Prokineticin ligands and receptors are expressed in the human fetal ovary and regulate germ cell expression of COX2

CONTEXT: Fetal ovarian development and primordial follicle formation underpin future female fertility. Prokineticin (PROK) ligands regulate cell survival, proliferation and angiogenesis in adult reproductive tissues including the ovary. However, their expression and function during fetal ovarian development remains unclear.

OBJECTIVE: To investigate expression and localization of the PROK ligands, receptors and their downstream transcriptional targets in the human fetal ovary.

SETTING: This study was conducted at the University of Edinburgh.

PARTICIPANTS: Ovaries were collected from 37 morphologically normal human fetuses.

DESIGN AND MAIN OUTCOME MEASURES: mRNA and protein expression of PROK ligands and receptors was determined in human fetal ovaries using qRT-PCR, immunoblotting and immunohistochemistry. Functional studies were performed using a human germ tumour cell line (TCam-2) stably transfected with PROKR1.

RESULTS: Expression of PROK1 and PROKR1 was significantly higher in mid-gestation ovaries (17-20 weeks) than at earlier gestations (8-11 and 14-16 weeks). PROK2 significantly increased across the gestations examined. PROKR2 expression remained unchanged. PROK ligand and receptor proteins were predominantly localised to germ cells (including oocytes within primordial follicles) and endothelial cells, indicating these cell types to be the targets of PROK signalling in the human fetal ovary. PROK1 treatment of a germ cell line stably-expressing PROKR1 resulted in ERK phosphorylation, and elevated COX2 expression.

CONCLUSIONS: Developmental changes in expression and regulation of COX2 and pERK by PROK1 suggest that PROK ligands may be novel regulators of germ cell development in the human fetal ovary, interacting within a network of growth and survival factors prior to primordial follicle formation.

Analgesic exposure in pregnant rats affects fetal germ cell development with inter-generational reproductive consequences

Analgesics which affect prostaglandin (PG) pathways are used by most pregnant women. As germ cells (GC) undergo developmental and epigenetic changes in fetal life and are PG targets, we investigated if exposure of pregnant rats to analgesics (indomethacin or acetaminophen) affected GC development and reproductive function in resulting offspring (F1) or in the F2 generation. Exposure to either analgesic reduced F1 fetal GC number in both sexes and altered the tempo of fetal GC development sex-dependently, with delayed meiotic entry in oogonia but accelerated GC differentiation in males. These effects persisted in adult F1 females as reduced ovarian and litter size, whereas F1 males recovered normal GC numbers and fertility by adulthood. F2 offspring deriving from an analgesic-exposed F1 parent also exhibited sex-specific changes. F2 males exhibited normal reproductive development whereas F2 females had smaller ovaries and reduced follicle numbers during puberty/adulthood; as similar changes were found for F2 offspring of analgesic-exposed F1 fathers or mothers, we interpret this as potentially indicating an analgesic-induced change to GC in F1. Assuming our results are translatable to humans, they raise concerns that analgesic use in pregnancy could potentially affect fertility of resulting daughters and grand-daughters.

Manipulations of germ-cell populations in the gonad of the fowl

1. Embryos of the domestic fowl have been partially sterilised by injecting the drug busulphan into 24-h incubated eggs. 2. Some of these embryos were injected with primordial germ cells (PGCs) after 55 h of incubation to attempt to repopulate the gonads. 3. Primordial germ cells transfected with a defective retrovirus containing the reporter gene lac Z were shown to settle in these sterilised gonads. 4. Quantitative histology of 6-d embryos showed that busulphan produced 75% sterilisation but that PGCs could repopulate these gonads. 5. The technique of producing such germ line chimaeras is of value in studying cell kinetics, gonad differentiation and the production of transgenics.

Germ Cell Transplantation in Felids: A Potential Approach to Preserving Endangered Species

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Spontaneous germ cell death by apoptosis in epididymis of the adult bat Artibeus lituratus

Many factors can lead cells to apoptosis during the various stages of cell life. This study was undertaken to characterize germ cell death in the epididymis of the adult Artibeus lituratus by histochemical and immunohistochemical techniques using light microscopy and transmission electron microscopy. The results showed that cells with a nuclear phenotype and ultrastructural characteristics of chromatin compaction were common in apoptosis. The Apoptag test confirmed that the suspected cells were apoptotic. It is suggested that immature germ cells, when released from the germinative epithelium, may be directed towards the epididymis instead of being disposed of in the testicle. Furthermore, intact immature cells can leave the testicle in the initial phases of apoptosis and complete this phenomenon in the epididymis.

