Morphometric parameters of neurons in reticular nuclei of the brain stem in kittens

Three types of Golgi-stained neurons were discovered in brain stem reticular nuclei of 30-day-old kittens: sparsely branching reticular neurons, those with densely branching dendritic trees, and giant multipolar neurons (Leontovich's classification). Adopting computerized morphometric techniques enabled 23 different parameters to be measured in cells of these types. The measurements taken from the neuronal groups investigated revealed statistically significant differences between them for most parameters. It was concluded from this that each of the neuronal types distinguished has its own morphological identity (or stability). Characteristics of structural differences and properties of differing cell types in reticular nuclei are discussed in relation to their functional properties.Institute of Higher Nervous Activity and Neurophysiology, Academy of Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 23, No. 4, pp. 399–409, July–August, 1991.     

Electrophysiological indices of degree of functional integration between gratified neural tissue and the host brain

Neocortex from 17–18-day rat embryos was transplanted into the barrel-field cortical area in adult rats. Neuronal response to deflecting the vibrissae was tested in the graft 3–8 months afterwards. Nine out of 11 grafts showed a response to sensory stimulation. Irregular and asynchronous discharges predominated in neuronal background firing activity in these grafts. Generalized slow-wave activity had much in common with that occurring on the diametrically opposite site on the intact host brain cortex. Hypersynchronous volleys were detected in neurons of unresponsive grafts. A predominance of waves within the delta range while other rhythms remained only faint, together with epileptiform sharp spikes were seen in generalized activity. Histological treatment of responding grafts revealed close fusion between tissue and host brain. Non-responsive grafts were surronded by a thick glial scar.Institute of Biophysics, Academy of Sciences of the USSR, Pushchino-on-Oka. Institute of Neurobiology and Brain Research, E. German Academy of Sciences, Magdeburg, E. Germany. Translated from Neirofiziologiya, Vol. 19, No. 4, pp. 498–504, July–August, 1987.     

GAD and GABA in an enriched population of cultured GABAergic neurons from rat cerebral cortex

To study various aspects of GABAergic metabolism in an easily accessible system, dissociated cells from postnatal rat cerebral cortex were cultured in a serum-based medium and characterized morphologically and biochemically. The majority (70–90%) of the neurons were GABAergic as determined by three double-labeling procedures. The specific activity of glutamine synthetase in the cultures was 4–5% of the levels in rat astrocyte cultures and intact rat brain, indicating that glia were a minor component. The developmental increase of GABA levels preceded the increase of GAD activity in both immunocytochemical and biochemical experiments. GABA turnover rates also increased with culture age and were 20–30% of GAD activity. Four anti-GAD antibodies, which recognize GAD subunits with differing molecular masses to varying degrees, were used to stain cultured neurons and make immunoblots. Immunoblots showed that the neurons contained two major subunits of GAD which differed in mass by 2 kDa. All four antibodies immunostained both neuronal perikarya and neurites but one antibody, which on the immunoblots predominantly labeled the GAD protein with the lower molecular weight, showed a somewhat more pronounced punctate staining, possibly indicating a principal localization to neurites.     

Colloids as mobile substrates for the implantation and integration of differentiated neurons into the mammalian brain

Neuronal degeneration and the deterioration of neuronal communication lie at the origin of many neuronal disorders, and there have been major efforts to develop cell replacement therapies for treating such diseases. One challenge, however, is that differentiated cells are challenging to transplant due to their sensitivity both to being uprooted from their cell culture growth support and to shear forces inherent in the implantation process. Here, we describe an approach to address these problems. We demonstrate that rat hippocampal neurons can be grown on colloidal particles or beads, matured and even transfected in vitro, and subsequently transplanted while adhered to the beads into the young adult rat hippocampus. The transplanted cells have a 76% cell survival rate one week post-surgery. At this time, most transplanted neurons have left their beads and elaborated long processes, similar to the host neurons. Additionally, the transplanted cells distribute uniformly across the host hippocampus. Expression of a fluorescent protein and the light-gated glutamate receptor in the transplanted neurons enabled them to be driven to fire by remote optical control. At 1-2 weeks after transplantation, calcium imaging of host brain slice shows that optical excitation of the transplanted neurons elicits activity in nearby host neurons, indicating the formation of functional transplant-host synaptic connections. After 6 months, the transplanted cell survival and overall cell distribution remained unchanged, suggesting that cells are functionally integrated. This approach, which could be extended to other cell classes such as neural stem cells and other regions of the brain, offers promising prospects for neuronal circuit repair via transplantation of in vitro differentiated, genetically engineered neurons.     

Convergence between projections from different thalamic nuclei to columns of the rat somatosensory cortex

Primuline fluorochrome retrograde transport technique was used to investigate sources of thalamocortical projections to a single rat somatosensory cortex column connected with the projection of the C₃ vibrissa. Labeled cells were identified in eight different thalamic nuclei: two specific, five nonspecific, and one association nucleus. Labeled neurons differed in the degree of stain accumulated as well as cell numbers and density of distribution from one nucleus to another, indicative of the different arborization patterns of their axons within the cortex. Highest numbers of heavily stained cells as well as highest density of distribution were observed in the ventral thalamic nucleus. The convergence seen between different thalamocortical inputs on to a single somatosensory cortex column explains the functional differences observed between neurons belonging to the same column and makes the formation of functionally distinct neuronal groupings appear possible on this structural basis.Neurocybernetics Research Institute, Rostov-on-Don. Translated from Neirofiziologiya, Vol. 21, No. 2, pp. 168–174, March–April, 1989.     

