Visual Experience-Dependent Expression of Fn14 Is Required for Retinogeniculate Refinement.

Sensory experience influences the establishment of neural connectivity through molecular mechanisms that remain unclear. Here, we employ single-nucleus RNA sequencing to investigate the contribution of sensory-driven gene expression to synaptic refinement in the dorsal lateral geniculate nucleus of the thalamus, a region of the brain that processes visual information. We find that visual experience induces the expression of the cytokine receptor Fn14 in excitatory thalamocortical neurons. By combining electrophysiological and structural techniques, we show that Fn14 is dispensable for early phases of refinement mediated by spontaneous activity but that Fn14 is essential for refinement during a later, experience-dependent period of development. Refinement deficits in mice lacking Fn14 are associated with functionally weaker and structurally smaller retinogeniculate inputs, indicating that Fn14 mediates both functional and anatomical rearrangements in response to sensory experience. These findings identify Fn14 as a molecular link between sensory-driven gene expression and vision-sensitive refinement in the brain.

Cannabinoid receptor CB2 modulates axon guidance.

Navigation of retinal projections towards their targets is regulated by guidance molecules and growth cone transduction mechanisms. Here, we present in vitro and in vivo evidences that the cannabinoid receptor 2 (CB2R) is expressed along the retino-thalamic pathway and exerts a modulatory action on axon guidance. These effects are specific to CB2R since no changes were observed in mice where the gene coding for this receptor was altered (cnr2 (-/-)). The CB2R induced morphological changes observed at the growth cone are PKA dependent and require the presence of the netrin-1 receptor, Deleted in Colorectal Cancer. Interfering with endogenous CB2R signalling using pharmacological agents increased retinal axon length and induced aberrant projections. Additionally, cnr2 (-/-) mice showed abnormal eye-specific segregation of retinal projections in the dorsal lateral geniculate nucleus (dLGN) indicating CB2R's implication in retinothalamic development. Overall, this study demonstrates that the contribution of endocannabinoids to brain development is not solely mediated by CB1R, but also involves CB2R.

A role for mDia, a Rho-regulated actin nucleator, in tangential migration of interneuron precursors.

In brain development, distinct types of migration, radial migration and tangential migration, are shown by excitatory and inhibitory neurons, respectively. Whether these two types of migration operate by similar cellular mechanisms remains unclear. We examined neuronal migration in mice deficient in mDia1 (also known as Diap1) and mDia3 (also known as Diap2), which encode the Rho-regulated actin nucleators mammalian diaphanous homolog 1 (mDia1) and mDia3. mDia deficiency impaired tangential migration of cortical and olfactory inhibitory interneurons, whereas radial migration and consequent layer formation of cortical excitatory neurons were unaffected. mDia-deficient neuroblasts exhibited reduced separation of the centrosome from the nucleus and retarded nuclear translocation. Concomitantly, anterograde F-actin movement and F-actin condensation at the rear, which occur during centrosomal and nuclear movement of wild-type cells, respectively, were impaired in mDia-deficient neuroblasts. Blockade of Rho-associated protein kinase (ROCK), which regulates myosin II, also impaired nuclear translocation. These results suggest that Rho signaling via mDia and ROCK critically regulates nuclear translocation through F-actin dynamics in tangential migration, whereas this mechanism is dispensable in radial migration.

Epilepsy gene LGI1 regulates postnatal developmental remodeling of retinogeniculate synapses.

Retinogeniculate connections undergo postnatal refinement in the developing visual system. Here we report that non-ion channel epilepsy gene LGI1 (leucine-rich glioma-inactivated), mutated in human autosomal dominant lateral temporal lobe epilepsy (ADLTE), regulates postnatal pruning of retinal axons in visual relay thalamus. By introducing an ADLTE-associated truncated mutant LGI1 (836delC) or excess full-length LGI1 into transgenic mice, we found that mutant LGI1 blocks, whereas excess LGI1 accelerates, retinogeniculate axon pruning. The normal postnatal single fiber strengthening was arrested by mutant LGI1 and, contrastingly, was enhanced by excess wild-type LGI1. The maximum response of the retinogeniculate synapses, conversely, remained the same in mature LGI1 transgenic mice, indicating that mutant LGI1 blocks, whereas excess wild-type LGI1 promotes, weak axon fiber elimination. Heterozygous deletion of the LGI1 gene, as found in ADLTE patients, inhibited postnatal retinogeniculate synapse elimination, an effect similar to the ADLTE truncated mutant LGI1. The results identify sensory axon remodeling defects in a sensory aura-associated human epilepsy disorder.

