Morphometric study of the primary ossification center of the frontal squama in the human fetus.

PURPOSES: Detailed morphometric data on the development of ossification centers in human fetuses is useful in the early detection of skeletal dysplasias associated with a delayed development of ossification centers and their mineralization. Quantitative analysis of primary ossification centers of cranial bones is sporadic due to limited availability of fetal material. MATERIAL AND METHODS: The size of the primary ossification center of the frontal squama in 37 human (16 males and 21 females) spontaneously aborted human fetuses aged 18-30 weeks was studied by means of CT, digital-image analysis and statistics. RESULTS: With neither sex nor laterality differences, the best-fit growth dynamics for the primary ossification center of the frontal squama was modelled by the following functions: y = 13.756 + 0.021 × (age)2 ± 0.024 for its vertical diameter, y = 0.956 + 0.956 × age ± 0.823 for its transverse diameter, y = 38.285 + 0.889 × (age)2 ± 0.034 for its projection surface area, and y = 90.020 + 1.375 × (age)2 ± 11.441 for its volume. CONCLUSIONS: Our findings for the primary ossification center of the frontal squama may be conducive in monitoring normal fetal growth and screening for inherited faults and anomalies of the skull in human fetuses.

Dynamic and Cell-Specific DACH1 Expression in Human Neocortical and Striatal Development.

DACH1 is the human homolog of the Drosophila dachshund gene, which is involved in the development of the eye, nervous system, and limbs in the fly. Here, we systematically investigate DACH1 expression patterns during human neurodevelopment, from 5 to 21 postconceptional weeks. By immunodetection analysis, we found that DACH1 is highly expressed in the proliferating neuroprogenitors of the developing cortical ventricular and subventricular regions, while it is absent in the more differentiated cortical plate. Single-cell global transcriptional analysis revealed that DACH1 is specifically enriched in neuroepithelial and ventricular radial glia cells of the developing human neocortex. Moreover, we describe a previously unreported DACH1 expression in the human striatum, in particular in the striatal medium spiny neurons. This finding qualifies DACH1 as a new striatal projection neuron marker, together with PPP1R1B, BCL11B, and EBF1. We finally compared DACH1 expression profile in human and mouse forebrain, where we observed spatio-temporal similarities in its expression pattern thus providing a precise developmental description of DACH1 in the 2 mammalian species.

Quantitative description of the morphology and ossification center in the axial skeleton of 20-week gestation formalin-fixed human fetuses using magnetic resonance images.

OBJECTIVES: Human tissues are usually studied using a series of two-dimensional visualizations of in vivo or cutout specimens. However, there is no precise anatomical description of some of the processes of human fetal development. The purpose of our study is to develop a quantitative description of the normal axial skeleton by means of high-resolution three-dimensional magnetic resonance (MR) images, collected from six normal 20-week-old human fetuses fixed in formaldehyde. METHODS: Fetuses were collected after spontaneous abortion and subsequently fixed with formalin. They were imaged using a 1.5 T MR scanner with an isotropic spatial resolution of 200 µm. The correct tissue discrimination between ossified and cartilaginous bones was confirmed by comparing the images achieved by MR scans and computerized axial tomographies. The vertebral column was segmented out from each image using a specially developed semi-automatic algorithm. RESULTS: Vertebral body dimensions and inter-vertebral distances were larger in the lumbar region, in agreement with the beginning of the ossification process from the thoracolumbar region toward the sacral and cephalic ends. CONCLUSION: In this article, we demonstrate the feasibility of using MR images to study the ossification process in formalin-fixed fetal tissues. A quantitative description of the ossification centers of vertebral bodies and arches is presented.

Positive reproductive family history for spontaneous abortion: predictor for recurrent miscarriage in young couples.

OBJECTIVE: The etiology of recurrent spontaneous abortions (RSA) in chromosomally normal parents is still unexplained. It is unclear whether or not some factors, such as spontaneous abortions (SA), which occur among extended family members can create a predisposition to RSA. Therefore, this study comprises two parts: (a) an epidemiological part, to evaluate the relationship between RSA in 567 couples and the frequency of SA among their first (I), second (II) and third (III) generation relatives, and (b) a genetic part, investigating whether parental and fetal chromosomal status may predispose to the occurrence of RSA. STUDY DESIGN: Couples (567) having one or more SA were analyzed in this retrospective case-control study. The family reproductive history data was collected from their medical charts. RESULTS: The total number of SA found in 567 couples was 1174, and the largest number occurred at 8-10 weeks of gestation. The majority of spouses had normal karyotypes (88.5% and 91%). Of the remainder, 65% of females and 76% of males expressed constitutional chromosomal variation, mostly pericentric inversion of chromosome 9. Cytogenetic analysis of aborted material showed some type of change in 40% of cases. The family reproductive history data indicated that SA among the couples' I, II and III generation relatives happened with a frequency two to three times higher than that of the general population (55.5, 47.6 and 32.6% for female relatives, and 45.8, 44.1 and 15.1% for male relatives). CONCLUSION: Positive reproductive family history for SA might be the causal factor for RSA and can also predetermine women that are of greater susceptibility to preterm pregnancy.

