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Physiological and morphological characterization of anaxonic non-spiking interneurons in the crayfish motor control system.

Spike activation of the motoneurons innervating uropod muscles in crayfish is controlled by anaxonic interneurons located within the terminal (the 6th abdominal) ganglion. These neurons do not generate spikes either spontaneously at the resting potential level or in response to current injection of either polarity. Yet the change in the membrane potential of these non-spiking interneurons caused an increase or decrease in the discharge frequency of motoneuron spikes, depending upon the direction of the membrane potential change. These non-spiking interneurons within the terminal ganglion presumably integrate various descending command signals and select the adequate information to be gated to the motoneurons.

Animals↗

Cerebrovascular muscle cultures. I. Isolation, growth and morphological characterization.

This study describes the development and establishment of pure cerebrovascular smooth muscle cells derived from dissociated cells of microvessels obtained from brains of rats. The cultured and propagated vascular smooth muscle cells display histochemical and ultrastructural characteristics of smooth muscle cells consistent with those reported in cultures of peripheral vessels. Thus, the cerebrovascular smooth muscle cultures provide a new model system for the investigation of their function, particularly in relation to the regulation of cerebral blood flow and blood pressure.

Animals↗

Growth cones isolated from identified Aplysia neurons in vitro: biochemical and morphological characterization.

The right upper quadrant (RUQ) cells (R3-R13) of Aplysia regenerating in dissociated cell culture form unusually large growth cones. The movement of these growth cones was observed by time-lapse phase microscopy and their ultrastructure was examined by transmission electron microscopy. Their behavior and ultrastructure have features that are typical of growth cones in vitro. Additionally, they contain neurosecretory granules similar to those found in these cells in vivo. Because RUQ growth cones are large, they can be isolated by manual dissection. RUQ cells were grown in the presence of [35S]methionine and the labeled proteins transported to the growth cones were analyzed by SDS-PAGE. These proteins were compared to those in RUQ cell bodies, RUQ neurites, and to those in the neurites and cell bodies of other identified neurons grown in vitro. Most proteins synthesized by RUQ cells in vitro are transported to their growth cones, including several glycoproteins and the precursor to the R3-R14 neuropeptide. Neuropeptides are also synthesized by a number of other Aplysia neurons growing in vitro. We examined R2, LPL1, R15, and left upper quadrant neurons and found that their precursor peptides, like those of R3-R14, are readily recognized as major cell-specific radiolabeled bands on SDS gels. The presence in regenerating growth cones of neuropeptides, neurosecretory granules, and glycoproteins known to be rapidly transported toward synapses in vivo supports the emerging view that the growth cone in vitro contains not only a motility apparatus but also a macromolecular assembly capable of forming an active synapse immediately upon or shortly after contacting targets.

Actins↗

Morphological characterization of beta-VLDL and acetylated-LDL binding and internalization by cultured pigeon monocytes.

The ultrastructure of binding, internalization, and translocation of beta-migrating very low density lipoprotein (beta-VLDL) and acetylated low density lipoprotein (Ac-LDL) by cultured pigeon monocytes was examined using lipoprotein-gold conjugates. Through morphometry, differences in the binding and uptake of beta-VLDL-gold and Ac-LDL-gold were documented. Cells exposed to either beta-VLDL-gold or Ac-LDL-gold for 2 hr at 4 degrees C had the label over noncoated regions of the plasma membrane. Upon warming the cells to 37 degrees C for 2 min, 35% of the surface-bound beta-VLDL-gold was within coated pits on the cell surface. Although coated pits occupied less than 2% of the surface, binding of beta-VLDL-gold was 53 times more concentrated in coated pits as compared to noncoated membrane regions. In contrast, Ac-LDL-gold neither bound to coated pits nor relocated into coated regions of the membrane upon warming to 37 degrees C. Both the beta-VLDL-gold and the Ac-LDL-gold were internalized when the cells were rewarmed at 37 degrees C. Most of the internalized gold particles for both lipoproteins were located in electron-lucent vesicles; however, 9% of the intracellular beta-VLDL-gold was observed within coated vesicles at early times. Upon prolonged rewarming (30-90 min), both lipoprotein-gold conjugates were within acid phosphatase-positive lysosomes. Ultimately 83% of the Ac-LDL-gold and 90% of the beta-VLDL-gold were within electron-dense and electron-lucent lysosomes. These results suggested that the receptor-mediated binding and internalization of beta-VLDL and Ac-LDL by pigeon monocyte macrophages proceeded by separate, distinct routes; beta-VLDL by both coated and noncoated pathways while Ac-LDL was internalized exclusively by noncoated mechanisms. Regardless of these internalization differences, both lipoproteins were delivered to lysosomes for degradation.

Acid Phosphatase↗

Biochemical, functional, and morphological characterization of a primary culture of rabbit proximal tubule cells.

