Intrauterine hypoxia-ischemia alters nitric oxide synthase expression and activity in fetal and neonatal rat brains.

The effects of intrauterine hypoxia-ischemia (HI) on nitric oxide synthase (NOS) activity and on expression of NOS isoforms were investigated in fetal and neonatal rat brains. Rat fetuses were subjected to either a 30-min intrauterine HI insult or a sham operation (SH) on gestational day 17 (G17). NOS activity in the homogenate of the rat brain was detectable on G17 and increased with age. NOS activity in the HI group was 20-30% higher than in the SH group from 6 to 48 h after the HI, but was 30% lower than in the SH group from postnatal day 8 to 14. Expression of the inducible NOS (iNOS) mRNA, as examined by RT-PCR, was increased as compared to the SH group from 6 to 24 h after the HI surgery. Expression of the constitutive neuronal NOS (nNOS) mRNA was reduced in the HI group from 24 h after the HI surgery up to postnatal day 14. Immunoblotting data have shown that alterations in NOS isoform protein expression caused by the intrauterine HI were consistent with the mRNA expression data. The overall results indicate that prenatal HI has long-lasting effects on function and expression of NOS in fetal and neonatal rat brains and that the altered NOS activity may be associated with prenatal HI-induced neurological abnormalities.

Why neurons die: cell death in the nervous system.

It is likely that humans are born with all of the nerve cells (neurons) that will serve them throughout life. For all practical purposes, when our neurons die, they are lost forever. During nervous system development, about one-and-a-half times the adult number of neurons are created. These "extra" neurons are then destroyed or commit suicide. This process of programmed cell death occurs through a series of events termed apoptosis and is an appropriate and essential event during brain development. Later in life, inappropriate neuronal cell death may result from pathological causes such as traumatic injury, environmental toxins, cardiovascular disorders, infectious agents, or genetic diseases. In some cases, the death occurs through apoptosis. In other cases, cell death is random, irreversible, and uncontrollable; to distinguish it from the controlled, planned cell death of apoptosis, we call this necrotic cell death. Understanding the difference between apoptotic and necrotic cell death is essential for designing therapies which will prevent or limit inappropriate cell death in the nervous system.

Mitochondrial manganese superoxide dismutase prevents neural apoptosis and reduces ischemic brain injury: suppression of peroxynitrite production, lipid peroxidation, and mitochondrial dysfunction.

Oxidative stress is implicated in neuronal apoptosis that occurs in physiological settings and in neurodegenerative disorders. Superoxide anion radical, produced during mitochondrial respiration, is involved in the generation of several potentially damaging reactive oxygen species including peroxynitrite. To examine directly the role of superoxide and peroxynitrite in neuronal apoptosis, we generated neural cell lines and transgenic mice that overexpress human mitochondrial manganese superoxide dismutase (MnSOD). In cultured pheochromocytoma PC6 cells, overexpression of mitochondria-localized MnSOD prevented apoptosis induced by Fe2+, amyloid beta-peptide (Abeta), and nitric oxide-generating agents. Accumulations of peroxynitrite, nitrated proteins, and the membrane lipid peroxidation product 4-hydroxynonenal (HNE) after exposure to the apoptotic insults were markedly attenuated in cells expressing MnSOD. Glutathione peroxidase activity levels were increased in cells overexpressing MnSOD, suggesting a compensatory response to increased H2O2 levels. The peroxynitrite scavenger uric acid and the antioxidants propyl gallate and glutathione prevented apoptosis induced by each apoptotic insult, suggesting central roles for peroxynitrite and membrane lipid peroxidation in oxidative stress-induced apoptosis. Apoptotic insults decreased mitochondrial transmembrane potential and energy charge in control cells but not in cells overexpressing MnSOD, and cyclosporin A and caspase inhibitors protected cells against apoptosis, demonstrating roles for mitochondrial alterations and caspase activation in the apoptotic process. Membrane lipid peroxidation, protein nitration, and neuronal death after focal cerebral ischemia were significantly reduced in transgenic mice overexpressing human MnSOD. The data suggest that mitochondrial superoxide accumulation and consequent peroxynitrite production and mitochondrial dysfunction play pivotal roles in neuronal apoptosis induced by diverse insults in cell culture and in vivo.