Natural and lesion-induced apoptosis in the dorsal lateral geniculate nucleus during development.

We investigated natural and lesion-induced apoptosis in the developing rat dorsal lateral geniculate nucleus (dLGN). These lesions involved: i) monocular enucleation, and ii) unilateral ablation of the visual cortex at different postnatal ages before eye opening. We identified dying cells as apoptotic with light and electron microscopy, using terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL), and immunohistochemistry for active caspase-3. In the dLGN of normal animals, TUNEL+cells were detected during the first postnatal week, with a peak at postnatal day (P) 1. Following enucleation at birth or at P7, the frequency of apoptotic cells in the contralateral dLGN increased significantly at postlesion day (PLD) 1 and returned to normal values by PLD7. In contrast to early lesions, enucleation at P14 did not induce significant changes in apoptosis in the dLGN. Cortical lesions performed at P0, P7 or at P14 induced the death of the overwhelming majority of cells in the ipsilateral dLGN, which led to a severe reduction in size of the nucleus by PLD7 and its complete elimination by adulthood. Double labeling with TUNEL and immunofluorescence for neuronal nuclear protein (NeuN) showed that in both normal and lesioned animals, apoptotic cells were mainly neurons. We suggest that: i) apoptosis in the dLGN occurs during the precritical period of neuronal maturation; ii) developing neurons in the dLGN are more dependent on the integrity of their connections with the visual cortex than with the retina for survival; and iii) lesion-induced apoptosis in the dLGN during development depends on the type and extent of the connectivity affected.

Molecular development of the lateral geniculate nucleus in the absence of retinal waves during the time of retinal axon eye-specific segregation.

When retinal waves are inhibited binocularly, eye-specific segregation of retinal axons is disrupted, and retinal axons from the two eyes remain intermingled in the lateral geniculate nucleus (LGN). This effect of binocular retinal wave inhibition is mediated by the lack of activity-dependent competition between retinal axons from the two eyes, but it is unknown whether this effect is also mediated by the developmental arrest of the LGN in an immature state. Here we find developmental markers of the LGN during eye-specific segregation. The expression levels of Purkinje cell protein 4 (PCP4/PEP19), transcription factor 7-like 2 (TCF7L2/TCF4) and LIM homeobox protein 9 (Lhx9) in the LGN change significantly during eye-specific segregation. Using PCP4, TCF7L2 and Lhx9 as developmental markers of the LGN, we examine whether LGN development is affected by binocular disruption of retinal waves during eye-specific segregation. Binocular injection of epibatidine strongly inhibits eye-specific segregation, whereas it does not affect the expression of PCP4, TCF7L2 and Lhx9. Furthermore, the expression of PCP4, TCF7L2 and Lhx9 is normal in binocularly enucleated animals and in mice treated with the monoamine oxidase A (MAOA) inhibitor, clorgyline. In addition, our experiments using LGN slice cultures show that the expression of PCP4 and TCF7L2 in LGN slices changes as in vivo. Our results suggest that LGN development proceeds, at least in part, even in the absence of retinal inputs. PCP4, TCF7L2 and Lhx9 should be useful to examine LGN development during eye-specific segregation in mice and in ferrets.

Neonatal nicotine exposure impairs nicotinic enhancement of central auditory processing and auditory learning in adult rats.

