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G R Holstein

Publications and source records attributed to G R Holstein.

28 records · Page 2Linked to original sources
PubMed1992

Monoclonal antibodies for ultrastructural visualization of L-baclofen-sensitive GABAB receptor sites.

Monoclonal antibodies were raised against the L-enantiomer of baclofen conjugated by glutaraldehyde to keyhole limpet hemocyanin. Hybridoma clones were selected for their stability and their production of high titers of antibodies directed against the p-chlorophenyl moiety of the L-baclofen molecule. The chosen antibody showed no cross-reactivity with conjugates of GABA and other neurotransmitters to human or bovine serum albumin. Specificity was further confirmed by the ability of L-baclofen-HCl to inhibit the binding of the antibody to L-baclofen-bovine serum albumin conjugate. Immunocytochemical studies were conducted on brain tissue from rats and monkeys injected with baclofen to localize baclofen-sensitive GABAB receptor sites. In these animals, the molecular layer of cerebellar cortex was clearly immunostained and the granular layer showed only some pale immunoreactivity. Ultrastructural observations were conducted in cerebellar cortex, as well as in the substantia nigra and the vestibular nuclei. Discrete labeling of neuronal profiles was observed in these structures, and both immunoperoxidase and colloidal gold methods were employed successfully. Material from saline-injected control animals showed no immunoreactivity at both light and electron microscopic levels. We conclude that the anti-L-baclofen antibody preferentially recognizes the p-chlorophenyl moiety of the baclofen molecule. Antibodies of such specificity are useful tools for the ultrastructural localization of baclofen-sensitive GABAB receptor sites. In general, antibodies directed against accessible moieties of specific neuroactive substances may serve as valuable markers for their sites of action.

Animals↗
PubMed1988

GABAergic elements in the neuronal circuits of the monkey neostriatum: a light and electron microscopic immunocytochemical study.

An antibody raised in rabbits against a GABA-BSA conjugate was used together with the PAP technique to label elements in the neostriatum of three Old World monkeys. Light microscopy revealed numerous immunoreactive medium-size neurons of various staining intensities, some of which had indented nuclei, as well as an occasional large cell. The neuropil showed a plexus of fine processes with frequent puncta. Ultrastructurally, the medium-size GABA-positive neurons were of two types: one with smooth nuclei and scanty cytoplasm, similar to spiny I cells, the other with invaginated nuclear envelopes and more abundant perikaryon, resembling the aspiny type. Correspondingly, labeled dendrites were either spiny or varicose. Some stained axons were myelinated, and the boutons had either large and ovoid, or small and pleomorphic vesicles. All of these boutons formed symmetric synapses, the former type with GABA-positive dendritic shafts but also with unlabeled dendrites; the latter type usually with GABA-negative elements, either dendrites, dendritic spines, or somata. Synapses were also observed between unreactive boutons and immunostained dendrites. Terminals with densely packed, small round vesicles that established asymmetric synapses with spines were always GABA-negative. Glial elements were consistently unlabeled, save for some astroglial endfeet. These findings provide positive evidence for the existence of two classes of GABAergic striatal neurons corresponding to a long-axoned spiny I type and an aspiny interneuron. Furthermore, the simultaneous labeling of GABA-immunoreactive presynaptic and postsynaptic profiles offers possible morphologic bases for the various kinds of intrastriatal inhibitory processes, including the feedforward, feedback, and "autaptic" types.

Animals↗
PubMed1988

Serotonin-immunoreactivity in the monkey lateral geniculate nucleus.

Serotonin-immunoreactivity in the monkey lateral geniculate nucleus appears as a plexus of fine, beaded fibers decreasing in density from magnocellular to parvocellular laminae. Ultrastructurally, these fibers show strictures and dilations, and are filled with dense round particles as well as granular material attached to outer mitochondrial membranes and microtubules. Most of the profiles followed in serial sections lack morphologically defined synapses. The few synapses observed are asymmetric, some with subjunctional dense bodies. This appearance suggests a possible excitatory effect mainly on interneurons which in turn would inhibit principal cells. Serotonin released non-synaptically may block the delivery of transmitters from retinal terminals and/or the receptors for such transmitters, thereby exerting a modulatory depressing action on principal cells.

