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Chapter Two – Serum osteocalcin is inversely associated with adiponectin, glucose,and serotonin in black bears ................................................................. 202.1. Introduction ......................................................................................................... 202.2. Methods and materials ....................................................................................... 212.2.1. Bear handling and sample collection ........................................................ 212.2.2. Biochemical assays .................................................................................. 222.2.3. Data representation and statistics ............................................................ 232.3. Results ................................................................................................................ 252.4. Discussion .......................................................................................................... 282.5. Tables and figures .............................................................................................. 34Chapter Three – Serum neuropeptide Y and IGF-1 favor reduced bone turnover inhibernating black bears ....................................................................... 483.1. Introduction ......................................................................................................... 483.2. Methods and materials ....................................................................................... 493.3 Results ................................................................................................................. 503.4 Discussion ........................................................................................................... 513.4.1. Maintenance of serum leptin concentrations several weeks intohibernation may prevent bone loss and help bears maintain nearnormalbody temperatures........................................................................ 513.4.2. Reduced IGF-1 and increased NPY in the serum of hibernating bearsfavor suppression of bone turnover ........................................................ 533.4.3. Summary .................................................................................................. 553.5. Tables and Figures ............................................................................................. 56iv
Chapter Four – Attempts to find an anti-apoptotic environment in hibernating bearserum remain inconclusive .................................................................... 634.1. Introduction ......................................................................................................... 634.2. Methods and materials ....................................................................................... 644.3. Results ................................................................................................................ 654.4. Discussion .......................................................................................................... 654.5. Tables and figures .............................................................................................. 67Chapter Five – Serum from hibernating bears protects cells from caspse-3 and -7activation ............................................................................................... 715.1. Introduction ......................................................................................................... 715.2. Methods .............................................................................................................. 725.2.1. Black bear handling and serum sample collection ................................... 725.2.2. Cell culture ............................................................................................... 725.2.3. MC3T3-E1 apoptotic response to seasonal bear serum .......................... 725.2.4. Caspase-3/7 activity of bear bone lysates ................................................ 735.2.5. MC3T3-E1 gene expression response to bear serum .............................. 745.2.6. Biochemical assays .................................................................................. 755.2.7. Data representation and statistics ............................................................ 755.3. Results ................................................................................................................ 755.4. Discussion .......................................................................................................... 785.5. Tables and figures .............................................................................................. 83v
Page 1 and 2: EXPLORATION OF THE ROLE OF SERUM FA
Page 3: Table of contentsIndex of Figures .
Page 7 and 8: Index of figures1.1. Simplified sch
