659 research outputs found
In vitro maturation of canine oocytes in a bi-phasic system using gonadotrophin and steroids
No full textCultivo in vitro de folículos pré antrais de gatas domésticas em meio suplementado ou não com Epidermal Growth Factor (EGF)
No full textO desenvolvimento in vitro de folículos pré antrais de gatas domésticas é positivamente influenciado
pela adição de EGF ao meio de cultivo, assim como a fase do ciclo estral da doadora
Spermatozoa co-culture does not improve oocyte nuclear maturation rates in dogs
No full textIn canids, in order to overcome the limited efficiency of oocyte in vitro maturation1, several attempts mostly investigating different supplementations of the culture media, culture periods, and variation of the conditions over the course of the culture period (so-called bi-phasic media)2 have been made. In vivo, as canine oocytes are ovulated immature and remain an extended period within the oviducts, spermatozoa are present before full maturation occurs. Taken this into account, it was interesting to verify whether the presence of spermatozoa in in vitro culture medium could trigger meiosis resumption and improve maturation rates. For this purpose, ovaries were collected from bitches at different reproductive status (various breeds, age 1-7 years) by routine ovariohysterectomy and sliced in PBS-PVA to release the cumulus-oocyte complexes (COCs). Grade I COCs (n. 405) were obtained and randomly allocated in four systems with the base medium (BM) consisting of TCM-199 with antibiotics, 10% FBS, 2.2 mg/ml sodium bicarbonate and 20 l/ml pyruvic acid. Hormones were supplemented at the dose of 10 i.u/ml hCG (Sigma Chemical Co., MO, USA), 1μg/ml P4 (Sigma), 1μg/ml E2 (Sigma) as follows: in system A (n.110) oocytes were matured for 48 h in BM with hCG and for additional 24 h in BM with P4; in system B oocytes (n.118) were matured for 48 h in BM with hCG, P4 and E2 and for 24 h in BM with P4; in system A-S (n.92) and B-S (n.85) oocytes were cultured in system A and B, respectively, but with the addition of spermatozoa (5 x 106 sperm/ml) at 48 h of culture. COCs were incubated at 38C, 5% CO2 in air. At the end of maturation period (72 h), oocytes were denuded within 0.2% hyaluronidase solution (Sigma) by repeated pipetting and then, were stained with Hoechst 33342 (Sigma) for evaluation of meiotic configuration. Data were analyzed by ANOVA at p0.05. The percentage of degenerated oocytes significantly increased with the presence of spermatozoa (9%, 7.6%, 32.6%, and 28.3%, for systems A, B, A-S, and B-S, respectively; p<0.001). The overall sperm penetration occurred both in immature and mature oocytes, however a higher proportion of matured oocytes (60.5%, 23/38) were penetrated compared to the immature ones (22%, 4/18; p=0.0006). We can speculate that the potential influence of sperm penetration could be on cytoplasmic maturation, rather than nuclear maturation. This aspect should be further investigated and the fact that different periods of co-incubation might influence the results.
[1] Luvoni GC, Chigioni S, Allievi E, Macis D. Factors involved in vivo and in vitro maturation of canine oocytes. Theriogenology 2005; 63: 41–59.
