THE UNIVERSITY OF MANITOBA STUDIES ON OVINE PLACINTAL ...

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-5- r'rhereas tryptophan is absent. a) The Alpha-Subunit: The alpha subunit of hCG is a glycopeptide $¡ith a molecular weight. of L4,9O0, of which the peptide portion represents I0r200 and the carbohydrate 4r200. The complete amino acid sequence of 89-92 amino acid residues for hCGlalpha has been proposed by Morgan and co¡,¡orke rs (L9?2, 1975) and by BahJ- and associates (BahI , L972bî Bahl et al , 1972¡ Bel-isario et al , 1973). It is shown in Figure l. Both groups of workers found heterogeneity at the amino terminus. Approxinately 30* of aLL hCG-alpha chains lacked the terminal tripept.ide, and another 10S the terminal d.ipeptide. Such heterogeneiÈy could represent precursor molecul_es at, various stages of cleavage but. could aríse from digest.ion during urinary excretion or even during purification of the hormone. hCG- aLpha has five disulfide bonds, but there is no evi
NHt-Aì a-Pro-Asp-Val -Gl n-Asp-Cys -Pro.rGl u-C1's-Thr-Leu-Gf n-Gl u-AìB Pro-Phe-Phe-Ser-Gl n -Pro-G1y-Al a-pro- I I e-Leu-Gl n-Cys-Met-Gty- 20 25 3ó Cys -Cys -Phe -Ss ¡-A¡g -Al a-Tyr-Pro -Thr-pro -Leu -Arg -Ser-Lys -Lys _ 35 40 4s Thr-Met-Leu-Val -Gl n -Lys -Asn-Val -Thr-Ser-G'l u-Ser-Thr_Cys _Cys_ 50s5-60 Va1 -Aì a-Lys-Ser-Tyr-Asn -Arg-Vaì -Thr-Val -Met-Gty-G1y_phe_Lys_ 6570'is Va'l -Gl u-Asn-Hi s-Thr-At a-Cys-Hl s-Cys-Ser-Thr-Cys-Tyr-Tyr-Hl s_ 80 85 90 Lys-Ser-C00H Figure l.Linear sequence of amino acids in the c-subunit of Human chorìon.ic gonadotropln. Based on Morgan et al (te7s)
Page 1 and 2: THE UNIVERSITY OF MANITOBA STUDIES
Page 3 and 4: To My parents and wife: Mn & Mrs. K
Page 5 and 6: CONTTNT S I. REVIEW 0N PLACENTAL PR
Page 7 and 8: V. RESULTS E. STUDIES ON THT BIOSYN
Page 9 and 10: Peptide Hormones: ACTH CG FSH GH LH
Page 11 and 12: LIST OF FIGURES FIGURE PAGE I Linea
Page 13 and 14: 22 Radioreceptor assay for oPRL us.
Page 15 and 16: Effect of PGF,. and lactogenic horm
Page 17 and 18: ABSTRACT STUDIES 0N 0VINE PLACENTAL
Page 19 and 20: eceptor-b i nd i ng studies show th
Page 21 and 22: -2- years have seen an increasing i
Page 23: -4- secretion of a peptide hor:none
Page 27 and 28: 6a NH, -Ser-Lys -G'l u.Pro -Leu -Ar
Page 29 and 30: -8- 197J-; Rayford et al | !9721 .
Page 31 and 32: -10- sialic acid díd not alter the
Page 33 and 34: -L2- and Biasucci (1969) denonstrat
Page 35 and 36: _14_ addition of CAMP (Handwerger e
Page 37 and 38: -16- conponents I the.faster haying
Page 39 and 40: _18_ by hcc, as evidenced by increa
Page 41 and 42: _20- (Mason et a1 , L973i Menon ¡
Page 43 and 44: 21a NH, -Vaì -G1 n-Thr-Val -Pro-Le
Page 45 and 46: -23- growth-promoting ef,fect, wher
Page 47 and 48: -25- diffusion teÊt. Josi¡novich
Page 49 and 50: -27- v¡hereas in the tissue extrac
Page 51 and 52: -29- found products of molecular we
Page 53 and 54: -3r- vÍii) S.ecreLion and metabol
Page 55 and 56: - 33 - metabolic factors may influe
Page 57 and 58: -35- Although hpI, si¡nuLates the
Page 59 and 60: -37- C. CIIHER ITUMAN PLACENTAL PEP
Page 61 and 62: _39_ stimulating activityf is found
Page 63 and 64: -4t-- (iii) Human Chorionic Cortlco
Page 65 and 66: - 43 - syncytiotrophoblast (Tatarin
Page 67 and 68: - 45 - Ib SUBPRTMATE PLACENTAT PR.O
Page 69 and 70: -47 - samples, signif,icant elevati
Page 71 and 72: -49- required for the chorignÍc ao
Page 73 and 74: B. (i) -51 - MONI(ET PLACENTAL I,AC
Page 75 and 76:
- 53 - brith 13 minutes for hpl,. T
Page 77 and 78:
55 Pregnarit Marê Sêrum Gonâdót
Page 79 and 80:
II b. NON - pRl¡late PT,ACENTAT.
