ALBERT

Notes: Early passage mouse embryo fibroblasts, mouse 3T3 cell lines, and early passage diploid human fibroblasts grew to higher cell densities in tissue culture medium supplemented with serum than in medium supplemented with defibrinogenated platelet-poor plasma (PPP). Unlike the mouse cells, the human fibroblasts displayed this differential growth response only in the presence of hypophysiologic concentrations of calcium. The addition of heat-treated extracts of human platelets to PPP-supplemented medium stimulated the replication of both the normal mouse cells and early passage human embryo fibroblasts.Human or mouse fibroblasts transformed by either retroviruses or by SV40, including SV40 infected “serum revertants” and “flat transformants,” grew to equal cell densities in medium supplemented with either serum or PPP. Infection of Balb/c-3T3 cells with SV40 rapidly induced them to grow in PPP-supplemented medium demonstrating that the ability of SV40-transformed cell lines to proliferate in PPP-supplemented medium does not arise from the cell culture selection procedures usually employed to obtain stable virus-transformed cell lines. 3T3 cells infected but not transformed by retroviruses do not replicate in PPP-supplemented medium demonstrating that reduction of the growth requirement for the platelet growth factor(s) by retroviruses is a transformation-specific response. Cell cultures that did not proliferate well in PPP-supplemented medium did not form tumors when inoculated into athymic nude mice. Many, although not all, of the lines which grew well in PPP medium were tumorigenic in nude mice. Together, these findings indicate that: (1) normal fibroblast-like cells display a growth requirement for factor(s) present in serum but not found in PPP; (2) this serum specific growth factor is derived from platelets; (3) a primary response to viral transforming genes is a reduction in the growth requirement for these platelet-derived factors; and (4) cells that have a reduced requirement for the platelet-derived growth factor are often tumorigenic.

Notes: The fine structure of the mid-gut musculature of the desert locust, Schistocerca gregaria is described and compared with that of the visceral muscles of other species. The gross morphology and fine structure of the nervous system which supplies the mid-gut muscle fibres is described.

Notes: The dorsal ventricular ridge (DVR) is a subcortical, telencephalic structure in reptiles and birds that protrudes into the lateral ventricle. The structure of DVR has been studied in the red-eared turtle (Pseudemys scripta elegans) in Nissl and Golgi preparations. The DVR in Pseudemys is divided into the anterior dorsal ventricular ridge (ADVR) and the basal dorsal ventricular ridge (BDVR) by the dorsal branch of the middle ventricular sulcus. The structure of ADVR has been examined in detail.The ADVR is divided into four regions with distinct boundaries termed dorsal area, medial area, ventral area and central area. Dorsal area, medial area and ventral area border on the lateral ventricle; central area lies deep to the other areas. Three classes of neurons are found in Golgi preparations of ADVR. Juxtaependymal cells have somata near the perikarya of ependymal cells; their dendrites are found primarily in a periventricular fiber zone. Aspiny neurons were observed only in the dorsal half of ADVR and appear to be restricted to deep regions of the ridge. These multipolar neurons are rarely encountered in Golgi preparations, and the observed distribution may not represent their actual distribution in ADVR. The majority of the cells observed in ADVR are spiny neurons with dendritic fields that range from stellate to double-pyramidal. Cells in this class may be subdivided on the basis of axonal morphology into at least two groups, but further studies are needed to determine the range of axonal morphology exhibited by these neurons.An analysis of the distribution of these cell types in Golgi material shows that dorsal area, medial area and ventral area are organized in four zones concentric with the ventricular surface. Central area apparently lacks a concentric pattern of organization. Zone 1 is a periventricular fiber band that contains juxtaependymal neurons and ascending dendrites of zone 2 spiny neurons, and it may serve as a structural substrate for segregated input onto these cell populations. Zone 2 contains clusters of spiny neurons with apposed somata, which vary in size and distribution between areas. Dendrites of zone 4 neurons are also found in the deep half of zone 2. Zone 3 is a cell-poor region which lies at the center of a region of overlapping dendritic fields of zone 2 and zone 4 neurons. Zone 4 contains predominantly spiny neurons (aspiny neurons are found only in the dorsal half of ADVR) which are either isolated or in small clusters with apposed somata. Dendrites of zone 2 cells extend superficially into zone 4, so that the deep portions of zone 4 may be a substrate for segregated input to zone 4 neurons. These zones are differentially elaborated in each area. Central area, by contrast, consists of scattered spiny and aspiny neurons among fibers connecting ADVR and the lateral forebrain bundle.A comparison of these findings with the ADVR of snakes (Ulinski, '78a,b) shows both similarities and differences in DVR organization in the two taxa. Although snakes lack areal divisions, ADVR is organized in four concentric zones (zones A-D). Zones A and B resemble zones 1 and 2 in turtles, consisting of a superficial fiber zone and a subjacent cell cluster zone. The clusters are smaller in snakes than in turtles. However, snakes lack a cell-poor band deep to zone B, and dendrites of cells in zone C enter zone A. Thus, there are differences in both areal and zonal dimensions of ADVR organization in turtles and snakes.

