Cooperative changes in GABA, glutamate and activity levels: the missing link in cortical plasticity

Different intracortical mechanisms have been reported to contribute to the substantial topographic reorganization of the mammalian primary visual cortex in response to matching lesions in the two retinas: an immediate expansion of receptive fields followed by a gradual shift of excitability into the deprived area and finally axonal sprouting of laterally projecting neurons months after the lesion. To gain insight into the molecular mechanisms of this adult plasticity, we used immunocytochemical and bioanalytical methods to measure the glutamate and GABA neurotransmitter levels in the visual cortex of adult cats with binocular central retinal lesions. Two to four weeks after the lesions, glutamate immunoreactivity was decreased in sensory-deprived cortex as confirmed by HPLC analysis of the glutamate concentration. Within three months normal glutamate immunoreactivity was restored. In addition, the edge of the unresponsive cortex was characterized by markedly increased glutamate immunoreactivity 2-12 weeks postlesion. This glutamate immunoreactivity peak moved into the deprived area over time. These glutamate changes corresponded to decreased spontaneous and visually driven activity in unresponsive cortex and to strikingly increased neuronal activity at the border of this cortical zone. Furthermore, the previously reported decrease in glutamic acid decarboxylase immunoreactivity was found to reflect decreased GABA levels in sensory-deprived cortex. Increased glutamate concentrations and neuronal activity, and decreased GABA concentrations, may be related to changes in synaptic efficiency and could represent a mechanism underlying the retinotopic reorganization that occurs well after the immediate receptive field expansion but long before the late axonal sprouting.     

Rapid down-regulation of tyrosine hydroxylase expression in the olfactory bulb of naris-occluded adult rats

In most sensory systems, afferent innervation regulates morphological and biochemical characteristics of target cells for a limited time during development. Sensory deprivation experiments in adult rats also have suggested a critical period for afferent influences on olfactory bulb structure and function. Previous odorant deprivation studies that employed unilateral naris closure in neonatal rats demonstrated down-regulation of the catecholamine biosynthetic enzyme tyrosine hydroxylase (TH) in dopamine neurons intrinsic to the olfactory bulb. Accompanying the altered biochemical parameters was a decrease in bulb size. To distinguish between deprivation-induced alterations in TH expression secondary to developmental sequelae and those occurring in mature neurons, the consequences of unilateral naris closure were assessed in young adult rats. In agreement with previous studies significant postnatal increases occurred in TH expression and total protein, an indication of bulb size. At 30 days post-closure, total protein was unaltered in the ipsilateral olfactory bulb but showed a small (12.9%), significant decline at 60 days. In contrast to the limited morphological consequences of odor deprivation, profound reductions occurred in TH expression. TH activity ipsilateral to the closure decreased significantly by 14 days post-closure and remained depressed for up to 6 months. In parallel with enzyme activity, TH immunoreactivity did not decline in the first few days post-closure. In situ hybridization revealed that TH mRNA levels decreased rapidly, i.e., by 2 days post-closure, reached a nadir at 1 month, and remained depressed for at least 6 months. The capacity of odor deprivation in the adult rat olfactory system to down-regulate TH expression suggests that this phenotypic alteration occurs independently of a presumed critical period. © 1996 Wiley-Liss, Inc.     

Metabolic and neurochemical plasticity of γ-aminobutyric acid-immunoreactive neurons in the adult macaque striate cortex following monocular impulse blockade: Quantitative electron microscopic analysis

The purpose of the present study was to examine the effects of retinal impulse blockade on γ-aminobutyric acid (GABA)-immunoreactive (GABA-IR) neurons in cytochrome oxidase (CO)-rich puffs of the adult monkey striate cortex. Specifically, we wished to know if changes occurred in their CO activity, GABA immunoreactivity, and synaptic organization. A double-labeling technique, which combined CO histochemistry and postembedding GABA immunocytochemistry on the same ultrathin sections, was used to reveal simultaneously the distribution of the two markers. We quantitatively compared changes in GABA-IR neurons of deprived puffs (DPs) with respect to non-deprived puffs (NPs) 2 weeks after monocular tetrodotoxin treatment. We found that the proportion of darkly CO reactive mitochondria in GABA-IR neurons of DPs drastically decreased to about half of those in NPs. There was a greater reduction of CO levels in GABA-IR axon terminals than in their cell bodies and dendrites. In contrast, most non-GABA-IR neurons displayed no significant change in their CO levels. Morphologically, GABA-IR neurons and axon terminals in DPs showed a significant shrinkage in their mean size. GABA immunoreactivity, as indicated by the density of immunogold particles in GABA-IR neurons, declined in DPs, and a greater decrease was also found in axon terminals than in cell bodies or dendrites. Moreover, the numerical density of GABA-IR axon terminals and synapses in DPs was significantly reduced without changes in that of asymmetric and symmetric synapses. Thus, the present results support the following conclusions: 1) Oxidative metabolism and neurotransmitter expression in GABA-IR neurons are tightly regulated by neuronal activity in adult monkey striate cortex; 2) GABA-IR neurons are much more vulnerable to functional deprivation than non-GABA-IR ones, suggesting that these inhibitory neurons have stringent requirement for sustained excitatory input to maintain their heightened oxidative capacity; and 3) intracortical inhibition mediated by GABA transmission following afferent deprivation may be decreased in deprived puffs, because the oxidative capacity and transmitter level in GABAergic neurons, especially in their axon terminals, are dramatically reduced. © 1996 Wiley-Liss, Inc.     

