Developing Rat Central Nervous System Research Articles

Alzheimer's disease cerebrospinal fluid antibodies display selectivity for microglia. Investigations with cell cultures and human cortical biopsies.

Previous investigations demonstrated that the cerebrospinal fluid (CSF) from Alzheimer's disease (AD) patients contains antibodies that recognize specific neuronal populations in the adult rat central nervous system (CNS). These findings suggest a pathogenic role for immunological aberrations in this disorder. To determine if antibodies may provide a means to differentially diagnose the dementias, CSF from a diversified dementia population was screened against the developing rat CNS and a cell culture system. Markings produced by AD CSF were distinctly different from those of vascular dementias (VAD) against the developing rat CNS. More importantly, some AD CSF recognized amoeboid microglia. The recognition of amoeboid microglia by antibodies in AD CSF is particularly interesting since these cells proliferate in response to nervous system disease and also engulf debris. A cell culture technique was developed to allow the rapid screening of CSF antibodies. Patient CSF produced five different types of markings in the cell culture: microglia, glioblasts, fibers, nonspecific, or negative. Correlations with these structures and the diagnosis of four different dementia populations revealed that, in comparison to the other groups, AD CSF displayed remarkable selectivity toward microglial cells. Cortical biopsies from patients suspected to have AD were incubated with the patient's own CSF and that of confirmed AD patients. Both CSF samples recognized microglial cells in the patient's cortical biopsy. The same CSF samples incubated against normal human cortical autopsy or a biopsy from a 3-mo-old child displayed negative immunoreactivity. These three approaches suggest that the presence of CSF microglial antibodies may be a means to distinguish AD patients from other dementias. The results add further support to the widely growing concept that inflammation and similar immune mechanisms may contribute to AD pathogenesis.

C-CAM expression in the developing rat central nervous system

C-CAM, a transmembrane glycoprotein belonging to the immunoglobulin superfamily, can mediate intercellular adhesion by homophilic, Ca 2+-independent binding. Immunohistochemical analysis of adult rat tissues has demonstrated that C-CAM is expressed in various epithelia, vessel endothelia, and hematopoietic cells. By molecular cloning and sequence analysis several isoforms differing both in the extracellular and the cytoplasmic domains have been found. Here we have analyzed the expression of C-CAM in the developing rat central nervous system. No neuronal expression was observed, but biochemical and immunohistochemical analyses demonstrated that C-CAM becomes expressed in the microvessels from embryonic day E-13; the intensity of the staining increased through day E-15 and then gradually decreased during the perinatal and early postnatal period. The expression of C-CAM in the walls of the microvessels was confirmed by in situ hybridization. Immunoelectron microscopy showed that C-CAM was localized both to the abluminal surface of the endothelial cells and to cellular processes of primordial pericytes where these two cell types are in contact with each other. No staining was found on the luminal endothelial cell surfaces or inter-endothelial cell contact areas. During the perinatal period C-CAM also became expressed on the opposite side of the pericytes and on other cells, possibly astrocytes, in contact with these areas of the pericytes. These observations suggest that C-CAM may be involved in heterotypic, homophilic adhesion between endothelial cells, pericytes and astrocytes, and in maturation of the vessel walls.

Cerebrospinal fluid microglial antibodies: Potential diagnostic markers for immune mechanisms in Alzheimer's disease

Hallmark lesions of Alzheimer's disease (AD) are filled with reactive immunocompetent microglia, suggesting that immunological abberations may participate in the pathophysiology of this disorder. Microglia may participate in the initial stages of neurodegeneration before the onset of dementia. If immune mediated processes are closely linked to neuronal breakdown it would be of importance to have a reliable means to detect these processes. Serum and cerebrospinal fluid (CSF) antibodies are discussed as such potential sources. The serendipitous use of the developing rat central nervous system (CNS) to screen CSF antibrain antibodies produced some unexpected findings. Firstly, CSF antibodies of AD and other dementia patients recognized distinctly different neuronal structures in the developing rat brain. Secondly, some AD CSF recognized fiber networks whereas others recognized amoeboid microglial cells. The same AD CSF which recognized amoeboid microglia cells also specifically marked activated microglia and neural macrophages in experimentally induced lesions. AD CSF microglial antibodies appear to be significant in view of the increasing association between microglia and neurogenerative processes in AD. In addition, CSF microglial antibodies are present in numerous at-risk descendants of familial AD patients. Some have subsequently developed the disorder. These findings together with the fact that microglial antibodies are usually found in the early stages of AD suggest that AD CSF microglial antibodies could be of value in detecting neurodegenerative processes before the onset of dementia. These findings add further support to the concept that inflammation and similar immune mechanisms may contribute to AD pathogenesis.