[2] Apparicio M, Mostachio GQ, Motheo TF, et al. Distribution of cortical granules and meiotic maturation of canine oocytes in bi-phasic systems. Reproduction, Fertility and Development 2015; 27:1082–1087
Viability of feline preantral follicles in vitro cultured with Insulin Growth Factor and Epidermal Growth Factor supplemented medium
No full textIn vitro culture of ovarian preantral follicles (PF) has emerged as a potential reproductive technology for the production of mature oocytes capable of fertilization [1]. Advances concerning the role of several growth factors (GF) on in vitro activation of primordial follicles have been described. The addition of EGF (Epidermal Growth Factor) and IGF (Insulin-like Growth Factor) in the in vitro culture of PF of different species has provided superior results of follicular development, antrum formation and proliferation of granulosa cells (GC) [2]. However, there are only few reports regarding the use of these factors on feline PF in vitro culture. We previously demonstrated that IGF-1 exerts a positive influence on follicular development [3] and has been recently shown that EGF sustains in vitro primordial follicle viability in the prepubertal cat ovary [4]. Thus, the aim of this study was to investigate the effect of a combination of IGF and EGF on in vitro growth and GC viability of PF collected from domestic cats. A total of 64 PF characterized by multi-layer granulosa cells and theca cells with 150–200 μm of diameter were isolated and individually cultured for 6 days (T6) in minimum essential medium (MEM; Sigma-Aldrich Co., St. Louis, LO, USA) supplemented with IGF+EGF (100 ng/ml each; Sigma) or without GF (control). Data from follicular and oocyte diameters were submitted to Kolmogorov–Smirnov. Mean values of the increase in follicular size were analysed by Student–Newman–Keuls test, and GC viability was analysed by chi-square test (P 0.05; Sigma). The GC of PF cultured with GF maintained a greater viability (fluorescein diacetate staining; Sigma) than those of PF cultured without (84.3% and 68.7% respectively; P<0.05). These data suggest that the addiction of IGF and EGF to the culture medium promotes better conditions to the in vitro development of preantral follicles of cats. It remains to investigate the precise role of the single growth factor to establish optimal culture conditions.
[1] Demeestere I, Centner J, Gervy Y, et al. Impact of various endocrine and paracrine factors on in vitro culture of preantral follicles in rodents. Reproduction 2005; 130:147-56.
[2] Silva J R V, Van Den Hurk R, Figueiredo J R. Ovarian follicle development in vitro and oocyte competence: advances and challenges for farm animals. Domest Anim Endocrinol 2016, 55:123-5.
[3] Alves EA, Padilha L, Savi PA, et al. In vitro survival of follicles recovered from queens at different stages of estrous cycle and cultured with IGF-1. Reprod Domest Anim 2012; 47 (Suppl. 6):109-12.
[4] Fujihara M, Comizzoli P, Keefer CL, et al. Epidermal growth factor (EGF) sustains in vitro primordial follicle viability by enhancing stromal cell proliferation via MAPK and PI3K pathways in the prepubertal, but not adult, cat ovary. Biol Reprod 2014; 90:86
In vitro survival of preantral follicles recovered from queens at different stages of estrous cycle and cultured with IGF-1
No full textOBJECTIVE AND METHODS. In vitro growth of preantral follicles may supply high numbers of competent oocytes that can be destined to in vitro embryo production. However, optimal culture systems that support follicular and oocyte development in vitro have not yet been defined. Ovarian follicular development is regulated mainly by endocrine, autocrine, and paracrine factors and follicles are exposed to specific hormonal environments during the different stages of the estrous cycle. The intraovarian insulin-like growth factor-I (IGF-1) regulates in vivo follicular development and there are important evidences that it stimulates in vitro growth of preantral follicles in domestic animals (1). Feline preantral follicles have been previously cultured in vitro (for review 2) and it has been suggested that IGF-1 enhances oocyte metabolism (3). However, no information are available on the response of preantral follicles recovered in different phases of the estrous cycle to cultural conditions. Thus, the aim of this study was to investigate the in vitro survival of preantral follicles recovered from ovaries of queens in follicular or luteal phase of the estrous cycle and cultured in presence of IGF-1.
Twelve queens were housed with 12 hours of light and submitted to estrous induction with IM injection of 100 UI eCG (Novormon®- Intervet) and 100 UI hCG (Vetecor®- Hertape Calier) 82 hours later (4). Six queens were spayed 96 h after eCG administration (follicular phase), the others 36 h after the hCG injection (luteal phase). Estrous phases were confirmed by vaginal cytology prior to the surgical procedure and macroscopic evaluation of ovarian structures after excision.
