Differential Expression of IGF Family Members in Heat-Stressed Embryos Produced In Vitro from OPU-Derived Oocytes of Nelore (Bos indicus) and Holstein (Bos taurus) Cows

Contents The IGF system is related to embryo quality. We aim to determine the effect of the heat stress on the mRNA expression of IGF1 and IGF2, IGFR1 and IGFR2, IGFBP2 and IGFBP4, and PAPPA in in vitro production (IVP) blastocysts from Nelore and Holstein after ovum pick up (OPU) to better understand the differences between these breeds.Oocytes from four Nelore and seven Holstein were collected in six OPU sessions. Following in vitro maturation and fertilization using six Nelore or Holstein sires, embryos were divided into control (cultured at 39 degrees C) and heat stress (HS; exposed to 41 degrees C for 9h). Blastocysts were submitted to RNA extraction. The IGF1 expression was higher in blastocysts under HS in both breeds, and the expression of IGFBP2 and IGFBP4 was higher in Holstein blastocysts under HS. The high PAPPA expression and the low expression of IGFBP2 and IGFBP4 are associated with a more efficient degradation of IGFBPs, which results in greater IGF bioavailability in Nelore blastocysts and may contribute to the superior HS tolerance in Nelore, when compared to Holstein.

Reference Gene Selection for Gene Expression Analysis of Oocytes Collected from Dairy Cattle and Buffaloes during Winter and Summer

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

In vitro Maturation of Oocytes from Santa Ines Ewes Subjected to Consecutive Sessions of Follicular Aspiration by Laparoscopy

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Meiotic arrest of sheep oocytes using roscovitine under different medium compositions

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Morphological effects of neem (Azadirachta indica A. Juss) seed oil with known azadirachtin concentrations on the oocytes of semi-engorged Rhipicephalus sanguineus ticks (Acari: Ixodidae)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Histochemical and morphological features of biopsied and stripped oocytes from the Brazilian endangered teleost pirapitinga, Brycon nattereri (Characiformes)

The aim of the present study was to characterize biopsied and stripped oocytes from the Brazilian endangered teleost pirapitinga (Brycon nattereri) using histochemical and morphological analyses. Biopsied oocytes had a mean diameter of 2.225 mm (modal diameter: 2.312 mm), complete vitellogenesis and a central or slightly eccentric nucleus. Neutral polysaccharides were detected in the follicular cells, zona radiata and yolk globules, while acidic polysaccharides were detected in the follicular cells and cortical alveoli. Ten out of the 19 females treated with two doses of carp pituitary extract (cPE) released oocytes, which were also analysed. Stripping occurred 292 +/- 39 degree-hours after the second dose of cPE and led to a mean spawning weight of 36.2 g, 10% spawning index, 241 oocytes/g of ova, 8222 oocytes/female and 23 oocytes/g of body weight. Stripped oocytes had a mean diameter of 2.33 mm and a mode at 2.375 mm, were weakly adhesive and coloration ranged from wine to brown. Under scanning electron microscopy, stripped oocytes exhibited a single funnel-shaped micropyle located at the animal pole and a zona radiata that measured 7.7 m in thickness with eight pore canals/m(2). Oocyte morphology in Brycon nattereri is similar to that found in other species of the genus, except for the larger size and weaker adhesiveness. These findings provide essential information for a better understanding of the reproductive biology of B. nattereri and the establishment of conservation measures for this threatened species.

