Ovulation and the role of the ovarian surface epithelium.

Epon sections from all tissue layers of the rabbit ovary, including the often neglected surface or germinal epithelium, were studied to elucidate the mechanism of follicle rupture. Scanning electron microscopy showed cells covering preovulatory follicles increased clearly in size up to 8 hours after human chorionic gonadotropin (HCG) injection. These showed an increasing number of large, intracellular structures with prominent rounded contours. Transmission electron microscopy showed large, electron dense, lysosomelike bodies in some cells 4 hours after HCG. These membrane-surrounded structures increased in size up to 8 hours after HCG, then decreased markedly. These obviously corresponded to the bodies found by light microscopy and scanning electron microscopy. Histochemistry revealed many represent lysosomes. During the last 2 hours before rupture, the dense bodies of the surface epithelium considerably decrease, signs of material emptying into vacuoles is found, and sometimes there is open communication from vacuoles towards the unerlying tunica albuginea. An extracellular edema appeared under the epithelium with degenerated fibroblasts and disintegrated collagen; these changes gradually proceeded inwards. Blood capillaries close to the membrana granulosa gradually showed small pores and close to ovulation the endothelial cells had gaps up to 3 mcm in diameter. The preovulatory follicles grew rapidly and an augmenting edema occupied the whole ovary. In the last hours before ovulation the membrana granulosa gradually dissociated. Whatever the net effect of the prostaglandins, prostaglandin F2alpha appears to be essential for follicle rupture since intrafollicular injections of antiserum blocked ovulation. Prostaglandin E1 promotes vascular permeability and lysosommal enzymes released extracellularly may be coupled to collagen degration. The enzyme synthesis and lysosomal growth in the surface epithelium of preovulatory follicles may be due to the high local concentration of sex steroids.     

Close relationship of mitochondria with intercellular junctions in the adrenaline cells of the mouse adrenal gland

Summary In adrenaline cells, junctional complexes formed by alternating gap junctions and attachment plaques were identified in close proximity to bilateral clusters of mitochondria. It is suggested that this proximity is related to a role of gap junctions in metabolic coupling.     

Fenestrated capillaries in ‘venous patch’ adventitia

Summary Fenestrated capillaries in venous patch adventitia are seen during the 2nd week after microsurgery. The fenestrae contain diaphragms. These capillaries are possibly transitory structures which appear during injury cicatrization. The importance of these capillaries during transcappillary exchange in injuried tissue is discussed.The technical assistance of M. Guerricabeitia is gratefully acknowledged.     

Biomolecular stability and life at high temperatures

It is not clear what the upper temperature limit for life is, or what specific factors will set this limit, but it is generally assumed that the limit will be dictated by molecular instability. In this review, we examine the thermal stability of two key groups of biological molecules: the intracellular small molecules/metabolites and the major classes of macromolecules. Certain small molecules/metabolites are unstable in vitro at the growth temperatures of the hyperthermophiles in which they are found. This instability appears to be dealt with in vivo by a range of mechanisms including rapid turnover, metabolic channelling and local stabilisation. Evidence to date suggests that proteins have the potential to be stable at substantially higher temperatures than those known to support life, but evidence concerning degradative reactions above 100 degrees C is slight. DNA duplex stability is apparently achieved at high temperature by elevated salt concentrations, polyamines, cationic proteins, and supercoiling rather than manipulation of C-G ratios. RNA stability seems dependent upon covalent modification, although secondary structure is probably also critical. The diether-linked lipids, which make up the monolayer membrane of most organisms growing above 85 degrees C are chemically very stable and seem potentially capable of maintaining membrane integrity at much higher temperatures. However, the in vivo implications of the in vitro instability of biomolecules are difficult to assess, and in vivo data are rare.     

Chromatin affinity-precipitation using a small metabolic molecule: its application to analysis of <Emphasis Type="Italic">O</Emphasis>-acetyl-ADP-ribose

In the cell, many small endogenous metabolic molecules are involved in distinct cellular functions such as modulation of chromatin structure and regulation of gene expression. O-acetyl-ADP-ribose (AAR) is a small metabolic molecule that is generated during NAD-dependent deacetylation by Sir2. Sir2 regulates gene expression, DNA repair, and genome stability. Here, we developed a novel chromatin affinity-precipitation (ChAP) method to detect the chromatin fragments at which small molecules interact with binding partners. We used this method to demonstrate that AAR associated with heterochromatin. Moreover, we applied the ChAP method to whole genome tiling array chips to compare the association of AAR and Sir2. We found that AAR and Sir2 displayed similar genomic binding patterns. Furthermore, we identified 312 potential association cluster regions of AAR. The ChAP assay may therefore be a generally useful strategy to study the small molecule association with chromosomal regions. Our results further suggest that the small metabolic molecule AAR associates with silent chromatin regions in a Sir2-dependent manner and provide additional support for the role of AAR in assembly of silent chromatin.     