Bcl-2 protects isolated plasma and mitochondrial membranes against lipid peroxidation induced by hydrogen peroxide and amyloid beta-peptide.

The bcl-2 protooncogene product possesses antiapoptotic properties in neuronal and nonneuronal cells. Recent data suggest that Bcl-2's potency as a survival factor hinges on its ability to suppress oxidative stress, but neither the subcellular site(s) nor the mechanism of its action is known. In this report electron paramagnetic resonance (EPR) spectroscopy analyses were used to investigate the local effects of Bcl-2 on membrane lipid peroxidation. Using hydrogen peroxide (H2O2) and amyloid beta-peptide (A beta) as lipoperoxidation initiators, we determined the loss of EPR-detectable paramagnetism of nitroxyl stearate (NS) spin labels 5-NS and 12-NS. In intact cell preparations and postnuclear membrane fractions, A beta and H2O2 induced significant loss of 5-NS and 12-NS signal amplitude in control PC12 cells, but not PC12 cells expressing Bcl-2. Cells were subjected to differential subcellular fractionation, yielding preparations of plasma membrane and mitochondria. In preparations derived from Bcl-2-expressing cells, both fractions contained Bcl-2 protein. 5-NS and 12-NS signals were significantly decreased following A beta and H2O2 exposure in control PC12 mitochondrial membranes, and Bcl-2 largely prevented these effects. Plasma membrane preparations containing Bcl-2 were also resistant to radical-induced loss of spin label. Collectively, our data suggest that Bcl-2 is localized to mitochondrial and plasma membranes where it can act locally to suppress oxidative damage induced by A beta and H2O2, further highlighting the important role of lipid peroxidation in apoptosis.

Should endolymphatic sac tumors be considered part of the von Hippel-Lindau complex? Pathology case report.

OBJECTIVE: Von Hippel-Lindau (vHL) disease is an inherited disorder characterized by numerous cystic and solid neoplasms. Because of the recent identification of the vHL gene, other investigators have demonstrated genetic mutations in this gene in several of the neoplasms associated with the disease. We describe a patient with an endolymphatic sac (ELS) tumor and vHL disease. The purpose of this study was to identify a similar genetic mutation within the vHL gene of the ELS tumor. METHODS: Using the patient's archival pathological slides, neoplastic cells were microdissected to yield a purely neoplastic cell population. The deoxyribonucleic acid of these cells was then extracted and amplified via polymerase chain reaction. After sufficient amplification, the specimen was analyzed on a single-strand conformation polymorphism gel system to detect putative changes in the base sequence. RESULTS: Single-strand conformation polymorphism gel system analysis yielded two bands representing the two single strands of deoxyribonucleic acid that were amplified. The upper band of the specimen was shifted down (compared with controls), representing a conformational change as a result of genetic mutation. CONCLUSION: ELS tumors are uncommon, and, to our knowledge, only seven cases associated with vHL disease have been reported in the literature. Although this association has been previously mentioned, no definitive studies have linked the two together. We report the eighth case of ELS tumor and vHL disease. We have demonstrated through molecular biological techniques, that, in our patient's tumor, a genetic mutation occurred, and that this mutation is similar to mutations previously reported in other neoplasms associated with vHL. We therefore suggest that ELS tumors be considered among the neoplasms associated with vHL.

Protein phosphorylation in response to PDGF stimulation in cultured neurons and astrocytes.

