GABAA Receptor Subtypes Expressed in Cerebellar Granule Cells: A Developmental Study

Abstract: The developmental properties of primary rat cerebellar granule cells have been characterised with respect to their expression of GABA_A receptor subtypes using both an immunological approach and radioligand binding assays. At day 1 in culture, the GABA_A receptor α1 subunit was detectable in immunoblots and increased in level up to day 9. The GABA_A receptor α6 subunit was not detectable at day 1; however, at days 3–5, a specific M_r 58,000 anti-α6 1–16 Cys immunoreactive species was present which further increased in level up to 9 days in culture. Similar qualitative results were obtained for the expression of the GABA_A receptor α6 subunit in age-matched rat cerebellar membranes. In parallel studies, it was found that although there was an overall increase in [³H]Ro 15–4513 binding sites with days in culture, the relative contributions of diazepam-sensitive and diazepam-in-sensitive [³H]Ro 15–4513 binding changed. A time-dependent enrichment of the diazepam-insensitive binding site up to a maximum of 74% of total [³H]Ro 15–4513 sites was found. This was concomitant with the appearance of the GABA_A receptor α6 subunit. These results are in agreement with the pharmacology described for α6βγ2 cloned receptors. They suggest a developmentally regulated expression of the GABA_A receptor α6 subunit gene at a time that is correlated in vivo with establishment of neuronal connections.     

A polymorphic alpha satellite sequence specific for human chromosome 13 detected by oligonucleotide primed in situ labelling (PRINS)

The centromeric alpha satellite DNA subfamilies from chromosomes 13 and 21 are almost identical in sequence and cannot be easily distinguished by mean of probes for Southern blot or in situ hybridisation. We have used the oligonucleotide-primed in situ (PRINS) labelling technique with primers defined from the alpha satellite sequence of chromosome 13. One primer was found to label specifically the centromeric region of chromosomes 13 and allowed the detection of a polymorphism between two chromosome 13 homologues in one individual.     

How does the G protein,Gi2, transduce mitogenic signals?

Serpentine receptors coupled to the heterotrimeric G protein, G_i2, are capable of stimulating DNA synthesis in a variety of cell types. A common feature of the G_i2-coupled stimulation of DNA synthesis is the activation of the mitogen-activated protein kinases (MAPKs). The regulation of MAPK activation by the G_i2-coupled thrombin and acetylcholine muscarinic M₂ receptors occurs by a sequential activation of a network of protein kinases. The MAPK kinase (MEK) which phosphorylates and activates MAPK is also activated by phosphorylation. MEK is phosphorylated and activated by either Raf or MEK kinase (MEKK). Thus, Raf and MEKK converge at MEK to regulate MAPK. G_i2-coupled receptors are capable of activating MEK and MAPK by Raf-dependent and Raf-independent mechanisms. Pertussis toxin catalyzed ADP-ribosylation of α_i2 inhibits both the Raf-dependent and-independent pathways activated by G_i2-coupled receptors. The Raf-dependent pathway involves Ras activation, while the Raf-independent activation of MEK and MAPK does not involve Ras. The Raf-independent activation of MEK and MAPK most likely involves the activation of MEKK. The vertebrate MEKK is homologous to the Ste11 and Byr2 protein kinases in the yeast Saccharomyces cerevisiae and Schizosaccharomyces pombe, respectively. The yeast Ste11 and Byr2 protein kinases are involved in signal transduction cascades initiated by pheromone receptors having a 7 membrane spanning serpentine structure coupled to G proteins. MEKK appears to be conserved in the regulation of G protein-coupled signal pathways in yeast and vertebrates. Raf represents a divergence in vertebrates from the yeast pheromone-responsive protein kinase system. Defining MEKK and Raf as a divergence in the MAPK regulatory network provides a mechanism for differential regulation of this system by G_i2-coupled receptors as well as other receptor systems, including the tyrosine kinases.     

THE POPULATION DYNAMICS OF NORTHERN SEA LIONS, 1975-1985

Abstract: Populations of northern sea lions ( Eumetopias jubatus ) in the vicinity of Marmot Island, Alaska declined during 1975–1985 at about 5% per year (Merrick et al. 1987). The cause of this decline is not known. A life table for the northern sea lion was calculated assuming that life spans follow a Weibull distribution. Samples of northern sea lions taken in the vicinity of Marmot Island, Alaska during 1975–1978 and 1985–1986 indicate that the average age of females older than 3 yr increased about 1.55 yr (SD = 0.35 yr) while the population was declining at about 5% per year. Fecundity rates decreased by 10% over the same period, but the decrease was not statistically significant (Calkins and Goodwin 1988). Possible causes of the population decline and the change in age structure were examined by writing the Leslie matrix population equation in terms of changes in juvenile and adult survival rates and fecundity, and examining the short–term behavior of the trajectories of the average age of adult females, total number of females, and total number of pups with respect to those changes in the vital parameters. From the observed rate of declines of adults and the changes in average age of adult females and fecundity, estimates of the changes in adult and juvenile survival were calculated; estimates of the standard deviations of these changes were estimated via a bootstrap procedure. One purpose of this exercise is to aid in setting priorities for research for determining the cause of the decline. An explanation for the observed declines in numbers of adult sea lions consistent with the observed fecundity rates, a rate of decrease of 5% in the number of adults, and the corresponding increase in average age (of females age 3 yr and older) was a 10%–20% decrease in the survival of juveniles (age 0-3 yr) coupled with an insignificant change in adult survival (0.03%, SD = 1%).     

