Motor patterns in the stomatogastric ganglion of the lobster Panulirus argus.

1. Acitivity patterns arising from the thirty cells of the stomatogastric ganglion of Panulirus argus are described for both a semi-intact preparation and an isolated one. 2. The thirty or so cells can be divided so far into two functional groupings: the gastric mill group, with at least ten motor elements, and the pyloric group with at least fourteen. There is some, but not extensive, interaction between groups. 3. The main gastric mill activity is arranged in two sets of elements, each of which is composed of reciprocating elements innervating antagonistic muscles. Thus alternation in activity between the single LC and the two LG neurones results in alternate closing and opening of the lateral teeth; alternation between the four GM and single CP units results in alternate protraction and retraction of the medial tooth. 4. The two sets are phased to each other in such a way that they cause gastric mill teeth to operate effectively to masticate food. 5. The main pyloric activity is arranged in a three-part cycle with each of three sets of units active in sequence. Activity in two PD and one AB unit is followed by bursts in IC and LP units followed in turn by activity in up to seven PY units. Activity in a single VD neurone is locked to this cycle in a more complex pattern.     

Eye stabilization by statocyst mediated oculomotor reflex inNautilus

Summary Nautilus pompilius can rotate its eye relative to its body so as to compensate for changes in its body orientation and maintain its eye fixed with respect to gravity. An ocular compensation reflex stabilizes its eye about the pitch axis against the rocking motions that occur as the animal swims by jet propulsion. The eye is not held absolutely still, as 1–2° spontaneous rotations occur even if the animal is clamped.If the animal is held in an unnatural orientation (rotated about the axis through its laterally directed eyes) the counterrotation of the eye is maintained for many minutes; it may compensate for 50–90% of imposed tilts up to ±30°. If the nautilus is tilted suddenly forward or backward, its compensation reflex is 50% complete within 0.3 s, and is complete in 1–2 s. The time course of the responses explains how the eye, during the 1 Hz rocking caused by swimming, can be held fixed in space to within 1–4°. The ocular compensation of each eye is mediated largely by the homolateral statocyst, as is shown by unilateral and bilateral ablation. The effect of ocular compensation is to keep image orientation on the retina fixed relative to gravity, and to prevent large rotational image motions from being caused by the nautilus's own swimming.This work would not have been possible without the fishing skill and the hospitality of the people of Bindoy, Negros Oriental, The Republic of the Philippines. We especially thank Mr. Wilson Vailoces who arranged for capture of experimental animals. We are indebted to Mr. Walter Schneider for his technical assistance on shipboard. This study, a part of the RV Alpha Helix Southeast Asian Expedition was supported by the National Science Foundation under grants of 74-01883 and OCE 74-02888 to the Scripps Institution of Oceanography and BM 575-01149 to Iowa State University. NIH Grant R01-EY01539 to P. Hartline supported data analyses.     

Human cytomegalovirus UL97 kinase activity is required for the hyperphosphorylation of retinoblastoma protein and inhibits the formation of nuclear aggresomes

Cells infected with human cytomegalovirus in the absence of UL97 kinase activity produce large nuclear aggregates that sequester considerable quantities of viral proteins. A transient expression assay suggested that pp71 and IE1 were also involved in this process, and this suggestion was significant, since both proteins have been reported to interact with components of promyelocytic leukemia (PML) bodies (ND10) and also interact functionally with retinoblastoma pocket proteins (RB). PML bodies have been linked to the formation of nuclear aggresomes, and colocalization studies suggested that viral proteins were recruited to these structures and that UL97 kinase activity inhibited their formation. Proteins associated with PML bodies were examined by Western blot analysis, and pUL97 appeared to specifically affect the phosphorylation of RB in a kinase-dependent manner. Three consensus RB binding motifs were identified in the UL97 kinase, and recombinant viruses were constructed in which each was mutated to assess a potential role in the phosphorylation of RB and the inhibition of nuclear aggresome formation. The mutation of either the conserved LxCxE RB binding motif or the lysine required for kinase activity impaired the ability of the virus to stabilize and phosphorylate RB. We concluded from these studies that both UL97 kinase activity and the LxCxE RB binding motif are required for the phosphorylation and stabilization of RB in infected cells and that this effect can be antagonized by the antiviral drug maribavir. These data also suggest a potential link between RB function and the formation of aggresomes.     

