RFRP3 influences basal lamina degradation, cellular death, and progesterone secretion in cultured preantral ovarian follicles from the domestic cat.

The hypothalamic neuropeptide RFRP3 can suppress hypothalamic GnRH neuron activation and inhibit gonadotropin release from the anterior pituitary. RFRP3 is also produced locally in the ovary and can inhibit steroidogenesis and follicle development in many vertebrates. However, almost nothing is known about the presence and regulatory action of RFRP3 in gonads of any carnivore species. Such knowledge is important for developing captive breeding programs for endangered carnivores and for inhibiting reproduction in feral species. Using the domestic cat as a model, our objectives were to (1) demonstrate the expression of feline RFRP3 (fRFRP3) and its receptor in the cat ovary and (2) assess the influence of fRFRP3 on ovarian follicle integrity, survival, and steroidogenesis in vitro. We first confirmed that fRFRP3 and its receptors (NPFFR1 and NPFFR2) were expressed in cat ovaries by sequencing PCR products from ovarian RNA. We then isolated and cultured preantral ovarian follicles in the presence of 10 or 1 µM fRFRP3 + FSH (1 µg/mL). We recorded the percentage of morphologically viable follicles (basal lamina integrity) over 8 days and calculated percentage survival of follicles on Day 8 (using fluorescent markers for cell survival and death). Last, we quantified progesterone accumulation in media. 10 µM fRFRP3 had no observable effect on viability, survival, or steroid production compared to follicles exposed to only FSH. However, 1 µM fRFRP3 decreased the percentage of morphologically viable follicles and the percentage of surviving follicles on Day 8. At the same time, 1 µM fRFRP3 increased the accumulation of progesterone in media. Our study shows, for the first time, direct action of RFRP3 on the follicle as a functional unit, and it is the first in a carnivore species. More broadly, our results support a conserved, inhibitory action of RFRP3 on ovarian follicle development and underscore the importance of comparative functional studies

UNIFORM SPERM MORPHOLOGY IN THE LEK-BREEDING WIRE-TAILED MANAKIN (PIPRA FILICAUDA)

Abstract ∙ When females copulate with multiple males, selection on spermatozoa can reduce variation in sperm morphology. We describe sperm morphology for a polygynous lek-breeding suboscine, the Wire-tailed Manakin (Pipra filicauda). Total sperm length averaged 41.5 ± 0.7 μm and the among-individual coefficient of variation in total sperm length was 1.8%. Variation was considerably lower than in the other manakin species with known sperm morphology, the Lance-tailed Manakin (Chiroxiphia lanceolata), despite similar promiscuity levels. This result highlights the need for further work on spermatozoa in lek-breeding species. Resumen ∙ Morfología uniforme en el esperma del Saltarín Uirapuru (Pipra filicauda), una especie con sistema de apareamiento de lek Cuando las hembras copulan con más de un macho, selección actuando al nivel del espermatozoide puede reducir la variación en la morfología del esperma. Aquí describimos la morfología del esperma para una especie poligínica de suboscín con sistema de apareamiento de lek, el Saltarín Uirapuru (Pipra filicauda). Los espermatozoides tuvieron una longitud total promedio de 41.5 ± 0.7 μm, y el coeficiente de variación para la longitud total fue de 1.8%. El nivel de variación fue menor que en la otra especie de saltarín estudiada al respecto, el Saltarín Lanceolado (Chiroxiphia lanceolata), aunque ambas especies tienen casi el mismo nivel de promiscuidad. Estos resultados sugieren la necesidad de más estudios sobre especies de aves con este sistema de apareamiento

Biobanking efforts and new advances in male fertility preservation for rare and endangered species