Children of women who smoke cigarettes during pregnancy display cognitive deficits in the auditory-verbal domain. Clinical studies have implicated developmental exposure to nicotine, the main psychoactive ingredient of tobacco, as a probable cause of subsequent auditory deficits. To test for a causal link, we have developed an animal model to determine how neonatal nicotine exposure affects adult auditory function. In adult control rats, nicotine administered systemically (0.7 mg/kg, s.c.) enhanced the sensitivity to sound of neural responses recorded in primary auditory cortex. The effect was strongest in cortical layers 3 and 4, where there is a dense concentration of nicotinic acetylcholine receptors (nAChRs) that has been hypothesized to regulate thalamocortical inputs. In support of the hypothesis, microinjection into layer 4 of the nonspecific nAChR antagonist mecamylamine (10 microM) strongly reduced sound-evoked responses. In contrast to the effects of acute nicotine and mecamylamine in adult control animals, neither drug was as effective in adult animals that had been treated with 5 days of chronic nicotine exposure (CNE) shortly after birth. Neonatal CNE also impaired performance on an auditory-cued active avoidance task, while having little effect on basic auditory or motor functions. Thus, neonatal CNE impairs nicotinic regulation of cortical function, and auditory learning, in the adult. Our results provide evidence that developmental nicotine exposure is responsible for auditory-cognitive deficits in the offspring of women who smoke during pregnancy, and suggest a potential underlying mechanism, namely diminished function of cortical nAChRs.

Differential effects of cortical neurotrophic factors on development of lateral geniculate nucleus and superior colliculus neurons: anterograde and retrograde actions.

Neurotrophins strongly affect visual system development and plasticity. However, the mode of delivery and targets of neurotrophin action are still under debate. For instance, cortical NT-4/5 (neurotrophin 4/5; Ntf4/5) was shown to rescue lateral geniculate nucleus (LGN) neurons from monocular deprivation-induced atrophy suggesting a retrograde action on thalamic afferents. It is still unclear whether LGN neurons respond to NT-4/5 and other neurotrophins during development in animals with normal vision. We now show that infusions of NT-4/5 and NGF (nerve growth factor) into visual cortex at the onset and the peak of the critical period accelerated LGN neuron growth. BDNF (brain-derived neurotrophic factor) was ineffective. The effects of neurotrophin on LGN development were clearly dissociated from the effects at cortical level because soma growth of cortical layer IV and VI neurons was strongly promoted by BDNF. NT-4/5 was only effective at the onset, but no longer at the peak of the critical period suggesting a switch in neurotrophin dependency for these cortical cell classes. To dissociate retrograde and anterograde effects of the TrkB ligands, we analyzed the stratum griseum superficiale (SGS) of the superior colliculus, a target of visual cortical efferents. Indeed, TrkB-expressing inhibitory SGS neurons responded to cortical NT-4/5 infusion with somatic growth. Strikingly, the TrkB-expressing excitatory tectothalamic calbindin neurons in the SGS did not respond. This demonstrated for the first time a selective cell type-specific anterograde action of NT-4/5 and suggested for the LGN that anterograde as well as retrograde effects contribute to soma size regulation. Strikingly, cortical infusion of the cytokine LIF, which affects development of visual cortex neurochemical architecture, transiently inhibited growth of neurons in LGN, cortical layer IV and VI and SGS. In summary, the study presents three important results. First, central neurons regulate soma size development in an age-and ligand-specific fashion. Second, NT-4/5 and NGF accelerate LGN development in rats with normal vision while LIF delays growth. Third, anterogradely transported NT-4/5 effectively promotes neuronal maturation. These differential actions on subcortical neurons may contribute to the different effects of neurotrophins on visual system development and plasticity.

Role of retinal afferents in regulating growth and shape of the lateral geniculate nucleus.

Segregated binocular maps in the dorsal lateral geniculate nucleus (LGN) develop from a stage where they initially overlap. Sophisticated computational models have been used to describe the dynamics of three-dimensional LGN shape changes that play a role in segregation. These models have revealed specific nuclear growth vectors associated with the process of ocular segregation in the LGN (Williams and Jeffery [2001] J Comp Neurol 430:332-342.). In this study, we use similar techniques to determine whether retinal innervation contributes to the dynamics of shape maturation in the ferret LGN. In this animal, 90% of the retinal innervation of the mature LGN comes from the contralateral eye. If one eye is removed before segregation, the projection from the remaining eye remains diffuse and nuclear growth is stunted. Here, we quantify this effect and show that removing the contralateral projection before segregation has a profound impact on LGN size but changes its ultimate shape by only 12%. The impact on shape on the other side of the brain where the ipsilateral projection is removed, which accounts for only 10% of its innervation in maturity, is less than 2%. Hence, retinal innervation plays a minor role in determining mature LGN shape. Although in both hemispheres, the ultimate shape of the nucleus is close to normal, removal of the larger projection disrupts normal growth vectors, with almost none being present in the 5 days after enucleation, when the normal nucleus expands markedly. Hence, the effect of enucleation is to delay shape maturation. Growth vectors absent after removal of the smaller projection are mainly confined to those in what would be the binocular region.