Storage of Steindachneridion parahybae oocytes at different temperatures

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

In vitro embryos production after oocytes treatment with forskolin

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Influence of lineage of oocyte donor on the yield and morphology of oocytes recovered by ultrasound-guided follicular aspiration in nellore cows

The objective of this study was to investigate the influence of lineage of oocytes donors on the number and quality of oocytes obtained through ultrasound-guided follicular aspiration in Nellore breed cows derived from two lineages of bulls (Karvadi; K and Taj-Mahal; T). Both maternal (Km and Tm) and paternal (Kp and Tp) lineages, as well as their combinations were investigated. Oocyte aspirations were repeatedly performed with an aspiration interval of 15 days in 56 donor females. Recovered cumulus oocyte-complexes (COCs) were counted, morphologically examined and classified into seven categories (grades from I to VII) according to the number of layers of the cumulus oocyte and cytoplasm appearance. The mean number of oocytes retrieved from donors of lineage Tp-Tm was significantly higher (28.23±1.92, P<0.05) than those obtained from lineages Kp-Tm, Kp-Km, and Tp-Km (21.34±1.32, 21.28±1.73, and 16.72±1.31, respectively). There was no significant difference in the mean number of recovered oocytes between donors of lineages Kp-Km and Kp-Tm, whereas animals of lineage Tp-Km yielded the lowest number of oocytes. Higher mean number of grade III oocytes was recovered from donors of lineage Kp than lineage Tp (10.11±0.66 versus 8.79±0.58, respectively), with more grade III oocytes being obtained in both lineages as compared to the others. Paternal lineage did not influence the quality of recovered oocytes in any other category, but both Kp and Tp yielded a great mean number of oocytes graded as I, II, and III (3.14±0.21; 4.93±0.33, and 10.11±0.66 versus 3.19±0.21, 5.59±0.44, and 8.79±0.58, respectively) than those classified as IV, V, VI, and VII. However, when considering the data from the maternal lineage significantly more oocytes (P<0.05) of grade I, II and III were obtained from Taj-Mahal (11.67 ± 0.67, 5.9±0.42 and 3.64±0.25, respectively) than for lineage Karvadi, with similar results for oocytes of grades IV, V, VI, and VII. Similarly to the paternal lineage, the number of oocytes of grade III was superior (P<0.05) when compared to other categories for both lineages. In conclusion, we demonstrate here a direct influence of lineage of oocyte donor on the production and quality of oocytes obtained through ultrasound-guided follicular aspiration in Nellore cows.

Heat stress induced alteration in bovine oocytes: functional and cellular aspects

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Spermathecae of the mangrove crab Ucides cordatus: a histological and histochemical view

Ucides cordatus is the most commercially, important mangrove crab in Brazil. In spite of its economic importance, there are few studies of its reproduction, in particular the female reproductive system. The present study describes the histology and histochemistry of the spermathecae of U. cordatus. Adult females were can lit monthly from July 2004 through June 2005, at Iguape, State of São Paulo. The crabs were anaesthetized, and their spermathecae removed and fixed in Davidson's fluid, following the histological routine for paraffin. The slides were stained with HE, xylidine Ponceau, PAS, alcian blue (pH 1.0 and 2.5), Sudan black B and picrosirius-haematoxylin. Histologically, the spermathecae possesses a capsule of conjunctive tissue, rich in collalgen fibres, which surrounds the secretory columnar epithelium. In the lumen, individual sperm packets are not observed; the spermatophores are intermixed with the seminal fluid and secretions of the spermathecae itself. A large proportion of the free spermatozoids and spermatophores are arranged in homogeneous masses in the proximal part of the spermathecae. The secretion produced by the columnar epithelium appears to promote the movement of the gametes to the fertilization chamber, in a ventral position, allowing fertilization of the oocytes. Histochemically, the secretion produced by the columnar epithelium was strongly positive for neutral polysaccharides, positive for acid polysaccharides, and weakly positive for proteins and lipids. Tills secretion forms a glycoprotein matrix which is associated with maintenance of the spermatophores, which can remain stored for long periods.

