THE UNIVERSITY OF MANITOBA STUDIES ON OVINE PLACINTAL ...

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220 These studies raise questions as to whether the suppression of prolactin secretion during pregnancy ìs due to the high level of opl fn the circulation, and that the inhibitory effect of opl on pituítary prolactin secretion is mediated by an increase in hypothalamic pIF activity. Whether progesterone also has a role in suppressing prolactin secretion dur.ing pregnancy in the sheep remains unclear. E. BIOSYNTHESIS OF OVINE PLACENTAL LACTOGEN BY OVINI CHORIONIC MEMBRANES in vitro It is apparent that, after gel fíltration of the incubation medium, two radioactive peaks of proteins of widely d.ifferìng molecular weight were precipitated by antÍsera to opl. Since the molecular weight of highly purified opl .is 19,500 to 2l ,500 as shown ìn Flgure .l4, the total counts precipitated by anti-opL in the or{ginal incubation medium as shown in Table vIII do refrect 3H-oPL ulon., The evidence presented suggests strongìy that the molecular weight of the radioactive proteins which emerged ìn the second peak is identical with that of opL. Accordingìy, it is Iíkely that the 3H-opL i, contrìbutíng sìgnificant.ly to the second radioactive peak.
221 The nature of thelarge nolecular weight species precìpiated by antí-oPL is not yet clear. An alternative explanation is that these proteins might be aggregated opl, or prohormone of oPL still attached to ribosomes, or there might be an oPL bindinq substance of ìarge molecular weight present in placental tissue, Our studÌes clearly indicate and support the fact that oPL is synthesized and is secreted by ovine chorionic membranes. F, BIOLOGICAL EFFECTS OF OVINE PLACENTAL LACTOGEN Bjoassay of Growth Promoting Activity of opl Bioassay of growth promoting activity using eplphyseai width increase in hypophysectomí zed rats ís indeed the most specific assay. Although opl promotes growth of hypophy_ sectomized rats, lts role in promoting growth of feta'l lambs remains undefjned, l^le have successfuìly demonstrated that opl binds specificaìly to fetal liver and that hìgh levels of opl are in the fetal circulation. Moreover Handwerger et al (.1976) have demonstrated that oPL increases somatomedin activity in hypophysectomi zed rat
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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
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V. RESULTS E. STUDIES ON THT BIOSYN
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Peptide Hormones: ACTH CG FSH GH LH
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LIST OF FIGURES FIGURE PAGE I Linea
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22 Radioreceptor assay for oPRL us.
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Effect of PGF,. and lactogenic horm
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ABSTRACT STUDIES 0N 0VINE PLACENTAL
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eceptor-b i nd i ng studies show th
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-2- years have seen an increasing i
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-4- secretion of a peptide hor:none
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NHt-Aì a-Pro-Asp-Val -Gl n-Asp-Cys
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6a NH, -Ser-Lys -G'l u.Pro -Leu -Ar
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-8- 197J-; Rayford et al | !9721 .
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-10- sialic acid díd not alter the
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-L2- and Biasucci (1969) denonstrat
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_14_ addition of CAMP (Handwerger e
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-16- conponents I the.faster haying
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_18_ by hcc, as evidenced by increa
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_20- (Mason et a1 , L973i Menon ¡
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21a NH, -Vaì -G1 n-Thr-Val -Pro-Le
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-23- growth-promoting ef,fect, wher
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-25- diffusion teÊt. Josi¡novich
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-27- v¡hereas in the tissue extrac
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-29- found products of molecular we
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-3r- vÍii) S.ecreLion and metabol
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- 33 - metabolic factors may influe
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-35- Although hpI, si¡nuLates the
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-37- C. CIIHER ITUMAN PLACENTAL PEP
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_39_ stimulating activityf is found
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-4t-- (iii) Human Chorionic Cortlco
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- 43 - syncytiotrophoblast (Tatarin
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- 45 - Ib SUBPRTMATE PLACENTAT PR.O
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-47 - samples, signif,icant elevati
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-49- required for the chorignÍc ao
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B. (i) -51 - MONI(ET PLACENTAL I,AC
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- 53 - brith 13 minutes for hpl,. T
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55 Pregnarit Marê Sêrum Gonâdót
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II b. NON - pRl¡late PT,ACENTAT.
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(iv¡ Inmuhologicat Mêthogs: - 59-
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_ 6t_ 1974; Fellows et a1, 1974¡ H
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63 Ordêr Rodêhtia (Tat, nicê, ro
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Guinea Pi g: 65 Hypophysectomy of t
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-67 Orilet Perissodactyla (Horse an
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-69 activity in the N-acetyl-lactos
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(Íi) coat: -71 The goat is a spont
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- 73- When serum from a pregnant go
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-75 Gusdon et al (1920)using . haer
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77 (c) Imnunologicaì Properties of
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-79 - SECTION II : OYINE REpROpucTI
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BI which contain' as lvell as the o
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83 ce11s usually contaln either num
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85 non-pregnant values did not diff
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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
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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
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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 .--.
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TABLE I TABLE OF PURIFICATION OF OV
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137 Figure lì. A, Poìyacrylamide
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Leaf blank to correct nr.rmbering
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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
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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: 219 5.Relationships amonq opRL. oGH
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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