Preantral follicles surrounded by complete basal membrane and containing more than one layer of granulosa cells were retrieved from excised ovaries and selected as previously described (5). A total of 72 follicles were cultured for 6 days at 38.5 ̊C and 5% CO2 in air in Minimum Essential Medium (Sigma Chemical Co., USA) with IGF-1 100 ng/ml (Sigma) or without (control). Before and after culture, follicular diameter was recorded and follicular viability was assessed by fluorescein diacetate (FDA, Sigma) staining. Increase (%) in diameter was analyzed by Tukey’s and Fisher’s test, viability rates by Chi-square test (P<0.05).
RESULTS. After 6 days of culture, preantral follicles retrieved during follicular phase showed a significant increase in the size and a higher viability rate than those retrieved in the luteal phase of the estrous cycle (18.8% vs.11.5%; P= 0.0001 and 75% vs. 62.5%; P=0.004). However, when IGF-1 was added to the culture medium, follicles retrieved in follicular or luteal phase showed similar increase in diameter (14.9% vs.13.4%; P>0.05) and viability (73.8% vs. 76%; P>0.05). Regardless of the stage of the estrous cycle, overall results showed that the increase in diameter was not different in follicles cultured with or without IGF-1 (14.6% vs. 15.8%; P>0.05), but follicular survival was enhanced when IGF-1 was added to the culture medium (75% vs. 69.4%; P=0.0001).
CONCLUSIONS: Present data suggest that in vitro survival of preantral follicles is affected by the estrous stage of the donor and IGF-1 improves survival of follicles retrieved in luteal phase of the estrous cycle. Thus, the hormonal environment of the follicles within the ovary might impact their potential development when isolated and cultured. Further investigations on growth factors are needed to evaluate their effect on follicles with reduced in vitro developmental capability.
REFERENCES
(1) Giudice LC. Insulin-like growth factors and ovarian development. Endocr Rev 1992; 13: 641-669.
(2) Jewgenow K, Paris MCJ. Preservation of female germ cells from ovaries of cat species. Theriogenology 2006; 66: 93-100.
(3) Jewgenow K. Impact of peptide growth factors on the culture of small preantral follicles of domestic cats. Theriogenology 1996; 45: 889-895.
(4) Villaverde AI, Melo CM, Martin I, Ferreira TH, Papa FO, Taconeli CA, Lopes MD. Comparison of efficiency between two artificial insemination methods using frozen-thawed semen in domestic cat (Felis catus): artificial insemination in domestic cats. Anim Reprod Sci 2009; 114: 434-442.
(5) Lima AK, Bezerra MB, Oliveira LC, Figueiredo JR, Silva LDM. Isolamento e caracterização de folículos ovarianos pré-antrais em gatas domésticas (Felis catus). Rev Bras Reprod Anim 2003; 27: 396-397
In vitro Survival of Follicles Collected from Domestic Cats' Ovaries at Different Stages of Oestrous Cycle and Cultured with IGF-1
No full textContents Optimal conditions for in vitro culture of feline ovarian follicles have not yet been defined. Follicular development is regulated by intraovarian growth factors, as insulin-like growth factor (IGF-1), and during the different stages of the oestrous cycle, follicles are exposed to specific hormonal environments. The aim of this study was to investigate the effect of IGF-1 on in vitro growth and granulosa cell (GC) viability of preantral follicles collected from domestic cats at follicular and luteal phases of the oestrous cycle. Oestrus and ovulation were induced in 12 cats. A total of 39 and 32 follicles collected at the follicular and luteal phases, respectively, were individually cultured in vitro for 6 days in minimum essential medium media supplemented with or without IGF-1 (100 ng/ml). Follicles collected during the follicular phase and cultured without IGF-1 displayed a significant increase in size and higher GC viability (46.5 +/- 22.1 mu m, 66.7%, respectively) than that of follicles collected at the luteal phase and cultured without IGF-1 (26.7 +/- 14.4 mu m, 50%, respectively; p < 0.05). In contrast, when IGF-1 was added to the culture medium, no differences were observed in size or GC viability between follicles collected at the two phases of the cycle. Nonetheless, follicles collected at the luteal phase and cultured with IGF-1 had a significant increase in their diameter and GC viability (31.9 +/- 15.9 mu m, 63.6%, respectively) than that cultured without IGF-1 (26.7 +/- 14.4 mu m, 50%, respectively; p < 0.05). These data suggest that in vitro growth and GC survival of feline preantral follicles are affected by the oestrous cycle phase, and the IGF-1 exerts a positive effect on follicles collected at the luteal phase.