Platelet-derived growth factor (PDGF) is an important growth factor for a variety of cells, including neurons and glial cells. PDGF signal transduction pathways have been studied primarily in mesenchyme-derived cells (such as fibroblasts and smooth muscle cells). However, little is known about these pathways in the central nervous system (CNS). It is believed that phosphorylation is a critical aspect of several steps in the signal transduction pathway. In this study, neurons and type 1 astrocytes in vitro were radiolabeled with 32P-orthophosphate (32P-Pi). The cells were lysed, and labeled proteins were separated by two-dimensional gel electrophoresis. Autoradiograms of PDGF-stimulated and control samples were compared. We found that in neurons and type 1 astrocytes in vitro, PDGF-BB greatly enhances protein phosphorylation while PDGF-AA has less of an effect on protein phosphorylation. Furthermore, because PDGF signal transduction pathways are likely to affect the cytoskeleton, we studied changes in actin-binding proteins induced by PDGF-BB. We found that PDGF-BB alters the expression, migration pattern and/or avidity of some actin-binding proteins in neurons. In conclusion, protein phosphorylation is up-regulated by PDGF in mouse cortical neurons and type 1 astrocytes in vitro. PDGF's effects on phosphorylation of cytoskeletal proteins might be a important mechanism by which PDGF affects the development and normal functions of central nervous system cells.

Expression of platelet-derived growth factor (PDGF) receptor alpha-subunit in mouse brain: comparison of Patch mutants and normal littermates.

1. Platelet-derived growth factor (PDGF) plays an important role not only in mesenchyme-derived tissues, but also in the mammalian central nervous system. The Patch mutant (Ph/+) lacks one copy of the PDGFR-alpha gene. However, it is not clear whether there are differences in expression of PDGF receptor alpha-subunit (PDGFR-alpha) in brain tissue of the Patch heterozygous (Ph/+) mutants compared to wild-type C57Bl (+/+) mice. 2. The level of PDGRF-alpha mRNA expression is slightly lower in Patch mutant than in normal littermate. 3. Protein and total RNA isolated from mouse brain tissue and primary type 1 astrocyte cultures were studied with Western and Northern blotting techniques. There was no measurable difference ir PDGFR-alpha protein expression between the Patch and wild-type mouse nervous system. Adjustment of transcriptional efficiency and messenger stability may contribute to this phenomenon, whose biological significance remains unclear. 4. Further, the expression of PDGRF-alpha protein and message in mouse brain tissues is developmentally regulated. Its level remains high during the embryonic period and declines below measurable levels in adult.

Amiodarone-induced lymphocyte toxicity and mitochondrial function.

Amiodarone is one of the most effective antiarrhythmic drugs available. However, its use is often limited by potentially life-threatening toxicities, including hepatotoxicity and pulmonary toxicity. We have used human lymphocytes as a system in which to study amiodarone-induced cytotoxicity. Using a tetrazolium dye reduction assay, we observed amiodarone-induced cytotoxicity with a lethal dose (LD)50 of 10.0 +/- 31.1 microM (mean +/- SD, n = 5) with a cellular concentration of 2.2 +/- 0.2 million/ml and of 55.5 and 39.2 microM with cellular concentrations of 8.9 and 7.2 million/ml, respectively, after only 2.75 h of drug exposure. Damage to mitochondria, but not other organelles, was observed with electron microscopy at an amiodarone concentration of 7.3 microM. Alterations in ATP synthesis and lactate dehydrogenase (LDH) release from cells had concentration-response curves similar to those for cytotoxicity. However, we did not observe extracellular accumulation of adenine nucleotides. These results suggest that amiodarone may have a direct toxic effect on mitochondria, beginning at < 10 microM, with membrane-damaging effects at higher drug concentrations.

Phosphorylation of F1F0 ATPase delta-subunit is regulated by platelet-derived growth factor in mouse cortical neurons in vitro.

The delta subunit of F1F0 ATPase (ATP synthase complex) is part of the stalk connecting the F1 and F0 moieties. Studies in Escherichia coli suggest that the analogous bacterial subunit, called epsilon, is essential for the ATPase assembly energy coupling. Platelet-derived growth factor (PDGF) is an important growth factor for various cell types, including neurons of the CNS. Using two-dimensional gel electrophoresis, microsequencing, western blot analysis, and immunoprecipitation techniques, we have found that PDGF induces phosphorylation of the delta subunit or a closely related peptide in cultured mouse cortical neurons.

