The fine structure of the infundibular process of the hedgehog

Summary The fine structure of the infundibular process of the hedgehog has been studied, using material fixed in osmium tetroxide and embedded in Vestopal W. The process resembles in general structure that of other mammals, but also shows features not previously described in other species.The nerve fibres contain a number of inclusions, namely: small vesicles, 300–500 Å in diameter; larger vesicles, up to 2000 Å in diameter, which contain a variable amount of osmiophilic material; hexagonal crystal-like bodies, approximately 1250 × 3000 Å in size, lying within a striated membranous sheath; and aggregate bodies made up of small electron dense granules, possibly derived from mitochondria.In addition complex multilamellate bodies occur in some nerve fibres, which apparently give rise to membranous vesicles. Pituicytes, of varying appearance, are often intimately related to the nerve fibres.The findings suggest that synthesis of material may occur in the distal part of the fibres of the hypothalamo-hypophysial tract.We are indebted to the Medical Research Council, who provided the electron microscope in the Department of Anatomy, University of Birmingham, which was used in this study.     

The neurohypophysis of the foetal monkey

Summary The neurohypophysis of foetal macaque monkeys has been studied by optical and electron microscopy. Abundant elementary neurosecretory granules are present in the infundibular process by the middle third of gestation. Most of these are of variable electron density, and surrounded by a membrane larger than the granule, so that they appear haloed. A few fibres contain membrane-bounded electron-dense granules which show no halo. Inclusions of synaptic vesicle size are rare. The infundibular stem contains a few fibres with typical inclusions smaller than 1000 Å in diameter; the latter resemble inclusions in fibres of the median eminence.Dedicated to Professor W. Bargmann on his 60th birthday.     

Studies into equine electrocardiography and vectorcardiography: I. Cardiac electric forces and the dipole vector theory

Theoretical consideration has been given in two horses to the properties of the electric field created by the equine heart acting as a simple electric generator. The principles of the vectorial theory have been applied to test the validity of application of the dipole concept. The cardiac electric forces, althrough complex in the immediate region of the heart, appear at the body surface in a similar form to those arising from a relatively immobile, single equivalent dipole. The potential value of the technique of vectorcardiography in cardiological investigations is briefly discussed.     

Differential expression of the receptors for epidermal growth factor and insulin in the developing human placenta

The detailed cellular distribution of epidermal growth factor (EGF) receptors and insulin receptors during the development of the human placenta was examined. We show that EGF receptors are expressed by villous cytotrophoblast cells in first trimester human placentae. However, where these cells proliferate to form extravillous cytotrophoblast cell columns, there is a dramatic decrease in EGF receptor expression. There is no such differential expression of insulin receptors on this cell population. In contrast, both EGF-and insulin-receptors are present throughout gestation on the microvillous membrane of the terminally differentiated and non-proliferative syncytiotrophoblast although, at term, EGF-but not insulin-receptors are also found on the basolateral membrane of this epithelium. We further show that EGF receptors isolated from first trimester and term human placentae have functional tyrosine kinase activities but differ in their extent of glycosylation. These results suggest that EGF receptors probably play several distinct functional roles in these epithelial cells depending on their proliferative capacity and differentiation status.     

The nonlinear characteristics of soft gels and hydrated connective tissues in ultrafiltration

A one-dimensional ultrafiltration problem of fluid flow through a soft permeable tissue or gel under high pressure and compressive strain is solved. A finite deformation biphasic theory is used to model the behavior of the soft porous permeable solid matrix. This theory includes a Helmholtz free energy function which depends on the three principal invariants (I, II, III) of the right Cauchy-Green tensor and which satisfies the Baker-Ericksen inequalities on the principal stresses and strains. The dependence of the porosity phi f and the solidity phi s on deformation is deduced and a generalization of the exponential strain-dependent functional form for the permeability, k = k0 exp (M epsilon), of Lai and Mow (Biorheology 103, 111-123, 1980) is proposed. In this one-dimensional problem, we show that the dependence of the permeability on phi f, phi s, and III is equivalent to its dependence on hydration as proposed by Fatt and Goldstick (J. Colloid Sci. 20, 962-988, 1965). The exact solution of the ultrafiltration problem is derived and asymptotic and numerical methods are used to evaluate it. For high pressures and finite strains, the solution provides some surprising effects. The theory predicts that a material starting with a homogeneous porosity will have a strongly non-homogeneous porosity throughout the column during ultrafiltration. The resulting change in pore size through the filtration column may be very important in understanding its filtration characteristics. It is also found that there is a long delay time, up to 10 to 15 min, before the filtration velocity reaches an equilibrium. In filtration experiments where the rate of mass transport across the tissue or column of gel is important, sufficient time must be allowed for the steady state to be reached.     