A phosphorylation epitope on MAP 1B that is transiently expressed in growing axons in the developing rat nervous system.

We have isolated a monoclonal antibody (150) that recognizes a phosphorylation epitope on the microtubule-associated protein (MAP) 1B. Immunoblot analysis of the developing rat central nervous system shows that monoclonal antibody 150 is directed against a protein of approximately 325 kDa (MAP 1B) that copolymerizes with microtubules through successive cycles of temperature-dependent assembly and disassembly. Furthermore, immunoprecipitated MAP 1B contains the epitope recognized by monoclonal antibody 150. Removal of phosphate from blotted proteins using alkaline phosphatase abolishes the binding of monoclonal antibody 150 to MAP 1B, indicating that the epitope is phosphorylated. In the developing rat nervous system, immunohistochemistry with monoclonal antibody 150 shows that the phosphorylation epitope on MAP 1B is transiently expressed in growing axons but not in dendrites. For instance, in the neonatal rat cerebellum, the parallel fibres of granule cells are stained only during elongation and not after synaptogenesis. The monoclonal antibody 150 epitope is also transiently expressed in radial glial fibres and in certain cell nuclei. All immunostaining of sections with monoclonal antibody 150 was completely abolished by alkaline phosphatase treatment. These observations and previous ones made by us in cell culture (Mansfield et al., J. Neurocytol., 20, 654-666, 1991) suggest that the phosphorylation epitope on MAP 1B recognized by monoclonal antibody 150, which has not been previously detected in vivo, may be important in axonogenesis.

Ethanol-induced changes in astrocyte gene expression during rat central nervous system development.

Disruption of spatial and temporal patterns of gene expression in cells of the developing brain could result in abnormal development. We report that briefly exposing neonatal rats to a moderate dose of ethanol on postnatal days 5 through 7 caused a large, specific increase in glial fibrillary acidic protein (GFAP) mRNA and GFAP. Astrocytes of the cerebral cortex were apparently more sensitive to this effect of ethanol than astrocytes in several other brain regions. As a first step in the characterization of an vitro model of ethanol's effect on GFAP gene expression, ethanol was added to the media of primary cultures of cortical astrocytes in a pattern of exposure and at concentrations equal to pups' peak blood levels. This resulted in an increase in GFAP mRNA whose magnitude and specificity mirrored that observed in the animal model. Together, these results suggest that even brief exposure to ethanol can alter gene expression in astrocytes, and forms the foundation for further characterization of an in vitro model that may be used to determine the mechanism of this effect.

Quantitation of NMDA receptor (NMDAR1) mRNA levels in the adult and developing rat CNS

A rapid and sensitive solution hybridization assay was used to quantitate N- methyl- d- aspartate (NMDA) receptor mRNA levels in the central nervous system (CNS) of rat, mouse and human. A riboprobe labelled with 32P was prepared from a plasmid containing a 1413 base sequence from the cDNA for the functional rat NMDA receptor subunit, NMDAR1. Using a full length sense transcript as the calibration standard, the assay reliably measures 8 pg of NMDAR1 mRNA. When expressed as pg of NMDAR1 mRNA/μg total cellular RNA, the highest levels in the adult rat CNS are in the olfactory bulb (20.9 pg/υg RNA) and the lowest levels are in the spinal cord (5.2 pg/υg RNA). Intermediate levels were found in frontal cortex, hippocampus, cerebellum and whole brain. In the mouse CNS the highest levels of NMDAR1 mRNA were found in the olfactory bulb (12.9 pg equivalents/υg RNA), followed closely by hippocampus, frontal cortex and cerebellum. Mouse spinal cord (4.4 pg equivalents/υg RNA) had the lowest levels of NMDAR1 mRNA. The NMDAR1 riboprobe hybridizes with the same size transcripts in Poly(A) + RNA from rat, mouse and human brain. In the developing rat, NMDAR1 mRNA levels in frontal cortex and hippocampus increased nearly 3 fold from postnatal day 3 to day 15 and approximately doubled from day 15 to day 67 (adult). Therefore, from postnatal day 3 to adult (day 67) frontal cortex and hippocampus levels of NMDAR1 mRNA increased nearly 6 fold. In contrast, cerebellum and brain stem showed no change in NMDAR1 mRNA levels when postnatal days 3 and 15 were compared while the levels were increased 2 fold when day 15 and day 67 were compared. These changes were not a function of alterations in total cellular RNA. These results demonstrate significant differences in frontal cortex and hippocampus when compared to cerebellum and brain stem in the postnatal development of NMDAR1 mRNA.