Platelet-derived growth factor receptors of mouse central nervous system cells in vitro.

This study evaluates the distribution of receptors for platelet-derived growth factor (PDGF) on central nervous cells maintained in vitro using colloidal gold-labeled immunocytochemical markers at the electron microscopic level. Platelet-derived growth factor receptors were found to be sparsely distributed over the surface of type 1 astrocytes, apparent type 2 astrocytes, and neurons. Receptors appeared to be preferentially associated with filopodia-like extensions of the cell membrane. The existence of functional receptors was confirmed using the impermeant, water-soluble affinity cross-linking agent bis(sulfosuccinimidyl)suberate to covalently link radiolabeled PDGF to its receptor. The PDGF/receptor complexes could also be immunoprecipitated with the same antibody used in immunocytochemical experiments. The improved resolution of these techniques allows definitive identification of PDGF receptors on cultured mammalian central nervous system cells other than oligodendrocytes. These data expand the range of possible roles of PDGF during nervous system development. Receptors for PDGF are likely to play a key role in the differentiation of cells in the central nervous system.

Acetylcholinesterase in the developing ferret retina.

The function of acetylcholinesterase (AChE) is to terminate the action of acetylcholine at the cholinergic synapse. Recent evidence suggests additional roles for acetylcholinesterase as a peptidase and/or a protease which is expressed by growing neurites as part of their invasion of developing neural structures. We report the localization of acetylcholinesterase in developing ferret retina. AChE histochemical staining is seen in the developing inner plexiform layer (IPL) of ferret retina at birth (post-natal day zero, PO), the earliest developmental stage examined. Transient expression is seen at the border between the ganglion cell layer and the nerve fiber layer at P14 and P21. A small amount of transient expression is seen in the outer plexiform layer (OPL) at this age as well. By P28, the transient expression in the OPL is at its peak, and is found at photoreceptor terminals and associated with apparent horizontal cell axons. Labeling is also seen intracellularly in the inner nuclear layer (INL), at the OPL/INL border, suggesting that horizontal cells are the source of the transient AChE expression in the OPL. Overt synaptic profiles also appear in the inner plexiform layer (IPL) at P21 and P28. About 2 days layer, the eyes open and the photoreceptor outer segments are fully developed. By 2 weeks later, at P42, the AChE staining pattern in the retina has taken on its adult appearance: no reaction product in the outer retina; intracellular reaction product in the Golgi apparatus of a subset of amacrine and displaced amacrine cells which manufacture AChE; and extracellular reaction product at both synaptic and non-synaptic sites in the IPL. These data are consistent with a role for AChE as a peptidase early in development, and as an enzyme essential in the termination of synaptic action at mature synapses.

Development of muscarinic acetylcholine receptors in the ferret retina.

The development of muscarinic acetylcholine receptor protein in the ferret retina was studied using biochemical, autoradiographic, and light and electron microscopic immunohistochemical techniques. The development of retinal muscarinic cholinergic receptor proteins involves transient shifts in their number and distribution, as well as changes in the relative abundance of two molecular weight variants. Receptor binding assays demonstrate changes in the number and affinity of retinal binding sites for the muscarinic cholinergic ligand [3H]quinuclidinylbenzilate ([3H]QNB). Light microscopic immunohistochemical studies reveal the presence of muscarinic acetylcholine receptor-like (mAChR-like) immunoreactivity in the adult inner plexiform layer. During development, the mAChR-like immunoreactivity appears in a number of other retinal layers. Electron microscopic immunohistochemical studies indicate that muscarinic acetylcholine receptor-like immunoreactivity is found at amacrine-amacrine cell contacts. Both autoradiographic and gel slice electrophoretic studies were carried out after labeling of developing and adult retinal muscarinic receptors with [3H]propylbenzilylcholine mustard ([3H]propylbenzilylcholine mustard ([3H]PrBCM), which irreversibly labels the muscarinic acetylcholine receptor. Polyacrylamide gel electrophoresis under reducing, denaturing conditions resolved two peaks of radioactivity corresponding to [3H]PrBCM-labeled protein; both were eliminated by pre- and co-incubation of labeled adult retinas with excess atropine. Combined with the results of earlier studies, these observations suggest that the subtypes, number and distribution of muscarinic receptor proteins changes during retinal synaptogenesis.