Nicotine exposure during a postnatal critical period alters NR2A and NR2B mRNA expression in rat auditory forebrain.

Chronic nicotine exposure (CNE) can alter brain development and is thought to produce deficits in auditory function. Previously, we found that CNE during the second postnatal week, but not before or after, increases the duration of excitatory postsynaptic potentials (EPSPs) mediated by N-methyl-D-aspartate receptors (NMDARs) in rat auditory cortex. It was proposed that a potential mechanism underlying increased EPSP duration could be over-stimulation of presynaptic nicotinic acetylcholine receptors, leading to prolonged glutamate release. Since glutamatergic activity regulates levels of postsynaptic NMDAR subunits, here we examine the effects of CNE on mRNA expression for the NR2A and NR2B subunits in auditory cortex and thalamus. Two days of CNE (postnatal days 8-9), produced no effects, but 5 days (postnatal days 8-12) enhanced cortical NR2A mRNA levels and reduced thalamic NR2B mRNA levels for up to 2 weeks. These effects are consistent with the hypothesis that CNE during a postnatal critical period disrupts auditory cortex development by over-stimulating glutamatergic synapses.

Onset of calbindin-D 28K and parvalbumin expression in the lateral geniculate complex and olivary pretectal nucleus during postnatal development of the rat.

The onset and distribution of calbindin (CB) and parvalbumin (PV) immunoreactivity were investigated in the lateral geniculate nuclear complex and the olivary pretectal nucleus (OPT) in developing rats. CB expression occurred early (before eye-opening) in the relay neurons of the intergeniculate leaflet, parvocellular portion of the ventral lateral geniculate nucleus and OPT relating to ambient vision mediated by W-like retinal ganglion cells. On the contrary, PV expression occurred late (after eye-opening) in the relay neurons of the magnocellular portion of the ventral lateral geniculate nucleus (VLGMC) and OPT relating to focal vision mediated by Y-like retinal ganglion cells. A unilateral eye enucleating experiment indicated that the VLGMC and OPT received dense input from PV-positive Y-like retinal ganglion cells. The results show the different onsets of CB and PV expressions in the retino-recipient thalamic and pretectal nuclei receiving inputs from different kinds of retinal ganglion cells.

Transient expression of synaptic zinc during development of uncrossed retinogeniculate projections.

The transition metal zinc is an essential dietary constituent that is believed to serve an important intercellular signaling role at certain excitatory synapses in the central nervous system. In the present study, we used histochemical techniques to investigate the distribution of synaptic zinc during postnatal development of retinogeniculate projections in rats. From postnatal day (P) 1 until P-21, the pattern of zinc histochemical staining in the dorsal lateral geniculate nucleus (LGNd) precisely matched the distribution of axon terminals from the ipsilateral eye that were labeled by anterograde transport of horseradish peroxidase. Regions of the LGNd that contained only crossed axons were devoid of zinc staining. Abnormalities in the distribution of uncrossed retinogeniculate projections in albino versus pigmented rats were paralleled by identical variations in localization of synaptic zinc. Unilateral enucleation on P-10 was followed within 5 days by loss of zinc staining in the LGNd ipsilateral to the removed eye without affecting staining in the contralateral nucleus. Finally, the ability to detect zinc histochemically in the LGNd ceased at approximately P-24. These findings provide evidence that zinc is sequestered within synaptic boutons of a subpopulation of retinal ganglion cells whose axons terminate on the ipsilateral side of the brain. The duration of zinc staining overlaps with the major period of axonal remodeling in the LGNd, suggesting that synaptically released zinc may play a role in postnatal refinement of retinogeniculate projections.