Mechanical forces in lymphatic vascular development and disease

The lymphatic vasculature is essential for fluid homeostasis and transport of immune cells, inflammatory molecules, and dietary lipids. It is composed of a hierarchical network of blind-ended lymphatic capillaries and collecting lymphatic vessels, both lined by lymphatic endothelial cells (LECs). The low hydrostatic pressure in lymphatic capillaries, their loose intercellular junctions, and attachment to the surrounding extracellular matrix (ECM) permit passage of extravasated blood plasma from the interstitium into the lumen of the lymphatic capillaries. It is generally thought that interstitial fluid accumulation leads to a swelling of the ECM, to which the LECs of lymphatic capillaries adhere, for example via anchoring filaments. As a result, LECs are pulled away from the vascular lumen, the junctions open, and fluid enters the lymphatic vasculature. The collecting lymphatic vessels then gather the plasma fluid from the capillaries and carry it through the lymph nodes to the blood circulation. The collecting vessels contain intraluminal bicuspid valves that prevent fluid backflow, and are embraced by smooth muscle cells that contribute to fluid transport. Although the lymphatic vessels are regular subject to mechanical strain, our knowledge of its influence on lymphatic development and pathologies is scarce. Here, we discuss the mechanical forces and molecular mechanisms regulating lymphatic vascular growth and maturation in the developing mouse embryo. We also consider how the lymphatic vasculature might be affected by similar mechanomechanisms in two pathological processes, namely cancer cell dissemination and secondary lymphedema.     

Potassium currents in cardiac cells

The kinetic properties of the inwardly rectifying K current and the transient outward current in cardiac cells were investigated. In sheep Purkinje fibers superfused with Na-free K-free solution, time-dependent changes in the conductance of the inward rectifier are described. In patch clamp experiments the inward rectifier inactivates during hyperpolarization, as can be seen by a decrease in the open state probability. Using whole cell clamp on ventricular myocytes it is demonstrated that the inactivation during hyperpolarization is due to blocking of the channel by external Na, Mg and Ca. The channels responsible for the transient outward current in cow, sheep and rabbit Purkinje fibers are identified using single channel recording. It is demonstrated that in all three preparations the channels are K-selective. The channel in cow Purkinje cells has a large conductance and is regulated by voltage and internal Ca concentration. The channels identified in the sheep and rabbit cells have a much smaller conductance.     

The lymphatic route. 1) Albumin and hyaluronidase modify the normal distribution of interferon in lymph and plasma

When human recombinant interferon-alpha 2 diluted in saline was injected s.c. into rabbits, the total amount recovered in thoracic lymph was less than 0.4%. Recoveries increased from 2- to 8-fold if interferon was injected in 4% albumin or with hyaluronidase, respectively. Albumin added to interferon acts as an interstitial fluid expander, thus favoring interferon absorption through lymphatics rather than blood capillaries. This strategy may increase the therapeutic index of interferon.     

Retinoic acid modulates gap junctional intercellular communication in hepatocytes and hepatoma cells

Gap junctional communication permits the direct exchange of small molecules and ions and has been implicated in tissue homeostasis/metabolite exchange. The lack of gap junctional intercellular communication (GJIC) plays important roles in the promotion and progression of carcinogenesis. In the present study, we demonstrate that treatment of human hepatoma Hep G2 cells with retinoic acid (RA) results in increased amounts and phosphorylation of connexins, their stabilisation in plasma membrane plaques and enhanced GJIC. In cultured fetal hepatocytes, which represent a non-transformed, proliferating and incompletely differentiated liver system, the effects of RA are limited to the establishment of connexin in areas of cell-cell contact and the improvement of GJIC. This suggests that modulation of cell-cell channel communication by RA occurs differently in these two experimental models: while RA is able to revert cell transformation in Hep G2 cells, in fetal hepatocytes it may induce the expression of a more differentiated phenotype. Received 19 June 2002; received after revision 29 July 2002; accepted 8 August 2002 RID="*" ID="*"Corresponding author.     

Effect of blood pH on anionic ferritin transport through rat aortic endothelium.

Acute lowering of blood pH between 7.4 and 6.9 in rats by ventilation with 10 or 20% CO2 does not increase the passage of ferritin molecules across the aortic endothelium. These results do not rule out alteration of endothelial permeability to anionic macromolecules in local circulatory disturbances when blood pH drops to levels much lower than 6.9.     

The relation between capillaries and neurons in the caudal neurosecretory system of Pomatomus saltatrix.