Correlation between geometric and electrophysiological characteristics of brainstem neurons in kittens

Unit activity was recorded extracellularly from the pontomedullary reticular nuclei of kittens aged 1–5 and 15–30 days, immobilized with diplacin. Properties of neurons located in the medial and lateral zones were compared. As regards the amplitude of spike potentials and types of spontaneous and evoked activity, the cells of the two groups were shown to differ. Tetanic stimulation with a frequency of 300 Hz caused a decrease in the medial zone but an increase in the lateral zone in the number of responding units compared with responses to single stimulation. In neurons of the medial zone intensification of spontaneous activity in the interval between stimuli was more marked and continued after the end of stimulation for a long time. It is suggested that units whose activity is recorded in the medial and lateral zones are mainly giant densely branched and reticular sparsely branched neurons respectively. The difference in the characteristics of activity is connected with the geometry of the dendrites and the foci of their maximal branching.Institute of Higher Nervous Activity and Neurophysiology, Academy of Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 14, No. 2, pp. 140–148, March–April, 1982.     

Survival of embryonic amygdala tissue grafted into various parts of the adult rat brain

It was found during the course of histological examination of preparations containing Nissl and Golgi stained neurons that portions of the embryonic amygdala can successfully survive in the intact adult rat brain. A number of parameters were used enabling development of the graft to be assessed objectively: parenchymal integration index, growth potential, cell density, and vascularization index. By comparing qualitative and quantitative findings of our analyses we showed that location within the host brain is one of the principal factors determining success in graft survival. Grafts transplanted into the cortex survived least well, ventricular cavity transplants fared better, and optimum results were observed with tissue grafted onto the subcortical structures. Normal nerve and glial cells were differentiated in grafts which had taken successfully: capillaries grew into the grafted tissue and common neuropil formed between the graft and the host brain. Structural integration between donor tissue and host brain provides a good model for studying both functional interaction and recovery of function impaired by damage to the host amygdala.Institute of Higher Nervous Activity and Neurophysiology, Academy of Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 19, No. 5, pp. 606–612, September–October, 1987.     

Effects of cold adaptation and noradrenaline on thermosensitivity of rat hypothalamic neurons studiedin vitro

The experiments performed on rat brain slices have shown that cold adaptation of an animal influences the thermosensitivity of hypothalamic medial preoptical neurons. The adaptation is followed by an increase in the proportion of 38–41°C-thermoresponsive neurons and by a decrease in the proportion of 35–38°C-thermoresponsive units. In control animals, noradrenaline (NA) increased the responses of hypothalamic neurons to the action of 35–38°C temperature and decreased them to the action of 38–41°C temperature. Cold adaptation prevented the effects of NA on neuronal thermosensitivity, which suggests that their NA sensitivity is modified by cold adaptation.Neirofiziologiya/Neurophysiology, Vol. 26, No. 3, pp. 171–176, May–June, 1994.     

Morphology and physiology of multiglomerular olfactory projection neurons in the spiny lobster

Whole-cell patch-clamp recording was used to characterize olfactory projection neurons in an isolated brain preparation of the spiny lobster, Panulirus argus. Responses to electrical stimulation of the olfactory afferents were recorded from projection neuron somata using biocytin-filled electrodes. All projection neurons were multiglomerular, innervating up to 80% of all olfactory lobe glomeruli, but the innervation was heterogeneous. Most neurons densely innervated only 3–4 glomeruli; the remaining glomeruli in their dendritic arbor were sparsely innervated, thereby creating two distinct patterns of intraglomerular branching. Projection neurons responded to orthodromic stimulation with an initial depolarization and spiking followed by a 1–3 s hyperpolarization. The inhibitory phase of the response was lower in threshold and longer in latency than the excitatory phase, a response pattern also reported in olfactory projection neurons of insects and vertebrates. The somata of the projection neurons supported voltage-activated currents and TTX-sensitive action potentials, suggesting that the soma, although spatially separated from the axon and dendrites, may play a significant functional role in these cells. Dye coupling between some projection neurons correlated with the presence of multiple amplitude action potentials, suggesting that at least some projection neurons may be coupled via gap junctions.     

