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We evaluated the influence of different media plus various concentrations of Glial cell line‐derived neurotrophic factor (GDNF) during the in vitro culture (IVC) of testicular tissues from prepubertal collared peccary. Testes from 5 individuals were collected, fragmented and cultured for 28 days (34°C and 5% CO2). Culture media were Dulbecco's modified essential medium (DMEM) or stem cell serum free media (StemPro‐34™ SFM), both supplemented with various concentrations of GDNF (0, 10, or 20 ng/mL). Fragments were cultured on the flat surface of 0.75% agarose gel and were evaluated every 7 days for fragment area, histomorphology, cellular viability, and proliferative activity. Data were expressed as mean ± standard error and analyzed by Kruskal–Wallis's and Tukey test. Fragments area decreased over the 28 days‐culture, regardless of the treatment. For morphology, the StemPro‐37 SFM medium plus 10 ng/mL GDNF provided higher scores at all time points in comparison to DMEM using any GDNF concentration (p <.05). After 28 days, similar cellular viability (~70%) was observed in all treatments (p >.05). For proliferating cell nuclear antigen assay, only DMEM plus 10 ng/mL GDNF improved (p <.05) cellular proliferation on Days 14 and 28. Looking at argyrophilic nucleolar organizing regions, after 28 days, there were no differences among treatments regarding cell proliferative capacity for both spermatogonia and Sertoli cells (p >.05). In summary, the DMEM and StemPro‐34 SFM are adequate medium for IVC of prepubertal peccary testicular tissue. Supplementation with GDNF, especially at a 10 ng/mL concentration, appears to be essential for the maintenance of cell survival and proliferation. [ABSTRACT FROM AUTHOR]

Rodents can present a large number of morphophysiological variations of the digestive system, allowing them to feed on different diets. In this context, the objective was to describe the morphology of the major salivary glands of Galea spixii. Gross dissection, light microscopy and transmission and scanning electron microscopy assessments of 12 specimens were carried out. The results demonstrate that the G. spixii has four pairs of major salivary glands (parotid, mandibular, zygomatic and sublingual). The glands are lobulated, composed acinar, consisting of mucous and serous acini, with the predominant parotid gland being serous and the zygomatic gland mucous. The mandibular and sublingual glands are mixed. The glands secrete acidic and neutral mucins and commonly present intercalary, striated and excretory ducts. The ultrastructure of the acinar cells suggests the presence of great cellular activity, with a cytoplasm taken by a rough endoplasmic reticulum, many mitochondria, vesicles of electrolucent nature and dispersed chromatin. In conclusion, the major salivary glands of G. spixii are constituted of mucous and serous acini, with serous acini predominant in the mandibular areas. The data also suggest that these rodents may adopt different types of diets, which may explain their adaptability to semi‐arid environments. [ABSTRACT FROM AUTHOR]

Fibroblast cycle synchronization in G0/G1 is an essential step for nuclear reprogramming by cloning or induced cells to pluripotency. Considering the diversity among rodents and the ecological and scientific importance of these animals, we compared the contact inhibition, serum starvation, and 10 µM of roscovitine as methods of synchronization of red-rumped agouti fibroblasts. The effects of each protocol were evaluated on the percentage of cycle phase, morphology, viability, and apoptosis levels. The results showed that culturing the cells to serum starvation for 24 h (75.9%), 48 h (81.6%), 72 h (86.2%), 96 h (84.0%), and 120 h (83.7%) yielded a significantly higher percentage of cells arrested in the G0/G1 (P < 0.05) phase than cells not subjected to any cell cycle synchronization method (31.4%). Also, this effect was not different between the times of 48 and 120 h (P > 0.05). A similar response was observed for cells cultured with roscovitine for 12 h (86.9%), 24 h (74.8%), and 48 h (81.7%), with a higher percentage of synchronized cells in G0/G1 compared to cells not submitted to any synchronization treatment (52.2%). Nevertheless, this effect was best evidenced at 12 h (P < 0.05). Also, the contact inhibition for 24–120 h could not synchronize cells in G0/G1, with values ranging from 70.9 to 77.9% (P > 0.05). Moreover, no difference was observed for morphology, viability, and apoptosis levels in any synchronization method (P > 0.05). Therefore, serum starvation is as efficient as roscovitine on cycle synchronization in G0/G1 of red-rumped agouti fibroblasts. [ABSTRACT FROM AUTHOR]