Understanding and sustaining biodiversity is a multi-disciplinary science that benefits highly from the creation of organized and accessible collections of biomaterials (Genome Resource Banks). Large cryo-collections are invaluable tools for understanding, cataloging, and protecting the genetic diversity of the world′s unique animals and plants. Specifically, the systematic collection and preservation of semen from rare species has been developed significantly in recent decades with some biobanks now being actively used for endangered species management and propagation (including the introduction of species such as the black-footed ferret and the giant panda). Innovations emerging from the growing field of male fertility preservation for humans, livestock species, and laboratory animals are also becoming relevant to the protection and the propagation of valuable domestic and wild species. These new approaches extend beyond the "classical" methods associated with sperm freezing to include testicular tissue preservation combined with xenografting or in vitro culture, all of which have potential for rescuing vast amounts of unused germplasm. There also are other options under development that are predicted to have a high impact within the next decade (stem cell technologies, bio-stabilization of sperm cells at ambient temperatures, and the use of genomics tools). However, biobanking efforts and new fertility preservation strategies have to expand the way beyond mammalian species, which will offer knowledge and tools to better manage species that serve as valuable biomedical models or require assistance to reverse endangerment

Production in vitro d'embryons de cervidés mise en évidence d'un effet paternel au début du développement de l'embryon caractérisation à l'aide du modèle bovin

TOURS-BU Sciences Pharmacie (372612104) / SudocPARIS-Museum-Bib zoologie mam. (751052312) / SudocSudocFranceF

Transfer of Galectin-3-Binding Protein via Epididymal Extracellular Vesicles Promotes Sperm Fertilizing Ability and Developmental Potential in the Domestic Cat Model

Key proteins transferred by epididymal extracellular vesicles (EVs) to the transiting sperm cells contribute to their centrosomal maturation and developmental potential. Although not reported in sperm cells yet, galectin-3-binding protein (LGALS3BP) is known to regulate centrosomal functions in somatic cells. Using the domestic cat model, the objectives of this study were to (1) detect the presence and characterize the transfer of LGALS3BP via EVs between the epididymis and the maturing sperm cells and (2) demonstrate the impact of LGALS3BP transfer on sperm fertilizing ability and developmental potential. Testicular tissues, epididymides, EVs, and spermatozoa were isolated from adult individuals. For the first time, this protein was detected in EVs secreted by the epididymal epithelium. The percentage of spermatozoa with LGALS3BP in the centrosome region increased as cells progressively incorporated EVs during the epididymal transit. When LGALS3BP was inhibited during in vitro fertilization with mature sperm cells, less fertilized oocytes and slower first cell cycles were observed. When the protein was inhibited in epididymal EVs prior to incubation with sperm cells, poor fertilization success further demonstrated the role of EVs in the transfer of LGALS3BP to the spermatozoa. The key roles of this protein could lead to new approaches to enhance or control fertility in clinical settings

Transfer of Galectin-3-Binding Protein via Epididymal Extracellular Vesicles Promotes Sperm Fertilizing Ability and Developmental Potential in the Domestic Cat Model

Key proteins transferred by epididymal extracellular vesicles (EVs) to the transiting sperm cells contribute to their centrosomal maturation and developmental potential. Although not reported in sperm cells yet, galectin-3-binding protein (LGALS3BP) is known to regulate centrosomal functions in somatic cells. Using the domestic cat model, the objectives of this study were to (1) detect the presence and characterize the transfer of LGALS3BP via EVs between the epididymis and the maturing sperm cells and (2) demonstrate the impact of LGALS3BP transfer on sperm fertilizing ability and developmental potential. Testicular tissues, epididymides, EVs, and spermatozoa were isolated from adult individuals. For the first time, this protein was detected in EVs secreted by the epididymal epithelium. The percentage of spermatozoa with LGALS3BP in the centrosome region increased as cells progressively incorporated EVs during the epididymal transit. When LGALS3BP was inhibited during in vitro fertilization with mature sperm cells, less fertilized oocytes and slower first cell cycles were observed. When the protein was inhibited in epididymal EVs prior to incubation with sperm cells, poor fertilization success further demonstrated the role of EVs in the transfer of LGALS3BP to the spermatozoa. The key roles of this protein could lead to new approaches to enhance or control fertility in clinical settings