The caudal neurosecretory system of Pomatomus saltatrix was examined. Particular interest was devoted to the perikarya of the neurosecretory cells. The majority of these cells were characterized by a close association with the capillary network. The cells appeared to ensheath the capillaries thus bringing the nucleus and perinuclear cytoplasm into close association with the capillaries.     

La barrière capillaire enzymatique pour la DOPA au niveau de quelques noyaux du tronc cérébral du rat

Summary An enzymatic barrier for Dopa exists at the level of the cerebral capillaries which changes Dopa into Dopamine by means of endothelial decarboxylase. It is abolished by a decarboxylase inhibitor. Its intensity is weaker in structures containing Dopamine (Striatum, Locus Niger). Neurones of the Raphé containing serotonin can be charged with catecholamines. There are close contacts between neurones and capillaries.     

Cell-to-cell coupling studied by diffusional methods in myocardial cells

Summary The diffusion of large molecular substances from cell to cell in multicellular and enzymatically isolated cell pairs is described. Permeability of the gap junctional membrane to these molecules and the critical diffusing diameter of the myocardial gap junctional channel are discussed.     

Cell-to-cell coupling studied by diffusional methods in myocardial cells

The diffusion of large molecular substances from cell to cell in multicellular and enzymatically isolated cell pairs is described. Permeability of the gap junctional membrane to these molecules and the critical diffusing diameter of the myocardial gap junctional channel are discussed.     

A simple method for observation of capillary nets in rat brain cortex

Summary By the authors' technic, the profiles of capillaries in rat brain cortex were clearly demonstrated. The capillaries formed complicated nets by sprouting of their finer branches and anastomosing with each other. Further, a kind of perivascular cells with yellow autofluorescent granules was distributed close to capillaries, arterioles or venules. They seemed to be a special form of macrophage in the brain cortex.     

The lymphatic route. 1) Albumin and hyaluronidase modify the normal distribution of interferon in lymph and plasma

Summary When human recombinant interferon-₂ diluted in saline was injected s.c. into rabbits, the total amount recovered in thoracic lymph was less than 0.4%. Recoveries increased from 2- to 8-fold if interferon was injected in 4% albumin or with hyaluronidase, respectively. Albumin added to interferon acts as an interstitial fluid expander, thus favoring interferon absorption through lymphatics rather than blood capillaries. This strategy may increase the therapeutic index of interferon.This work was supported by Ministero Pubblica Istruzione and Progetto Finalizzato Oncologia, contract No. 84.00461.44, CNR, Roma.     

A simple method for observation of capillary nets in rat brain cortex.

By the authors' technic, the profiles of capillaries in rat brain cortex were clearly demonstrated. The capillaries formed complicated nets by sprouting of their finer branches and anastomosing with each other. Further, a kind of perivascular cells with yellow autofluorescent granules was distributed close to capillaries, arterioles or venules. The seemed to be a special form of macrophage in the brain cortex.     

Effect of blood pH on anionic ferritin transport through rat aortic endothelium

Summary Acute lowering of blood pH between 7.4 and 6.9 in rats by ventilation with 10 or 20% CO₂ does not increase the passage of ferritin molecules across the aortic endothelium. These results do not rule out alteration of endothelial permeability to anionic macromolecules in local circulatory disturbances when blood pH drops to levels much lower than 6.9.Supported by The Medical Research Council of Canada grant No. MA-5958.Dr J.R. Fraser was a recipient of a Summer Research Scholarship from McGill University during the completion of this study.The authors thank Colin Bier for the statistical analysis.     

The relation between capillaries and neurons in the caudal neurosecretory system ofPomatomus saltatrix

Summary The caudal neurosecretory system ofPomatomus saltatrix was examined. Particular interest was devoted to the perikarya of the neurosecretory cells. The majority of these cells were characterized by a close association with the capillary network. The cells appeared to ensheath the capillaries thus bringing the nucleus and perinuclear cytoplasm into close association with the capillaries.Supported byA. D. Williams No. 3558 (505), Medical College of Virginia. The authors are grateful to Dr.H. R. Seibel for reading this report.     

Ultrastructural and seasonal aspects of the kidney lymphatic system of hibernating animals

The kidney lymphatic system of bat, dormouse and marmot consists of intraparenchymal (interlobar, arcuate, interlobular) and extraparenchymal (capsular) vessels sharing common ultrastructural aspects. We did not observe medullary lymphatics. The qualitative and quantitative seasonal changes in the ultrastructure of the lymphatic endothelium represent not only a species-linked feature but also (and mainly) an evident seasonal fluctuation in lymph formation. Furthermore, these ultrastructural changes emphasize the important role played by the different mechanisms involved in the translymphatic movement of proteins and interstitial fluid with particular regard to the 'vesicular route' and intraendothelial channels.