Effects of monocular enucleation on parvalbumin in rat visual system during postnatal development.

PURPOSE: To evaluate the hypothesis that the expression of the calcium-binding protein parvalbumin (PV) in a subpopulation of gamma-aminobutyric acid (GABA)ergic neurons is an appropriate molecular marker for the effect on ocular dominance plasticity of monocular deprivation during the postnatal sensitive period. METHODS: Long-Evans rats underwent monocular enucleation immediately before eye opening (postnatal day [P] 14). Immunohistochemical analysis using anti-PV antibody was performed on the superior colliculus (SC) and lateral geniculate nucleus (LGN) at P45. In the visual cortex (VC) developmental changes in immunoreactivity were also examined at the ages of P17, P20, P27, and P45. Northern blot analysis for PV mRNA was also performed at P45. Changes in PV expression in the visual system of these rats were evaluated by use of a computer-based quantitative technique. RESULTS: PV-immunoreactive neurons were present in the SC and VC, whereas only a few were found in the LGN. The monocular enucleation at the onset of the sensitive period markedly reduced PV immunoreactivity in the neuropil of the SC, contralateral to the enucleated eye when examined one month later. No consistent and significant change in PV immunoreactivity was found in either the LGN or the VC. The number of PV-immunoreactive neurons in the VC rapidly decreased to the adult level during the middle of the sensitive period. The expression of PV mRNA in these central visual structures was not affected by early monocular enucleation. CONCLUSIONS: Expression of PV is developmentally regulated, and marked changes in its protein expression in the SC can be induced by monocular enucleation. Contrary to the original hypothesis, monocular enucleation did not consistently affect the expression of PV in the rat VC. The expression of PV is probably regulated by multiple factors, not merely by binocular competition.

Specification of retinogeniculate X and Y axon arbors in cats: fundamental differences in developmental programs.

Monocular enucleation at E36, followed by intracellular labeling of single, physiologically identified X and Y axons, demonstrates fundamental differences in their termination patterns within the lateral geniculate nucleus (LGN). X axons have arbors that appear normal in their dorsoventral extent, though some are located in inappropriate regions of the LGN. Y axons have arbors that are either abnormally tall, spanning the entire extent of the LGN, or of normal height but located in inappropriate regions of the LGN. These termination patterns resemble patterns seen after monocular enucleation at E44, and reinforce the conclusion that X and Y axons differ fundamentally in the cues that constrain the dorsoventral extents of their arbors.

Competition in retinogeniculate patterning driven by spontaneous activity.

When contacts are first forming in the developing nervous system, many neurons generate spontaneous activity that has been hypothesized to shape appropriately patterned connections. In Mustela putorius furo, monocular intraocular blockade of spontaneous retinal waves of action potentials by cholinergic agents altered the subsequent eye-specific lamination pattern of the lateral geniculate nucleus (LGN). The projection from the active retina was greatly expanded into territory normally belonging to the other eye, and the projection from the inactive retina was substantially reduced. Thus, interocular competition driven by endogenous retinal activity determines the pattern of eye-specific connections from retina to LGN, demonstrating that spontaneous activity can produce highly stereotyped patterns of connections before the onset of visual experience.

Development of primary visual projections occurs entirely postnatally in the fat-tailed dunnart, a marsupial mouse, Sminthopsis crassicaudata.