Low concentrations of hydrogen sulphide alter monoamine levels in the developing rat central nervous system.

The central nervous system is one of the primary target organs for hydrogen sulphide (H2S) toxicity; however, there are limited data on the neurotoxic effects of low-dose chronic exposure on the developing nervous system. Levels of serotonin and norepinephrine in the developing rat cerebellum and frontal cortex were determined following chronic exposure to 20 and 75 ppm H2S during perinatal development. Both monoamines were altered in rats exposed to 75 ppm H2S compared with controls; serotonin levels were significantly increased at days 14 and 21 postnatal in both brain regions, and norepinephrine levels were significantly increased at days 7, 14, and 21 postnatal in cerebellum and at day 21 in the frontal cortex. Exposure to 20 ppm H2S significantly increased the levels of serotonin in the frontal cortex at day 21, whereas levels of norepinephrine were significantly reduced in the frontal cortex at days 14 and 21, and at day 14 in the cerebellum.

Expression of s-laminin and laminin in the developing rat central nervous system.

The extracellular matrix component, s-laminin, is a homologue of the B1 subunit of laminin. S-laminin is concentrated in the synaptic cleft at the neuromuscular junction and contains a site that is adhesive for motor neurons, suggesting that it may influence neuromuscular development. To ascertain whether s-laminin may also play roles in the genesis of the central nervous system, we have examined its expression in the brain and spinal cord of embryonic and postnatal rats. S-laminin was not detectable in synapse-rich areas of adults. However, s-laminin was present in discrete subsets of three laminin-containing structures: (1) In the developing cerebral cortex, laminin and s-laminin were expressed in the subplate, a transient layer through which neuroblasts migrate and cortical afferents grow. Both laminin and s-laminin disappeared as embryogenesis proceeded; however, laminin was more widely distributed and present longer than s-laminin. (2) In the developing spinal cord, laminin was present throughout the pia. In contrast, s-laminin was concentrated in the pia that overlies the floor plate, a region in which extracellular cues have been postulated to guide growing axons. (3) In central capillaries, s-laminin appeared perinatally, an interval during which the blood-brain barrier matures. In contrast, laminin was present in capillary walls of both embryos and adults. To extend our immunohistochemical results, we used biochemical methods to characterize s-laminin in brain. We found that authentic s-laminin mRNA is present in the embryonic brain, but that brain-derived s-laminin differs (perhaps by a posttranslational modification) from that derived from nonneural tissues. We also used tissue culture methods to show that glia are capable of synthesizing "brain-like" s-laminin, and of assembling it into an extracellular matrix. Thus, glia may be one cellular source of s-laminin in brain. Together, these results demonstrate that s-laminin is present in the developing central nervous system, and raise the possibility that this molecule may influence developmental processes.

On the ontogenetic and sequential characteristics of bombesin-induced grooming in the infant rat

This paper examined the grooming effects of central injections of the tetradecapeptide bombesin (BN) in the infant rat. Anaesthetized rat pups of 1–20 days of age ( n = 7/dose/age) were injected intracerebroventricularly (i.e.v.) with either BN (0.01–1.0 μg) or saline (control condition) and placed in heated test cages. Following recovery from the anaesthetic, the subsequnnt behaviour displayed by the pups was videotaped for the next 60 min. These videotaped responses were then scored for a variety of grooming and other behaviours. Pups of all ages (1–20 days) groomed in response to BN with the 1-day-old pups the least and 20-days-olds the most sensitive to BN. In the 10-days-olds, scratching, in an immature, non-contactt form, was elicited by BN. This was replaced by mature, contact type scratching in the sequence of grooming behavious at 20 days of age. Scratching activities appeared to form a subsystem connected to but nonetheless separate from washing behaviours and this was particularly distinguishable at 20 days of age. Changes in BN-induced grooming appeared to reflect the maturation of the motor capabilities of the developing rat. In conclusion, these results indicate that BN binding sites in the developing rat central nervous system (CNS) are pharmacologically functional from an early stage in ontogeny when there is little or no measurable amount of the peptide itself present in the CNS.

