Two types of luliberin-immunoreactive perikarya in the preoptic area of the rat

Summary At the light microscopic level, following immunostaining with a single antiserum against luliberin (LRF), two types of hormone-producing perikarya in the preoptic area are demonstrated. The two cell types differ in their morphological features: a bipolar, smooth-contoured cell type can be differentiated from an irregularly contoured unipolar type. Intermediate forms between both cell types occurring in the same area are not observed. Electron microscopically, both cell types contain labeled granules of similar size and immunoreactivity. It is dicussed whether the uneven surface of the one cell type is due to areas of synaptic contacts, and whether both cell types are integrated in different neuronal and functional circuits. Moreover, at the ultrastructural level, from the irregularly contoured LRF-producing perikarya a further positively stained cell type, probably a glial cell, can be differentiated. The specific labeling of the latter is caused by its content of immunoreactive lysosomal bodies. Differentiation between the labeled glial cells and the irregularly contoured LRF-producing perikarya is not possible at the light microscopic level.Supported by the Deutsche Forschungsgemeinschaft (Grant Nr. Kr 569/3) and by the Stiftung Volkswagenwerk     

Isolation and maintenance of differentiated exocrine gland acinar cells in vitro

Summary Methods have been developed for isolating and maintaining differentiated rat exorbital lacrimal, parotid, and pancreatic acinar cells for up to 1 month in culture. The dissociated cells retained their differentiated morphology when cultured as suspension cultures at 35°C with the appropriate secretagogue (exorbital lacrimal, 10⁻⁶ M carbamyl choline; pancreas 10⁻⁵ M carbamyl choline; parotid, 10⁻⁶ M isoproterenol). Under these conditions the cells remained viable and differentiated for up to 4 weeks in culture and continued to incorporate³H-leucine at rates similar to those of freshly isolated cells. If secretagogue was omitted from the medium, the cells rapidly degenerated. These results indicate that differentiated from the medium, the cells rapidly degenerated. These results indicate that differentiated exocrine gland acinar cells may be maintained in vitro and utilized as a model system for the study of secretory processes.     

Bacteriopheophytin c in reaction center complexes of green photosynthetic bacteria

Two reaction center complexes prepared from cytoplasmic membranes of Chlorobium limicola f. thiosulfato-philum were compared by absorption and CD spectrophotometry. Bacteriopheophytin c (670 nm), which is optically active in one complex but not in the other, may serve as a secondary electron acceptor in the reaction center.     

The photosynthetic electron transfer chain of Chromatium vinosum chromatophores flash-induced cytochrome b reduction

Reduction of a cytochrome b following excitation by a single, short, near-saturating light flash has been demonstrated in Chromatium vinosum chromatophores. The extent of reduction is increased by addition of antimycin. The cytochrome has an α-band maximum at 562 nm in the presence of antimycin.The cytochrome b reduction is most readily observed in the presence of antimycin at high redox potential when cytochrome c-555 is oxidised before excitation. Under these conditions the half-time for reduction is about 20 ms, and the extent is about 0.5 mol of cytochrome b reduced per mol of reaction center oxidised. This extent of reduction is observed on the first flash-excitation from the dark-adapted state, and there was no indication that the reaction center quinone acceptor complex acted as a two-electron accumulating system. With cytochrome c-555 reduced before excitation, the extent of cytochrome b reduction is approximately halved. The factors which result in substoichiometric cytochrome b reduction are not yet understood.Agents which appear to inhibit primary acceptor oxidation by the secondary acceptor (UHDBT, PHDBT, DDAQQ, HOQNO, o-phenanthroline), inhibit reduction of the cytochrome b. DBMIB inhibits cytochrome b reduction but does not appear to inhibit primary acceptor oxidation.These observations confirm that a cytochrome b receives electrons delivered from the primary acceptor complex, and indicate that the photoreduced cytochrome b is reoxidised via an antimycin-sensitive pathway.     

