. Introduction 1 1 2 C Materials and Methods________________________ 1 1 3 D. Results ________________________________________ 1 1 9 E Discussion _____________________________________ 1 3 9 F. References____________________________________ 146 VI. EVIDENCE FOR A BRAIN FACTOR THAT STIMULATES SYNTHESIS OF PUPARIAL HYDROCARBONS IN DIAPAUSING FLESH FLIES________________________ 1 5 1 A. Results and Discussion _________________________ 1 5 1 B. References____________________________________ 159 SUMMARY. 161 7. Net transpirational water loss from eggs of E. tiaratum as early embryos at 20°C (A) and at 25°C (B) and as pharate first instar larvae at 25°C (C) 9 5 8. Percent change in weight in response to long term exposure to three hydrating atmospheres at 20°C for early embryonic eggs 9 7 9. Rates of water vapor exchange for eggs of E. tiaratum as early embryos at 20°C (A) and at 25°C (B) and as pharate first instar larvae at 25°C ( C) 9 9 10. Total ion count (TlC)-gas chromatograms (abundance versus carbon number) of hydrocarbons from empty puparia of nondiapausing (A) and diapausing (B) S. crassipalpis. Both groups were reared at 20°C. indicates the C29 alkane unique to diapause _________ 129 11. Mass spectrum (abundance versus mass/charge ratio) of C29 indicates a straight-chain alkane _________________ 1 3 1 12. Formation of CO and alkane from [1-14C], [9,10-3H] octadecanal by the 105,000 x g supernatant and resuspended microsomal pellet.________________________ 133 13. Radio-gas chromatography by the alkane generated from [1-14C], [9,10-3H]octadecanal by the microsomal preparation___________________________ 135 14. Effects of the cofactors, ATP, CoA and NADH on the formation of alkane from octadecanal and octodecanoic acid catalyzed by microsomes of flesh fly pupae_________________________________________ 1 3 7 xi LIST OF TABLES TABLE 1. A comparison of wet weight, percent body water, net transpiration rate, 24 h equilibrium weight, CTT, and Ea below and above the CTT for temperate zone and tropical fly pupae________________________ 2. A comparison of wet weight and weight change (%) at saturation for eggs of six species exhibiting different developmental patterns__________________ 3. Quantities of epicuticular hydrocarbon and other lipids extracted from the surface of S. crassipalpis at different developmental stages_________________________ 1 4. A comparison of net transpiration rate (-kt) at 20°C and quantities of hydrocarbon (pg/empty puparium) for pupae of S. crassipalpis_____________________________ 1 LIST OF FIGURES FIGURE PAGE 1. Total body water pool expressed in mg or % for nondiapausing (A) and diapausing (B) groups of flesh flies (S. crassipalpis) at different stages of development 4 5 2. Net transpiration at 20°C from nondiapausing (A) and diapausing (B) S. crassipalpis pupae into dry air_____________________________________________ 47 3. Percent change in weight in response to long term exposure to hydrating (av 0.85) and dehydrating (av 0.33) atmospheres at 20°C for nondiapausing (A) and diapausing (B) S. crassipalpis pupae 4 9 4.