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Univ Estadual Paulista, Dept Med Vet Prevent & Reprod Anim, Setor Reprod & Obstet Vet, FCAV,UNESP, BR-14884900 Jaboticabal, SP, BrazilUniv Milan, Dipartimento Sci Vet Salute Prod Anim & Sicurezza, Milan, ItalyUniv Estadual Paulista, Dept Med Vet Prevent & Reprod Anim, Setor Reprod & Obstet Vet, FCAV,UNESP, BR-14884900 Jaboticabal, SP, Brazi
Correction to : Cost-Effectiveness of Parent–Child Interaction Therapy in Clinics versus Homes : Client, Provider, Administrator, and Overall Perspectives (Journal of Child and Family Studies, (2018), 27, 10, (3329-3344), 10.1007/s10826-018-1159-4)
No full textThe original version of this article unfortunately contained a mistake. The Author contributions section currently states: “A.F.: designed and conducted data analyses for this study, supervised by B.Y. with input by T.F. A.F. and T.F. assisted B.Y. in writing and revising the manuscript.” In fact, B.Y. and T.F. assisted A.F. in writing and revising the manuscript. Although T.F. and B.Y. were both involved in the writing, primary credit should go to A.F. (Alexis French, the first author). The authorship order above is accurate.</p
MALDI imaging mass spectrometry (MALDI-IMS): a new approach for spatial identification of proteins in feline oviducts
No full textThe knowledge and understanding of many reproductive processes have improved significantly over the years, especially after the development of tools for protein identification1. The traditional approach for spatial identification of proteins requires a prior knowledge of the target entities and narrows the number of targets that can be monitored simultaneously1. For that reason, MALDI imaging mass spectrometry (MALDI-IMS) once it is able to directly scan the target tissue, is considered an outstanding approach for molecular profiling and imaging. With the purpose to identify the factors involved in early stages of embryo development in the domestic cat, this technology was used for the first time to analyse the spatial distribution of proteins in oviducts of queens. Reproductive tracts were collected from 2 queens (cross-breed, 2 and 4 years old) undergoing routine ovariohysterectomy. The oviducts were carefully dissected, divided into three segments (distal to the ovary, proximal to the ovary and the mid-section between the two), snap-frozen in liquid nitrogen and stored at -80oC until use. Then, they were sectioned (11 μm) in a cryostat and fixed on ITO (indium tin oxide) conductive glass slides while serial sections were collected on microscope slides for haematoxylin and eosin staining. For MALDI-IMS, samples were coated with a thin homogeneous layer of CHCA (α-Cyano-4-hydroxycinnamic acid) matrix using a nebulization device. MALDI images were acquired on an Autoflex III Smartbeam instrument (Bruker Daltonics) with 400 shots/spectrum, over the mass range of m/z 2 to 20 kDa in the positive ion mode. The spatial resolution of images was 80 μm. The molecular images obtained in this study show that protein distribution was different in the oviductal infundibulum, ampulla, and isthmus identified by histology. Mass spectra was characterized by abundant ions of m/z 1259, 4939, 4960 and 10626. Based on the literature2 these ions might correspond to keratin, thymosin β10, thymosin β4 and S100, respectively. S100 proteins are calcium modulated proteins implicated in a variety of cellular activities, including cell differentiation and regulation of cell motility. Keratin and thymosins are involved in the biological response to tissue damage. In women, this defence system is adapted to operate in the genital tract, being altered according to the hormonal and cellular changes that occur during menstruation (characterized by cell death and necrosis)3. Protein attributions described herein must be confirmed by MS/MS experiments, but our results indicate that it is possible to use this technology to elucidate key molecular processes involved in the reproductive physiology of carnivore species.