Suppression of visual cortical evoked responses following deprivation of pattern vision in adult mice

The effect of visual loss on the adult neocortex can have significant impact on the success of a visual implant. Recent research has shown that the adult neocortex retains substantial plasticity following a disruption to its afferent input. The result of these changes may hamper the development of a visual prosthesis if visual sensation cannot be effectively restored by stimulation of the surviving elements of the visual pathway. In order to evaluate further the visual performance of the mammalian adult brain following visual loss, especially the dominant form of blindness in humans, namely loss of pattern vision, we examined the cortical evoked potential of adult mice following 7, 30 and 120 days of visual deprivation via bilateral eyelid suture. Cortical potentials were elicited with a flash visual stimulus or by electrical stimulation of the retina. We found that after 7 days deprivation there was a potentiation of the evoked response while at 30 and 120 days deprivation the visual evoked responses were significantly reduced. Increasing the visual stimulus intensity reduced the effects. The electrical evoked potential demonstrated a corresponding reduction in stimulus threshold at 7 days and a corresponding rise (40–50%) after 30 and 120 days. These findings suggest that the adult brain exhibited significant experience-dependent modifications following visual loss, and the impact depended on the duration of deprivation. Such reduction in visual responsiveness, especially with electrical activation, will need to be taken into account in the development of a visual implant.     

Differential effects of neurotrophins on ocular dominance plasticity in developing and adult cat visual cortex

In the present study we examine the influence of neurotrophins on experience-dependent synaptic rearrangement in developing and adult visual cortex. Brain-derived neurotrophic factor (BDNF) or nerve growth factor (NGF) was continuously infused into cortical area 18, and the functional architecture of the cortex was examined by use of optical and electrophysiological recording techniques. In kittens, BDNF infusion during monocular deprivation (MD) reversed the normally occurring ocular dominance (OD) shift towards the non-deprived eye so that the deprived eye dominated the BDNF-treated cortex after MD. Under conditions of equal activation of thalamocortical synapses, i.e. when animals were either subject to binocular deprivation (BD) or reared without deprivation, BDNF infusion did not disrupt binocularity of cortical units, but reversed the natural OD bias towards the contralateral eye in favour of the ipsilateral eye. In addition, BDNF treatment in kittens led to a loss of the orientation selectivity of cortical units irrespective of rearing conditions. In adult animals, BDNF influenced neither OD distributions nor orientation selectivity. The effect of NGF was markedly different. It was ineffective in kittens but in adult animals it caused a shift of OD towards the deprived eye when MD was combined with NGF infusion. However, in this case orientation selectivity was preserved. Thus, both neurotrophins have profound activity- and age-dependent effects on the functional architecture of the visual cortex. Moreover, our results indicate that simple substitution of neurotrophins in excess is unlikely to compensate for deprivation effects by preserving or restoring the normal functional architecture of the cortex.     

Reversal of the physiological effects of monocular deprivation in adult dark-reared cats

If kittens are dark-reared for 4 months and subsequently monocularly sutured, cells in area 17 become dominated by the experienced eye. We now find that the effects of monocular deprivation in adult dark-reared cats can be reversed by suturing the experienced eye and allowing the cat to use the deprived eye, an effect that has previously been shown only in young kittens. The presence of continuous or nearly continuous visual experience during infancy is required for the critical period to exhaust itself--brief periods of visual experience will not suffice.     