Page 81 and 82:
(iv¡ Inmuhologicat Mêthogs: - 59-
Page 83 and 84:
_ 6t_ 1974; Fellows et a1, 1974¡ H
Page 85 and 86:
63 Ordêr Rodêhtia (Tat, nicê, ro
Page 87 and 88:
Guinea Pi g: 65 Hypophysectomy of t
Page 89 and 90:
-67 Orilet Perissodactyla (Horse an
Page 91 and 92:
-69 activity in the N-acetyl-lactos
Page 93 and 94:
(Íi) coat: -71 The goat is a spont
Page 95 and 96:
- 73- When serum from a pregnant go
Page 97 and 98:
-75 Gusdon et al (1920)using . haer
Page 99 and 100:
77 (c) Imnunologicaì Properties of
Page 101 and 102:
-79 - SECTION II : OYINE REpROpucTI
Page 103 and 104:
BI which contain' as lvell as the o
Page 105 and 106:
83 ce11s usually contaln either num
Page 107 and 108:
85 non-pregnant values did not diff
Page 109 and 110:
8rl Leaf blank to corect nr.rmberin
Page 111 and 112:
- 89- luteolysis when supesistpessd
Page 113 and 114:
9t SECTION IV¡ MtTttODs AND IIATER
Page 115 and 116:
- 93- (.TCA) solution were added co
Page 117 and 118:
-95 - (ÞPRL' NIH-P-B3) , and all q
Page 119 and 120:
97 buffer, the eluates were pooled,
Page 121 and 122:
Char4ctêf ization Pfocedures l. Ch
Page 123 and 124:
t0t (c) À.nalvticaL .{rod.iu¡n do
Page 125 and 126:
103 B : MFTÍÍOD OF DETECTION AND
Page 127 and 128:
AFsa)¡- ProceiiureS 105 Specific b
Page 129 and 130:
107 ', C : IIEBI{OÐ OF DEVELOPMENT
Page 131 and 132:
109 D. RTNG PREGNôXCY (l) Maternâ
Page 133 and 134:
l1ì (wtlvo'). After the hornogenat
Page 135 and 136:
(4) Tissúe distributiori df radloi
Page 137 and 138:
l'1 5 (2) Procg.ssjnq, gf. incuÞ¡
Page 139 and 140:
1.l7 for 30 min. Then the precipita
Page 141 and 142:
(2) Assav of ìuteo.trgpic effec.t
Page 143 and 144:
EXPERIMENT # TI (7 days after HCG)
Page 145 and 146:
123 to progesterone (l/300 djlution
Page 147 and 148:
SECTION V : RESULTS (r) LACTOGEN DU
Page 149 and 150:
127 Fractioñâtión of oPL. from c
Page 151 and 152:
129 Figure 6. carborymethyì(cM,)-c
Page 153 and 154:
t 3't óo0 ,oo 800 900 FRACIION r,t
Page 155 and 156:
E o ø ô¡ t- z ao É, o '133 .--.
Page 157 and 158:
TABLE I TABLE OF PURIFICATION OF OV
Page 159 and 160:
137 Figure lì. A, Poìyacrylamide
Page 161 and 162:
Leaf blank to correct nr.rmbering
Page 163 and 164:
14r Anal.vtical Sodi um Dodecyl Sql
Page 165 and 166:
o 9 x F- -)- 9 rU I æ -t f () LT'
Page 167 and 168:
¿ o s : t45 Figure 15b. Displaceme
Page 169 and 170:
HoR MoNE coNc€NrRArto ln ß/mt )
Page 171 and 172:
Tissue used for binding ôtudies Li
Page 173 and 174:
Noñ. p..s'ro¡r {N,p.l Pr.eñonr (
Page 175 and 176:
¡( É T 2, \ ¡a.2¡./ir ,..r.¡.
