The mare as a model for luteinized unruptured follicle syndrome: intrafollicular endocrine milieu

Bashir, Shah Tauseef; Gastal, M.O.; Tazawa, Shuji P.; Tarso, S. G. S. de; Hales, D. B.; Cuervo‐Arango, J.; Baerwald, Angela

doi:10.1530/rep-15-0457

Cited by 32 publications

(16 citation statements)

References 78 publications

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“…This technique could be helpful for repeated in vivo sampling of the follicle wall and FF of the same individual simultaneously, without disturbing ovarian function. Furthermore, this technique would provide a better understanding of physiological and pathological mechanisms involved in ovarian function, such as follicle selection and dominance, and formation of the hemorrhagic anovulatory follicle (HAF) or luteinized unruptured follicle (LUF) syndrome, and ovarian cyst in different species [ 23 – 27 ].…”

Section: Introductionmentioning

confidence: 99%

In vivo antral follicle wall biopsy: a new research technique to study ovarian function at the cellular and molecular levels

Ishak

Bashir

Dutra

et al. 2018

Reprod Biol Endocrinol

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BackgroundIn vivo studies involving molecular markers of the follicle wall associated with follicular fluid (FF) milieu are crucial for a better understanding of follicle dynamics. The inability to obtain in vivo samples of antral follicle wall (granulosa and theca cells) without jeopardizing ovarian function has restricted advancement in knowledge of folliculogenesis in several species. The purpose of this study in mares was to develop and validate a novel, minimally invasive in vivo technique for simultaneous collection of follicle wall biopsy (FWB) and FF samples, and repeated collection from the same individual, during different stages of antral follicle development. We hypothesized that the in vivo FWB technique provides samples that maintain the normal histological tissue structure of the follicle wall layers, offers sufficient material for various cellular and molecular techniques, and allows simultaneous retrieval of FF.MethodsIn Experiment 1 (ex vivo), each follicle was sampled using two techniques: biopsy forceps and scalpel blade (control). In Experiment 2 (in vivo), FWB and FF samples from 10-, 20-, and 30-mm follicles were repeatedly and simultaneously obtained through transvaginal ultrasound-guided technique.ResultsIn Experiment 1, the thickness of granulosa, theca interna, and theca externa layers was not influenced (P > 0.05) by the harvesting techniques. In Experiment 2, the overall recovery rates of FWB and FF samples were 97 and 100%, respectively. However, the success rate of obtaining samples with all layers of the follicle wall and clear FF varied according to follicle size. The expression of luteinizing hormone receptor (LHR) was mostly confined in the theca interna layer, with the estradiol-related receptor alpha (ERRα) in the granulosa and theca interna layers. The 30-mm follicle group had greater (P < 0.05) LHR expression in the theca interna and ERRα in the granulosa layer compared to the other groups. The overall expression of LHR and ERRα, and the intrafollicular estradiol were higher (P < 0.05 – P < 0.0001) in the 30-mm follicle group.ConclusionThe in vivo technique developed in this study can be repeatedly and simultaneously used to provide sufficient FWB and FF samples for various cellular and molecular studies without jeopardizing the ovarian function, and has the potential to be translated to other species, including humans.Electronic supplementary materialThe online version of this article (10.1186/s12958-018-0380-8) contains supplementary material, which is available to authorized users.

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Section: Introductionmentioning

confidence: 99%

In vivo antral follicle wall biopsy: a new research technique to study ovarian function at the cellular and molecular levels

Ishak

Bashir

Dutra

et al. 2018

Reprod Biol Endocrinol

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“…In the present study, the pig model allowed the carrier in both mono-and polyovulator animals as well as in living large animals. We believe the proposed methodology is likely to support future intrafollicular attempts to promote follicle and/or oocyte growth and to develop treatments to address follicle diseases such as luteinized unruptured follicle in mares and, potentially, in women as well [5,40]. (Hayward, California, USA).…”