Prenatal expression of purinergic receptor P2X3 in human dorsal root ganglion.

The dorsal root ganglion (DRG) is consisted of neurons that relay multiple types of spinal sensory stimuli to the central nervous system. Several neuroactive molecules may be involved in sensory modulation especially pain processing at the DRG, including the purinergic receptor P2X3 and calcitonin-gene-related peptide (CGRP). P2X3 receptor has been considered a promising pharmaceutical target for the development of new pain medicine. Currently, litter is known about the expression of P2X3 in the human DRG. The present study characterized the localization of P2X3 in prenatal human DRG obtained from fetuses at 4-8 gestational months, by comparing to CGRP expression as well as binding pattern of isolectin-B4 (IB4), a marker of small DRG neurons presumably relevant to nociception. P2X3 immunoreactivity (IR) appeared in most neuron-like perikarya, with their numerical density reduced during the gestational period studied. P2X3 IR was co-labeled very commonly with IB4 binding and infrequently with CGRP IR and was not colocalized with IR for the gliocyte marker glutamine synthetase. Together, the data show an early and broad expression of P2X3 in prenatal human DRG neurons, pointing to a biological role of purinergic signaling during the development of spinal sensory system.

The expression of thyroid hormone transporters in the human fetal cerebral cortex during early development and in N-Tera-2 neurodifferentiation.

Associations of neurological impairment with mutations in the thyroid hormone (TH) transporter, MCT8, and with maternal hypothyroxinaemia, suggest that THs are crucial for human fetal brain development. It has been postulated that TH transporters regulate the cellular supply of THs within the fetal brain during development. This study describes the expression of TH transporters in the human fetal cerebral cortex (7­20 weeks gestation) and during retinoic acid induced neurodifferentiation of the human N-Tera-2 (NT2) cell line, in triiodothyronine (T3) replete and T3-depleted media. Compared with adult cortex, mRNAs encoding OATP1A2, OATP1C1, OATP3A1 variant 2, OATP4A1, LAT2 and CD98 were reduced in fetal cortex at different gestational ages, whilst mRNAs encoding MCT8, MCT10, OATP3A1 variant 1 and LAT1 were similar. From the early first trimester, immunohistochemistry localised MCT8 and MCT10 to the microvasculature and to undifferentiated CNS cells. With neurodifferentiation, NT2 cells demonstrated declining T3 uptake, accompanied by reduced expressions of MCT8, LAT1, CD98 and OATP4A1. T3 depletion significantly reduced MCT10 and LAT2 mRNA expression at specific time points during neurodifferentiation but there were no effects upon T3 uptake, neurodifferentiation marker expression or neurite lengths and branching. MCT8 repression also did not affect NT2 neurodifferentiation. In conclusion, many TH transporters are expressed in the human fetal cerebral cortex from the first trimester, which could regulate cellular TH supply during early development. However, human NT2 neurodifferentiation is not dependent upon T3 or MCT8 and there were no compensatory changes to promote T3 uptake in a T3-depleted environment.

The immunolocalization and possible role of c-Met (MET, hepatic growth factor receptor) in the developing human fetal mandibular condyle.

The c-Met system is involved in skeletogenesis and is expressed in the cartilage of growth plates. However, the localization and role of c-Met during endochondral ossification of developing mandibular condyles or during intramembranous ossification has not yet been elucidated. In this study, c-Met was examined immunohistochemically in the mandibles of human fetuses during weeks 9 and 16 of pregnancy. c-Met was immunolocalised in the whole area of the developing mandible, although to different extents. In the intramembranous bone, mesenchymal cells showed a weak immunopositivity. Osteoprogenitor cells demonstrated a moderate immunopositivity for c-Met, while osteoblasts and osteocytes showed a very strong immunolabelling of c-Met. In the developing mandibular condyles, c-Met immunopositivity increased gradually throughout the proliferative layer towards the pre-hypertrophic cell layer, whereas the cells of the hypertrophic layer were weakly immunopositive. These findings have demonstrated, for the first time, the prominent immunolocalization of c-Met in osteogenic and chondrogenic tissues of developing human mandibles, which indicates possible functions for this receptor during mandibular development.