Platelet-derived growth factor (PDGF) receptors in the developing mouse optic pathway.

The molecules which control the patterns of cell division, growth, and precise interconnections characteristic of the central nervous system still remain largely unidentified. The protein platelet-derived growth factor (PDGF) has been shown to mediate interactions among glial cells in vitro. More recent evidence has indicated that PDGF may also be involved in controlling communication between neurons and glial cells and among neurons. The presence of receptors for PDGF on neurons of the developing nervous system is an essential piece of evidence in this chain of events. Ganglion cells are labeled with antibodies to PDGF receptor only during the period of active process outgrowth. These findings suggest that PDGF is used as a mediator of intercellular signaling during neuronal development.

Expression of platelet-derived growth factor and its receptor in rat neuronal and astroglial cultures.

Platelet-derived growth factor (PDGF) is a potent mitogen and differentiation signal for glial cells in the central nervous system. Several lines of evidence have indicated that neurons are one source of PDGF. We present data suggesting that they may respond to a PDGF signal as well. Immunofluorescence techniques have been used to demonstrate the presence of PDGF and its receptor in both neurons and glial cells isolated from developing rat brain and maintained in vitro. Similar findings hold either in medium supplemented with serum or under serum-free conditions. PDGF receptor beta subunit (PDGF-Rbeta)-like immunoreactivity was found encrusting the surface of cultured neurons. Virtually all neurons in this study contained PDGF-Rbeta on the surface of the perikaryon and on neuronal processes down to their fine distal tips. PDGF receptor was found on neurons cultured in serum-containing or serum-free medium, and in cultures with few glial cells as well as neuronal-glial co-cultures. Neurons and glial cells also contain PDGF subunits (designated PDGF-A and PDGF-B), revealed by immunofluorescence staining. Isoform-specific antibodies stain cultured neurons and glia in either serum-containing or serum-free medium. The identity of stained cells was confirmed using double- or triple-labeling immunofluorescence procedures for simultaneous localization of PDGF isoforms, PDGF receptor, phosphorylated neurofilament protein, and/or glial fibrillary acidic protein. These studies indicate that both neurons and glia contain platelet-derived growth factor and carry the PDGF receptor on their cell surface in vitro. While the localization of PDGF and its receptor have previously been considered separately, the present study demonstrates that both the growth factor and the receptor are expressed by the same population of neurons, indicating that PDGF may act us an autocrine factor regulating neuronal differentiation.

Choline acetyltransferase expression studied with an oligonucleotide probe.

1. The localization of choline acetyltransferase messenger RNA has been studied using a digoxigenin-tailed complementary oligodeoxynucleotide probe for in situ hybridization. 2. Putative cholinergic cells of the rat and ferret spinal cord and the ferret retina were labeled. 3. This technique affords superior resolution compared to radioactively labeled probes, with apparently equal sensitivity.

Developmental distribution of platelet-derived growth factor in the mouse central nervous system.

