Carolyn J.P. Jones

An ultrastructural and ultrahistochemical study of the human placenta in maternal pre-eclampsia

At the electronoptical level the placental villi of the pre-eclamptic woman are characterized by focal syncytial necrosis, loss and distortion of microvilli, dilatation of syncytial rough endoplasmic reticulum, decreased syncytial pinocytotic activity, a reduced number of syncytial secretory droplets, cytotrophoblastic hyperplasia, degeneration of occasional cytotrophoblastic cells, thickening of the trophoblastic basement membrane and the presence of small fetal capillaries with bulbous endothelial cells. Ultrahistochemical studies show reduced alkaline phosphatase and dehydrogenase activity in the syncytiotrophoblast but an increased acid phosphatase activity. It is suggested that all the observed morphological abnormalities, with the exception of cytotrophoblast cell degeneration, are explicable solely on the basis of utero-placental ischaemia and that no other aetiological factor need be invoked: the cause of the cytotrophoblast cell degeneration is, however, unknown. The ultrastructural findings indicate that there is a decrease in the transfer and synthetic activity of the trophoblast and that cellular respiration is also probably depressed. Nevertheless, there is some evidence that changes of a compensatory nature, designed to limit the effects of the tissue damage, are brought into play.

Ultrastructure of the normal human placenta

In this review, the ultrastructure of the normal human chorionic villus is examined and illustrated. The outer covering of trophoblast, comprising the generative cytotrophoblast and the multinucleated syncytiotrophoblast which is derived from it, is described, including such features as the microvillous surface, vesicles and vacuoles, endoplasmic reticulum and secretory droplets. The structure, composition, development and inclusions of the trophoblastic basement membrane are discussed, and the ultrastructure of the various components of the stroma, including reticulum cells, fibroblasts, Hofbauer cells, capillaries and the non-cellular matrix are illustrated and described, with special reference to their inter-relationships and function.

Fibrillin microfibrils are stiff reinforcing fibres in compliant tissues.

Fibrillin-rich microfibrils have endowed tissues with elasticity throughout multicellular evolution. We have used molecular combing techniques to determine Youngs modulus for individual microfibrils and X-ray diffraction of zonular filaments of the eye to establish the linearity of microfibril periodic extension. Microfibril periodicity is not altered at physiological zonular tissue extensions and Youngs modulus is between 78 MPa and 96 MPa, which is two orders of magnitude stiffer than elastin. We conclude that elasticity in microfibril-containing tissues arises primarily from reversible alterations in supra-microfibrillar arrangements rather than from intrinsic elastic properties of individual microfibrils which, instead, act as reinforcing fibres in fibrous composite tissues.

Fibrillin microfibrils are reduced in skin exhibiting striae distensae

Striae distensae (striae: stretch marks) are a common disfiguring condition associated with continuous and progressive stretching of the skin—as occurs during pregnancy. The pathogenesis of striae is unknown but probably relates to changes in those structures that provide skin with its tensile strength and elasticity. Such structures are components of the extracellular matrix, including fibrillin, elastin and collagens. Using a variety of histological techniques, we assessed the distribution of these extracellular matrix components in skin affected by striae. Pregnant women were assessed for the presence of striae, and punch biopsies were obtained from lesional striae and adjacent normal skin. Biopsies were processed for electron microscopy, light microscopy and immunohistochemistry. For histological examination, 7 μm frozen sections were stained so as to identify the elastic fibre network and glycosaminoglycans. Biopsies were also examined with a panel of polyclonal antibodies against collagens I and III, and fibrillin and elastin. Ultrastructural analysis revealed alterations in the appearance of skin affected by striae compared with that of normal skin in that the dermal matrix of striae was looser and more floccular. Light microscopy revealed an increase in glycosaminoglycan content in striae. Furthermore, the number of vertical fibrillin fibres subjacent to the dermal–epidermal junction (DEJ) and elastin fibres in the papillary dermis was significantly reduced in striae compared with normal skin. The orientation of elastin and fibrillin fibres in the deep dermis showed realignment in that the fibres ran parallel to the DEJ. However, no significant alterations were observed in any other extracellular matrix components. This study identifies a reorganization and diminution of the elastic fibre network of skin affected by striae. Continuous strain on the dermal extracellular matrix, as occurs during pregnancy, may remodel the elastic fibre network in susceptible individuals and manifest clinically as striae distensae.

Syncytial knots, sprouts, apoptosis, and trophoblast deportation from the human placenta.

The syncytiotrophoblast (STB) that forms the epithelial covering of the placental villous tree has a unique cell biology on account of its syncytial nature. The tissue is in a terminally-differentiated, postmitotic state, and expands through the recruitment by fusion of underlying progenitor cytotrophoblast cells. This process occurs from the time of implantation until term, and so its nuclei will be of various ages, producing a spectrum of contrasting appearances; whilst some are euchromatic, others display dense condensations of heterochromatin, the latter often aggregating to form clusters referred to as syncytial knots. These appearances have led to the suggestion that knots are apoptotic, and a hypothesis has developed that the nuclei are transcriptionally inactive and transit through the STB before being shed into the maternal circulation. Here, we review the evidence for this hypothesis, looking at the morphology of the nuclei, their number throughout gestation, evidence of transcriptional activity, and trophoblast deportation. We conclude that there is little evidence to support the concept that turnover of syncytial nuclei takes place in the normal placenta, or that this occurs through an apoptotic-related process. Instead, we suggest that a proportion of syncytial nuclei are transcriptionally active, that epigenetic modifications underlie the changes in chromatin appearance, and that syncytial nuclei continue to accumulate until term. We recognize that apoptotic changes can occur in pathologic pregnancies, but consider the deportation of trophoblast that has been linked to preeclampsia to be most likely of necrotic origin following ischemic injury.

