2011 (SBTE) 25th Annual Meeting Proceedings - International ...

More documents

Recommendations

Info

Acta Scientiae Veterinariae, 2011. 39(Suppl 1): Abstracts - 25th Annual Meeting SBTE-Brazil. August 2011. A179 EMBRYOLOGY, BIOLOGY OF DEVELOPMENT AND PHYSIOLOGY OF REPRODUCTION DIMENSIONS, STEROIDOGENESIS AND VASCUL ASCULARIZA ARIZATION OF THE BOVINE CORPUS LUTEUM IN RESPONSE TO MANIPULATION OF THE PRE-OVULATOR ORY FOLLICLE GROWTH Fernando Silveira Mesquita 1 , Moana Rodrigues França 2 , Saara Carolina Scolari 3 , Valdir Pavanello Jr. 4 , Fabiana Fernandes Bressan 5 , Flávio Vieira Meirelles 6 , Paula de Carvalho Papa 7 , Guilherme de Paula Nogueira 8 , Luciano Andrade Silva 9 , Claudia Maria Bertan Membrive 10 & Mário Binelli 11 1,2,3,11 DEPARTAMENTO DE REPRODUÇÃO ANIMAL-FMVZ-USP, PIRASSUNUNGA, SP, BRAZIL. 4,7 DEPARTAMENTO DE CIRURGIA-FMVZ-USP, SÃO PAULO, SP, BRAZIL. 5,6,9 DEPARTAMENTO DE CIÊNCIAS BÁSICAS-FZEA-USP, PIRASSUNUNGA, SP, BRAZIL. 8 DEPARTAMENTO DE APOIO DE PRODUÇÃO E SAÚDE ANIMAL-UNESP, ARAÇATUBA, SP, BRAZIL. 10 FACULDADE DE ZOOTECNIA - UNESP, DRACENA, SP, BRAZIL. In cattle, fluctuations in plasma concentrations of estradiol and progesterone (P4), which characterize the peri-ovulatory phase of the estrous cycle, influence oocyte maturation, transport of gametes, fertilization and early embryonic development. This work aimed to study the effects of manipulations of the pre-ovulatory follicle (POF) growth on growth, P4 production and expression of steroidogenesis- and angiogenesisrelated genes of the subsequent corpus luteum (CL). Cyclic, non-pregnant Nelore cows received two injections of PGF2α (PGF; 0.5 mg; i.m.) 14 days apart. Ten days later (day -10; D-10), cows received a P4-releasing device along with estradiol benzoate (2 mg; i.m.). In order to modulate the growth of the POF and alter post-ovulatory P4 production, on D-10 animals received PGF (high post-ovulatory P4 group; HP; n = 10) or not (low post-ovulatory P4 group; LP; n = 10). Progesterone-releasing devices were removed and PGF injected on D-2.5 for the HP group and on D-1.5 for the LP group. Ovulation was induced with GnRH (buserelin; 10 µg; i.m.) on D0. Growth and ovulation of the POF and CL formation and vascularization were assessed by Doppler ultrasonography from D-2 to D7. On D7, plasma was obtained for measurement of P4 concentration, cows that ovulated were slaughtered (HP, n = 7 and LP, n = 6) and CLs were dissected and stored. Relative concentrations of transcripts related to steroidogenesis (STAR, CYP11A1, HSD3B) and angiogenesis (VEGF, FLT1, KDR) were determined by TaqMan qPCR, using tubulin as the endogenous control gene. Differences between group means were determined by student’s t test. Maximum diameter of the POF (mean ± standard error of the mean; 13.01±0.35 vs. 10.98±1.07 mm; P = 0.06), weight (3.10±0.32 vs. 1.91±0.33g), diameter (17.75±0.76 vs. 13.75±1.14 mm), volume (1659.94±220.53 vs. 801.99±191.11 mm 3 ) and vascularization of the CL on D7 (80.00±3.54 vs. 56.25±8.00%) and P4 concentration (4.34±0.13 vs. 2.74±0.35 ng/mL) on D7 were greater in HP vs. LP (P < 0.05). Progesterone concentrations were positively correlated with diameter, volume and weight of the CL (r = 0.61, 0.56 e 0.5, respectively). Weight of the CL was positively correlated with its diameter (r = 0.84) and with POF diameter (r = 0.73). Luteal gene expression of CYP11A1, STAR (P < 0.05) and VEGF (P = 0.07) on the HP group