We have examined the development of retinal projections in a diminutive polyprotodont marsupial, the fat-tailed dunnart, Sminthopsis crassicaudata. Here, we document the most immature mammalian visual system at birth described to date. At postnatal day (P) 0, the retinal ganglion cell layer has yet to form, and axons have not entered the optic stalk. By P4, the retinal ganglion cell layer could be distinguished at the posterior pole, and the front of growing axons extended one-third the length of the optic stalk, a distance of approximately 150 microm; a few pioneer growth cones had grown beyond the main axon group but had still to reach the midline. Axons had decussated at the optic chiasm by P10 to penetrate the base of the contralateral optic tract and, by P15, had reached the dorsal lateral geniculate nucleus (dLGN), superior colliculus (SC), and accessory optic system (AOS); ipsilaterally projecting axons matured slightly later. From P20, axons had reached the caudal SC both contralaterally and ipsilaterally and terminated throughout the depth of the retinorecipient layers. After P30, the projections gradually refined. Within the rostral dLGN, segregation into four contralateral and four ipsilateral bands occurred by P50, approximately 5 days after eye opening. The projection to the ipsilateral SC underwent refinement by P50, becoming restricted to its rostral pole, and presented as discrete patches within the stratum opticum. At birth, the dunnart visual system is comparable to early to midembryonic stages [embryonic day (E) 12, E14, E19, E24, and E30, respectively] in the mouse, rat, ferret, cat, and monkey. The extreme immaturity of the neonatal dunnart together with the observation that the entire development of the primary optic pathway occurs postnatally over a protracted period make this marsupial especially valuable for investigating factors that control pathway formation in the early developing mammalian primary visual system.

Radiation-induced, lamina-specific deletion of neurons in the primate visual cortex.

To examine the early determinants of cortical cytoarchitecture, we deleted specific neuronal classes in the primate visual cortex by ionizing irradiation at selected prenatal stages. Multiple doses of X-rays were delivered to the macaque monkey brain between embryonic day (E) 80 and E90 to block the division of cells destined to populate the superficial cortical layers, between E70 and E79 to eliminate neurons destined for the middle layers; and between E33 and E40 to delete neurons destined for the lateral geniculate nucleus (LGN) that project to the cortex. All animals were killed after birth, and their brains were processed for histological and electron microscopic analyses. Cell density and number in the LGN and visual cortex were determined by using three-dimensional, computer-aided morphometry. In animals irradiated with low doses (total of approximately 200 cGy) during the genesis of the LGN but before the onset of corticogenesis (E33-40), the LGN was reduced in both volume and number of neurons. Area 17 in these animals displayed only slight changes in cortical thickness, cell density, and area-specific cytoarchitectonic features, whereas the total surface devoted to area 17 was significantly diminished. In contrast, animals irradiated with low doses during the period of corticogenesis, after the completion of the LGN genesis, showed no significant change in the volume of the LGN or in the number of its cells. Moreover, in these animals, the surface of area 17 was not significantly altered, although the cortical layers generated at the time of irradiation had a significantly lower density and total number of cells, whereas the layers generated before and after the period of irradiation were spared. In contrast, cases exposed to high doses of X-ray (total > 300 cGy) showed more severe effects, including all layers. However, layers normally generated during irradiation were depleted and consisted of cell-sparse strata populated by densely packed neuropil (axons, small dendrites, dendritic spines, and synaptic boutons). These cell-sparse strata were situated deeper in the early irradiated animals than in the later irradiated animals, and their laminar position changed abruptly at the area 17/18 border. These results show that low doses of irradiation in a slowly developing primate brain can be used effectively to eliminate targeted classes of neurons before they reach their final position, providing an opportunity to examine the role of cell-cell interactions in the formation of circuitry and the role of specific cell classes in cortical development.

Developmental shift of synaptic vesicle protein 2 from axons to terminals in the primary visual projection of the hamster.

