THE UNIVERSITY OF MANITOBA STUDIES ON OVINE PLACINTAL ...

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-36- lipid stores (picard et al, 1968; Fairweather, 197I). Prolonged infusion of hpl, into norrnal or hypopituitary subjects resulted in increased free fatty acid levels and irnpaired insulin leve1 following glucose administrat.ion (Beck and Daughaday, 1962i Samaan et al , 196g). However, the changes in blood sugar response are disputed by Josimovich and Mintz (1968), and the changes in free fatty acid levels were not observed by Kalkhoff et al (1969). Studies on animals treated vrith hpl, and in vitro experiments confirm the increased secretíon of insulin (Martin and Friesen, 1969; Lopez et a1 , 1971) and the release of free fatty acids from adipose tissue (Turtle and Kipnis, 1967; Genazzíni et a1, 1969; Felber et al , L9Z2; Mochizuki et al , 1975). The benefits to the fetus of this mobilizat,ion of substrates are illustrated by experiments in which hpL was given to pregnant rats resulting in increased fetal Ìíeight, glycogen, protein, and lipid content (Mochizuki et, al , 1973). No such response bras observed when hpl. was administered directly to the fetus. Finally, there is some evidence (Spellacy et al , 1971ar Kim and Felig, I97!.; ¡.,aube et aI , 1972; Gaspard et, al, l9?5) that. the leve1 of maternaL blood sugar provides a negative feedback system for controlling hPt secretion rate.
-37- C. CIIHER ITUMAN PLACENTAL PEPTTDE TIORM<strong>ON</strong>E S AND' SECRETED PROTETNS. (i) HumanChorionic Thyrotropin (hCT): A thyrotropic substance was first isolated from term human placentas by Hennen in 1965. L,ike pituítary TSH, hCT stimuLates thyroid hormone secretion when injected into mice and increases uptake of inorganic phosphate ínto phosphorylatêd compounds. hcT has been found irnmunoJ-ogically to be much more closely related Èo bovine and porcine TSH than to human TSH, so that its biological- activity is more effectively neutralized by antibovine TSH than by antihuman TSH (Hennen et al, 1974; Hennen, 1975¡ Hennen and Pierce, 1969). hCT is a glycoprotein and has a moLecular weight of 28,000 as determined by gel filtration (Hershman and Staines, r.97l-) . Hor¡rever, untike TSH, vrhich has an alpha- chaín identical- in peptide sequence to that of hCG, hCT probably does not contaj.n the same alpha-chain, as demonstrated by the failure to cÌoss-react !Íith hcc or hcc-alpha subunits in homologous radioimmunoas say systems (Nistula et aI , 1973t Tojo, 1975). Analysis of purified hCT by Tojo et a1 (1973) indicates an overall amino acid composition quite dist,inct from the hCG, and a much lower sugar content (3.58 versus 30t). The prímary sequence of hCT has not been determined. Un1ike the plasma concentrations of hTSH which remained within non-preçJnant Iimits, hCT became detectable early in pregnancy with an average level of 7 yg/ml plasma
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THE UNIVERSITY OF MANITOBA STUDIES
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To My parents and wife: Mn & Mrs. K
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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 and 24: -4- secretion of a peptide hor:none
Page 25 and 26: NHt-Aì a-Pro-Asp-Val -Gl n-Asp-Cys
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: -35- Although hpI, si¡nuLates the
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
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- 89- luteolysis when supesistpessd
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9t SECTION IV¡ MtTttODs AND IIATER
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- 93- (.TCA) solution were added co
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-95 - (ÞPRL' NIH-P-B3) , and all q
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97 buffer, the eluates were pooled,
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Char4ctêf ization Pfocedures l. Ch
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t0t (c) À.nalvticaL .{rod.iu¡n do
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103 B : MFTÍÍOD OF DETECTION AND
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AFsa)¡- ProceiiureS 105 Specific b
Page 129 and 130:
107 ', C : IIEBI{OÐ OF DEVELOPMENT
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109 D. RTNG PREGNôXCY (l) Maternâ
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l1ì (wtlvo'). After the hornogenat
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(4) Tissúe distributiori df radloi
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l'1 5 (2) Procg.ssjnq, gf. incuÞ¡
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1.l7 for 30 min. Then the precipita
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(2) Assav of ìuteo.trgpic effec.t
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EXPERIMENT # TI (7 days after HCG)
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123 to progesterone (l/300 djlution
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SECTION V : RESULTS (r) LACTOGEN DU
Page 149 and 150:
127 Fractioñâtión of oPL. from c
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129 Figure 6. carborymethyì(cM,)-c
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t 3't óo0 ,oo 800 900 FRACIION r,t
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E o ø ô¡ t- z ao É, o '133 .--.
Page 157 and 158:
TABLE I TABLE OF PURIFICATION OF OV
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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
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o 9 x F- -)- 9 rU I æ -t f () LT'
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¿ o s : t45 Figure 15b. Displaceme
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HoR MoNE coNc€NrRArto ln ß/mt )
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Tissue used for binding ôtudies Li
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Noñ. p..s'ro¡r {N,p.l Pr.eñonr (
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¡( É T 2, \ ¡a.2¡./ir ,..r.¡.
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P 80 \ x.. ¡,azrot¡j .1.¿¡i¡ .
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o c o U o òs -o c2o ao eÀo o 'o .
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F 2 o o o\ ¡¡ig ô 2 Þ 0 (¡ l5e
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! o 't6 l HORMO¡¡ E COr{CENr FAIt
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163 Figure 24 shows the sensltivity
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165 D. STUDIES ON THE SECRETION OF
Page 189 and 190:
E .Dc À o - art & 167 Flgure 26. P
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lJhen blood samples were taken four
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Sheep no. l{6 25í 960 464 5?0 680
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't73 fetal cotylêdons at various d
Page 197 and 198:
TABLE VI OPI CONCENTRATTON IN FETAL
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l. Súrglcat Method 't77 Figure 3l
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= >€ ,3 ; I 179 rwt 76-0450 o----
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dUrlng PrêOnancy in thè Ewes tBr
Page 205 and 206:
! ñ' l83 NoRMON! C oNCENTßArlO þ
Page 207 and 208:
l ol
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I I I "+ _t rl \l :- 'i rl -l l87 a
Page 211 and 212:
l89 Flgure 37. Percentage ot l2sl-o
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19t column is shown in Figure 3BA.
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- 193 3H -couNdro _________SPECtFtC
Page 217 and 218:
TABLE IX /- BIOASSAY OF qROIItIfH p
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z o 197 Figure 40, Radioreceptor as
Page 221 and 222:
5A L lNl Pgtzq cô-tlzt Pgfl6 + cl
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SFCT.IoN II : pl.scussr0N 4d ã 1È
Page 225 and 226:
Ch aracteri zati on 203 Analytical
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205 size, corresponding to the mole
Page 229 and 230:
207 the specific binding of oPL to
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209 than with prolactin-like effect
Page 233 and 234:
2t1 the jncrease in oPRL binding si
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213 The resuìts obtained indicate
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215 on the role af PL in the fetus.
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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
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223 data argue for a close interact
Page 247 and 248:
]. TABLE OF CHEMICAL PROPERTIES GUI
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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
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233 Our studies on the blnding of o
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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
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SECTICN IX PUBLICATIONS A) Manuscr
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SECTION X: BIBLIOGRAPHY Abramovich,
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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
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?57 Knopp,R,H,, H,J. Ruder,E. Herra
Page 281 and 282:
259 Manku,M.S., and D.F. Horrobin,
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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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