Page 177 and 178:
P 80 \ x.. ¡,azrot¡j .1.¿¡i¡ .
Page 179 and 180:
o c o U o òs -o c2o ao eÀo o 'o .
Page 181 and 182:
F 2 o o o\ ¡¡ig ô 2 Þ 0 (¡ l5e
Page 183 and 184:
! o 't6 l HORMO¡¡ E COr{CENr FAIt
Page 185 and 186:
163 Figure 24 shows the sensltivity
Page 187 and 188:
165 D. STUDIES ON THE SECRETION OF
Page 189 and 190:
E .Dc À o - art & 167 Flgure 26. P
Page 191 and 192:
lJhen blood samples were taken four
Page 193 and 194:
Sheep no. l{6 25í 960 464 5?0 680
Page 195 and 196:
't73 fetal cotylêdons at various d
Page 197 and 198:
TABLE VI OPI CONCENTRATTON IN FETAL
Page 199 and 200:
l. Súrglcat Method 't77 Figure 3l
Page 201 and 202:
= >€ ,3 ; I 179 rwt 76-0450 o----
Page 203 and 204:
dUrlng PrêOnancy in thè Ewes tBr
Page 205 and 206:
! ñ' l83 NoRMON! C oNCENTßArlO þ
Page 207 and 208:
l ol
Page 209 and 210:
I I I "+ _t rl \l :- 'i rl -l l87 a
Page 211 and 212:
l89 Flgure 37. Percentage ot l2sl-o
Page 213 and 214:
19t column is shown in Figure 3BA.
Page 215 and 216:
- 193 3H -couNdro _________SPECtFtC
Page 217 and 218:
TABLE IX /- BIOASSAY OF qROIItIfH p
Page 219 and 220:
z o 197 Figure 40, Radioreceptor as
Page 221 and 222:
5A L lNl Pgtzq cô-tlzt Pgfl6 + cl
Page 223 and 224:
SFCT.IoN II : pl.scussr0N 4d ã 1È
Page 225 and 226:
Ch aracteri zati on 203 Analytical
Page 227 and 228:
205 size, corresponding to the mole
Page 229 and 230:
207 the specific binding of oPL to
Page 231 and 232:
209 than with prolactin-like effect
Page 233 and 234:
2t1 the jncrease in oPRL binding si
Page 235 and 236:
213 The resuìts obtained indicate
Page 237 and 238:
215 on the role af PL in the fetus.
Page 239 and 240:
Surgi ca1 method- 217 Our studies o
Page 241 and 242:
219 5.Relationships amonq opRL. oGH
Page 243 and 244:
221 The nature of thelarge nolecula
Page 245 and 246:
223 data argue for a close interact
Page 247 and 248:
]. TABLE OF CHEMICAL PROPERTIES GUI
Page 249 and 250:
cor,t coAT (bPL) (cpl) 3. TABLE OF
Page 251 and 252:
À) E 229 Figure 43. The secretion
Page 253 and 254:
231 H. POSSIBLE ROLT OF OVINE PLACE
Page 255 and 256:
233 Our studies on the blnding of o
Page 257 and 258:
235 through its binding to cotpus l
Page 259 and 260:
237 Furthermore, oPL neasurement in
Page 261 and 262:
239 circulationand alìantoic f'lul
Page 263 and 264:
SECTICN IX PUBLICATIONS A) Manuscr
Page 265 and 266:
SECTION X: BIBLIOGRAPHY Abramovich,
Page 267 and 268:
Beas,F., Saìinas, 4., GonzalezrF.,
Page 269 and 270:
247 Brody,S,, and Carlstrom,G.,J.Cl
Page 271 and 272:
249 Crosignanì ,P.G. , Nencioni ,
Page 273 and 274:
25t Friesen,H.G., Endocrinoìogy 76
Page 275 and 276:
253 Grinwich,D.L., M. Hichens, and
Page 277 and 278:
25s Hum, V.G., Knipfel , J.G., and
Page 279 and 280:
?57 Knopp,R,H,, H,J. Ruder,E. Herra
Page 281 and 282:
259 Manku,M.S., and D.F. Horrobin,
Page 283 and 284:
261 , :iir .,.-;;r-,:, Morgan, F.J.
Page 285 and 286:
263 Pierce,G.B., Midþley,A.R. and
Page 287 and 288:
?65 Shepard, T.H.., Gen. Comp. Endo
Page 289 and 290:
267 Tsuruhara,T., Dufau,M.L., Hickm
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