In vitro production and cryotolerance of prepubertal and adult goat blastocysts obtained from oocytes collected by laparoscopic oocyte-pick-up (LOPU) after FSH treatment.
This study compares the developmental capacity and cryotolerance of embryos produced from oocytes of stimulated prepubertal and adult Sarda goats. Twelve prepubertal and 13 adult goats were each given 110 and 175 IU FSH, respectively, and cumulus-oocyte complexes (COCs) were collected by laparoscopic oocyte-pick-up (LOPU). After in vitro maturation, fertilisation and culture (IVMFC), blastocysts were vitrified, warmed and blastocoel re-expansion and gene expression were evaluated. Prepubertal goats produced a higher COCs number than adults (mean +/- s.e.m., 89.67 +/- 5.74 and 26.69 +/- 3.66, respectively; P < 0.01). Lower developmental competence was demonstrated in the prepubertal oocytes as shown by a higher number of COCs discarded before IVM (21.1% and 14.7% for prepubertals and adults, respectively; P < 0.01) and IVF (23.4% v. 9.1%; P < 0.01) and by the lower cleavage (55.6% and 70.3%, respectively; P < 0.01) and blastocyst rates (24.2% and 33.9%, respectively; P < 0.05). Compared with the adult, prepubertal vitrified/warmed blastocysts showed significantly (P < 0.05) lower in vitro viability, as determined by the re-expansion rate (62.5% and 40.3%). No differences were observed in the time required for blastocoel re-expansion or in cyclin B1, E-cadherin, Na/K ATPase, HSP90beta and aquaporin 3 messenger RNA quantity. These results show that in vitro-produced embryos produced from prepubertal goat oocytes have a lower developmental rate and cryotolerance compared with their adult counterparts. However, we can assume that the quality of re-expanded embryos does not differ between the two groups.
Leoni
Giovanni Giuseppe
GG
Department of Physiological, Biochemical and Cellular Sciences, Sassari, Italy. gioleoni@uniss.it
Succu
Sara
S
Satta
Valentina
V
Paolo
Mereu
M
Bogliolo
Luisa
L
Bebbere
Daniela
D
Spezzigu
Antonio
A
Madeddu
Manuela
M
Berlinguer
Fiammetta
F
Ledda
Sergio
S
Naitana
Salvatore
S
D000367:Age Factors; D000818:Animals; D001755:Blastocyst / Q000502:physiology*; D049109:Cell Proliferation; D002470:Cell Survival; D002970:Cleavage Stage, Ovum; D015925:Cryopreservation / Q000662:veterinary*; D046149:Embryo Culture Techniques / Q000662:veterinary; D005300:Fertility Agents, Female / Q000008:administration & dosage*; D005307:Fertilization in Vitro / Q000662:veterinary*; D005640:Follicle Stimulating Hormone / Q000008:administration & dosage*; D018507:Gene Expression Regulation, Developmental; D006041:Goats / Q000196:embryology*; D010535:Laparoscopy / Q000662:veterinary*; D054315:Oocyte Retrieval / Q000662:veterinary*; D010060:Ovulation / Q000187:drug effects*; D013997:Time Factors
D005300:Fertility Agents, Female; D005640:Follicle Stimulating Hormone