title: Preliminary study in immature canine oocytes stained with brilliant cresyl blue and obtained from bitches with low and high progesterone serum profiles.
abstract: This study was conducted: (i) to observe the features and levels of blue colour impregnation in morphologically selected immature canine cumulus oocyte complexes (COCs) stained with the brilliant cresyl blue (BCB) dye, as indicators of quality, and integrity of nuclear oocyte chromatin configuration before in vitro maturation (IVM); (ii) to observe the relationship between the influence of serum progesterone (SP) concentrations from ovary donors and BCB staining of immature dog oocytes. The results showed that out of 138 canine COCs, germinal vesicle (GV) stage prevailed in BCB+ oocytes at percentages of 67.4% (60/89), which were statistically higher than those observed in BCB+/- (52.2%; 23/44) and BCB- (20%; 1/5) oocytes (p = 0.023). Oocytes BCB+ were interpreted as those having completed their growth and therefore possessing the capacity to mature and develop in vitro. Ooplasm and cumulus cells (CCs) of canine oocytes were BCB staining independent. Ooplasm blue colour staining reaction varied between grown oocytes, revealing different levels of glucose-6-phosphate dehydrogenase activity among and within oocytes. Additionally, SP profile of ovary donors was not a relevant indicator for selection of oocytes screened with the BCB stain. Similar numbers of high quality oocytes were observed to be BCB+, BCB+/- and BCB- between groups of females with SP varying from 0 to 2.5 ng/ml (n = 5), and those with SP varying from 2.6 to 16.7 ng/ml (n = 4) (p = 0.680). It may be inferred that bitches with low and high SP profiles have grown oocytes in their ovaries, as determined by the BCB absorbance in their ooplasms.
authors.affiliation: Laboratory of Embryology and Biotechnics of Reproduction, Faculty of Veterinary Medicine, UFRGS, Porto Alegre, RS, Brazil. berenice@portoweb.com.br
authors.forename: B A
authors.initials: BA
authors.lastname: Rodrigues
authors.forename: P
authors.initials: P
authors.lastname: Rodriguez
authors.forename: A E F
authors.initials: AE
authors.lastname: Silva
authors.forename: L F
authors.initials: LF
authors.lastname: Cavalcante
authors.forename: C
authors.initials: C
authors.lastname: Feltrin
authors.forename: J L
authors.initials: JL
authors.lastname: Rodrigues
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D004396:Coloring Agents / Q000737:chemistry; D004285:Dogs; D005260:Female; D009865:Oocytes / Q000166:cytology* / Q000502:physiology; D010078:Oxazines / Q000737:chemistry*; D011374:Progesterone / Q000097:blood*; D013194:Staining and Labeling
chemical_list: D004396:Coloring Agents; D010078:Oxazines; C006797:Brilliant Cresyl Blue; D011374:Progesterone