In vitro analysis of the oligodendrocyte lineage in mice during demyelination and remyelination

A demyelinating disease induced in C57B1/6N mice by intracranial injection of a coronavirus (murine hepatitis virus strain A59) is followed by functional recovery and efficient CNS myelin repair. To study the biological properties of the cells involved in this repair process, glial cells were isolated and cultured from spinal cords of these young adult mice during demyelination and remyelination. Using three-color immunofluorescence combined with [3H]thymidine autoradiography, we have analyzed the antigenic phenotype and mitotic potential of individual glial cells. We identified oligodendrocytes with an antibody to galactocerebroside, astrocytes with an antibody to glial fibrillary acidic protein, and oligodendrocyte-type 2 astrocyte (O-2A) progenitor cells with the O4 antibody. Cultures from demyelinated tissue differed in several ways from those of age-matched controls: first, the total number of O-2A lineage cells was strikingly increased; second, the O-2A population consisted of a higher proportion of O4-positive astrocytes and cells of mixed oligodendrocyte-astrocyte phenotype; and third, all the cell types within the O-2A lineage showed enhanced proliferation. This proliferation was not further enhanced by adding PDGF, basic fibroblast growth factor (bFGF), or insulin-like growth factor I (IGF-I) to the defined medium. However, bFGF and IGF-I seemed to influence the fate of O-2A lineage cells in cultures of demyelinated tissue. Basic FGF decreased the percentage of cells expressing galactocerebroside. In contrast, IGF-I increased the relative proportion of oligodendrocytes. Thus, O-2A lineage cells from adult mice display greater phenotypic plasticity and enhanced mitotic potential in response to an episode of demyelination. These properties may be linked to the efficient remyelination achieved in this demyelinating disease.     

Polyclonal activation of the murine immune system by an antibody to IgD. X. Evidence that the precursors of IgG1-secreting cells are newly generated membrane IgD+B cells rather than the B cells that are initially activated by anti-IgD antibody

Injection of BALB/c mice with an affinity-purified goat antibody to mouse IgD (GaM delta) stimulates T cell-independent B cell activation as well as later T cell activation. Activated T cells then induce polyclonal differentiation of B cells into IgG1-secreting cells, which results in an approximately 100-fold increase in serum IgG1 level. It is not known whether the same B cells that are initially activated by GaM delta are the progenitors of the IgG1-secreting cells. To investigate this issue a system was developed in which CB20 mice, which are congenic to BALB/c mice but express Ig of the beta allotype rather than the BALB/c alpha allotype, were injected with GaM delta and simultaneously or subsequently also received BALB/c B cells. The IgG1 response generated by the donor BALB/c B cells was quantitated by an assay specific for IgG1 of the alpha allotype. Our experiments with this system indicate that: 1) BALB/c B cells transferred 2 days after CB20 mice were injected with GaM delta generate a much larger IgG1 response than do BALB/c B cells transferred simultaneously with GaM delta antibody; 2) B cells that express membrane IgD generate the great majority of this response; 3) differences in the magnitudes of the responses of BALB/c B cells transferred at different times after CB20 mice were injected with GaM delta antibody cannot be explained by differences in homing of the donor B cells to the host spleen or by short survival of donor BALB/c B cells after their transfer; and 4) the response made by donor BALB/c B cells transferred 2 days after CB20 mice were injected with GaM delta is proportionate to donor cell representation in the host spleen 1 day after their transfer, whereas the response made by donor cells transferred simultaneously with GaM delta is disproportionately small. These observations suggest that most of the IgG1 antibody made by GaM delta-injected mice is generated by newly produced, mIgD+ B cells that appear approximately 2 days after GaM delta injection, rather than by those B cells that are present in the spleen at the time of GaM delta injection, and support the view that signals that induce B cell secretion of Ig require an interaction with at least partially activated Th cells.     

Purification of the 110-kilodalton glycoprotein receptor for mouse hepatitis virus (MHV)-A59 from mouse liver and identification of a nonfunctional, homologous protein in MHV-resistant SJL/J mice.

The receptor for mouse hepatitis virus strain A59 (MHV-A59) is a 110- to 120-kilodalton (kDa) glycoprotein which is expressed in MHV-susceptible mouse strains on the membranes of hepatocytes, intestinal epithelial cells, and macrophages. SJL/J mice, which are highly resistant to MHV-A59, were previously shown to lack detectable levels of receptor by using either solid-phase virus receptor assays or binding of a monoclonal anti-receptor antibody (MAb) which blocks infection of MHV-susceptible mouse cells. This MAb was used for affinity purification of the receptor glycoprotein from livers of MHV-susceptible Swiss Webster mice. The MHV receptor and an antigenically related protein of 48 to 58 kDa were copurified and then separated by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The first 15 amino acids of the receptor were sequenced, and a synthetic peptide of this amino acid sequence was prepared. Rabbit antiserum made against this peptide bound to the MHV receptor glycoprotein and the 48- to 58-kDa protein from livers of MHV-susceptible BALB/c mice and Swiss Webster mice and from the intestinal brush border of BALB/c mice. In immunoblots of intestinal brush border and hepatocyte membranes of MHV-resistant SJL/J mice, the antibody against the amino terminus of the receptor identified proteins that are 5 to 10 kDa smaller than the MHV receptor and the 48- to 58-kDa related protein from Swiss Webster or BALB/c mice. Thus, SJL/J mice express a protein which shares some sequence homology with the MHV receptor but which lacks virus-binding activity and is not recognized by the blocking anti-receptor MAb. These results suggest that resistance of SJL/J mice to MHV-A59 may be due to absence or mutation of the virus-binding domain in the nonfunctional receptor homolog in SJL/J mice.