Animals↗
PubMed1986

Synapses between GABA-immunoreactive axonal and dendritic elements in monkey substantia nigra.

gamma-Aminobutyric acid (GABA)-positive elements in the monkey substantia nigra pars reticulata (SNr) were identified using the peroxidase-antiperoxidase immunocytochemical technique with a direct GABA antibody. Light-microscopic examination revealed fine-caliber immunostained fibers piercing the cerebral peduncle and entering the SNr, where they formed a dense plexus. Ultrastructural observations included boutons with densely packed ovoid synaptic vesicles forming synapses with labeled and unlabeled dendritic elements. Most terminals contained small dark mitochondria, although some endings held larger, paler and fewer of these organelles. The synapses observed between GABA-immunoreactive profiles may represent the morphologic basis for disinhibition of pars compacta neurons reported to follow neostriatal or pallidal stimulation.

Aging↗
PubMed1985

Early postnatal development of the monkey visual system. II. Elimination of retinogeniculate synapses.

Profiles of retinal terminals, and of their synaptic and non-synaptic contacts, were measured in electron micrographs from magnocellular and parvocellular laminae of the dorsal lateral geniculate nucleus (LGNd) in newborn, 1-,4-,8- and 17-week-old rhesus monkeys. Morphologic criteria, i.e., the presence of pale mitochondria and large round vesicles, were used to identify the profile of retinal origin. Size-frequency histograms were stereologically reconstructed and used to calculate the density of retinal boutons and synaptic and non-synaptic plaques. The density values were adjusted for laminar growth to yield estimates of total numbers of these elements. Numerical estimates indicate bouton proliferation during the first week, followed by substantial reductions in bouton number accompanied by profound decreases in synapse number and cumulative synaptic area. In magnocellular layers, the reduction in synapse number is more pronounced after the eighth week, whereas the decrements in both features in the parvocellular laminae occur before this time. This synapse elimination process may be due entirely to retinal bouton retraction in parvocellular layers, but involves additional retinal synapse loss in the magnocellular segment. The parvocellular division shows a further size contraction of the remaining synapses. Immature synapses predominate in the LGNd throughout the 4-month period, and no quantitative evidence for direct transformation of immature to mature contacts is obtained. Non-synaptic junctions are stable in number but of increasing size in magnocellular layers, whereas substantial increases in number and area are found in parvocellular laminae. The preceding modifications in synaptic organization of the monkey LGNd occurring during the initial postnatal period may provide morphologic bases for the physiological and behavioral changes observed in this species during the same interval. Our data underscore the conclusion that synaptic reorganization occurs over a prolonged period, probably extending beyond 4 months, and involving the process of synapse elimination.

Animals↗
PubMed1983

The influence of rod light and dark adaptation upon rod-cone interaction.

The influence of a rod-detected mask of illuminance IR on the threshold illuminance of a cone-detected test flash (ICth) was assessed while rods were recovering from the effects of a bleach, and when rods were selectively light adapted. Providing that IR was restricted to within 2 log10 units of rod mask threshold (IRth), results show that ICth/IC0 = K (square root (IR/IRth) + D), where IC0 is cone absolute threshold, D is a dark noise term and K is a proportionality constant. These data were used to obtain 'equivalent background functions' or 'Crawford (1947) transforms' (illuminance of a background field plotted against time in the dark). The same Crawford transform was obtained when either IRth or ICth (in the presence of a fixed illuminance IR) were used as equating variables. All of the foregoing results could be predicted by considering both the influence of light adaptation on rods (see Fain, 1976) and the model developed by Bauer, Frumkes & Nygaard (1982). Under dark-adapted conditions, 40' and 60' diameter rod masks of equal illuminance have very similar influences on ICth. When rods are selectively light adapted 60' masks have smaller influences on ICth. The foregoing results were used to extend the model developed in the previous paper. We suggest that rod adaptation has a distinct influence on neural loci designated E (the excitatory spatial summator) and I (the inhibitory spatial summator), and that E represents a site for both adaptation and spatial summation.

Adaptation, Ocular↗
PubMed1979

Rod-cone interrelationships at light onset and offset.

The spatial constraints needed to obtain rod-cone interaction under steady adaptation levels apply to transient conditions of adaptation. The influence of cones upon rod threshold, however, is qualitatively different at light onset as opposed to light offset.

Adaptation, Ocular↗