Three dimensional organization of mammalian adrenal cortex

Summary The adrenal cortex of different mammals was studied by SEM in order to demonstrate its actual three-dimensional organization. In the rat, as well as in the cat and pig, the adrenal cortex appeared as a tunnelled continuum of polyhedral cells arranged in plate-like structures (laminae). This laminar arrangement was more evident in the inner fasciculate and reticular zones where the cortex revealed a striking similarity to liver tissue. The polyhedral cells of all cortical zones possessed regular facets populated by small pits, larger invaginations and numerous microvilli with the exception of very short and smooth areas probably corresponding to attachment zones and/or gap junctions. This cellular architecture produced a labyrinthic system of intercellular channels or lacunae in which the capillaries were suspended.The pericapillary areas of this labyrinth contained microvilli, amorphous material, a delicate net of fibrils and occasional cells. The intercellular compartment of this lacunar system was mainly bordered by numerous microvilli arising from endocrine cells.The luminal surface of the capillary wall showed not only irregularly protruding margins (interpretable as endothelial junctions) but also clearly overlapping and flattened endothelial extensions.In all the animals and areas of the adrenal cortex examined, the endothelial wall was provided with abundant clusters of small fenestrations (about 50 nm in diameter) generally arranged in sieve plates. Larger fenestrations were noted mainly in the fasciculate and reticular zones of the cat and pig and occasionally in the rat.A final point related to the nature and significance of sinusoidal fenestrations was the occurrence of irregularly shaped and intracapillary located cells mainly noted in the deeper zones of the fasciculate and reticular zones of the gland. These elements — possessing the surface characteristics of macrophages — were observed, with their irregular and slender evaginations, in close proximity to the large fenestrations in a manner reminiscent of Kupffer cells within the lumen of liver sinusoids.     

Experimental study of changes in osteoblastic shape induced by calcitonin and parathyroid extract in an organ culture system

Summary Neonate rat endocranial osteoblasts were cultured on their bone surfaces in control medium (CC) or medium to which either parathyroid extract (PTE) or calcitonin (CT) had been added for 2, 4, 8 or 24 h. Some were cultured for 24 h in CC, then for 2, 4, 8 or 24 h in either CT or PTE medium; or for 24 h in PTE, then for 2, 4, 8 or 24 h in either CC or CT; or 24 h in CT and 2, 4, 8 or 24 h in CC. The dorsal ruffling of the cells in CC was found to be suppressed by later culturing with PTE and the disoriented cells reorganized to form arrays of parallel cells. The effects of PTE were also reversed by CC or CT: the osteoblasts in the second culture (CC) lost elongation and order, and proceeded through a proliferative phase before exhibiting the ruffling form similar to a single CC 24 h culture. PTE-cultured osteoblasts showed an increase in cell overlap and contact so that a more competent barrier was formed separating the bone from the medium. In control or CT medium, however, intercellular gaps were greater than in vivo.We are grateful for the expert technical assistance of Elaine Bailey, for laboratory facilities kindly provided by Dr. Martin Evans, and for financial support from the Medical Research Council     

Fine Structure of Oocyst Transformation and the Sporozoites of Leucocytozoon dubreuili*

SYNOPSIS. The sporogonic stages of Leucocytozoon dubreuili in the midgut and salivary glands of the simuliid vectors was studied by electron microscopy. Young uninucleate oocysts have a pellicle that initially resembles that of the ookinetc. Numerous electron-dense bodies and microtubules in the peripheral cytoplasm may be involved in the formation of the cyst wall. The dense bodies appear to give rise to the amorphous material of the wall. The tubules which run circumferentially beneath the oocyst's boundary probably serve as a skeletal support for the cell surface during deposition of the wall material. A subcapsular “space” which provides area for expansion of the developing sporozoites is formed in early multinucleate oocysts. The subcapsular “space” appears to be formed through a condensation of the peripheral cytoplasm, resulting in an osmotic gradient across the oocyst's limiting membrane. Consequently water diffuses out, creating a fluid-filled space. Sporozoite formation begins with localized thickenings on the oocyst's limiting membrane. Subsequent extension of the thickened regions into the subcapsular “space” marks the onset of sporozoite budding. The process is highly synchronized, and culminates with the production of up to 150 sporozoites about the sporoblastoid body. The structure of sporozoites from mature oocysts and of the salivary glands of the vector is basically similar, although salivary gland sporozoites are more elongate and have numerous electron-dense micronemes. The paired rhoptries in the latter sporozoites are more elongate and uniformly electron-dense than in oocyst sporozoites.