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G Prusky

Publications and source records attributed to G Prusky.

10 recordsLinked to original sources
PubMed1999

Intracellular filling and reconstruction of identified neurons in fixed rat brain slices.

A simple technique for the morphological characterization of pre-labelled neurons in fixed brain slices is described. Neurons are retrogradely-labeled with a carbocyanine dye and the tissue is fixed and sliced. Individual labeled cells from the interior of a slice are then visualized on an upright fluorescence microscope and impaled with a micropipet containing rhodamine or fluorescein-peroxidase conjugates. The cells are filled by iontophoresis, postfixed, and the peroxidase is oxidized into a permanent, opaque reaction product. The dendritic morphology of the neurons is then reconstructed under camera lucida and quantified.

3,3'-Diaminobenzidine↗
PubMed1997

Retinal activity regulates developmental switches in functional properties and ifenprodil sensitivity of NMDA receptors in the lateral geniculate nucleus.

Previous studies have shown that marked changes occur in the kinetic properties of N-methyl-D-aspartate (NMDA) receptors during development of the visual pathways. In the lateral geniculate nucleus (LGN) of the ferret, excitatory postsynaptic currents (EPSCs) induced by activation of NMDA receptors display a very slow decay time during the first postnatal month, then become shorter in duration following eye-opening (around postnatal day 32; P32). In view of the critical role that NMDA receptors play in activity-dependent refinement of visual connections during development, we have examined the mechanisms that underlie these changes and how they are regulated. To examine the role of retinal activity, whole-cell recordings were conducted in the LGN slice preparation obtained from normal ferrets and ferrets treated with continuous intraocular application of tetrodotoxin (TTX) from P25 until the time of recording. Blockade of ganglion cell activity with TTX prevented the changes in decay rate of the postsynaptic current induced by NMDA receptors. Treated animals older than P40 had NMDA-EPSCs markedly longer in duration than normal animals at a similar age, resembling responses present in normal newborn animals. To examine whether changes in subunit composition of the NMDA receptor may contribute to the maturation of its kinetic properties, we used the antagonist ifenprodil, which produces selective inhibition of heteromeric NMDA receptors containing the NR-2B subunit. Ifenprodil induced profound inhibition of NMDA receptor activity in normal young animals and TTX-treated mature animals, but substantially less inhibition in normal mature animals. These findings indicate that retinal activity is required for the developmental switch from a juvenile form of the NMDA receptor to a more mature form, possibly affecting NR2 subunit expression.

Action Potentials↗
PubMed1996

Morphology of identified corticospinal cells in the rat following motor cortex injury: absence of use-dependent change.

After unilateral injury to the forelimb area of the motor cortex, rats are reported to show relatively increased use of the ipsilateral forelimb during exploratory behavior and a concomitant transient increase in dendritic growth in neurons in the undamaged motor cortex. To identify the specific population of cells in motor cortex that undergo such use-dependent change, we examined the morphology of corticospinal pyramidal cells in the intact hemisphere 18 days following unilateral motor cortex damage. Corticospinal neurons in the motor cortex were retrogradely labeled with injections of the fluorescent tracer, DiO, into the cervical enlargement of the spinal cord. Seven days later, the rats received a lesion in the forelimb area of the contralateral motor cortex and 18 days following the lesion, limb use was assessed in two behavioral tests after which the rats were sacrificed. Under fluorescent light, corticospinal cells were visualized and injected with a horseradish peroxidase-fluorescein conjugate that was then reacted with diaminobenzidine. The labeled cells were reconstructed and the number and centrifugal order of the branches were analyzed. The increased use of the paw contralateral to the intact motor cortex was not associated with an increase in dendritic arborization in corticospinal motor neurons in the intact motor cortex. The results are discussed in light of methodological and theoretical considerations relevant to the study of neural plasticity in the motor system.

Animals↗
PubMed1990

The distribution of M1 and M2 muscarinic acetylcholine receptor subtypes in the developing cat visual cortex.