Retinal lesions induce fast intrinsic cortical plasticity in adult mouse visual system

Neuronal activity plays an important role in the development and structural–functional maintenance of the brain as well as in its life‐long plastic response to changes in sensory stimulation. We characterized the impact of unilateral 15° laser lesions in the temporal lower visual field of the retina, on visually driven neuronal activity in the afferent visual pathway of adult mice using in situ hybridization for the activity reporter gene zif268. In the first days post‐lesion, we detected a discrete zone of reduced zif268 expression in the contralateral hemisphere, spanning the border between the monocular segment of the primary visual cortex (V1) with extrastriate visual area V2M. We could not detect a clear lesion projection zone (LPZ) in areas lateral to V1 whereas medial to V2M, agranular and granular retrosplenial cortex showed decreased zif268 levels over their full extent. All affected areas displayed a return to normal zif268 levels, and this was faster in higher order visual areas than in V1. The lesion did, however, induce a permanent LPZ in the retinorecipient layers of the superior colliculus. We identified a retinotopy‐based intrinsic capacity of adult mouse visual cortex to recover from restricted vision loss, with recovery speed reflecting the areal cortical magnification factor. Our observations predict incomplete visual field representations for areas lateral to V1 vs. lack of retinotopic organization for areas medial to V2M. The validation of this mouse model paves the way for future interrogations of cortical region‐ and cell‐type‐specific contributions to functional recovery, up to microcircuit level.     

Metabolic activity in SmI cortical barrels of adult rats is dependent on patterned sensory stimulation of the mystacial vibrissae

Cytochrome oxidase (CO) histochemistry was used to examine the effect of sensory deprivation on metabolic activity in the somatosensory cortex (SmI) of adult rats. Chronic trimming of one or several rows of mystacial vibrissae resulted in a decrease in CO reactivity in the corresponding barrels in layer IV. Reduced CO staining also was observed in cortical laminae superficial and deep to the affected layer IV barrels, suggesting that patterned deflections of the whiskers are important for maintaining the metabolic activity of neurons at least 3 and perhaps 4 synapses removed from the periphery.     

Membrane characteristics of visual cortical neurons in in vitro slices

In in vitro slices prepared from the visual cortex of cats a total of 15 neurons were studied for their membrane properties by passing currents through an intracellular microelectrode and by electrical stimulation of the white matter. Passive membrane properties (mean ±S.D.): resting membrane potential, 58 ± 10mV; input resistance, 42.1 ± 34.0MΩ; time constant, 9.0 ± 3.6ms. Some neurons showed a linear relationship between intensity of injected DC current and frequency of thereby induced discharges.     

A comparison of the number of neurons in individual laminae of cortical areas 17, 18 and posteromedial suprasylvian (PMLS) area in the cat

The binocular region of area 17 (17B) has a greater number of neurons under a given unit of cortical surface (NC, number per column) than either the monocular region of area 17 (17M), area 18 or the posteromedial suprasylvian area (PMLS). The latter three areas follow the general principle of basic uniformity in the number of neurons under given units of cortical surface formulated by Rockel et al. This basic uniformity is maintained in layers I and II. The NC of other layers varies. The greatest differences are found in layer IV where the NC of each area is significantly different from that of each other area. Though the difference between layer IV of 17M and of either 18 and PMLS is largely offset by changes in the third layer, layers V (PMLS) and VI (18 and PMLS) also contribute to the compensation. The compensation between 18 and PMLS is due entirely to changes in layer VI. It is important to note that there are statistically demonstrable interindividual differences in the neuronal NC of the cats used in the present study. We suggest that these may be due to age, breeding, or rearing conditions, probably the latter.     

Cellular localization of the neural cell adhesion molecule L1 in adult rat neuroendocrine and endocrine tissues: comparisons with NCAM.

The tissue distribution and cellular localization of the neural cell adhesion molecule L1 was determined by immunocytochemistry at the optical and ultrastructural levels in adult rat neuroendocrine tissues and pancreatic endocrine cells. L1 was found to be abundant in the neurohypophysis but undetectable in the rest of the pituitary gland. It was barely detectable in the normal rat endocrine pancreas, but a rat pancreatic insulinoma cell line was found by immunofluorescence to express low levels of L1. In the adrenal medulla, it was present on a sub-population of chromaffin cells and its density appeared to be lower on surfaces exposed to the extracellular matrix. Double immunolabelling showed this sub-population to consist of noradrenergic chromaffin cells. Adrenergic chromaffin cells were found not to express L1. In addition, the tissue distribution and cellular localization of NCAM mRNAs was determined by in situ hybridization, extending our previous studies on the cellular expression of NCAM proteins in endocrine and neuroendocrine tissues. This confirmed that the NCAM message has a wider cellular distribution than L1 within the hypophysis and the adrenal gland. In addition to secretory cells, L1 immunoreactivity was detected in glial cells, in particular in the pituicytes of the neurohypophysis, which further distinguishes them from astrocytes, their counterparts in the central nervous system. These data are discussed in terms of the different embryological origins of the various endocrine tissues examined and also in terms of the specific design constraints imposed on these tissues during their development.     

