THE UNIVERSITY OF MANITOBA STUDIES ON OVINE PLACINTAL ...

More documents

Recommendations

Info

$il\VOLVEMENT OF RETII\OIC ACID II{ - MSpace at the University of ...$

(l ) Cutture method lt4 CHoRIoNIC MEMBRANES, in Vltro Ovjne chorlonic membranes were obtained lmmediateìy after surgery of pregnant ewes (26,35,55 days of gestation). Chorionic membranes from several ewes of same gestationaì perjod were pooled and cut into fragments, rinsed with ice-cold.Hank,s balanced salt solution' Approximately r00 mg fragnrents were pìaced into a 20 rnr grass flask and incubated for 30 min in l0 ml of Du.lbecco,s Modified Eagìe Medium(DMEM) obtaíned from Gibco, New york. Each flask ç6¡_ tained 100 U of PenicilIin-Streptomycin so.lution. After 30 nin prei ncubat I on"the medium was repìaced with 10 mr of Durbeccors Modified Eagle Medium (without leucîne) containing 50 uCl,¿ml of 3H_leucine (specific activity, 70-100 Cilmmol , New Englancl Nuclear Corporation). The incubation was carried out at 37 C in an atmosphere of gsi¿ O? and 5% C1r. After 24 hours of incubation, the medium and the tissues were frozen at -20 C immediateìy unti.l use. Flasks containïng chorionic membranes were set up at least in duplicate and analyzed separateìy.
l'1 5 (2) Procg.ssjnq, gf. incuÞ¡rjgI, fp.djg The frozen 24 h incubation medium and tÍssue fragments were thawed and centrlfuged at 3,000 x g. The supernatant was further centrif,uged at 50,000 x g for 30 min and the c.lear supernatant was frozen at -20 C until use. (3) Radioimmunoassa.y for opl content The oPL content in the media was determined by the doub.le antibody radÌoimmunoassay as described previously. (4) Determination of the s.yntheSis and Secretion of tipl The newly synthesized opl was determined by measuring the Íncorporation of 3H-leucine into opl molecule. The quantities of 3H-oPL in the medium was determined by inrmunoprec i pi tati on using specific antiserum to opl prepared in rabbits as described previous'ry. (i) Method of determination of the equivalence zone for opl antiserum. The equivalence zone for maximal precipitatíon of radio_ active oPL was determined as fo ows. 0.r mr of cord opl in various concentration ( 10 to l0O,000ng/ml) was added to test tubes containing 300 ul of phosphate buffered saline(pgS) , pH 7.6,25 ul opL antiserum, and 20,000 cpm of l2SI-opL. The finaì volume was approximately 525 ul . The mixture was then ìncubated for I h at 37 C, and then for an. additional 16 h at 4 C. After overnight incubation (.16 h),2 ml of PBS were added into the reaction nixture, and then centrifuged at 3,000 x g for 30 min. The pellets were washed and re-ientrifuged twice
Page 1 and 2:
THE UNIVERSITY OF MANITOBA STUDIES
Page 3 and 4:
To My parents and wife: Mn & Mrs. K
Page 5 and 6:
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 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 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
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: (4) Tissúe distributiori df radloi
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 and 174: Noñ. p..s'ro¡r {N,p.l Pr.eñonr (
Page 175 and 176: ¡( É T 2, \ ¡a.2¡./ir ,..r.¡.
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
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 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
show all

THE UNIVERSITY OF MANITOBA STUDIES ON OVINE PLACINTAL ...

You also want an ePaper? Increase the reach of your titles

Delete template?

Save as template?