was 3.2, 2.1 and 2.5 times greater than that of the LP group. In conclusion, stimulation of POF growth resulted in a larger CL that produced more P4. It is suggested that an increase in the differential gene expression of steroidogenic enzymes and angiogenesis-related molecules by the CL is associated with a higher functional capacity of this organ. [Supported by CNPq (481087/2010-9) and FAPESP (2010/ 01284-4)]. Keywords: bovine, corpus luteum, progesterone. A180 EMBRYOLOGY, BIOLOGY OF DEVELOPMENT AND PHYSIOLOGY OF REPRODUCTION EFFECT OF DEXAMETHASONE AND TRIIODOTHYRONINE ON IN VITRO BOVINE EMBRYO Nathália Nogueira da Costa 1 , Marcela da Silva Cordeiro 2 , Thiago Velasco Guimarães Silva 2 , Priscila Di Paula Bessa Santana 1 , Rafael Vilar Sampaio 1 , Danuta Carolina Sastre 1 , Bruno Baraúna da Silva 1 , Andre Luiz Alves de Sá 1 , Carlos Leonardo de Aragão Araújo 1 , Simone do Socorro Damasceno Santos 1 , Moysés dos Santos Miranda 1 & Otávio Mitio Ohashi 1 1 UNIVERSIDADE FEDERAL DO PARÁ, BELEM, PA, BRAZIL. 2 INSTITUTO FEDERAL DE EDUCAÇÃO, CIÊNCIA E TECNOLOGIA DO PARÁ, ABAETETUBA, PA, BRAZIL. The embryos produced in vitro differ in many aspects from those in vivo produced and in most cases this difference is related to the conditions and composition of the culture media (Khurana & Niemann, 2000, Biology of Reproduction, 62: 847-856). Therefore, the aim of this study was to evaluate the effect of the hormone triiodothyronine (T3) and the synthetic glucocorticoid, dexamethasone (Dexa) on IVC, since they are important regulators of metabolism, cell growth and differentiation (Aranda & Pascual, 2001, Physiological Reviews, 81:1269- 1304). For that COCs were selected and incubated for IVM at 38.5°C and high humidity for 18 h. For IVF, the sperm of a single bull was thawed and co-incubated with the COCs under the same conditions described for IVM. After approximately 30 h of incubation, presumptive zygotes were subjected to repeated pipetting to remove remaining cumulus cells. Then, they were randomly distributed among the following groups: Control, without hormones; Dexa (0.1 ug / mL), T3 (50 nM T3), T3 + Dexa (0.1 ug / mL Dexa and 50 nM T3). All groups were cultured in SOF medium supplemented with 6 mg / mL BSA and 10% FCS. Cleavage rate and the rate of blastocyst and kinetics of development were analyzed on the Day 2 and Day 8 of culture. The results were analyzed using the ANOVA test, adopting P < 0.05. The rate of cleavage and blastocyst (P > 0.05) were similar between groups, control (79% and 52% respectively), Dexa (75% and 49% respectively), T3 (80% and 46% respectively) and Dexa + T3 (81% and 53% respectively). There was no difference in the kinetics of development between the embryos cultured with or without hormones, and observed a predominance of hatched embryos in all groups (control group, 69.2%; Dexa, 70%, T3 71% and T3 + Dexa 73%, P > 0.05). Thus, these results suggest that T3 and Dexa used alone and / or combined during IVC, showed no effect on in vitro development of bovine embryos. Keywords: in vitro production embryo, dexamethasone, triiodothyronine. s426