Page 9 and 10: 6.17. Longitudinal analysis of OPG
Page 11 and 12: 6.49. Longitudinal analysis of ColI
Page 13 and 14: A.2. Complete list of BSALP correla
Page 15 and 16: List of abbreviationsα-MEM. Alpha-
Page 17 and 18: PCR. Polymerase chain reactionPer1
Page 19 and 20: G2/M checkpoint. The point in the c
Page 21 and 22: Species names13-lined ground squirr
Page 23 and 24: Chapter One - Background and signif
Page 25 and 26: 1.3 Bone turnover is unbalanced dur
Page 27 and 28: pro-apoptosis regulator BAX. An imp
Page 29 and 30: low-dose administration of parathyr
Page 31 and 32: decrease in bone trabecular thickne
Page 33 and 34: experience periodic arousals in whi
Page 35 and 36: ears must have evolved a mechanism
Page 37 and 38: 1.7.6 Hypothesis 2: OCN interacts w
Page 39 and 40: ain, heart, and femoral muscle of h
Page 41 and 42: Hypothesis 3a: Serum concentrations
Page 44: were released. Behavior indicating
Page 47 and 48: 2.3 ResultsSerum activity of the bo
Page 49 and 50: (p
Page 51 and 52: post-hibernation sampling in the be
Page 53 and 54: also unclear whether ucOCN may affe
Page 55 and 56:
atios of intact to fragmented OCN m
Page 57 and 58:
Table 2.3—Bone marker “seasonal
Page 59 and 60:
Table 2.7—Chemistry panel “seas
Page 61 and 62:
ABNormalized BSALPBSALP (U/L)CO N D
Page 63 and 64:
Total Calcium (mg/dL)A1.21.151.11.0
Page 65 and 66:
ABTotal OCN (ng/mL)150100500O N D J
Page 67 and 68:
Adiponectin (ng/mL)AB40003500300025
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ABNormalized InsulinInsulin (μU/mL
Page 71 and 72:
surrounding and encompassing hibern
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decreased from 788+30 ng/mL in preh
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120-122]. Most small hibernators ar
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ears. Similarly, serum NPY increase
Page 79 and 80:
Table 3.2—Serum factor longitudin
Page 81 and 82:
Leptin (ng/mL)ABO N D J F M A M O N
Page 83 and 84:
Norepinephrine (ng/mL)A807060504030
Page 85 and 86:
IGF‐1 (ng/mL)AN D J F M A MB10009
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2008. Samples were collected as des
Page 89 and 90:
sampling points. It is possible to
Page 91 and 92:
C‐Terminal PTH (Relative)AB504030
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Chapter Five — Serum from hiberna
Page 95 and 96:
Synergy HT Multi-Detection Micropla
Page 97 and 98:
gene specific primers and 12.5 μL
Page 99 and 100:
involved in the reduced caspase-3/7
Page 101 and 102:
hibernation cycles without problem
Page 103 and 104:
filtered seasonal bear serum and fo
Page 105 and 106:
which is dependent upon caspase-3 a
Page 107 and 108:
Caspase‐3/7 ActivityO N D J F M A
Page 109 and 110:
6Caspase‐3/7 Activity54321**0Vehi
Page 111 and 112:
ABCaspase‐3/7 Activity9876543210*
Page 113 and 114:
eyond the G1/S checkpoint, thus ind
Page 115 and 116:
ears, it is possible that tissue se
Page 117 and 118:
PTH1R, RANKL, Runx2, Smurf1, TLR4,
Page 119 and 120:
factors Runx2 and OCN also increase
Page 121 and 122:
Table 6.3—Gene expression 3 hour
Page 123 and 124:
Table 6.5—Gene expression 6 day d
Page 125 and 126:
Cyclin D1 Gene ExpressionO N D J F
Page 127 and 128:
Smurf1 Gene ExpressionO N D J F M A
Page 129 and 130:
BAK Gene ExpressionO N D J F M A MM
Page 131 and 132:
M‐CSF Gene ExpressionO N D J F M
Page 133 and 134:
Per1 Gene ExpressionO N D J F M A M
Page 135 and 136:
Akt Gene ExpressionO N D J F M A MM
Page 137 and 138:
Bcl‐2 Gene ExpressionO N D J F M
Page 139 and 140:
Cyclin D1 Gene ExpressionO N D J F
Page 141 and 142:
OPN Gene ExpressionO N D J F M A MM
Page 143 and 144:
2PTH1R Gene Expression1.510.50O N D
Page 145 and 146:
TLR4 Gene ExpressionO N D J F M A M
Page 147 and 148:
Bcl‐2 Gene ExpressionO N D J F M
Page 149 and 150:
OCN Gene ExpressionO N D J F M A MM
Page 151 and 152:
PTH1R Gene ExpressionO N D J F M A
Page 153 and 154:
TLR4 Gene ExpressionO N D J F M A M
Page 155 and 156:
condition in renal patients in whic
Page 157 and 158:
hibernation. This report brings to
Page 159 and 160:
14. Kaneps, A.J., S.M. Stover, and
Page 161 and 162:
41. Chowdhury, I., B. Tharakan, and
Page 163 and 164:
68. Ashe, M.C., et al., Bone geomet
Page 165 and 166:
95. Perrien, D.S., et al., Aging al
Page 167 and 168:
122. Lesser, R.W., et al., Renal re
Page 169 and 170:
151. Confavreux, C.B., R.L. Levine,
Page 171 and 172:
178. Toribara, T.Y., A.R. Terepka,
Page 173 and 174:
206. Luo, X.H., et al., Adiponectin
Page 175 and 176:
233. Florant, G.L., et al., Fat-cel
Page 177 and 178:
259. Bhasin, S., et al., Older men
Page 179 and 180:
284. Miura, M., et al., A crucial r
Page 181 and 182:
Appendix A—Complete tables of cor
Page 183 and 184:
Table A.5—Complete list of ionize
Page 185 and 186:
Table A.7—Complete list of adipon
Page 187 and 188:
Table A.12—Complete list of norep
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