Expression of P30, a protein with adhesive properties, in Schwann cells and neurons of the developing and regenerating peripheral nerve.

P30 is a heparin-binding protein with adhesive and neurite outgrowth-promoting properties present at high levels in the developing rat central nervous system (Rauvala, H., and R. Pihlaskari. 1987 J. Biol. Chem. 262:16625-16635). Partial sequencing of p30 has revealed homology or identity with HMG-1 (Rauvala, H., J. Merenmies, R. Pihlaskari, M. Korkolainen, M.-L. Huhtala, and P. Panula. 1988. J. Cell Biol. 107:2292-2305), a 28-kD protein that was originally purified from the thymus (Goodwin, G.H., C. Sanders, and E. W. Johns. 1973. Eur. J. Biochem. 38:14-19) which binds DNA in vitro. We have analyzed the distribution of p30 in the developing rat peripheral nervous system (PNS). P30 was detected by immunohistochemistry and Western blot analysis using antibodies raised against intact p30 and against a synthetic peptide corresponding to the amino terminus of the p30 molecule. P30 was localized to nonnuclear compartments of neurons and peripheral glial cells (Schwann cells). P30 immunoreactivity of PNS neurons persisted into adulthood. In contrast, Schwann cell staining decreased after the second postnatal week and was not detectable in adult animals. Neuron-Schwann cell contact was correlated with diminished p30 levels in Schwann cells. Schwann cells of the normal adult sciatic nerve did not express p30; however, when deprived of axonal contact by nerve transection, the Schwann cells of the distal nerve stained intensely for p30. In addition, when Schwann cells and dorsal root ganglion neurons were grown in coculture, Schwann cells that were associated with neurites were not as intensely stained by anti-p30 as Schwann cells that were not in contact with neurons. The pattern of p30 expression during development and regeneration, and its apparent regulation by cell-cell contact suggests that p30 plays a role in the interaction between neurons and Schwann cells during morphogenesis of peripheral nerves.

Growth factors and transcription factors in oligodendrocyte development.

O-2A progenitor cells, the precursors of oligodendrocytes in the central nervous system (CNS), probably originate in the subventricular germinal zones of the developing CNS, and subsequently migrate away from there to populate the rest of the CNS with oligodendrocytes. We are trying to understand how the O-2A progenitor cells interact with their changing environment as they migrate, and how this influences each stage of their development into mature, myelinating oligodendrocytes. In this article we summarize evidence that platelet-derived growth factor (PDGF) is important for stimulating O-2A progenitor cell proliferation in vivo, and describe our efforts to map the distribution of PDGF and its receptors in the developing rat CNS by in situ hybridization and immunohistochemistry. These studies suggest that, in the CNS, PDGF alpha-receptor subunits may be restricted to O-2A lineage cells that have started to migrate away from the subventricular zones towards their final destinations. Many neurons express the A and/or B chains of PDGF, and astrocytes express the A chain, but it is not yet clear which of these cell types might be the major source of PDGF for O-2A lineage cells in vivo. O-2A progenitor cells can be purified and maintained in a proliferating state in vitro by culturing in the presence of PDGF and bFGF. Under these conditions, the POU transcription factor SCIP/Tst-1 is expressed at a high level; when oligodendrocyte differentiation is initiated by withdrawing the growth factors, SCIP/Tst-1 mRNA is rapidly down-regulated, followed by a decline in SCIP/Tst-1 protein and sequential activation of myelin-specific genes.(ABSTRACT TRUNCATED AT 250 WORDS)

Antibodies in the cerebrospinal fluid of some Alzheimer's disease patients recognize amoeboid microglial cells in the developing rat central nervous system

Previous investigations have demonstrated that the cerebrospinal fluid from Alzheimer's disease patients contains antibodies that recognize specific neuronal populations in the adult rat central nervous system. These findings suggest a pathogenic role for immunological aberrations in this disorder. In the present report the investigation of antibodies in the cerebrospinal fluid of Alzheimer's disease patients was extended to developing rat central nervous system. The antibody in cerebrospinal fluid from Alzheimer's disease patients recognized entirely different types of antigens in the developing rat central nervous system as compared to adult rat central nervous system. One of the most remarkable differences was the recognition of amoeboid microglial cells. Diverse morphological forms of amoeboid microglial cells were observed, located mainly in the cavum septum pellucidum and in the corpus callosum. Electron microscopy revealed that the cerebrospinal fluid antibody from the Alzheimer's disease patients recognized specific membrane receptors in the macrophagic microglia. The unexpected recognition of amoeboid microglia by antibodies in Alzheimer's disease-cerebrospinal fluid is particularly interesting since these cells proliferate in response to nervous system disease and also engulf debris. The results add further support to the concept that inflammation and similar immune mechanisms may contribute to Alzheimer's disease pathogenesis.