IKK-i and TBK-1 are enzymatically distinct from the homologous enzyme IKK-2: comparative analysis of recombinant human IKK-i, TBK-1, and IKK-2

NF-kappaB is sequestered in the cytoplasm by the inhibitory IkappaB proteins. Stimulation of cells by agonists leads to the rapid phosphorylation of IkappaBs leading to their degradation that results in NF-kappaB activation. IKK-1 and IKK-2 are two direct IkappaB kinases. Two recently identified novel IKKs are IKK-i and TBK-1. We have cloned, expressed, and purified to homogeneity recombinant human (rh)IKK-i and rhTBK-1 and compared their enzymatic properties with those of rhIKK-2. We show that rhIKK-i and rhTBK-1 are enzymatically similar to each other. We demonstrate by phosphopeptide mapping and site-specific mutagenesis that rhIKK-i and rhTBK-1 are phosphorylated on serine 172 in the mitogen-activated protein kinase kinase activation loop and that this phosphorylation is necessary for kinase activity. Also, rhIKK-i and rhTBK-1 have differential peptide substrate specificities compared with rhIKK-2, the mitogen-activated protein kinase kinase activation loop of IKK-2 being a more favorable substrate than the IkappaBalpha peptide. Finally, using analogs of ATP, we demonstrate unique differences in the ATP-binding sites of rhIKK-i, rhTBK-1, and rhIKK-2. Thus, although these IKKs are structurally similar, their enzymatic properties may provide insights into their unique functions.     

THE OSMOTIC PROPERTIES OF LIVING CELLS (EGGS OF ARBACIA PUNCTULATA)

We have attempted to answer the question: How nearly ideal, as an osmometer, is the unfertilized Arbacia egg? The following conclusion have been reached: 1. Volumes can be measured accurately over a wide range of pressures since the cell is in general spherical and does not suffer deformation from its own weight or other factors. 2. The product of volume and pressure is approximately constant, if allowance be made for osmotically inactive cell contents. It is computed that from 7 to 14 per cent of cell volume is occupied by osmotically inactive material. 3. Evidence is presented that no appreciable escape of cell contents occurs while the cell is in hypotonic sea water; that, therefore, the semipermeability of the membrane is approximately perfect, so long as injury to the cell is avoided. 4. In comparison with osmotic pressure the influence of other forces, such as elasticity or surface tension, on cell volume must in these experiments be slight.     

Preferential uptake of d-α-aminoadipate from a racemic mixture by an Alcaligenes denitrificans

During growth on racemic α-aminoadipate as a sole source of carbon and nitrogen an Alcaligenes denitrificans displayed a two-phase (diauxic) growth not shown by a Pseudomonas putida. Oxidation studies in the presence of chloramphenicol showed that in both growth phases whole cells of A. denitrificans had the ability to oxidize either isomer of α-aminoadipate. These data negated as the physiological basis for diauxic growth a proposal that one isomer was repressing the synthesis of enzymes which transport or degrade the other enantiomer. Growth studies with various ratios of the d- and l-isomers of α-aminoadipate, differentiating one isomer from the other by radioactivity, indicated that diauxic growth resulted from a preferential uptake of d-α-aminoadipate over the l-isomer.     

Optic nerve responses to visual stimuli in squid

Summary Single unit optic nerve responses were studied in isolated eye-optic lobe preparations ofLoligo opalescens. Units could be classified as fast or slow. Fast units were invariably receptor cell axons; slow units might be either receptor cell axons or centrifugal axons.On, on-off, andoff units could be found within these classes. A given unit was not stable with respect to these latter attributes which depended greatly on the history and level of illumination. The curve which relates quantal content of a brief flash to number of spikes in the response has a logarithmic phase, but it saturates as brightness is increased further. An inhibitory component has been demonstrated following the response to a flash. It is probably responsible for the non-monotonic relationship between frequency and light intensity which is observed for sustained stimuli. Background light or previous illumination can lead to a facilitation of the response to a flash.The authors share equally in credit and responsibility for this paper and for the research reported here. We thank Dr. A. Hurley and Dr. T. H. Bullock for valuable comments and suggestions and L. Ball and S. St. John for technical assistance. This research was supported by PHS grants NS 09342 to GDL and EY 29405 to PHH, by a grant from the Sloan Foundation to the group in Neurosciences at UCSD and NSF grants GB 24816 and GD 28838 to the Scripps Institution of Oceanography for operation of the Alpha Helix Research Program.