THE UNIVERSITY OF MANITOBA STUDIES ON OVINE PLACINTAL ...

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152 'late in pregnancy were used. Again maximar Þinding of opl was obtarned after incubation at 4 C for 24 h, with the exception of,the ovary, where blnding of opl at ZZ C oî 37 C was slighily greater. Dur.ing routine assays, 24 h incubation periods at 4 C were used for all tissue receptors because this gave the highest level of blnding wlthin a convenient tirne period. oPL usl Adipose Ti ssues Figure 19 shows the competition by pitultary hormones for oPL binding sites in liver and adìpose tissue obtarned from the non- pregnant ewe. PituÍtary growth hormones from various specres competed with oPL for binding sites in liver to varying degrees(fig. tSA). Human placental lactogen competed only minlmally.and other pituitary hormones not at all. Similarìy, the results ln Fig. .l9B for adipose tissue(ì00,000 x g fractions) show that growth honnones inhiblted the binding of the tracer ìn a paraller manner to that of opl except that 2 to 5 fo]d lncreases in concentration were required. scatchard ana.lysis of the data obtained with opl are dispìayed in the figure and the nunber of binding sites (n) and the affinity constant (fa) ot opL for each ti ssue is indicated_ for oPL usi Figure 20 shows the inhiblilon of binding of 125I-opL to the corpus luteum and to the ovary (.|00,000 x g fractions) by opL and three ovine pituitary hormones (pRL, GH, and LH). Corpora .lutea
¡( É T 2, \ ¡a.2¡./ir ,..r.¡. \. .l53 htH hFSH oLH llon".nn*on ll l'o0o lo'ooo xo,.,,on. n.,r,nr bTSH Icc pACTH p lñ¡ùlin Figure '19. Radioreceptor assay for opl using pellets (IOO,OOO x g fractions) from the liver(A) and adipose tissue(B) of non_pregnant ewes' using 500 ug protein per assay tube. The anount of radioactivity bound in the absence of oPL was taken to be equal to 1001. The specific blndlng of l2SI-opL to ilver and adlpose ilssue was zzl. and '16%, respectlve'ly. The lnset depicts a Scatchard analysis of the data. hPRI bPRT o?Rt pPRl 'PRI
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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
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: Noñ. p..s'ro¡r {N,p.l Pr.eñonr (
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
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Ch aracteri zati on 203 Analytical
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205 size, corresponding to the mole
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207 the specific binding of oPL to
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209 than with prolactin-like effect
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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
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219 5.Relationships amonq opRL. oGH
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221 The nature of thelarge nolecula
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223 data argue for a close interact
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]. TABLE OF CHEMICAL PROPERTIES GUI
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cor,t coAT (bPL) (cpl) 3. TABLE OF
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À) E 229 Figure 43. The secretion
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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
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237 Furthermore, oPL neasurement in
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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.,
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247 Brody,S,, and Carlstrom,G.,J.Cl
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249 Crosignanì ,P.G. , Nencioni ,
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25t Friesen,H.G., Endocrinoìogy 76
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253 Grinwich,D.L., M. Hichens, and
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25s Hum, V.G., Knipfel , J.G., and
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?57 Knopp,R,H,, H,J. Ruder,E. Herra
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259 Manku,M.S., and D.F. Horrobin,
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261 , :iir .,.-;;r-,:, Morgan, F.J.
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263 Pierce,G.B., Midþley,A.R. and
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?65 Shepard, T.H.., Gen. Comp. Endo
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267 Tsuruhara,T., Dufau,M.L., Hickm
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