Context: Several assisted reproduction techniques have been proposed for collared peccaries due to the interest in its conservation. However, there is little information about the requirements for sperm capacitation, an initial step in in vitro fertilisation. Aims: We aimed to determine the optimal conditions for collared peccary sperm capacitation by comparing Tyrode's albumin lactate pyruvate (TALP) or the same media plus heparin, caffeine, or their combination at different exposure times (1, 3, and 6 h). Methods: The samples were evaluated for kinetic parameters, membrane functionality and integrity, mitochondrial activity, morphology, DNA integrity, reactive oxygen species (ROS) expression levels, capacitation, and sperm binding ability using swine oocytes. Key results: Samples incubated with caffeine or heparin had a higher percentage of capacitated spermatozoa. The maximum percentage of capacitation was achieved after 3 h of incubation with either agent. Moreover, spermatozoa subjected to heparin capacitation showed better motility than when subjected to caffeine, and lower ROS expression levels after 1 h. No differences were observed among incubation times for the binding ability. Conclusions: In summary, collared peccary spermatozoa can be capacitated with caffeine or heparin; however, heparin better maintains sperm motility and ROS expression levels. The co-incubation of gametes in a medium with heparin for 3 h could be efficient for in vitro fertilisation in collared peccaries. Implications: This result will contribute to the development of assisted reproduction techniques for conservation and productivity of collared peccaries. In vitro fertilisation can contribute to the conservation and knowledge of the reproductive characteristics of collared peccary. There is little information about the requirements for sperm capacitation in the collared peccary, an initial step in in vitro fertilisation. We compared different capacitating media and incubation times and we observed that heparin improved sperm motility and promoted the capacitation of collared peccary spermatozoa. This result will contribute to the development of assisted reproduction techniques for conservation and productivity of collared peccaries. [ABSTRACT FROM AUTHOR]

Background: Hystricomorpha rodents display a similar placentation model to humans. The present study was carried out considering the scarcity of information concerning the placental development in agouti. Objective: Describe the microscopy of the placenta, subplacenta and yolk sac of agoutis in early pregnancy and report on the inversion of the yolk sac. Methods: Fifteen females between the 14th-32nd day of gestation were used following euthanasia. Gestational buttons were collected, fixed, processed, stained to optical microscopy or immunohistochemistry. Results: Chorioallantoic placenta (CP) ranged from conical to a half-sphere, as follows: from the 14th to 17th day, the CP displays an inverted "V" shape, predominantly formed by cytotrophoblasts; from 20 to 22 days, formed almost entirely by cytotrophoblasts; at 28 days, a half sphere, with distinct lobes and interlobular area, numerous maternal gaps delimited by syncytiotrophoblasts and trophoblast giant cells; at 32 days, globose and undergoing the maturation process. Subplacenta, located between decidua and CP, initially presents septa consisting of simple columnar epithelium and after 17 days, comprising stratified epithelium. Visceral yolk sac (VYS) is attached to two CP projections between 14 and 17 days, formed by a simple cubic epithelium and inverted. Between 20 and 22 days, the epithelium displays apical villous projections with cytoplasmic vacuoles and a vascularized mesoderm. After the 24 th day, the VYS near the placenta is pleated, very vascularized and villous, with decreased villi sizes further away from the placenta. Conclusion: The agouti CP displays similar characteristics to other hystricomorpha, including placenta lobulation, a subplacenta and an inverted vitelline placenta. [ABSTRACT FROM AUTHOR]