Whole inactivated SIV vaccine grown on human cells fails to protect against homologous SIV grown on simian cells

Several groups have reported protection against experimental SIV infection in macaques immunized with a whole inactivated virus vaccine. The aim of the current study was to investigate whether five macaques vaccinated with whole inactivated SIV and previously shown to be protected against challenge with two divergent strains of SIV grown on human cells could resist challenge with a subsequent homologous SIV grown on macaque cells. We show here that this same vaccine did not protect when the challenge virus was grown on primary cells of monkey origin.     

Sequence analysis of sheep T-cell receptor β chains

The nucleotide sequences reported in this paper have been submitted to the GenBank database and have been assigned the accession numbers M94181-M94183.     

Simulated annual plankton production in the northeastern Pacific Coastal Upwelling Domain

A microcomputer simulation model is presented that describesthe generalized plankton production dynamics, in the surfacemixed layer, of the Juan de Fuca Eddy located on the southwesternBritish Columbia continental shelf. The Juan de Fuca Eddy simulationmodel evaluates how the annual biomass production of diatoms,copepods and euphausiids is forced by plankton feeding interactions,seasonal variability in upwelling, water temperature and solarradiation, and generalized fish predation. The model estimatesannual primary production of 345 g C m^–2 year^–1and secondary production of 19.4 g C m^–2 year^–1for copepods and 6 g C m^–2 year^–1 for euphausiids,during 1985–89; -90% of the annual plankton productionwas generated during the April-October upwelling season. Perturbationsof 22 abiotic and biotic parameters, one at a time by ±10%of nominal values, indicated that oceanic variability (e.g.upwelling rate) most strongly affected primary production. Conversely,zooplankton production was most sensitive to variability inbiological parameters describing zooplankton grazing potentialand growth (e.g. gross growth efficiency). Simulated seasonalbiomass patterns of diatoms, copepods and euphausiids were foundto closely match empirical data. However, euphausiid biomassproduction in the Juan de Fuca Eddy alone was unable to meetthe demands of estimated pelagic fish consumption. Local Eddyeuphausiid populations had to be supplemented, from regionaleuphausiids. by a mechanism that is proposed to be linked tothe seasonal pattern and intensity of positive Ekman transport(upwelling).     

A new classification for the subgenera of the genus Acanthophthirius Perkins,with descriptions of twelve new taxa (Acarina,Trombidiformes, Myobiidae)

The myobiid genus Acanthophthirius Perkins, to date comprising four subgenera, is reviewed and divided into just two subgenera, the nominate subgenus and Myotimyobia Fain. The male genital shield and female opisthogastric sclerites, which are here considered to be part of female genitalia, are adopted as the criteria for dividing the subgenera. These structures are essentially the same in form and position in the subgenus Acanthopthirius in its revised sense, while they are both more heterogeneous in species of the redefined subgenus Myotimyobia. The subgenera Acanthophthirius Fain and Chiromyobia Fain are thought to represent species-groups in the nominate subgenus, named respectively the etheldredae and miniopteri groups. The following one new subspecies and 11 new species are described: A. (A.) womersleyi eptesicus, A. (A.) glauconycteris, A. (A.) mauritaniensis, A. (A.) philetoris, A. (A.) otonycteris, A. (A.) steatocaudatus, A. (A.) nyctophilis, A. (M.) vagus, A. (M.) longus A. (M.) baueris, A. (A.) hesperopteris (female only) and A. (M.) pixonixeos (female only). A. (M.) hanensis is synonymised with A. (M.) namurensis, and A. (M.) capacini is emended to A. (M.) capaccinii and relegated to a subspecies of A. (M.) myotis. All the known and new species are assigned to their respective subgenera and shown in a table. Incongruent host-relationships of some of the mites are clarified, although not completely solved, by the introduction of the new classification for the subgenera.     

Contrasting effects of PACAP and carbachol on [Ca]i and inositol phosphates in human neuroblastoma NB-OK-1 cells

The effects of PACAPs on [Ca²⁺]_i were compared to those of carbachol in human neuroblastoma NB-OK-1 cells. PACAP(1–27) and PACAP(1–38) increased [Ca²⁺]_i in a biphasic manner: a transient rise and a secondary plateau. The transient phase reflected the mobilization of [Ca²⁺]_i pool(s) via the inositol phosphate pathway. The modest sustained plateau required extracellular Ca²⁺. Carbachol also increased [Ca²⁺]_i in a biphasic manner, but it mobilized intracellular Ca²⁺ pool(s) with a higher efficacy than PACAPs, then greatly increased Ca²⁺ entry, this being accompanied by a more marked and prolonged elevation of IP₃ and IP₄ than with PACAPs. It is likely that cAMP-mediated phosphorylations due to PACAPs facilitated desensitization at the PACAP receptor-phospholipase C level, so that there was less Ca²⁺ handling through PACAP receptors than with muscarinic M₁ receptors.