Immunoblotting and immunohistochemical techniques have been used to characterize the developmental changes in the distribution and relative quantity of platelet-derived growth factor (PDGF), an important mitogen and growth regulator for glial (and possibly neuronal) cells. PDGF exists as a dimer of two chains, A and B, and antibodies which are relatively specific for one chain or the other can be used to localize PDGF isoforms during development. We have also studied the distribution of PDGF receptor beta subunit (PDGF-R beta)-like immunoreactivity using an antibody probe. All 3 isoforms of PDGF are found in neural structures during development, beginning at about the midpoint of embryogenesis. Immunoblotting studies confirm the presence of PDGF isoforms in brain during embryonic and postnatal development, with the distribution and relative abundance of each isoform appearing to be independently regulated. Similarly, immunoblotting studies have verified the relative abundance and specificity of PDGF receptor beta subunit. The immunohistochemical findings confirm and extend these biochemical observations. Each PDGF chain (A and B) has a discrete localization during nervous system development, and the immunohistochemical distribution of PDGF-R beta is distinct from each of the PDGF isoforms. PDGF A-chain (localized with an antibody to PDGF(AA) dimers) appears to be found in growth cones of developing neurons in mid-embryonic brain development. By 11.5 days post-conception (embryonic day 11.5, E11.5) to E12, PDGF isoforms are found in apparent neurons in the basal plate (future ventral horn) of spinal cord. PDGF-R beta-like immunoreactivity is localized to the boundary cap region of the developing spinal cord at the same age. Similarly, at E13.5, all 3 PDGF isoforms are found, to varying extents, within cells of the dorsal root ganglia and trigeminal ganglia. At the same developmental stage, PDGF receptor protein is most prevalent in the nerves accompanying these structures. By E15, both PDGF isoform and PDGF receptor beta subunit immunoreactivity have declined to near-background levels in the sensory ganglia, while in the spinal cord and developing forebrain, levels of all PDGF-related proteins remain high.

Development of the lateral geniculate nucleus: interactions between retinal afferent, cytoarchitectonic, and glial cell process lamination in ferrets and tree shrews.

We have studied the relationship of retinal afferents, glial cell processes, and neuronal cytoarchitectonics in the lateral geniculate nucleus (LGN) of two species: tree shrews (Tupaia belangeri) and ferrets (Mustela putoris). Both species are relatively immature at birth, allowing the development of these features to be studied in the perinatal period. Retinal afferents, visualized by intraocular injection of a wheat germ agglutinin/horseradish peroxidase conjugate (WGA-HRP), are apparently the first elements of the developing LGN to exhibit a characteristic layered pattern in tree shrews and ferrets. Some radial glia still remain in the LGN of both species as the retinal afferents are in the process of segregating. Glial cell processes were visualized immunohistochemically with antibodies to glial fibrillary acidic protein (GFAP) or vimentin. In both the ferret and tree shrew, layering of glial cell processes is first seen as the overlap of retinal terminal fields diminishes. In the tree shrew LGN, these bands of dense glial cell staining are seen in apparent future cellular layers, whereas in the ferret, glial cell banding appears in interlaminar zones. If one or both eyes are removed at birth in tree shrews (before LGN cell layers are formed), the glial cell pattern seen 1 week later is in accord with the distribution of surviving nerve cells. The glial processes do not appear to invade regions left by degenerating retinal terminals or dying LGN cells. Several days after the appearance of layered glial cell processes (in the tree shrew) or at about the same time as glial layering (in the ferret), the first interlaminar spaces develop between neuronal cells, marking the beginning of cytoarchitectonic lamination, with its distinctive alternating cell-rich and cell-poor zones. Over the next several weeks, LGN neurons in both species continue to segregate into characteristic layers until the final, adult pattern of neuronal lamination is evident; as this process is completed, glial cell lamination disappears. These observations suggest that glial cells may be involved in establishing the neuronal layers that characterize the mature LGN of many species.

Vimentin: changes in distribution during brain development.