Reversible biochemical changes in the developing rat central nervous system following ultrasound exposure.

Rat embryos in utero at 10 days of gestation (gd) were exposed to the intensity of 2.5 W/cm2 (SATA) of 1 MHz continuous-wave ultrasound for 5 min. The right uterine horn was exteriorized, exposed to the ultrasound beam, and then reinserted into the abdominal cavity. Embryos from the unexposed left uterine horn served as controls. The cerebral hemispheres of each fetus were removed and analyzed for enzymatic activity at four different stages of development. There was an increase in the temperature at the surface of the intact uterus during the ultrasound exposure. No gross anatomical malformations were observed in the exposed embryos. There were no differences in brain weight or protein content between exposed and control fetuses. However, the ultrasound exposure caused a transient decrease in acetylcholine esterase and 2',3'-cyclic nucleotide phosphohydrolase activities of the insonated fetal brains compared to the control brains. Because of the relatively high temporal-average intensity, the relatively large volume of tissue exposed, and the minimal sound path through tissue, the biochemical changes observed in this experiment would be unlikely to occur in clinical examinations.

Biochemical changes in the developing rat central nervous system due to hyperthermia

Rat embryos at 10 days of gestation were exposed to 43 degrees C for 8 minutes by submerging the exteriorized right uterine horn in heated saline solution and then reinserting the uterine horn into the abdominal cavity. At 15 days, the fetuses were removed, and cells from the cerebral hemispheres were dissociated and grown as primary cultures. Embryos from the left uterine horn served as controls. No morphological changes were observed between the cultures of cells from control and heat-exposed embryos at different days in culture. However, exposure of embryos to hyperthermia at 10 days significantly affected the developmental pattern of activities of acetylcholine esterase associated with cholinergic neurons and of 2',3'-cyclic nucleotide phosphohydrolase associated with oligodendrocytes and myelin membrane formation. These results suggest that hyperthermia at 10 days of gestation in the rat may lead to an impairment in the development of neurons and oligodendrocytes in the central nervous system.

Developmental Expression of Insulin-Like Growth Factor-II Receptor Immunoreactivity in the Rat Central Nervous System*

A polyclonal antiserum raised against the insulin-like growth factor-II (IGF-II) receptor has been used to map the distribution of this receptor in the developing rat central nervous system (CNS). Transiently high levels of receptor immunoreactivity were found in the developing brain, particularly in the cortex and hypothalamus. The amount of receptor immunostaining in these areas decreases toward the time of birth, and levels are approximately equivalent to those in the adult by postnatal day 7. The choroid plexus, cerebral vasculature, ependymal cells, retina, and pituitary contain high levels of receptor immunoreactivity throughout embryogenesis and adulthood. Some mesodermally derived tissues, such as bone, also demonstrate transient expression of IGF-II receptor during fetal development. These data are consistent with potential roles for IGF-II in CNS development, in the development of specific mesodermal tissues, and in specific regions of the postnatal CNS.

Differential regulation of three sodium channel messenger RNAs in the rat central nervous system during development.

The levels of the mRNAs encoding sodium channels I, II and III in various regions of the developing rat central nervous system (from embryonal day 10 to postnatal day 90) have been examined by blot hybridization analysis with specific probes. The three sodium channel mRNAs exhibit different temporal and regional expression patterns. The expression of sodium channel I mRNA rises after a lag phase to adult levels during the second and third postnatal weeks with stronger increases in caudal regions of the brain and in spinal cord. Sodium channel II mRNA increases steadily until the first postnatal week, keeping high adult levels in rostral regions of the brain or reaching low adult levels after the second postnatal week in most caudal regions of the brain and in spinal cord; cerebellum shows low levels during the first two postnatal weeks but high adult levels. In all regions, sodium channel III mRNA attains maximum levels around birth and decreases during the first and second postnatal weeks to reach variable low adult levels. These results suggest that sodium channel III is expressed predominantly at fetal and early postnatal stages and sodium channel I predominantly at late postnatal stages, whereas sodium channel II is expressed throughout the developmental stages studied with greater regional variability.