The binding site characteristics and ontogenesis of [3H]pirenzepine ([3H]PZ) (M1 receptor) and [3H]oxotremorine-M ([3H]OXO-M) (M2 receptor) binding sites were investigated in the cat visual cortex. Scatchard analysis of [3H]PZ binding in adult cat visual cortex revealed a single site with a Kd of 17.3 nm and a Bmax of 352.45 fmol/mg protein. [3H]OXO-M also bound to a single site with a Kd of 7.1 nM and a Bmax of 256.39 fmol/mg protein. Receptor autoradiography revealed that [3H]PZ binding sites were present only in telencephalic structures while [3H]OXO-M sites were distributed heterogeneously throughout the brain. [3H]PZ binding sites in adult visual cortex were present in the superficial and deep cortical layers with the densest labeling in layer I and a distinct band in layer V. [3H]OXO-M sites also avoided the middle cortical layers, but were most prominent in layers V and VI with less pronounced binding in layers I and II. Deafferentation of extrinsic inputs to the visual cortex did not reduce [3H]PZ nor [3H]OZO-M binding, but neuron-specific excitotoxic lesions of visual cortex abolished both populations of binding sites. This indicates that both populations of binding sites are located on cells intrinsic to the cortex. In early postnatal life, both [3H]PZ and [3H]OXO-M binding sites were localized to intermediate cortical layers. Following this, the laminar distribution of both populations redistributed; each with its own idiosyncratic profile. By postnatal day 49, [3H]PZ binding sites redistributed into the superficial and deep layers, the pattern of adult animals, while [3H]OXO-M sites maintained a pattern similar to younger animals, with substantial binding persisting in layer IV. As late as postnatal day 70, well after [3H]PZ binding sites had achieved their mature laminar pattern, [3H]OXO-M binding sites in visual cortex had not achieved their characteristic adult pattern. In addition, the normal laminar redistribution of both [3H]PZ and [3H]OXO-M binding sites during postnatal development of the cat visual cortex was prevented by eliminating cortical afferents in early postnatal life. This indicates that muscarinic receptor rearrangement in development is dependent upon cortical input or output.

Animals↗
PubMed1989

Differential effects of quinolinic acid lesions on muscarinic acetylcholine receptors in cat visual cortex during postnatal development.

Quinolinic acid (QA) lesions of neurons in cat visual cortex were combined with conventional in vitro autoradiographic methods in order to define the cellular locus of the muscarinic acetylcholine receptor (mAChR). Animals of various postnatal ages had QA unilaterally injected into the visual cortex. Four to fourteen days later they were sacrificed and processed for electron microscopy (EM) or in vitro autoradiography. QA lesions at the various postnatal ages were found to eliminate intrinsic cortical neurons and their processes while leaving intact glia, fibers of passage and axon terminals from outside the lesion zone. Autoradiograms of visual cortex labelled with [3H]QNB (which labels M1 and M2 subtypes) showed an age-dependent loss of binding sites, with the greatest decreases occurring after 65 days postnatal. Examined separately, only the M1 mAChRs labelled with [3H]pirenzepine exhibited these age-dependent alterations. The results indicate a differential distribution of the M1 mAChRs during postnatal development. The loss of receptors late in postnatal life following QA suggests a dominantly neuronal locus; the relatively small loss early in postnatal life suggests a locus on other cellular elements.

Aging↗
PubMed1988

Development of phorbol ester (protein kinase C) binding sites in cat visual cortex.

Tritiated phorbol-12,13-dibutyrate [( 3H]PDBu), a phorbol ester, was utilized to autoradiographically localize protein kinase C (PKC) in the cat visual cortex. Thin, slide-mounted sections of adult cat brain were used to characterize binding of [3H]PDBu. This was found to be saturable, reversible, and more readily displaced by phorbol ester than by synthetic diacylglycerols. Binding sites displayed a tissue concentration of 20 pmol/mg protein, and a dissociation constant of 8.0 nM. [3H]PDBu was slow to associate with its receptor, requiring 9.5 h to reach equilibrium. Autoradiograph revealed that PKC is heterogeneously distributed in the cat brain, and displays a laminar-specific pattern in the visual cortex. This laminar distribution undergoes marked changes during the first two months of postnatal life. In the visual cortex of neonatal kittens, [3H]PDBu binding is confined to layers I and V. Layer III acquires high levels of binding by postnatal day 15, layer II by 28 days, and layer VI becomes labelled by 40 days of age. Adult animals exhibit high levels of binding in all laminae except layer IV. Age-dependent changes in PKC's laminar distribution do not seem to be correlated with specific anatomical, neurochemical, or behavioural events during development. PKC appears to be associated with cell bodies or processes intrinsic to the visual cortex, and is probably not located on the terminals of cortical afferents.

Aging↗
PubMed1988

Surgical undercutting prevents receptor redistribution in developing kitten visual cortex.