[1] Lagarreigue M, Lavigne R, Guevel B, et al. Matrix-Assisted Laser Desorption/Ionization Imaging Mass Spectrometry: A Promising Technique for Reproductive Research. Biology of Reproduction 2012; 86(3): 1–11. [2] McDonnell LA, Walch A. Stoeckli M, Corthals GL. MSiMass list: a public database of identifications for protein MALDI MS imaging. Journal of proteome research 2013, 13 (2), 1138-1142. [3] Horne AW, Stock SJ, King AE. Innate immunity and disorders of the female reproductive tract. Reproduction 2008; 135(6): 739-749
Review of the Monograph T.F. Khaidarov “Age of ‘Black Death’ in Golden Horde and Adjacent Regions” Kazan: Marjani Institute of History of Academy of Sciences, 2018. 304 P.
No full textThe author examines the monographic work of T.F. Khaidarov, devoted to the study of the second plague pandemic in the Golden Horde and neighboring territories, which was published in 2018. Researchers have already studied some aspects of the course of the Black Death in Eastern Europe. Before the publication of the monograph, T.F. Khaidarov had published several articles devoted to this problem. Despite this, the monograph is the first experience of a comprehensive study of the manifestations of the second plague pandemic in all their diversity in Eastern Europe. The objectives set in the book required the synthesis of various natural sciences (biology, climatology, medicine) and social (history, historical source study, social anthropology) disciplines. In the monograph phenomena of different scale are investigated. On the one hand, the epidemics in Eastern Europe were considered in the context of the worldwide course of the pandemic. On the other hand, from the researcher’s view did not escape the local phenomena: the specific dating of a particular outbreak of the epidemic, the problem of the perception of the disease by society, the features of its description, etc. In the review, an assessment is given to this "total" study of the pandemic. The advantage of this approach is that it overcomes the boundaries between different scientific disciplines, allows us to fill the knowledge that is fragmentary in each separate science. The drawback of this approach is that it leads to the absolutization of the plague factor in history. The study of Black Death becomes the main link in the whole multidisciplinary research, turns into a kind of paradigm. Sometimes this leads to incorrect reading of historical sources and scientific works. Despite the revealed shortcomings, it should be recognized that the monograph of T.F. Khaidarov is a serious step forward in the study of the problem at the present stage of the development of science
Viability and growth of feline preantral follicles in vitro cultured with insulin growth factor and epidermal growth factor supplemented medium
No full textIn vitro culture of ovarian preantral follicles has emerged as a reproductive technology aimed at obtaining large amount of oocytes for in vitro embryo production. The addition of growth factors (GF) in the in vitro culture of preantral follicles of different species has provided superior results of follicular development, antrum formation and proliferation of granulosa cells. However, there are only few reports regarding the use of these factors on feline preantral follicle in vitro culture. Thus, the aim of this study was to investigate the effect of a combination of IGF-1 and EGF on in vitro viability and growth of preantral follicles and enclosed oocytes collected from domestic cats. A total of 64 follicles characterized by multilayer granulosa cells were isolated and individually cultured for 6 days (T6) in minimum essential medium supplemented with IGF-1+ EGF (100 ng/ml each) or without (control). A higher percentage of follicles were viable after culture with GF than without, and an increase in size when IGF-1+ EGF were added to the medium (170 ± 32.4 μm (T0) vs. 201 ± 22.3 μm (T6); p .05). These data suggest that the addition of IGF-1 and EGF to the culture medium promotes the in vitro development of preantral follicles of cats
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