This paper examines both the anatomical changes in the distribution of vimentin intermediate filament protein and the biochemical changes in vimentin and its degradative enzyme during postnatal brain development in the tree shrew (Tupaia belangeri). A pattern of multiple immunoreactive bands at birth (postnatal day 0, or P0) was revealed in nitrocellulose blots of polyacrylamide gels ("Western blots"). These multiple bands gradually disappear during development, and in the adult a single band at the published molecular weight for vimentin (57 kD) is seen. This pattern of bands probably reflects shifts in the activity of a calcium-activated vimentin protease. The changes in the anatomical distribution of vimentin-immunoreactive (vimentin+) cells and their fine processes parallel the biochemical shifts seen in immunoblots. We have examined the neocortex, lateral geniculate nucleus (LGN), and hippocampus in detail. During the first postnatal week, vimentin+ glia, especially radial glia, are prominent in both neocortex and hippocampus. In contrast, only a few vimentin+ radial glia remain in the thalamus at this age. Vimentin+ glia appear to coincide with bundles of axons and often seem to outline subdivisions of thalamic nuclei. Additionally, cellular layers of the lateral geniculate nucleus (LGN) appear to stain with antibodies to vimentin several days before the characteristic neuronal cell layers appear in this area. During the second postnatal week, vimentin+ cells appear in "patches" throughout the cortex. Some subdivisions of the thalamus and hippocampus (as defined by cytoarchitectonic differences in the adult) are distinct when the tissue is stained with an antibody to vimentin, even though a conventional Nissl stain at this age shows no apparent delineation in these same regions. Finally, in the adult, only a few vimentin+ cells remain, primarily in the white matter. Taken together, these results suggest that the remodeling of vimentin+ intermediate filaments in immature glial cells (including radial glia) is paralleled by the action of the enzyme which breaks down these filaments. The apparent activity of this enzyme is high early in development as radial and other glia are rapidly dividing and undergoing morphological changes, with a decrease in activity in the juvenile and adult brain, as immature glial cells are supplanted by mature forms.

Glial cells develop a laminar pattern before neuronal cells in the lateral geniculate nucleus.

The lateral geniculate nucleus, which lies between the retina and the striate cortex in the visual pathway of mammals, is often made up of several distinctive cell layers, or laminae. We have used immunohistochemical methods to localize two glial cell intermediate filament proteins, glial fibrillary acidic protein and vimentin, and have found that layering of glial cells is evident before neuronal cell layers develop in the lateral geniculate nucleus. The correlation between glial cell lamination and neuronal lamination is consistent with the suggestion that glia are guiding neurons not only during the early postmitotic migratory phase of development but also during the later formation of functional divisions such as layers and nuclei.

Development of acetylcholinesterase activity in the lateral geniculate nucleus.

The pattern of acetylcholinesterase activity in the tree shrew (Tupaia belangeri) lateral geniculate nucleus (LGN) undergoes a number of striking changes during postnatal development. The adult tree shrew LGN is made up of six cellular layers divided by relatively cell-free interlaminar zones. At birth, however, the nucleus appears unlaminated when processed with conventional Nissl-staining techniques. The cellular lamination appears during the first postnatal week. The eyes open much later, typically at the end of the third week after birth. In the adult tree shrew, acetylcholinesterase (AChE) activity is found throughout the nucleus (both within and between the six cellular layers). In most sections examined, reaction product is slightly more intense in the lateral cell layers (4, 5, and 6). This is in sharp contrast to the pattern at birth (postnatal day zero, or P0). The detectable AChE activity at this age is apparently found in inchoate layers 1-2 and 4-5. Within these pairs, areas innervated by the ipsilateral eye (i.e., incipient layers 1 and 5) appear to contain more reaction product. From P0 to P4, the density of AChE activity increases in layers 1-2 and 4-5 and becomes detectable in the barely evident layers 3 and (usually) 6 at this age. By the middle of the second postnatal week, after laminae are clearly apparent with a Nissl stain, AChE activity has increased and is mainly associated with each cellular layer in the nucleus. During the third week after birth this pattern undergoes a radical shift. The most intense AChE activity is now in the interlaminar zones. Finally, as the adult pattern emerges, AChE activity increases in the cellular layers and all areas of the nucleus exhibit relatively high levels of AChE activity. Superimposed on this changing laminar pattern of AChE activity are changes related to the retinotopic map within the nucleus. Portions of the LGN representing central vision develop their characteristic pattern of activity several days ahead of the regions representing more peripheral visual field locations. AChE activity is also found transiently in the optic tract near the LGN during the first 3 postnatal weeks. Two (possibly three) groups of AChE-carrying fibers can be traced from the optic chiasm to their apparent sites of termination (or origin) in the parabigeminal nucleus, ventral lateral geniculate nucleus, and dorsal LGN. The activity present in the optic tract disappears shortly after eye opening.(ABSTRACT TRUNCATED AT 400 WORDS)