Induced spawning of hatchery-raised Brazilian catfish, cachara Pseudoplatystoma fasciatum (Linnaeus, 1766)

In the months of January 2001 and 2002, female cachara Pseudoplatystoma fasciatum were selected during their first and second gonadal maturation (2 years and 7 months old and 3 years and 7 months old, respectively) with an of oocyte diameter of 937.5 mum (82.5% with central nuclei and 17.5% with peripheral nuclei). Nine females in first maturation received two doses of carp pituitary extract (CPE), 0.5 mg/kg and 5.0 mg/kg; seven received two doses of human chorionic gonadotropin (hCG), 5 and 10 IU/g; five received doses of 0.5 CPE mg/kg and 5 hCG IU/g (CPE+hCG); and four received 0.9% saline (saline). Nine females from CPE and seven from hCG presented oocytes with the same diameter at the moment of oocyte release (100% with germinal vesicle breakdown and fertilization rate of 53.44 +/- 18.3 and 54.81 +/- 11.8%; larvae number of 165,330 +/- 94.1 and 158,570 +/- 20.6, respectively). The five females from CPE+hCG did not respond to the hormonal treatment. The four females from the saline group did not ovulate. In January 2002, 6 of 15 selected females that were going through the second reproductive cycle received CPE (five received hCG and four received saline), showing oocyte diameters similar to the ones in the first maturation. At stripping, CPE females had an oocyte diameter of 1062.5 mum (the hCG females had oocyte diameters ranging from 937.5 to 1125.0 mum; fertilization rates of 56.08 +/- 30.9 and 81.90 +/- 17.3%; 364,547 +/- 244 and 633,129 +/- 190, larvae, respectively). The fertilization rates and larvae number were higher in the second gonad maturation, both for CPE and hCG. (C) 2004 Elsevier B.V. All rights reserved.

Chronic exposure to sub-lethal concentration of a glyphosate-based herbicide alters hormone profiles and affects reproduction of female Jundia (Rhamdia quelen)

This work was carried out to verify the effect of a glyphosate-based herbicide on Jundia hormones (cortisol, 17 beta-estradiol and testosterone), oocyte and swim-up fry production. Earthen ponds containing Jundia females were contaminated with glyphosate (3.6 mg/L); blood samples were collected from eight females from each treatment immediately before, or at 1, 10, 20 30 and 40 days following contamination. A typical post-stress rise in cortisol levels was observed at the 20th and 40th days following exposure to glyphosate. At the 40th day, 17 beta-estradiol was decreased in the exposed females. A similar number of oocytes were stripped out from females from both groups, however, a lower number of viable swim-up fry were obtained from the herbicide exposed females, which also had a higher liver-somatic index (LSI). The results indicate that the presence of glyphosate in water was deleterious to Rhamdia quelen reproduction, altering steroid profiles and egg viability. (c) 2006 Elsevier B.V. All rights reserved.

Dose inseminante para fertilização artificial de ovócitos de dourado

Objetivou-se determinar a dose inseminante adequada para uso na fertilização artificial de ovócitos de dourado (Salminus brasiliensis). Os ovócitos foram distribuídos em delineamento inteiramente casualizado, e fertilizados com uma das relações espermatozoides/ovócito 6,0×10³; 6,0×10(4); 6,0×10(5); 6,0×10(6) ou 3,0×10(7), cada uma com quatro repetições. Considerou-se unidade experimental uma incubadora de volume útil de 2,5 L, contendo 2,0 mL de ovócitos não-hidratados. As taxas de fertilização foram mensuradas 8 horas após o início da fertilização. Com intuito de verificar possíveis efeitos da diluição seminal na movimentação dos espermatozoides, realizou-se a mensuração do tempo de duração da motilidade espermática dos espermatozoides de dourado, ativados por meio de diferentes relações de diluição: 6,8×10-5; 6,8×10-4; 6,8×10-3; 6,8×10-2; 3,4×10-1 e 1,0 mL de sêmen por mL de água. O tempo de duração da motilidade foi avaliado em delineamento inteiramente casualizado composto de seis tratamentos e três repetições. As taxas de fertilização apresentaram relação quadrática com o número de espermatozoides por ovócito. As relações de diluição do sêmen tiveram efeito inversamente proporcional sobre a duração da motilidade espermática. A relação que proporcionou melhores taxas de fertilização artificial de ovócitos de dourado (Salminus brasiliensis) foi de 30.722 espermatozoides por ovócio.