THE UNIVERSITY OF MANITOBA STUDIES ON OVINE PLACINTAL ...

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SECTIr)N VIII 240 SIGNIFICANCE OF PRESENT STUDIES l. l.lith the identification and Jsoration of opl from sheep placentas we have demonitrated unequlvocally that pìacental lactogen is present in non-primates. 2. l^Je have established a method for the pur.ification of opl, By empìoying this method, one can purlfy opL ín .large quantities and in high purity for chemlcal and biologìcal studies. 3. l.le have developed a specifìc rad i oirmu noa ssay for opL. By emptoylng this assay, one can study the physloìogy of opl in the sheep. 4. }le have demonstrated for the first time that opl can maintain the lntegr'lty of LH-receptors in the corpora lutea. 5. Binding studles of opL using anima'l and human tissues show that ' oPL and hGH bind to the same receptor sites in both animal and human tissues, suggesting that the blnding sites in opl and hGH molecule are very simi'lar. Thus, further structurai anaìysis on the active sites of opl and hGH will elucidate the peptide(s) which may be responsible for the bindlng for growth. promoting activity. Hence, structural analysls may have potentìaì impl icatìons for clinical use.
SECTICN IX PUBLICATIONS A) Manuscrìpts: 241 'I . Friesen, H.G. , Shiu, R.p.C., Tsushima, T,, Robertson,M,C., Keìly,p., Chan, J., peeters,S., and Carr,D., in III lnternational Syrnpos i um on Ear.ly Diabetes, Canary Island, December 1974,pubìished by Academic press lnc. ,p279,1975 2. Chan, J,S.D., Robertson,H.A., and H.G. FrÍesen, Endocrinoì ogy 9B :65,ì 976 . 3. 81ank,M.S., Chan,J.S.D., and H.G. Friesen, J. Steroid Biochem B:403,.l977 4. Chan, J.S.D., Robertson, H.A., and H.G. Friesen, tndocr.inology .l02:632,1979 5, Chan,J.S.[J., Robertson,H.A., and H.G. Friesen, Endocrinology 102:l 606,'l 978 ' 6. Carnegie, J.A., Chan,J.S,D., Robertson,H.A.,Friesen,H,G., and M.E. McCully, J.Reprod Fert (pub.l ication pending) 7. Robertson,H.A., Chan, J.S.D., and H.G. Frlesen, J Reprod Fert (publication pending) B. Robertson,H.A., Chan,J.S,D,, Hackett,A.J., Marcus,G.R., and H.G. Friesen, Animal Reprod Sclence (pubìication pendi ng )
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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
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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
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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
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! ñ' l83 NoRMON! C oNCENTßArlO þ
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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
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: 239 circulationand alìantoic f'lul
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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