THE UNIVERSITY OF MANITOBA STUDIES ON OVINE PLACINTAL ...

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- 72- (1972) reported that when goat plasma samples taken during pregnancy were exa¡rlined for prolactin by radioimmunoassay and for total lactogenic act,ivity by a rabbit mammary gland culture assay, high levels of Lactogenic activity were detected in the second. and 1ast third of pregnancy when the concentration of immunoreactive pituitary prolactin was low or even absent. Co-culture experiments (Forsyt.h, :-g72l demonstrated that this mam¡notropic substance (caprine placental lactogen, cpL) is secret.ed by fetal cotyledons, and could be detected in the maternal circulat,ion from the 9th week (64 days) of gestation until term. Horúever, it is not clear which cells of the placental cotylêdons are secreting placental lactogen and also it is not clear at v¡hat stages of pregnancy this hormone is first secretêd. UsÍng radioreceptorassay for prolactín (RRA_PRL) and RRA-GH, Kelly et at, (19?6) and Currie et. al (l9ZZ) estimated the serum levels of caprine placental lactogen (cPL) throughout pregnancy. In their studies, they found that lactogenic and GH-l_ike act.ivities increased from less than 100 ng,/ml (ovine proLactin and grob¡t.h hormone equivalents) about 60 days after mating t,o reach peak levels (400-1600 nglmt) b et¡.reen days ll0 and 130 of pregnancy. The levels of both activities increased in essentially the s arne fashion but during the last 15 days of pregmancy, lactogenic activity decLined l-ess than Gu-like activity This probably reflected the increased secretion of pituitary prolêctin near parturition.
- 73- When serum from a pregnant goat or a simple alkaline extrAct of placental cotyLedons hras f,ractionated on Sephadex G-100r J.actogenic and GH-like activities eluted lvíth a rBolecuLar weight of 20¡000 to 22:OOO. More recently, crissom et al (I9?Z) reported that they have purified caprine placental 1actogen from goat placentas. They found thatcpt has an isoelectric point of 6.4 and it.s molecular Weight estimatêd by gel-filtration was 22,OOO daltons. The amino acid conposition of cPL resembles that of ovine and bovine placental Lactogens. Furthermore, cpl, is as potent as ovine prolactin when tested in the radioreceptorassay for prolactin.
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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 ¡
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: (Íi) coat: -71 The goat is a spont
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)
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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 .--.
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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
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
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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
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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
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TABLE IX /- BIOASSAY OF qROIItIfH p
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z o 197 Figure 40, Radioreceptor as
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5A L lNl Pgtzq cô-tlzt Pgfl6 + cl
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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 ,
Page 273 and 274:
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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