Notes: The American cockroach has a total of 368 muscles inserting on the post-coxal segments of its legs. By using a narrow morphological definition for delimiting individual muscles, it is shown (i) that the protrochanteral musculatures (23 muscles/leg) differ from the essentially identical meso- and metatrochanteral musculatures (24 and 26 muscles/leg) in number and disposition of extensors and in having a completely different flexor composition, and (ii) that the musculatures of the more distal segments of the legs are completely serially homologous, there being 2 muscles for moving each femur, 23 for each tibia, 7 for each first tarsomere, and 5 for each of the paired pretarsal claws. In all six legs, the trochanteral and tibial musculatures each contain single slender muscles that may be acting proprioceptively to measure the angular displacements between, respectively, the coxas and trochanters, and the femurs and tibias. Neurological and phylogenetic considerations are used to demonstrate why a narrow morphological definition should be employed, and why the widely used functional definition of Snodgrass ('35) is not only fallacious on evolutionary grounds, but also leads to making erroneous conclusions regarding the manner in which insect musculature is controlled by the insect central nervous system. Finally, it is hypothesized that the physiological limitations imposed by having an open circulatory system and the problems inherent in the neural control of large muscles may have been major evolutionary factors in forcing insects to use many slender muscles to control their body movements.

Notes: End-plate distributions have been determined for three frog muscles of different morphology in order to relate end-plate topography to spatial muscle structure and nerve branching. Koelle's cholinesterase technique was applied, both on whole muscles and frozen sections. The end-plates of the short parallel-fibered cutaneus pectoris muscle appeared to be located in short bands along the nerve branches. The nerve tree is restricted to a zonal area across the middle part of the muscle. Depending on the way the nerve branches, the end-plate bands form innervation patterns, varying from one single continuous band to multiple distributed bands. In the latter case one frequently observes that different end-plate bands do not run across the same longitudinal muscle fiber area, although the respective nerve branches run parallel across this area. The long parallel-fibered sartorius muscle has a wider nerve tree and exhibits the same phenomenon for close parallel nerve branches, but end-plate bands along parallel nerve branches far apart cover the same muscle fiber area. The end-plate distribution in the bipennate, short-fibered gastrocnemius is zonal throughout the muscle except in certain compartments containing tonic fibers. The end-plate zone centers around the inner aponeurosis about half-way between the muscle tendon junctions of the fibers and is visible only at the muscle surface where muscle fibers run over their entire length at that surface. The results are of general use in the electrophysiology of neuromuscular transmission because they illustrate how in certain twitch muscles neuromuscular morphology may help to localize end-plates.

Notes: Histological observations using specialized techniques reveal neurosecretory cells in 18 centers throughout the rind (cortex) of the central nerve mass or synganglion of Dermacentor variabilis. Many cells contribute to complicated networks of neurosecretory pathways and tracts in pre- and post-esophageal portions of the synganglion. The four types of neurohemal-neuroendocrine associations found in Dermacentor resemble structures found in soft ticks (Argasidae) and in other Arachnida, but are more diverse than those described from any other single species. Neurosecretory terminals are distributed diffusely and in two concentrated associations within the perineurium of the synganglion and major peripheral nerves. Terminals are also distributed in the perineurial layers of lateral segmental organs which lie in the general hemocoel at the level of the pedal nerves. A retrocerebral organ complex surrounds the esophagus at its junction with the midgut. The complex includes dorsal and ventro-lateral lobes (containing neurosecretory terminals and intrinsic secretory cells) and the proventricular (neurohemal) plexus. This plexus seems to be a modified (concentrated) cardioglial association. Cardioglial associations are also formed by the neurosecretory innervation of vascular walls of the dorsal aorta and circulatory sinuses which envelope the synganglion and major peripheral nerves. Inferential considerations of neurosecretory and endocrine interactions in the Acari are based on these anatomical and histological data which also provide the basis for evolutionary considerations of anatomical relationships and specializations in the neurosecretory systems of other Arachnida.