Acta Scientiae Veterinariae, 2011. 39(Suppl 1): Abstracts - 25th Annual Meeting SBTE-Brazil. August 2011. A181 EMBRYOLOGY, BIOLOGY OF DEVELOPMENT AND PHYSIOLOGY OF REPRODUCTION EFFECT OF CELL QUANTIT ANTITY IN OBTAINING MICE CHIMERAS BY DEMI-BLAST ASTOCY OCYST STS S AGGREGA GGREGATION Caio Pontes Godói, Pablo Diego Moço, Bruno Cazari, Patrícia Tiemi Mihara, Patrícia Villela Silva, Isabele Picada Emanuelli & Marcelo Fábio Gouveia Nogueira UNESP, ASSIS, SP, BRAZIL. Eight-cell-stage to pre-compaction morula are the most used embryonic stages to aggregation, because the embryos, in these early stages, synthesize cell adhesion molecules that increase the aggregation chances among them (Vestweber et al., 1987; Developmental Biology 124:451-6). Although post-compaction embryos produce reduced aggregation rates, they are not refractory to this process (Nogueira et al., 2010; Transgenic Research 19:344-5). Based on this evidences, the aim of this study was to evaluate, in mice, the influence of cell quantity in the chimerism rate of split blastocysts. Embryos, with preferentially different phenotypes, were obtained from C57BL/6/EGFP and/or Swiss Webster strains. Females ranging from 21 to 45 days old were superstimulated and mated according to Mancini et al. (2008; Transgenic Research 17:1015). Eight-cell-stage embryos (8C) and pre-compaction morula (PCM) were recovered (2 to 2.5 days post coitum) and had their zona pellucida removed using pronase treatment (2 mg/mL for 15 min), while blastocysts (recovered 3.5 dpc) were split with microblade controlled by micromanipulator in an inverted microscope (NK2, Eppendorf, Germany and Eclipse Ti, Nikon, Japan, respectively). The aggregation groups were: a control (C) with two pre-compaction whole embryos 8C and/or PCM and two experimental with post-compaction embryos, i.e., two (2DB) or four (4DB) demi-blastocysts. The structures (2 or 4) of the groups were sticked each other with the use of phytohemagglutinin (1 mg/mL) and cultured in vitro by 24 h (37ºC, 5% CO2 and saturated humidity). After culture, the presence of chimeric embryos was verified by detection of a single, cohesive cell mass or a structure in “8 shape” with more than half of its total diameter aggregated. For the 4DB group, a successful aggregation was considered when, at least, two of four demi-blastocysts have aggregated. The results were analyzed using Chisquare test, Fisher’s exact test and Kruskal-Wallis (to comparison among groups, between groups and among median of group replicates, respectively) and significance was considered when P < 0.05. The aggregation rates for the groups C, 2DB and 4DB were, respectively, 77.3a; 8.3b e 36.4%c (P < 0.001). The increasing of the aggregation technique efficacy, in post-compaction stages, would be particularly interesting in farm animals (e.g., bovine species), where it is not feasible to obtain, in vivo, pre-compaction stages embryos (as eight cells) and when only trophectoderm aggregation is desired. It was concluded that cell increasing (from 2 to 4 demi-blastocysts) improved the chimerism rate, but not enough to be similar to control group. Keywords: aggregation chimerism, demi-blastocyst, mice. A182 EMBRYOLOGY, BIOLOGY OF DEVELOPMENT AND PHYSIOLOGY OF REPRODUCTION EFFECT OF DIFFERENT MEDIA ON BONE MORPHOGENETIC PROTEIN 15 (BMP-15) AND GROWTH DIFFERENTIATION TION FACT CTOR 9 (GDF-9) MRNA EXPRESSION DURING OOCYTE IN VITRO MATUR TURATION TION (IVM) Paula Fernanda de Lima 1 , Mariana Fernandes Machado 1 , Ester Siqueira Caixeta 1 , Felipe Morales Dalanezi 1 , Christopher Price 2 & José Buratini Junior 1 1 UNESP, BOTUCATU, SP, BRAZIL. 