Primary cultures of renal rabbit proximal tubule cells were initiated from a pure suspension of proximal tubule fragments. Proximal tubule cells were grown in a hormone-supplemented, serum-free medium containing low concentrations of antibiotics. Confluent monolayers exhibited multicellular dome formation, indicating the presence of transepithelial solute and water transport. Ultrastructural examination revealed a monolayer of polarized epithelial cells with tight junctions and sparse membraneous microvilli facing the culture medium. Time course biochemical characterization was performed using a palette of 12 enzymes, representative of important metabolic functions or pathways. Brush-border-associated enzymes (gamma-glutamyl transpeptidase and alanine aminopeptidase) were moderately reduced throughout the culture whereas alkaline phosphatase was markedly decreased at confluency. Mitochondrial and lysosomal marker enzymes were well preserved over the culture period. Glutathione-S-transferase activity remained stable during the 16-day culture period investigated. Glycolysis enzyme activities (lactate dehydrogenase and hexokinase) were enhanced, as a function of culture age. Na(+)-K(+)-ATPase activity rise was concomitant with the increase of glycolysis marker enzymes. In contrast, the gluconeogenesis marker enzyme, glucose-6-phosphatase, fell dramatically to reach a low level equivalent to 4% of the activity measured in isolated proximal tubules. Primary cultures exhibited several differentiated functions of the proximal tubule cell: (a) PTH alone was able to induce a significant stimulation of adenylate cyclase activity, unlike isoproterenol, thyrocalcitonin, and arginine vasopressin, and (b) sodium-dependent alpha-methylglucoside (AMG) transport was detected. This AMG uptake was selectively inhibited by phlorizin (5 X 10(-3) M), which is a competitive inhibitor of glucose uptake at the apical membrane. Complete characterization made it possible to investigate hitherto unexplored aspects of in vitro cultured proximal tubule cells. This primary culture model could provide a useful and reliable tool to investigate in vitro renal proximal tubule function, under normal conditions or after a drug-induced toxicity.

Adenylyl Cyclases↗

Ascaris lumbricoides suum: morphological characterization of apparent cuticular pores by ionic permeability and electron microscopy.

The cuticle of the parasitic roundworm Ascaris lumbricoides suum has been found to contain apparent pores, using scanning and transmission electron microscopy. However, X-ray spectral analysis and dot mapping analysis of diffusing divalent cations in the cuticle found these ions to be randomly distributed on the pseudocoelomic surface of the cuticle. This indicates that the pores seen with the electron microscope were not true structural pores and that the cuticle is homogeneously permeable to ions.

Animals↗

Isolation of follicular dendritic cells from human tonsils and adenoids. I. Procedure and morphological characterization.

Follicular dendritic cells have been isolated from human tonsils and adenoids and characterized at the ultrastructural level. Follicles were dissected and digested with different hydrolytic enzymes. The cells were separated by sedimentation at unit gravity. By this procedure we obtained follicular dendritic cells enveloping lymphocytes with their cytoplasmic extensions in a way analogous to that described for isolated thymic nurse cells. The ultrastructural features of isolated follicular dendritic cells are similar to those observed in situ. Prolonged enzymatic action caused loss of the enveloped lymphocytes.

Adenoids↗

Epithelial carcinoma of the ovary in the reproductive years: clinical and morphological characterization.

In the 10-year interval from 1972 through 1981, 247 patients were treated for epithelial ovarian carcinoma at New York University Medical Center. Of this group, 31 patients (12.6%) were below age 40, and this group had distinctive features favoring longer survival. Of these patients 12 (38.7%) had borderline tumors, compared to 9 of 216 patients (4.2%) over 40 years of age (P less than 0.001). Stage I disease and well differentiated lesions were also common in the younger group. Early symptomatology and frequent gynecologic examinations during the childbearing years may also contribute to early detection of these tumors. The median survival in this group (7 years) was better than that reported for all ages. After careful evaluation, conservative surgical management can be considered for patients with Stage Ia disease which is either borderline or grade 1 carcinoma.

Adult↗

Principles for the morphological characterization of transmitter-identified nerve cell groups.

Principles for a morphometric description of transmitter identified nerve cell groups have been introduced, exemplified on the 5-HT nerve cell group of nucleus raphe dorsalis (B7 cell group), using immunocytochemical procedures to visualize 5-HT. Both cell body parameters (mean diameter, mean perimeter, mean area, shape factor) and cell group parameters (number of cells, mean free distance among cells, volume fraction, gravity centre, major axis slope with respect to the midline) have been considered. These parameters have been obtained by the use of a semiautomatic image analyzer (Kontron MOP AMO2) plugged into an Apple II computer. By the use of this system and of suitable computer programs, it is possible to give a Cartesian representation of 5-HT nerve cell bodies in a coronal section under study. The present work has mainly one aim. To detect whether subgroups exist within a transmitter-identified cell group. Two approaches have been introduced to obtain, on objective grounds, evidence whether or not a group consists of subgroups. The first of these approaches is based on differences in cell body density within the cell group, while the second approach is based on frequency distribution analysis. This second approach is mainly sensitive to shape changes of the cell group.

Animals↗

Striatal interneurones: chemical, physiological and morphological characterization.

The neostriatum is the largest component of the basal ganglia, and the main recipient of afferents to the basal ganglia from the cerebral cortex and thalamus. Studies of the cellular organization of the neostriatum have focused upon the spiny projection neurones, which represent the vast majority of neurones, but the identity and functions of interneurones in this structure have remained enigmatic despite decades of study. Recently, the discovery of cytochemical markers that are specific for each of the major classes of striatal interneurones, and the combination of this with intracellular recording and staining, has revealed the identities of interneurones and some of their functional characteristics in a way that could not have been imagined by the classical morphologists. These methods also suggest some possible modes of action of interneurones in the neostriatal circuitry.

Animals↗