Germ Cell Dynamics during the Annual Reproductive Cycle of Dendropsophus minutus (Anura: Hylidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Mixed germ cell tumor of the pituitary-hypothalamic region presenting as craniopharyngioma: Case report and review of the literature

Craniopharyngiomas and germ cell tumors (GCT) may affect the pituitary-hypothalamic region during childhood. Although different in origin, their clinical and radiological features may be similar. In this article we present a 5-year-old girl with clinical and radiological findings (computer tomography calcification) that were initially considered as craniopharyngioma. However clinical outcome, blood and cerebral spinal fluid tumoral markers, and results from anatomopathology and immunohistochemistry disclosed a mixed GCT. This case report highlights that some clinical features and radiological findings of pituitary-hypothalamic tumors may be misdiagnosed as craniopharyngioma mainly when there is a mature teratoma with cartilaginous tissue differentiation. Copyright© ABE&M.

An overview of functional and stereological evaluation of spermatogenesis and germ cell transplantation in fish

Although there are almost thirty-thousand species of fish living in a great variety of habitats and utilizing vast reproductive strategies, our knowledge of morphofunctional and quantitative aspects of testis structure and spermatogenesis is still incipient for this group of vertebrates. In this review, we discuss aspects that are important to better understanding of testis structure and function, and of the development of germ cells (GC) during spermatogenesis. To achieve this, we have recently completed a number of studies presenting morphometric and functional data related to the numbers of GC and Sertoli cells (SC) per each type of spermatogenic cyst, the number of spermatogonial generations, the SC efficiency, and the magnitude of GC loss that normally occurs during spermatogenesis. We also investigated SC proliferation and the relationship of this important event to early spermatogenic cysts. The available data strongly suggest that SC proliferation in sexually mature tilapia is the primary factor responsible for the increase in testis size and for determination of the magnitude of sperm production. The influence of temperature on the duration of spermatogenesis in tilapia was also evaluated and we have used this knowledge to deplete endogenous spermatogenesis in this teleost, in order to develop an experimental system for GC transplantation. This exciting technique results in new possibilities for investigation of spermatogenesis and spermatogonial stem cell biology, creating also an entirely new and promising scenario in biotechnology - transgenic animal production and the preservation of the genetic stocks of valuable animals or endangered species. © Springer Science+Business Media B.V. 2008.

Female germ cell renewal during the annual reproductive cycle in Ostariophysians fish

The objective was to characterize female germ cell renewal during the annual reproductive cycle in two species of ostariophysian fish with distinct reproductive strategies: a siluriform, Pimelodus maculatus, in which oocyte development is group synchronous and the annual reproductive period is short; and a characiform, Serrasalmus maculatus, with asynchronous oocyte development and a prolonged reproductive period. These reproductive strategies result in fish determinate and indeterminate fecundity, respectively. Annual reproductive phases were determined by biometric and histologic analysis of gonads and interpreted according to new proposals for phase classification and stages of oocyte development (with special attention to germinal epithelium activity). Histologically, there were two types of oogonia in the germinal epithelium: single oogonia and those in mitotic proliferation. Oogonial proliferation and their entry into meiosis resulted in formation of cell nests (clusters of cells in the ovarian lamellae). Morphometric analysis was used to estimate germ cell renewal. Based on numbers of single oogonia in the lamellar epithelium, and nests with proliferating oogonia or early prophase oocytes throughout the annual reproductive cycle, oogonial proliferation and entrance into meiosis were more intense during the regenerating phase and developing phase, but decreased sharply (P < 0.05) during the spawning-capable phase. Oogonial proliferation gradually recovered during the regressing phase. We concluded that, independent of species or features of the reproductive cycle, germ cell renewal occurred during the regenerating phase, ensuring availability of eggs for the spawning event. © 2013 Elsevier Inc.

Genomic screening of testicular germ cell tumors from monozygotic twins

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Should extragonadal germ cell tumors be included in studies of families with testicular germ cell tumors?

Abstract Background Family history is among the few established risk factors for testicular germ cell tumor (TGCT). Approximately 1.4% of newly diagnosed TGCT patients report a positive family history of TGCT. Sons and siblings of TGCT patients have four- to six fold and eight- to tenfold increase in TGCT risk, respectively. In twins of men with TGCT the relative risk of testicular cancer is 37.5 (12.3-115.6). Nevertheless, information about the occurrence of TGCT in relatives of patients with extragonadal germ cell tumor is limited. Case report A 24 year-old male patient was diagnosed with a mediastinum tumor and was submitted to image-guided biopsy, which revealed a seminoma. Two months later, his non-identical asymptomatic twin brother was submitted to an elective ultrasound of the testes, which showed a left testicular mass of 4.2 cm. This patient underwent orchiectomy revealing a seminoma of the left testis. There are no other cases of seminoma or other types of cancers reported in first-degree relatives in this family. Conclusions Although familial aggregations of TGCT have been well described, to the best of our knowledge, no data concerning the association of gonadal and extragonadal germ cell tumor in relatives has been previously reported. Further investigation on this association is warranted and may help in improving our knowledge of familial pattern inheritance.