2 UNIVERSITY OF MONTREAL, MONTREAL, CANADA. N BMP-15 and GDF-9 are oocyte secreted factors (OSF) that control cumulus-oocyte complex (COC) maturation. They bind to receptors in cumulus cells and regulate indirectly oocyte quality. Considering that the oocyte retains transcriptional activity during IVM (Mano et al. BMC Genomics 2011, 12:151), we tested the hypothesis that different media currently used in IVM would determine different levels of BMP-15 and GDF-9 mRNA expression in the oocyte. More specifically, we tested the effects of the addition of fetal calf serum (FCS), bovine serum albumin (BSA) and BSA plus estradiol to the base medium on BMP-15 and GDF-9 mRNA expression during IVM. Immature COCs (grades 1 and 2) were obtained from 3-8 mm follicles from abattoir ovaries (predominantly Bos indicus). Four groups of 20 COCs per treatment were cultured for 22 h in TCM 199 bicarbonate plus BSA (0.4%), or BSA (0.4%) plus estradiol (1µg/mL), or FCS (10%). After culture, oocytes were denuded and stored at -80°C. Four groups of 20 immature oocytes were also included in the analysis to verify the effect of maturation on gene expression. Total RNA was extracted from pools containing 20 oocytes with RNeasy ® (Qiagen). Reverse transcription was performed with Sensiscript ® (Qiagen). mRNA expression of both target genes was investigated by real time RT-PCR and normalized by the expression of cyclophilin A (CYC-A). Expression values were determined by the Pfaffl equation. Effects of treatments were tested by ANOVA and groups were compared by Tukey-Kramer HSD. Differences were considered significant when P < 0.05. mRNA abundance for both BM-15 and GDF-9 did not vary with different maturation media. Interestingly, GDF-9 mRNA expression was higher in the immature group compared with all others. In conclusion, addition of BSA or FCS determined the same levels of mRNA expression for BMP-15 and GDF-9 and the addition of estradiol did not alter the expression pattern. The suppressing effect of maturation on GDF-9 mRNA expression is interesting and deserves further investigation. [Financial support FAPESP]. Keywords: bmp-15, gdf-9, bovine. s427
Page 1 and 2:
Proceedings of the 25 th Annual Mee
Page 3 and 4:
Acta Scientiae Veterinariae. 39 (Su
Page 5:
Page 8 and 9:
Page 10 and 11:
Page 12 and 13:
Page 14 and 15:
Page 16 and 17:
Page 18 and 19:
Page 20 and 21:
Page 22 and 23:
A262 Bovine Oocyte Vitrification: E
Page 25 and 26:
C.A. Rodr drigues igues, R.M. Fer e
Page 27 and 28:
Page 29 and 30:
Page 31 and 32:
Page 33 and 34:
Page 35:
Page 38 and 39:
L.F. Nasser asser, L. Pen enteado e
Page 40 and 41:
Page 42 and 43:
Page 44 and 45:
Page 46 and 47:
R.B. Lôbo, D. Nkr uman, D.A. Gross
Page 48 and 49:
R.B. Lôbo, D. Nkr uman, D.A. Gross
Page 51 and 52:
M.E.F .F. Oliv liveir eira. 2011. E
Page 53 and 54:
Page 55 and 56:
Page 57:
Page 60 and 61:
A.L. Gusmão usmão. 2011. Estado d
Page 62 and 63:
Page 64 and 65:
Page 66 and 67:
J. R. Figueir igueiredo edo, A.P.R.
Page 69 and 70:
E.L.A. Motta, M. Nichi & P.C. Ser e
Page 71 and 72:
Page 73 and 74:
Page 75 and 76:
Page 77:
Page 80 and 81:
E.L.Gastal, M.O. Gastal, A. Wischra
Page 82 and 83:
Page 84 and 85:
Page 86 and 87:
Page 88 and 89:
Page 90 and 91:
Page 92 and 93:
Page 95 and 96:
D.P .P.A.F .A.F. Braga & E. Bor org
Page 97 and 98:
Page 99 and 100:
Page 101 and 102:
Page 103:
Page 106 and 107:
F.F .F. Bressan, F. Per erecin, eci
Page 108 and 109:
Page 110 and 111:
Page 112 and 113:
Page 114 and 115:
Page 116 and 117:
Page 119 and 120:
C.E. Ambrósio mbrósio, C.V. Wenc