Section: Confocal Microscope Fluorescence Imaging Of Fwb Samples and mentioning

confidence: 99%

“…The current efforts to overcome fertility problems through assisted reproductive technologies (ARTs) are still unsatisfactory because of the insufficient maturation of the oocytes and/or the 49 existing follicular diseases [1][2][3][4][5][6]. Furthermore, the ability to assess the quality of the oocytes 50 mainly, if not only, through in vitro manipulations constitutes a serious limitation [7,8].…”

mentioning

confidence: 99%

A Nano-Platform for in situ Investigations of Ovarian Follicles

Feugang

Ishak

Eggert

et al. 2020

Preprint

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Background: Despite the growing array of assisted reproductive techniques, there is still a lack of rapid, non-cytotoxic, and minimally invasive in situ approaches for further enhancements through cell targeting. Here we synthesized clinically relevant liposome nanoparticles for real-time cellular targeting and drug (doxorubicin) delivery, using pigs and mares as animal models. In Experiment 1, fluorescently labeled and doxorubicin-loaded (without fluorescent probe) liposomes were injected in cultured pig ovarian follicles to assess plasma membrane binding and intracellular doxorubicin delivery. In Experiment 2, fluorescent liposomes were in vivo injected into small and large ovarian follicles of living mares to assess their binding capability to follicular cells. Twenty-four hours post-injection, all cultured pig follicles were collected while mare samples (i.e., follicle wall fragments, granulosa cells and follicular fluids) were harvested through follicle wall biopsy (FWB), and follicle aspiration and flushing techniques using transvaginal ultrasound-guided approach.Results: All injected follicles were healthy and samples were subjected to fluorescence imaging before and after fixation. Findings revealed successful intrafollicular migration and binding of liposomes to all follicle cell layers (granulosa, theca interna, and theca externa) regardless of the follicle size. The intracellular delivery of doxorubicin was confirmed with the staining of nuclei of follicle cells. Conclusions: This study demonstrates the promising combination of the FWB technique and nanotechnology tools for real-time monitoring of intrafollicular treatment, follicular health and oocyte development, which in turn has the potential to help understand the mechanisms of ovulatory dysfunction and to select high-quatity oocytes for assisted reproduction techniques.

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“…As an example, the single-chain EquiPureLH (Venaxis) was used in mares in combination with EquiPureFSH for superovulation treatment [32], or in combination with a pituitary FSH (eFSH; Bioniche Animal Health) to study their effect in follicle and oocyte development [60]. Also, LH has been administered in mares as a model to treat the luteinizing unruptured follicle syndrome in humans [61]. Furthermore, the human-indicated Luveris (hLH; Merck) has been used for early embryonic development treatments in rabbits [62] combined with Gonal-F (see the FSH section), and in mice [63] combined with Pregnyl (hCG; Organon) (Table 1).…”

Section: Recombinant Hormones In Reproductionmentioning

confidence: 99%

Trends in recombinant protein use in animal production

et al. 2017

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Recombinant technologies have made possible the production of a broad catalogue of proteins of interest, including those used for animal production. The most widely studied proteins for the animal sector are those with an important role in reproduction, feed efficiency, and health. Nowadays, mammalian cells and fungi are the preferred choice for recombinant production of hormones for reproductive purposes and fibrolytic enzymes to enhance animal performance, respectively. However, the development of low-cost products is a priority, particularly in livestock. The study of cell factories such as yeast and bacteria has notably increased in the last decades to make the new developed reproductive hormones and fibrolytic enzymes a real alternative to the marketed ones. Important efforts have also been invested to developing new recombinant strategies for prevention and therapy, including passive immunization and modulation of the immune system. This offers the possibility to reduce the use of antibiotics by controlling physiological processes and improve the efficacy of preventing infections. Thus, nowadays different recombinant fibrolytic enzymes, hormones, and therapeutic molecules with optimized properties have been successfully produced through cost-effective processes using microbial cell factories. However, despite the important achievements for reducing protein production expenses, alternative strategies to further reduce these costs are still required. In this context, it is necessary to make a giant leap towards the use of novel strategies, such as nanotechnology, that combined with recombinant technology would make recombinant molecules affordable for animal industry.

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The mare as a model for luteinized unruptured follicle syndrome: intrafollicular endocrine milieu

Cited by 32 publications

References 78 publications

In vivo antral follicle wall biopsy: a new research technique to study ovarian function at the cellular and molecular levels

In vivo antral follicle wall biopsy: a new research technique to study ovarian function at the cellular and molecular levels

A Nano-Platform for in situ Investigations of Ovarian Follicles

Trends in recombinant protein use in animal production

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