The comparative aspects of hystricomorph subplacenta: potential endocrine organ.

BACKGROUND: The placenta of hystricomorph rodents, lagomorphs and some primates includes an unusual structure, termed a subplacenta, which essentially consists of trophoblastic cells located deep to the central implantation site within the area of decidualization. It has been suggested that the subplacenta is functionally important, although considerable controversy remains on the issue. In this context, our objective was to compare the architecture and structure of the subplacentas of different hystricomorph species, to investigate the possibility that it is active in hormone synthesis. METHODS: In total, the placentas of 3 capybaras (Hydrochaeris hydrochaeris), 2 pacas (Agouti paca), 5 agoutis (Dasyprocta leporina), 5 rock cavies (Kerodon rupestris) and 3 guinea pigs (Cavia porcellus) at different stages of pregnancy (early, middle and near term) were used for gross and microscopic examination. This included the preparation of latex injection casts, immunohistochemistry for steroidogenic enzymes, scanning and transmission electron microscopy. Tissue steroid concentrations were also determined. RESULTS: The gross morphology and microvascular arrangement of the subplacentas were similar among the hystricomorphs studied including ultra-structural verification of cytotrophoblast and syncytiotrophoblast in all species. In guinea pigs, trophoblast cells exhibited characteristics consistent with intense metabolic and secretory activity in general. However, immuno-histochemical evidence also indicated that subplacental trophoblast expressed key steroidogenic enzymes, mainly in the chorionic villus region, consistent with tissue steroid concentrations. CONCLUSIONS: The subplacentas within placentas of hystricomorph rodent species are structurally similar and, in guinea pigs, have potential for steroid hormone secretion from, at least the early stages of pregnancy.

The Tumor Microenvironment: Focus on Extracellular Matrix.

The extracellular matrix (ECM) regulates the development and maintains tissue homeostasis. The ECM is composed of a complex network of molecules presenting distinct biochemical properties to regulate cell growth, survival, motility, and differentiation. Among their components, proteoglycans (PGs) are considered one of the main components of ECM. Its composition, biomechanics, and anisotropy are exquisitely tuned to reflect the physiological state of the tissue. The loss of ECM's homeostasis is seen as one of the hallmarks of cancer and, typically, defines transitional events in tumor progression and metastasis. In this chapter, we discuss the types of proteoglycans and their roles in cancer. It has been observed that the amount of some ECM components is increased, while others are decreased, depending on the type of tumor. However, both conditions corroborate with tumor progression and malignancy. Therefore, ECM components have an increasingly important role in carcinogenesis and this leads us to believe that their understanding may be a key in the discovery of new anti-tumor therapies. In this book, the main ECM components will be discussed in more detail in each chapter.

Resiliency of equid H19 imprint to somatic cell reprogramming by oocyte nuclear transfer and genetically induced pluripotency†.

Cell reprogramming by somatic cell nuclear transfer and in induced pluripotent stem cells is associated with epigenetic modifications that are often incompatible with embryonic development and differentiation. For instance, aberrant DNA methylation patterns of the differentially methylated region and biallelic expression of H19-/IGF2-imprinted gene locus have been associated with abnormal growth of fetuses and placenta in several mammalian species. However, cloned horses are born with normal sizes and with no apparent placental anomalies, suggesting that H19/IGF2 imprinting may be epigenetically stable after reprogramming in this species. In light of this, we aimed at characterizing the equid H19 gene to determine whether imprinting is altered in somatic cell nuclear transfer (SCNT)-derived conceptuses and induced pluripotent stem cell (iPSC) lines using the mule hybrid model. A CpG-rich region containing five CTCF binding sites was identified upstream of the equine H19 gene and analyzed by bisulfite sequencing. Coupled with parent-specific and global H19 transcript analysis, we found that the imprinted H19 remains monoallelic and that on average the methylation levels of both parental differentially methylated regions in embryonic and extra-embryonic SCNT tissues and iPSC lines remained unaltered after reprogramming. Together, these results show that, compared to other species, equid somatic cells are more resilient to epigenetic alterations to the H19-imprinted locus during SCNT and iPSC reprogramming.

Optimization of Canine Placenta Decellularization: An Alternative Source of Biological Scaffolds for Regenerative Medicine.

Due to the scarcity of tissues and organs for transplantation, the demand for bioengineered tissues is increasing with the advancement of technologies and new treatments in human and animal regenerative medicine. Thus, decellularized placental extracellular matrix (ECM) has emerged as a new tool for the production of biological scaffolds for subsequent recellularization and implantation for recovery of injured areas or even for replacement of organ and tissue fractions. To be classified as an ideal biological scaffold, the ECM must be acellular and preserve its proteins and physical features to be useful for cellular adhesion. In this context, we developed a process of decellularization of canine placentas with 35 and 40 days of gestation using dodecyl sulfate sodium under immersion and agitation in sterile conditions. Before use of this scaffold in recellularization processes, the decellularization efficiency needs to be confirmed by the absence of cellular content and an irrelevant amount of reminiscent DNA. Both vasculature architecture and ECM proteins, such as collagen types I, III, and IV, laminin, and fibronectin, were preserved with our method. In this way, we established a new biological scaffold model that could be used for recellularization in regenerative medicine of tissues.