Synaptic vesicle protein 2 is an integral synaptic vesicle membrane glycoprotein which is present in all synapses for which it has been examined. We used an anti-synaptic vesicle protein 2 monoclonal antibody to examine synaptic vesicle protein 2 localization in the developing hamster retinofugal pathway. From postnatal day 0 to day 1, a period of elongation of retinal ganglion cell axons to their central targets, fiber fascicles in the optic tract over the lateral geniculate nucleus were intensely synaptic vesicle protein 2-immunoreactive. Adjacent to the optic tract, single fibers could be seen. We also observed a marked immunostaining in growth cones and fiber fascicles in retinal explants in culture. By postnatal day 2, the staining of single fibers had ended, and by postnatal day 5, during the formation of terminal arbors, numerous fine puncta of synaptic vesicle protein 2 immunoreactivity were distributed within the neuropil of the lateral geniculate nucleus. In the adult, the optic tract was devoid of synaptic vesicle protein 2 staining, while the neuropil contained distinct immunoreactive profiles, particularly in the outer shell of the lateral geniculate. These synaptic vesicle protein 2-positive profiles closely resembled the grape-like clusters and large swellings of two known retinal axon terminal types. Eye removal resulted in the rapid disappearance of these synaptic vesicle protein 2-labelled terminal profiles contralateral to the enucleation. A similar pattern of synaptic vesicle protein 2 immunoreactivity was observed in the superior colliculus. From postnatal day 0 to day 2, retinal fiber fascicles in the stratum griseum superficiale/stratum opticum were darkly stained for synaptic vesicle protein 2. By postnatal day 5, the immunoreactivity shifted to the neuropil and from postnatal day 6 onwards, the synaptic vesicle protein 2 immunoreactivity was more intense in the stratum griseum superficiale than in the optic fibre layer. This study demonstrates dense synaptic vesicle protein 2-labelling of elongating axons both in vivo and in vitro. However, coincident with the transition from retinal ganglion cell axon elongation to terminal arborization, synaptic vesicle protein 2 is progressively restricted to synaptic terminals and becomes undetectable in axons. This study is the first to document an axonal localization of synaptic vesicle protein 2 during development and raises the question as to its role during axonal elongation.

Dendritic development in the dorsal lateral geniculate nucleus of ferrets in the postnatal absence of retinal input: a Golgi study.

In order to determine the ongoing role of retinal fibers in the development of dorsal lateral geniculate nucleus (dLGN) neurons during postnatal development, the development of dLGN neurons in the postnatal absence of retinal input was studied in pigmented ferrets using the Golgi-Hortega technique. The development of four dLGN cell classes, defined on the basis of somatic and dendritic morphology, was described previously in normal ferrets (Sutton and Brunso-Bechtold, 1991, J. Comp. Neurol. 309:71-85). The present results indicate that the morphological development of dLGN neurons is strikingly similar in normal and experimental ferrets. The exuberant dendritic appendages that appear after eye opening in normal ferrets are overproduced and eliminated in the postnatal absence of retinal input; however, the final reduction of these transient appendages is delayed. Because exuberant appendages develop in the absence of retinal input, their production cannot depend upon visual experience. Differences in cell body size between normal and experimental ferrets are apparent only after neurons can be classified at the end of the first postnatal month. Cell body size is markedly reduced for class 1 neurons; class 2 cells also are reduced in size but to a far lesser extent. As there is a general trend for class 1 neurons to have the functional properties of Y-cells, it is likely that the dLGN neurons most affected by the absence of retinal input also are Y-cells.

Transition from developing to mature patterns of acetylcholinesterase activity in rat visual cortex: implications for the time-course of geniculocortical development.

Patterns of acetylcholinesterase (AChE) histochemical staining in cortical area 17 differ in infant and mature rats. In infants, intense AChE activity is seen as a band corresponding to layer IV and deep layer III of the visual cortex, and this staining is associated with terminal fields of geniculocortical neurons. In adult animals, AChE activity is densest in deep layer IV and layer V and is associated with projections originating in the basal forebrain. The present study investigated the transition from developing to mature patterns of AChE staining in visual cortex. Unilateral lesions were placed in either the lateral geniculate body or the basal forebrain of rats postnatal days 8 (P8) to adulthood; the effects of these lesions on patterns of AChE activity in visual cortex were studied with histochemical techniques and optical densitometry. Lesions involving the lateral geniculate body markedly reduce AChE activity in visual cortex of P12 rats, had moderate effects in P20 rats, and had no apparent effect on AChE activity of visual cortex of rats aged P40 and older. Lesions of basal forebrain had little effect on AChE activity in visual cortex of P12 animals, increasing effect in P15-35 rats, and eliminated much of AChE staining in visual cortex of adults. The period of transition from developing to mature patterns of AChE activity in visual cortex of animals bilaterally enucleated at birth was not different from the period of transition in normally sighted animals. These data indicate that mature patterns of AChE activity in visual cortex are not achieved until well into the second month of life. If transient AChE expression is characteristic of geniculocortical neurons during the period of time in which axons are proliferating within visual cortex, then these data indicate that geniculocortical connections may be forming well into the second month of life in the rat.