Recent studies have shown that several receptor populations in cat visual cortex undergo alterations in their laminar distributions during postnatal development (Shaw et al., 1984a,b; 1986b). These redistributions occur during the first few months of postnatal life, coincident with the physiologically defined critical period for cortical plasticity. In the present communication, we demonstrate that receptor redistributions can be prevented from occurring, or progressing once started, by surgically isolating the visual cortex at appropriate postnatal ages. These data suggest that the maturation of the chemical circuitry of the visual cortex is dependent on factors of extrinsic origin.

Animals↗
PubMed1987

Benzodiazepine ([3H]flunitrazepam) binding in cat visual cortex: ontogenesis of normal characteristics and the effects of dark rearing.

[3H]Flunitrazepam (FNZ) binding sites were characterized in homogenates of cat visual cortex during normal postnatal development and following dark rearing from birth. In parallel experiments, the distribution and density of [3H]FNZ binding sites were examined by in vitro autoradiographic or 'scrape' methods. In homogenates, Bmax measurements showed low early values, rising to a peak in receptor density at about 60 days postnatal, followed by a decline in adulthood. At all ages, gamma-aminobutyric acid (GABA) altered the Kd, but not the Bmax of [3H]FNZ binding sites. Kd values showed a general increase with age, parallelled by an increased sensitivity to GABA. Receptor autoradiography revealed that the highest density of [3H]FNZ binding sites was in layer IV of cats of all ages. Deafferentation of extrinsic inputs to the visual cortex by surgical undercutting did not alter this pattern of laminar distribution, indicating that the receptors were associated with intrinsic cortical elements rather than subcortical inputs. Dark rearing had no effect on [3H]FNZ laminar distribution in the visual cortex. The Bmax was higher at 30 days postnatal, but did not differ significantly thereafter. Modulation by GABA was concomitantly higher at 30 days, but lower than normal in dark-reared animals at ages greater than 30 days postnatal. The results are discussed in relation to the normal and abnormal development of GABA receptors in the cat visual cortex.

Animals↗
PubMedDate not supplied

The role of spontaneous retinal activity before eye opening in the maturation of form and function in the retinogeniculate pathway of the ferret.

During early mammalian development, inputs from the two retinas intermix within the lateral geniculate nucleus (LGN), then segregate during the first postnatal week into layers that receive input from a single retina. Functionally, the LGN also changes markedly during the first postnatal month; early geniculate responses to retinal input are mainly excitatory, then inhibitory circuits mature within the LGN. These remarkable changes in form and function of the retinogeniculate pathway occur at a time when patterned visual activity is not present, but retinal ganglion cells already manifest spontaneous action potential activity. To examine the role of early retinal activity in these critical developmental processes, we placed the slow release polymer Elvax embedded with tetrodotoxin (TTX) into the vitreous chamber of one or both eyes of neonatal ferrets. Animals receiving monocular injection of TTX had the other eye treated with Elvax containing control citrate buffer. Intraocular injection of horseradish peroxidase was made at the end of the period of TTX treatment to reveal the retinal terminals in the LGN. Chronic monocular or binocular blockade of retinal activity during the first postnatal week did not prevent eye-specific segregation, although it made the boundaries between layers less distinct. Retinal terminals ended preferentially in the appropriate layer, but a large number of terminals were also present in the inappropriate layer. Further segregation was achieved during the second postnatal week of activity blockade, when most retinal terminals ended preferentially in the appropriate geniculate layer and sharper layer boundaries were present. However, a small but significant number of terminals still extended into the inappropriate layer. Together, these findings indicate that monocular as well as binocular blockade of retinal activity resulted in some anomalous retinogeniculate projections and delayed eye-specific patterning, but segregation was largely intact at the end of the second postnatal week. We also report here that intraocular tetrodotoxin had a marked effect on the maturation of intrinsic geniculate circuits prior to eye opening. Whole-cell patch-clamp recordings in the LGN slice preparation revealed that activity blockade prevented the maturation of the slow, but not the fast, hyperpolarizing potential of LGN neurons during the first postnatal month and up to P38, the oldest age studied. In conclusion, these results indicate that spontaneous retinal activity modulates the time course of binocular segregation but does not alone account for the segregation of retinogeniculate terminals. However, early retinal activity plays an important role in developing the intrinsic circuitry of the LGN.

Action Potentials↗