Effects of growth differentiation factor-9 and FSH on in vitro development, viability and mRNA expression in bovine preantral follicles

Vasconcelos, G. L. de; Saraiva, M.V.A.; Costa, Jonathan L.; Passos, M. J.; Silva, A W B; Rossi, R.O.D.S.; Portela, A. M. L. R.; Duarte, A.B.G.; Magalhães-Padilha, D.M.; Campelo, Cláudio Cabral; Figueiredo, J.R.; Hurk, R. van den; Silva, J R V

doi:10.1071/rd12173

Cited by 28 publications

(17 citation statements)

References 74 publications

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“…In accordance with the present results, Vasconcelos et al (2012) also demonstrated that the presence of GDF-9 in culture medium of bovine preantral follicles was not accompanied by an enhancement of the number of follicles showing antrum formation. The formation of antrum after culture of goat follicles even in the control medium could be due to the presence of insulin, since previous studies have shown that insulin stimulated the formation of antrum in cultured bovine preantral follicles (Itoh et al 2002).…”

Section: Discussionsupporting

confidence: 80%

“…The signals for antrum formation were not well understood, but several studies have shown that antrum formation was an event independent of gonadotropins (Gulyas et al 1977;Halpin et al 1986;Hillier et al 1994;Cain et al 1995). Recently, administration in culture medium of FSH did not stimulate the expression of mRNA for proteoglycans involved in antrum formation, i.e., HAS-1, HAS-2, perlecan and versican, in developing bovine follicles (Vasconcelos et al 2012).…”

Section: Discussionmentioning

confidence: 99%

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Effects of GDF-9 and FSH on mRNA expression for FSH-R, GDF-9 and BMPs in in vitro cultured goat preantral follicles

Leitão

Costa

Brito

et al. 2014

Braz. arch. biol. technol.

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The aim of the present study was to determine the role of GDF-9 and/or FSH on the growth and mRNA expression for FSH-R, GDF-9, and BMPs in goat secondary follicles after culture in vitro. Goat secondary follicles (~200µm) were isolated and cultured for six days in minimum essential medium (MEM) supplemented with GDF-9 (200 ng/mL), FSH (50 ng/mL) or both. At the beginning and end of culture, the follicular diameter was evaluated and compared. The levels of mRNA for reduced the expression of BMP-2 and -15 in caprine preantral follicles after their culture, but FSH either alone or in association with GDF-9 did not control the expression of

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Section: Discussionsupporting

confidence: 80%

Section: Discussionmentioning

confidence: 99%

Effects of GDF-9 and FSH on mRNA expression for FSH-R, GDF-9 and BMPs in in vitro cultured goat preantral follicles

Leitão

Costa

Brito

et al. 2014

Braz. arch. biol. technol.

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“…SCP3 ‐positive oocytes were previously considered to be direct evidence of post‐natal oogenesis through differentiation of germline stem cells in mice (Pan et al., ). GDF9 is expressed in bovine oocytes (Vasconcelos et al., ), and it has been used as marker to identify oocytes differentiated from stem cells cultured in vitro (Linher et al., ). Both BMP2 and BMP4, as well as follicular fluid, are currently found to stimulate expression of ZPA in OLCs.…”

Section: Discussionmentioning

confidence: 99%

Bovine ovarian stem cells differentiate into germ cells and oocyte‐like structures after culture in vitro

Souza

Costa

Cunha

et al. 2016

Reprod Domestic Animals

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Stem cells have been isolated from ovaries, and their ability to differentiate into oocytes in vitro has been demonstrated for mice and human, but not for bovine species. The aims of this study were to isolate germline stem cells from bovine ovaries and to evaluate the effects of bone morphogenetic proteins (BMPs) 2 and 4, and follicular fluid on the differentiation of these stem cells into oocyte-like structures. The ovarian stem cells were isolated and cultured in α-MEM supplemented with BMP2, BMP4 or follicular fluid. On days 0 and 14, cells were evaluated for their morphological appearance, viability, expression of alkaline phosphatase and for markers of germ cell formation (VASA and DAZL) and oocyte development (GDF9, ZPA and SCP3) by qPCR. Levels of mRNA were analysed using ANOVA and Bonferroni test (p < .05). The results showed that at day 0, ovarian stem cells expressed specific markers of pluripotency (OCT4, SOX). In addition, these cells were positive for alkaline phosphatase, which is a marker commonly used to identify primordial germ cells (PGCs). After the period of differentiation, cells had morphological features that resemble PGCs and oocyte-like cells (OLCs). An increase, ranging from five to 14 times, in the expression of VASA was observed in cells cultured in medium supplemented with BMPs and follicular fluid, while the increase in DAZL expression ranged from four to six times. In addition, OLCs had an increase in expression of mRNAs for GDF9, ZPA and SCP3 that ranged from two to eight times. In conclusion, OLCs can be differentiated in vitro from ovarian stem cells and BMPs and follicular fluid are effective in stimulating the expression of mRNAs for germ cell and oocyte markers.