Canine Placenta Recellularized Using Yolk Sac Cells with Vascular Endothelial Growth Factor.

Regenerative medicine has been growing because of the emergent need for tissues/organs for transplants and restorative surgeries. Biological scaffolds are important tools to try to solve this problem. The one used in this reserach was developed by an acellular biological scaffold from canine placenta with a rich source of cellular matrix. After decellularization, the cellular matrix demonstrated structural preservation with the presence of important functional proteins such as collagen, fibronectin, and laminin. We used cells transduced with vascular endothelial growth factor (VEGF) to recellularize this scaffold. It was succeeded by seeding the cells in nonadherent plaques in the presence of the sterelized placenta scaffold. Cells were adhered to the scaffold when analyzed by immunocytochemistry and scanning electron microscopy, both showing sprouting of yolk sac VEGF (YSVEGF) cells. This recellularized scaffold is a promissory biomaterial for repairing injured areas where neovascularization is required.

Morphological characteristics of mule conceptuses during early development.

Hybrids between species are often infertile and extremely rare among mammals. Mules, i.e. crossing between the horse and the donkey, on the other hand are very common in agricultural and leisure practices due to their enhanced post-natal physical characteristics that is believed to occur for outbreeding or hybrid vigor. Since no reports are availableon the effects of hybrid vigor during early development, this study focused on characterizing the intrauterine development of mule conceptuses during critical embryo-to-fetus transition period. Nine embryos and fetuses of early gestation, obtained after artificial insemination and transcervical flushing, were evaluated by means of gross anatomy and histology and compared to data available for the equine. We found that some events, such as C-shape turning, apearence of branchial archs, limb and tail buds, formation of primary and secondary brain vesicles, heart compartmentalization, and development of somites, occurred slightly earlier in the mule. Nonetheless, no major differences were observed in other developmental features, suggesting similarities between the mule and the horse development. In conclusion, these data suggest that the effect of hybrid vigor is present during intrauterine development in the mule, at least with regard to its maternal parent.

Organogenesis of the Musculoskeletal System in Horse Embryos and Early Fetuses.

Musculoskeletal system development involves heterotypical inductive interactions between tendons, muscles, and cartilage and knowledge on organogenesis is required for clarification of its function. The aim of this study was to describe the organogenesis of horse musculoskeletal system between 21 and 105 days of gestation, using detailed macroscopic and histological analyses focusing on essential developmental steps. At day 21 of gestation the skin was translucid, but epithelial condensation and fibrocartilaginous tissues were observed on day 25 of pregnancy. Smooth muscle was seen in lymphatic and blood vessel walls and the beginning of cartilaginous chondrocranium was detected at day 30 of gestation. At day 45, typical chondroblasts and chondrocytes were observed and at day 55, mandibular processes expanded toward the ventral midline of the pharynx. At day 75, muscles became thicker and muscle fibers were seen developing in carpal and metacarpal joints with the beginning of the ossification process. At day 105, major muscle groups, similar to those seen in an adult equine, were observed. The caudal area of the nasal capsule and trabecular cartilages increased in size and became ossified, developing into the ethmoid bone. The presence of nasal, frontal, parietal, and occipital bones was observed. In conclusion, novel features of equine musculoskeletal system development have been described here and each process was linked with an early musculoskeletal event. Data presented herein will facilitate a better understanding of the equine muscular system organogenesis and aid in the detection of congenital deformities. Anat Rec, 299:722-729, 2016. © 2016 Wiley Periodicals, Inc.

Microvascularization of corpus luteum of bovine treated with equine chorionic gonadotropin.

This study aimed to evaluate the morphological changes in microvascular density and corpus luteum (CL) vascularization in cows treated with eCG during stimulatory and superovulatory protocols. Sixteen cows were synchronized and divided into three groups: control (n = 6), stimulated (n = 4) and superovulated (n =6), one was submitted to estrous synchronization (ES) and received no eCG (control), and those that were submitted to ES and received eCG before or after follicular deviation (superovulation and stimulation of the dominant follicle, respectively). Ovulation was synchronized using a progesterone device-based protocol. After six days of ovulation, the cows were slaughtered and the ovaries and CL were collected. The CLs were processed and photomicrographs were taken under light microscopy to assess the vascular volume density (Vv) by stereology, and scanning electron microscopy (SEM) was used to perform ultrastructural analysis of the microvasculature. The Vv in stimulated and superovulated cows significantly increased (P ≤ 0.0001) when compared to control, indicating that the eCG is able to induce angiogenic activity in bovine CL. However, no significant differences were observed between stimulated and superovulated cows. The SEM demonstrated ratings indicative of angiogenesis, marked by several button-shaped projections in the capillaries, and the presence of more dilated capillaries in CL treated with eCG. These morphological findings are evidence of an angiogenic effect of the eCG treatment in CL of cows.