Expression of insulin-like growth factor binding protein-4 and -5 mRNAs in adult rat forebrain

Accumulating evidence indicates that the insulin-like growth factors (IGFs) can act as neurotrophic factors. A family of at least six IGF binding proteins (IGFBPs) has been characterized. The IGFBPs prolong the half-life of IGFs in plasma and may modulate IGF action in a cell- or tissue-specific fashion. Two recently characterized IGFBPs, IGFBP-4 and -5, have been shown by northern blot hybridization to be expressed in rat brain, but their cellular sites of synthesis are poorly characterized. Because IGFBP-4 and IGFBP-5 could potentially modulate IGF actions in the brain, we used in situ hybridization histochemistry and ³⁵S-labeled IGFBP-4 and IGFBP-5 riboprobes to localize sites of IGFBP-4 and -5 mRNA expression in adult rat brain. The two IGFBP mRNAs are abundantly expressed within discrete regions of brain. The expression patterns of the two genes are largely nonoverlapping. Notably, IGFBP-4 mRNA is highly expressed within hippocampal and cortical areas, whereas IGFBP-5 mRNA is not detected above background in these areas. Within the hippocampus, abundant IGFBP-4 mRNA expression is detected in pyramidal neurons of the subfields of Ammon's horn and the subiculum and in the granule cell layer of the anterior hippocampal continuation. In the cortex, IGFBP-4 mRNA is widely expressed in most areas and layers. In contrast, IGFBP-5, but not IGFBP-4, mRNA is detected within thalamic nuclei, leptomeninges, and perivascular sheaths. The distinct expression patterns of IGFBP-4 and -5 mRNAs within the brain suggest that these IGFBPs may modulate paracrine/autocrine actions of the IGFs in discrete brain regions or compartmentalization of the IGFs within the brain. © 1994 Wiley-Liss, Inc.     

Tenascin-C expression and axonal sprouting following injury to the spinal dorsal columns in the adult rat

We have examined the expression and distribution of the extracellular matrix molecule tenascin-C in and around lesions of the thoracic dorsal columns in adult rats 3 days to 8 weeks after injury, using in situ hybridization, immunofluorescence, electron microscopy and immunoelectron microscopy. Numerous tenascin-C mRNA+ cells were present in and around the lesion at 3 days; fewer were present at 14 days and almost none 30 days after injury. Most tenascin-C mRNA+ cells in the spinal cord around the lesion were GFAP+, but most of those within the lesion were not, suggesting that tenascin-C is produced in the injured spinal cord by a subpopulation of astrocytes and by other cells that invade the lesion; these cells may include meningeal cells, macrophages, and Schwann cells. From 3 to 30 days after injury, heavy tenascin-C immunoreactivity was present at the lesion site (especially transections), and there was lighter immunoreactivity around the lesion and in the degenerating dorsal column. The heaviest immunoreactivity was associated with collagen fibrils in areas of expanded extracellular space and with basal laminae (covering Schwann cells and some astrocytes) but tenascin-C was also found close to the surfaces of some OX-42+ macrophages/microglia, leptomeningeal cells, and capillaries. Neurofilament (NF)+ axons grew into the highly tenascin-C-immunoreactive lesion sites, indicating that tenascin-C does not prevent axonal growth into these areas. However, such axons were not coated with tenascin-C except where directly exposed to the extracellular space. J. Neurosci. Res. 49:433–450, 1997. © 1997 Wiley-Liss, Inc.     

The Distribution of Brain-derived Neurotrophic Factor and its Receptor trkB in Parvlbumin-containing Neurons of the Rat Visual Cortex

We analysed the distribution of brain-derived neurotrophic factor (BDNF) and its receptor trkB in the adult rat visual cortex, paying particular attention to a GABAergic neuronal subpopulation—the parvalburnin-positive cells. We found expression of trkB in the cell body and apical dendrite of pyramidal neurons and in the cell body of non-pyramidal neurons. Double labelling experiments revealed extensive colocalization of parvalbumin and trkB immunoreactivity in non-pyramidal neurons. Interestingly, the trkB-positive pyramidal neurons appeared surrounded by parvalbumin-labelled boutons. The use of double immunohistochemistry and in situ hybridization histochemistry showed that parvalbumin-positive neurons express trkB mRNA. BDNF rnRNA was found in several cells. Coexpression of BDNF mRNA and parvalbumin immunoreactivity was extremely rare. These data strongly suggest that BDNF synthesized by cortical neurons acts as a postsynaptically derived factor for parvalbumin-positive neurons in the adult rat visual cortex.