Growth hormone and its receptor in human ovaries from fetuses and adults.

OBJECTIVE: To investigate the presence of growth hormone (GH) and its receptor (GH-R) in early developing follicles. DESIGN: Immunocytochemical and in situ hybridization study. SETTING: Major tertiary care and referral academic centers. PATIENT(S): Ten ovarian samples from adults/girls aged 6-38 years and from 10 fetuses of women undergoing second and third trimester pregnancy terminations. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Immunocytochemistry and in situ hybridization on paraffin sections of human ovaries from fetuses and women/girls. RESULT(S): The proteins and the mRNA transcripts for GH and GH-R were detected in oocytes, granulosa (GC), and stroma cells from both sources (fetuses and women/girls), with low staining intensity only in a portion of the fetal GC. CONCLUSION(S): This is the first report of the expression of GH-R in human ovaries from fetuses as well as women/girls and of GH in human fetal ovaries. Be that as it may, further experiments should be conducted to elucidate if indeed GH is involved in the initiation of human primordial follicular growth.

Emergence of a glomus-like body in the human yolk sac: a microanatomical analysis of its structure.

Since the last decade the Yolk sac (YS) has been a topic of increasing interest due to the growing use of high-resolution sonography in early determination of pregnancy. Human YS shape and diameter are indicators of viability of pregnancy during the early embryonic period. Nevertheless, the major interest concerns the vital function it plays in early embryo growth and development. Two compartments are recognized in this organ: the yolk sac proper and the vitelline stalk. In this study we report the identification and partial characterization of a glomus-like body in the wall of the secondary YS in humans. A detailed structural description is also presented on the time course of formation of this new structure, at precisely sequential stages between 4-8 wk post-conception. The significance of this new compartment on the YS function is analyzed. Light and scanning electron microscopy were used to investigate the microstructure of the YS and the vitelline stalk during the first 8 wk of development. Ten YSs were collected from embryos (aged between 24-50 days) obtained from emergency salpingectomies due to tubal ectopic pregnancy. From 5 wk onward a new structure was observed in the YS located near the apex of the pear-shaped yolk vesicle and closed to the connecting stalk. We designate this differentiation as glomus-like body. This structure is 1-1.5 mm long and merged from a pocket-like structure of the extraembryonic splanchnic mesoderm of the YS wall. It likely represents an area of convergence of the vascular network of the YS wall. Our findings underline the remarkable complexity of the human secondary yolk sac during early development. The detailed description of the microanatomy of this vital organ is of theoretical and practical interest in order to unravel the mechanisms used by the yolk sac to transport nutrients to the embryo.

Microscopical survey of the development and differentiation of the epithelium of the uterine tube and uterus in the human fetus.

The development and differentiation of the coelomic epithelium lining the paramesonephric ducts in human fetus, that gives rise to the female genital organs, have been ultrastructurally examined. The epithelium appeared pseudostratified, consisting of basal, microvillous and ciliated cells. In younger fetuses (12th gestational week) ciliogenic elements could be detected mainly on the developing tubal fimbriae, but most of the cells showed microvilli and often single cilia. In the subsequent phases of development, morphodynamics of cell renewal were documented by aspects of apoptosis. Fully ciliated cells were numerous on the fimbriae and at the utero-tubal junction, but not in the uterus; however, these were less abundant than those showing microvillous. In older fetuses (31st gestational week) microapocrine secretion by microvillous cells, in the form of droplets contacting cilia, could be observed. In the same fetuses the ectocervix was covered by a mature squamous epithelium, made up of polygonal flat desquamating cells, showing labyrinthine surface microplicae. Our observations demonstrated that ciliation in the human female genital organs, like that of other systems, is neither simultaneous nor uniform, and ciliated cells are gathered preferentially in strategic sites, to mediate germ cell migration and blastocyst implantation in adult life. These ultrastructural data seem to indicate that the female genital tract epithelium, at least in its general features, is sketched since fetal life, and cell morphodynamics, including microvillous and ciliated cell differentiation, as well as the secretory activity, are the morphological expression of the complex molecular mechanisms, involved in developmental biology and reproductive physiology.