Page 121 and 122:
Page 123:
Page 127 and 128:
J.C. Fer erreir eira, F.S. Ignácio
Page 129 and 130:
Page 131 and 132:
Page 133:
Page 136 and 137:
R.C. Uliani, L.A. Silv ilva, M.A. A
Page 138 and 139:
Page 140 and 141:
F.S. Ignácio, J.C. Fer erreir eira
Page 142 and 143:
F.S. Ignácio, J.C. Fer erreir eira
Page 145 and 146:
L.A. Silva. 2011. Local Effect of t
Page 147 and 148:
Page 149 and 150:
Page 151 and 152:
Page 153 and 154:
Page 155 and 156:
Page 157 and 158:
M.M. Franc anco, A. Pellegr ellegri
Page 159 and 160:
M.M. Franc anco, A. Pellegr ellegri
Page 161 and 162:
B. Str troud & G.A. Bó. 2011. The
Page 163 and 164:
Page 165 and 166:
Page 167 and 168:
Page 169 and 170:
W.W .W. Tha hatcher cher. 2011. Tem
Page 171 and 172:
Page 173 and 174:
Page 175 and 176:
Page 177 and 178:
Page 179 and 180:
Page 181 and 182:
Page 183 and 184:
Page 185 and 186:
Page 187 and 188:
Page 189 and 190:
Page 191:
Page 195 and 196:
O. Sandra. 2011. Deciphering early
Page 197 and 198:
Page 199 and 200:
Page 201 and 202:
Page 203 and 204:
Page 205 and 206:
R.C. Cheb hebel. el. 2011. Use of A
Page 207 and 208:
Page 209 and 210:
Page 211 and 212:
Page 213 and 214:
Page 215 and 216:
Page 217 and 218:
Page 219 and 220:
Page 221 and 222:
Page 223 and 224:
Page 225 and 226:
Page 227 and 228:
Page 229 and 230:
Page 231 and 232:
Page 233 and 234:
Page 235 and 236:
Page 237 and 238:
Page 239 and 240:
Page 241 and 242:
Page 243:
Page 246 and 247:
A.C.A. Net eto, A.R. Galdos aldos,
Page 248 and 249:
A.C.A. Net eto, A.R. Galdos aldos,
Page 250 and 251:
P.C .Cha havett ette-P e-Palmer alm
Page 252 and 253:
Page 254 and 255:
Page 256 and 257:
Page 258 and 259:
Page 260 and 261:
Page 262 and 263:
Page 264 and 265:
Page 266 and 267:
E.H. Bir irgel Junior unior, F.V .V
Page 268 and 269:
Page 270 and 271:
Page 272 and 273:
Page 274 and 275:
Page 276 and 277:
P. Humblot. 2011. Reproductive Tech
Page 278 and 279:
Page 280 and 281:
Page 282 and 283:
Page 284 and 285:
Page 286 and 287:
J.A. Piedrahita. 2011. Application
Page 288 and 289:
Page 290 and 291:
Page 292 and 293:
Page 294 and 295:
Page 296 and 297:
O.E. Smith, B.D. Murphy & L.C. Smit
Page 298 and 299:
Page 300 and 301:
Page 302 and 303:
Page 304 and 305:
Page 307 and 308:
D. Salamone alamone, R. Bevacqua, F
Page 309 and 310:
Page 311 and 312:
Page 313 and 314:
Page 315:
Page 318 and 319:
E.A. Maga & J.D. Mur urray. 2011. G
Page 320 and 321:
Page 322 and 323:
Page 325:
Page 328 and 329:
C.G. Gutier utierrez, S. Fer errar
Page 330 and 331:
Page 332 and 333:
Page 334 and 335:
Page 336 and 337:
Page 338 and 339:
Page 340 and 341:
B. Gasparrini. 2011. Ovum pick-up a
Page 342 and 343:
Page 344 and 345:
Page 346 and 347:
Page 348 and 349:
Page 350 and 351:
Page 352 and 353:
Page 354 and 355:
Page 356 and 357:
Page 359 and 360:
Acta Scientiae Veterinariae, 2011.
Page 361 and 362:
Page 363 and 364:
Page 365 and 366:
Page 367 and 368:
Page 369 and 370:
Page 371 and 372:
Page 373 and 374:
Page 375 and 376:
Page 377 and 378:
Page 379 and 380:
Page 381 and 382:
Page 383 and 384:
Page 385 and 386:
Page 387 and 388:
Page 389 and 390:
Page 391 and 392:
Page 393 and 394:
Page 395 and 396:
Page 397 and 398: Acta Scientiae Veterinariae, 2011.
Page 447: Acta Scientiae Veterinariae, 2011.
Page 489: Acta Scientiae Veterinariae, 2011.
show all

2011 (SBTE) 25th Annual Meeting Proceedings - International ...

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

Delete template?

Save as template?