Conservation Biology and Reproduction in a Time of Developmental Plasticity

The objective of this review is to ask whether, and how, principles in conservation biology may need to be revisited in light of new knowledge about the power of epigenetics to alter developmental pathways. Importantly, conservation breeding programmes, used widely by zoological parks and aquariums, may appear in some cases to reduce fitness by decreasing animals’ abilities to cope when confronted with the ‘wild side’ of their natural habitats. Would less comfortable captive conditions lead to the selection of individuals that, despite being adapted to life in a captive environment, be better able to thrive if relocated to a more natural environment? While threatened populations may benefit from advanced reproductive technologies, these may actually induce undesirable epigenetic changes. Thus, there may be inherent risks to the health and welfare of offspring (as is suspected in humans). Advanced breeding technologies, especially those that aim to regenerate the rarest species using stem cell reprogramming and artificial gametes, may also lead to unwanted epigenetic modifications. Current knowledge is still incomplete, and therefore ethical decisions about novel breeding methods remain controversial and difficult to resolve

Three-dimensional culture of endometrial cells from domestic cats: A new in vitro platform for assessing plastic toxicity.

Plastic polymers can be combined with additives that modify physical properties and stability of the material. However, the biocompatibility of those additives is not well known. The objective of the study was to characterize the impact of zinc stearate-a common additive-through the development of a novel three-dimensional (3-D) in vitro platform with endometrial cells from domestic cats. Epithelial and stromal cells from adult uteri were isolated and cultured in medium supplemented with 3% Matrigel for two weeks in plastic tissue culture dishes that had been identified as polystyrene with and without zinc stearate by Raman, FTIR, and X-ray fluorescence spectroscopies. Three-dimensional cell structures that were obtained were measured and categorized by shape. Cell viability, proliferation, differentiation, organization, and apoptosis then were assessed by immuno-staining. Results indicated that zinc stearate did not affect 3-D endometrial cell structure morphology, viability, or cellular composition. This first study of a new in vitro platform will be useful for studies testing the influence of other additives, drugs, or exogenous hormones

Reproductive biotechnologies for endangered mammalian species

Assisted reproductive techniques (gamete cryopreservation, artificial insemination, embryo transfer, and in vitro fertilization) allow to propagate small fragmented populations of wild endangered species or domestic breeds. There are the best way for producing several offspring from selected genitors in order to avoid inbreeding depression. However, few mammalian species have been well studied for their reproductive biology whereas huge differences have been observed between these species. Furthermore, materials, methods and experimental designs have to be adapted for each case and each limiting factor (wildness, poor quantity of biological material, disparate locations). Genome resource banking is currently arising and the most applied reproductive biotechnology remains artificial insemination. Assisted reproductive techniques currently developed in domestic species (intra-cytoplasmic sperm injection, nuclear transfer) may offer new opportunities for the propagation of endangered species.Biotechnologies de la reproduction appliquées aux mammifères en voie de disparition. Les techniques de reproduction assistée (cryoconservation des gamètes, insémination artificielle, transfert embryonnaire, fécondation in vitro) permettent d'accroître des populations, parfois dispersées géographiquement, d'espèces sauvages ou domestiques en voie de disparition. Ces méthodes sont le meilleur moyen pour produire plusieurs descendants à partir de géniteurs sélectionnés de façon à éviter la consanguinité. La biologie de la reproduction est connue pour peu d'espèces de mammifères alors que de grandes différences ont pourtant été mises en évidence entre ces espèces. En outre, les matériels, les méthodes, ainsi que les schémas expérimentaux doivent être adaptés à chaque cas et pour chaque facteur limitant (animaux sauvages, peu de matériel biologique, populations dispersées). Des banques génétiques sont actuellement en voie de constitution mais la technique la plus utilisée reste l'insémination artificielle. Cependant, des biotechnologies de la reproduction en cours de mise au point chez les animaux domestiques (injection intra-cytoplasmique de spermatozoïde, transfert nucléaire) pourraient offrir de nouvelles possibilités pour l'accroissement de populations menacées