A neuroectoderm-associated ganglioside participates in fibronectin receptor-mediated adhesion of germinal cells to fibronectin

Previous studies with a rat neural cell line have shown that the D1.1 ganglioside, an O-acetylated derivative of GD 3, is involved in cellular adhesion to fibronectin. In vivo, D1.1 is present on germinal cells of the developing rat central nervous system, but not on postmitotic cells that migrate away from the germinal zones. These observations suggest that D1.1 could participate in adhesive interactions within germinal zones and that the loss of D1.1 could be involved in the decision to begin migration. In support of this hypothesis, immunofluorescence histochemistry shows that both fibronectin and fibronectin receptor are colocalized with D1.1 in the ventricular zones of the embryonic rat brain and in the external granule cell layer of the postnatal cerebellum. Dishes coated with monoclonal antibody against D1.1 were used to isolate D1.1-positive germinal cells from Embryonic Day 14 cerebrum and from Postnatal Day 6 cerebellum. These cells are able to adhere to fibronectin-coated dishes by a mechanism that is inhibitable by a synthetic hexapeptide containing the arg-gly-asp cell recognition sequence of fibronectin. Adhesion is also partially inhibited by antibody against fibronectin receptor and is slowed by anti-D1.1 antibody, implicating both the receptor and the ganglioside in the adhesion process. During 3 days in culture these D1.1-positive, fibronectin receptor-positive cells exhibit a neuronal phenotype, as judged by morphology and staining with tetanus toxin. This further confirms the neuroepithelial origin of the cells. The cells do not synthesize detectable amounts of fibronectin, thus leaving unidentified the source of the fibronectin seen in the germinal zones in tissue sections. Immunoprecipitation experiments show that the fibronectin receptors present on these cells are heterodimers. Under nonreducing conditions, the immunoprecipitates contain an α-subunit of 150–160 kDa and a β-subunit of 115–125 kDa.

Expression of pp60 c- src in adult and developing rat central nervous system

Brains of adult and fetal (E13–E19) rats were assayed by region for the presence of the proto-oncogene product pp60 c- src . pp60 c- src was abundant in the adult brain with the highest levels found in the cerebral hemispheres and localized to the cortical cellular layers. In the embryonic nervous system the levels of pp60 c- src activity were much higher throughout the brain than those observed in the adult. The expression of pp60 c- src was developmentally regulated, but demonstrated a regionally distinct pattern of expression. In the cortex src activity steadily rose during gestation, while in the basal forebrain and midbrain maximal activity was observed at E17 which then declined to adult levels. The data demonstrate that pp60 c- src is differentially expressed in regions of the brain, both during development and in the adult.

Monoclonal antibody g10 against microtubule-associated protein 1x distinguishes between growing and regenerating axons

Monoclonal antibody G10 binds to an epitope on a microtubule-associated protein and specifically labels elongating axons in the developing rat central nervous system. In this paper the expression of this microtubule-associated protein epitope has been studied in developing and regenerating rat peripheral neurons and sprouting and transplanted neurons in the central nervous system. During development bright immunofluorescent fibres were seen from embryonic day 12 (the earliest age studied) to birth, but immuno reactivity declined postnatally and was only barely detectable in adult sciatic nerves. In contrast to the situation in developing axons, immunostaining could not be demonstrated in axons regenerating after crushing of the sciatic nerve nor in axons growing from grafts of septal tissue into the adult hippocampus, despite good evidence for extensive reinnervation in both cases. Immunostaining was only seen at early stages within the septal grafts, and not as axons grew out to reinnervate the host. No staining was seen in areas of deafferented hippocampus where collateral sprouting takes place. These results suggest a molecular difference between the cytoskeletons of developing and regenerating or sprouting nerves.

Neurotoxicity of N-methyl- d-aspartate is markedly enhanced in developing rat central nervous system

The neurotoxic lesion produced by direct injection of 25 nmol of N-methyl- d-aspartate (NMDA) into the corpus striatum of 7-day-old rats was compared to the effects of injecting 75 nmol into the striatum or hippocampus of adults. The area of histopathology in the immature striatum was 21 × larger than the striatal lesions in adults. Damage from NMDA injected into the immature striatum also extended into the dorsal hippocampus and produced an area of destruction which was 16 × larger than observed after direct injection into the adult hippocampus. Several studies have implicated excessive N-methyl- d-aspartate receptor activation in the pathogenesis of hypopoxic-ischemic and hypoglycemic injury and our results suggest that this neurotoxic mechanism is extremely active in the immature brain.