Elimination of the non-specific binding of avidin to tissue sections

SummaryA simple procedure is described for eliminating non-specific staining with avidin—peroxidase conjugates. Murine ovaries were embedded in either paraffin wax or epoxy resin and, after blocking endogenous peroxidase activity, were treated with 10 µg/ml biotinylatedPisum sativum agglutinin. Avidin—peroxidase conjugates (5 µg/ml), diluted in standard 0.05m tris-buffered saline, pH 7.6, containing 0.139m NaCl, produced considerable background coloration and intense mast cell staining in controls without the lectin. This background diminished as the ionic strength of the buffer was raised. At 0.125m Tris-buffered saline (containing 0.347m NaCl) the background was completely unstained, with elimination of all binding to mast cells and only minimal loss of specific lectin binding.

The Q-switched neodymium: YAG laser and tattoos: A microscopic analysis of laser-tattoo interactions

The Nd:YAG laser effectively removes or lightens amateur and professional tattoos. The biomechanics of the removal of tattoo particles at the cellular level are incompletely understood. We examined biopsies obtained from 35 amateur and professional tattoos (including coloured tattoos), treated on three or more occassions with the Nd:YAG laser. Biopsies taken immediately after laser treatment showed vacuolation with complete clearance of tattoo particles in the most superficial layers of the dermis, as assessed by light and electron microscopy. We propose that the ‘disappearance’ of the tattoo particle arises from the formation of atomic species and gaseous products, which are rapidly dissolved in the extracellular fluid. Residual fragmented particles that are commonly found in the mid‐and lower dermis are rephagocytosed. The interaction between the Nd:YAG laser and black tattoo particles at 1064nm, and red tattoo particles at 532nm, appears to be specific, as there was little evidence of thermal damage to adjacent cells or stromal collagen.

Adhesion Molecules in Human Trophoblast – A Review. I. Villous Trophoblast

In the placental villus, cells attach to basement membrane via integrin alpha6beta4 and adhere both laterally and apically to their neighbours. The most prominent adhesive specialisation seen using the electron microscope is the desmosome, which connects cytotrophoblast cells (CTB) laterally and also contributes to the attachment of CTB to the overlying syncytium. However, numerous cadherins and other junctional proteins are also present in the corresponding plasma membrane domains, indicating a multiplicity of adhesive interactions. Integrins, tight junction components and cadherins are all found in the syncytial microvillous membrane, perhaps reflecting its ability to form intersyncytial bridges. There is a wide gulf to be filled between molecular anatomy and functional studies, with much to be learned about the role of adhesion molecules in regulating villous epithelial integrity, homeostasis and growth.

MUC1, glycans and the cell-surface barrier to embryo implantation.

As it approaches the maternal surface, the attaching embryo encounters the epithelial glycocalyx, which contains the mucin, MUC1. A high density of MUC1 at the cell surface can inhibit cell adhesion. This raises the possibility of the existence of a uterine barrier to implantation that might allow maternal rejection of poorer quality embryos. To investigate the mechanism of implantation, human embryos were incubated with endometrial epithelial monolayers. Hatched blastocysts were found to attach readily to the epithelial surface. MUC1 was lost from epithelial cells beneath and near to the attached embryo, while normal expression persisted in neighbouring cells.

Clinical features of photodamaged human skin are associated with a reduction in collagen VII

Chronically sun‐exposed or photodamaged human skin is characterized by a number of clinical features, including wrinkles. However, little is known about the molecular mechanisms that underlie these features. We investigated the hypothesis that the mechanism of wrinkle formation may involve loss of anchoring fibrils, composed mainly of collagen VII, which are important in maintaining dermal‐epidermal junction integrity. Ten volunteers with moderate to severe photodamage of dorsal forearm skin were recruited to the study. Using immunohistochemistry, transmission electron microscopy and in situ hybridization, we compared collagen VII protein and mRNA content of photodamaged forearm skin with that of sun‐protected hip and upper inner arm skin from the same subjects. Numbers of anchoring fibrils per linear μm of basement membrane (mean ± SEM) were significantly lower in photodamaged skin (1·79±0·10) as compared with sun‐protected hip (2·28±0·11) and upper inner arm skin (2·21±0·10) (P<0·01), and similarly keratinocyte expression of collagen VII mRNA, quantitated as number of positively stained keratinocytes per high power field, was significantly reduced in photodamaged skin (6·3±2·5) as compared with sunprotected hip (20·0±5·6) and upper inner arm skin (17·7±4·9) (P<0·001). Semiquantitative assessment of immunohistochemical staining for collagen VII showed a non‐significant reduction in photodamaged skin as compared with sun‐protected skin. We propose that reduced content of collagen VII in photodamaged skin contributes to wrinkle formation by weakening the bond between the dermis and epidermis.