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“…Since, rRNA and mRNA have a short life in the environment (Pecson et al, 2006;Sander et al, 2011;Vasconcelos et al, 2012), they can be used for detecting viable hookworm ova from treated wastewater and sludge samples. There is, however, no information regarding the status of rRNA and mRNA in ova inactivated during their incubating phase.…”

Section: Molecular Viability Pcrmentioning

confidence: 99%

“…Molecular viability PCR uses species-specific ribosomal RNA (rRNA) and messenger RNA (mRNA) precursors required for basic cell function or life within the cell, as indicators of viable ova (Widmer et al, 1999;Kobayashi et al, 2009;Vollmer et al, 2010;Sander et al, 2011;Vasconcelos et al, 2012). Since, rRNA and mRNA have a short life in the environment (Pecson et al, 2006;Sander et al, 2011;Vasconcelos et al, 2012), they can be used for detecting viable hookworm ova from treated wastewater and sludge samples.…”

Section: Molecular Viability Pcrmentioning

confidence: 99%

Detection of viable hookworm ova from wastewater and sludge

Gyawali¹

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Infectious helminths are a worldwide major public health problem. In terms of morbidity, approximately 3 × 10 9 people are infected by soil-transmitted helminths (STHs) influencing rates of malnutrition and failure to thrive. All of this increases the global disease burden (GDB) by up to 5.9 × 10 6 Daily Adjusted Life Years (DALYs). Helminth infections are more often seen as a major public health issue for developing countries however, concerns have been expressed in many of the developed nations because of the increased use of treated wastewater and sludge with no clear way of knowing the infection potential that can be attributed from such water.Guidelines have been established to minimise the potential public health risk associated with wastewater and sludge reuse. For example, the WHO guideline specified ≤ 1 ova (Ascaris lumbricoides) per L liquid (wastewater) or 4 g dry solid (sludge) for unrestricted use. Various wastewater treatment processes have been recommended to remove the helminths ova from wastewater depending upon its reuse. However, detection methods used to quantify viable helminths ova from wastewater has limitations. Therefore there is always a potential public health risk associated with reuse of wastewater and sludge.In this research, a real-time PCR method was developed. The new PCR method is rapid and specific. The method was found to be able to detect less than one ovum in one litre treated wastewater and approximately four ova in one litre raw wastewater and ~ 4 g sludge.The real-time PCR method was modified to a quantitative PCR (qPCR) method and further used to quantify hookworm ova from wastewater and sludge. The qPCR had estimated an average of 1.1, 8.6 and 67.3 ova for treated wastewater that was seeded with 1, 10 and 100 ova, respectively. The gene copy numbers obtained for 1, 10 and 100 ova by qPCR varied significantly (P < 0.05) within the tested samples indicating that absolute quantification of ova may not be accurate. Despite the difficulty quantifying accurate numbers of hookworm ova, the lower limit of quantification (LLOQ) of the qPCR method was 30 gene copies. This was a lot less than the gene copies produced by one ovum. Therefore, the qPCR method has potential to use for complying with wastewater guidelines.. iiAlthough the overall aim of this research was to develop a sensitive and specific method for quantitative detection of viable hookworm ova from wastewater, the importance of recovering the ova from wastewater and sludge samples for accurate detection was identified. While determining appropriate recovery rates was not an aim of this research, a suitable method that could be used to standardise research outcomes for further study was established. Therefore, ova recovery rate by different rapid methods was evaluated for further experiments. The result indicated that the ova recovery rate was higher for the treated wastewater (0.2 -50%) than the raw wastewater (0.3 -35%) and sludge (0.02 -4.7%) samples. A significant difference (P < 0.05) was observed between...

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Effects of growth differentiation factor-9 and FSH on in vitro development, viability and mRNA expression in bovine preantral follicles

Cited by 28 publications

References 74 publications

Effects of GDF-9 and FSH on mRNA expression for FSH-R, GDF-9 and BMPs in in vitro cultured goat preantral follicles

Effects of GDF-9 and FSH on mRNA expression for FSH-R, GDF-9 and BMPs in in vitro cultured goat preantral follicles

Bovine ovarian stem cells differentiate into germ cells and oocyte‐like structures after culture in vitro

Detection of viable hookworm ova from wastewater and sludge

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