Notes: The antennae of the sawyer beetles Monochamus notatus and M. scutellatus were examined with the light and scanning electron microscopes to determine the types, number, distribution and innervation of the sense organs. Nine types of sensilla are described. Both short, thin-walled pegs (sensilla basiconica) and reversely curved thick-walled hairs (sensilla trichodea) are chemoreceptors. There are three types of long, thick-walled hairs (sensilla chaetica) which may be mechanoreceptors. One of these is modified in males to form unique snail-shaped pegs. A few dome-shaped organs, probably campaniform sensilla, were found.In addition to sense organs, many glands occur in association with the sensilla, and the antennae are well supplied with dermal glands connected by canals to small pores on the surface.

Notes: The fiber composition of the distal accessory flexor muscle (DAFM) and the branching pattern of its excitor axon were compared in several species of crabs, in the lobster and the crayfish. The muscle is composed exclusively of long sarcomere (〉 6 μm) fibers and therefore of the slow type. In all the crab species, except one, there is a distal to proximal gradient of fibers with increasing sarcomere lengths; this gradient is reverse in lobsters and crayfish. A proximal to distal gradient of increasing fiber diameters occurs in the DAFM of all crab species but not in the lobster and crayfish, in which all the fibers are approximately equal in diameter. The single excitatory axon traverses the width of the DAFM and gives off primary branches on either side in the lobster and crayfish but on only one side in crabs. The hypothesis that the axonal branching pattern may govern the regional distribution of fibers with differing sarcomere lengths in proposed.

Notes: The host-virus interactions of Simian virus 40 (SV40) and polyoma virus (Py) with cell lines established from a teratocarcinoma were studied. The cells utilized in this study were the multipotential stem cell of the teratocarcinoma, embryonal carcinoma, and differentiated cells derived from embryonal carcinoma. Several lines of differentiated cells were established in vitro which included parietal yolk sac, epithelial, and spindle cell types. Embryonal carcinoma cells are not susceptible to infection by either SV40 or Py virus. However, differentiated cells are susceptible to infection by these viruses. The differentiated cells are permissive for Py virus replication and nonpermissive for SV40. Several continuously growing cell lines have been established from the SV40 infected cultures which express T antigen in 100% of the cells. The results indicate that undifferentiated embryonal carcinoma cells and their differentiated progeny respond quite differently to challenge with these two oncogenic DNA viruses.

Notes: The multiplication rate of sparse cultures of chick embryo cells is only slightly lower at pH 6.9 than at pH 7.4. There is, however, a marked reduction in the multiplication rate of the pH 6.9 cultures before they reach confluency. Cultures at pH 7.4 continue to multiply beyond confluency with only a slight decrease in the multiplication rate.Eighty to ninety percent of the glucose taken up by the cells growing at each pH is converted to lactic acid which is released into the medium. Metabolic reduction in pH of the medium is almost entirely accounted for by the amount of lactic acid produced by the cells. Neither the intracellular nor extracellular accumulation of lactic acid nor the accompanying reduction in pH is sufficient to explain density dependent inhibition of the rate of multiplication of chick cells.The rate of lactic acid production and the multiplication rate of chick cells are independent of glucose concentration in the range of 2-16 mM. In view of the kinetic parameters for the uptake of glucose, this shows that glycolysis is not limited by the rate of glucose uptake and that depletion of glucose from the medium cannot account for the onset of density dependent inhibition of multiplication. However, when cells reach very high population densities, conventional glucose concentrations of 5 mM can be depleted overnight by chick cells. Since the multiplication rate of cells is dependent on glucose concentration when it falls below 2 mM, depletion of glucose may cause some growth inhibition in crowded cultures supplied with conventional medium.