Effects of lingual nerve section on morphometric characteristics of reticular nucleus neurons in the kitten brain stem

Computerized morphometric techniques were used to investigate each of 23 parameters in three types of brain stem reticular nucleus neurons in Golgi-stained frontal slices from the brain of 30-day-old kittens after uni- and bilateral lingual nerve section 5–7 days after birth. Particular statistically significant differences in some parameters were discovered in all types of cell. Certain group-specific differences in parameters could be most frequently distinguished in each category: distribution of loose dendritic endings through the dendritic area in reticular neurons, length of dendritic segments in branching cells, and distribution of foci of dendritic arborization in giant multipolar neurons. Unilateral lingual nerve section results in quantitatively more marked deviation from the normal state. It was only under these circumstances, moreover, that differences in overall length of dendrites could be seen, which could indicate a difference in the surface area of the cell.Institute of Higher Nervous Activity and Neurophysiology, Academy of Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 23, No. 4, pp. 409–418, July–August, 1991.     

Quantitative morphological characteristics of developing neurons of the sensory trigeminal nuclei in kittens

Five types of neurons were distinguished in the sensory nuclei of the trigeminal nerve, stained by Golgi's method, in kittens aged 1–5 days and 30 days: reticular and short-dendritic cells (with few branches), and multipolar giant cells, arborescent, and bushy neurons (densely branching). Yet another special type of cell, with a few short dendrites and one long dendrite, was distinguished in preparations from the brain of newborn kittens. Analysis of the dimensions of the bodies, the number, length, and ramification of the dendrites, and the total ramification of the cell yielded quantitative morphological characteristics of these neurons at different times of development. These types of neurons differed in their qualitative and quantitative parameters and in the features of their maturation.Bushy neurons underwent regressive changes during development. Foci of maximal ramification of dendrites of densely branched neurons changed their location during the first months of life relative to the cell body, moving into the more distal regions of the dendrites. Differences in orientation of dendrites with foci of maximal ramification were found relative to neighboring brain formations, which depended on the types of cells and the animal's age. The high level of maturity of trigeminal neurons at birth was demonstrated.Institute of Higher Nervous Activity and Neurophysiology, Academy of Sciences of the USSR, Moscow. Brain Institute, Academy of Medical Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 14, No. 6, pp. 592–600, November–December, 1982.     

Neurotransmitter enzyme activities in neurons purified by bulk-isolation

Neurons, purified by bulk-isolation procedures from 10 day-old rat brain, are 80–90% homogeneous. Contaminants are primarily blood vessels and occasional oligodendroglia. All sizes and shapes of neurons are obtained, as they are isolated from all areas of the brain, except the cerebellum. Neurotransmitter enzyme activities involved in the metabolism of catecholamines, acetylcholine, and GABA are found at twice the level in these purified neurons when compared to that found in whole brain tissue. However, acetycholinesterase is found at a similar activity as in whole brain tissue, suggesting its localization in other cell types as well. Thus purified rat neurons are a good model system for studying neuronal function.     

Neural stem cells transplanted into intact brains as neurospheres form solid grafts composed of neurons, astrocytes and oligodendrocyte precursors

Neural stem cells (NSCs) are tissue-specific stem cells with self-renewal potential that can give rise to neurons and glia in vivo and in vitro. The aim of this study was to transplant NSCs as whole neurospheres into intact brain and assess the fate and phenotype of their progeny generated in vivo. We isolated NSCs from E14 foetal rat forebrains and cultured them in basic fibroblast and epidermal growth factor-supplemented serum-free medium in the form of neurospheres in vitro. Neurospheres were transplanted into the intact brains of 2 Wistar rats and after a period of 3 weeks, grafted brains were examined immunohistochemically. Neurospheres formed solid grafts that were found in the lateral ventricle and in the velum interpositum under the hippocampus. The majority of cells in the transplanted tissue were identified as beta-III-tubulin(+), NeuN(+), PanNF(+) and synaptophysin(+) neurons and were accumulated throughout the graft centre. GFAP(+) astrocytes were scattered throughout the entire graft and astrocyte processes delimited the outer and perivascular surfaces. A great number of NG2(+) oligodendrocyte precursors was detected. Nestin(+) endothelial cells were found to line capillaries growing in the transplant. These data indicate that nestin(+) NSCs prevailing in neurospheres differentiate following transplantation into nestin(-) neuronal and glial cells which confirms the multipotency of NSCs. Three weeks posttransplantation neuronal and astrocyte cells reached terminal differentiation (formation of synaptic vesicles and superficial and perivascular limiting membranes) while elements of oligodendroglial cell lineage remained immature. Grafting stem cells as non-dissociated neurospheres provide cells with favourable conditions which facilitate cell survival, proliferation and differentiation. However, in the intact brain, grafted neurosphere cells were not found to integrate with the brain parenchyma and formed a compact structure demarcated from its surroundings.     

Transferrin and Transferrin Receptor Function in Brain Barrier Systems

1. Iron (Fe) is an essential component of virtually all types of cells and organisms. In plasma and interstitial fluids, Fe is carried by transferrin. Iron-containing transferrin has a high affinity for the transferrin receptor, which is present on all cells with a requirement for Fe. The degree of expression of transferrin receptors on most types of cells is determined by the level of Fe supply and their rate of proliferation.2. The brain, like other organs, requires Fe for metabolic processes and suffers from disturbed function when a Fe deficiency or excess occurs. Hence, the transport of Fe across brain barrier systems must be regulated. The interaction between transferrin and transferrin receptor appears to serve this function in the blood–brain, blood–CSF, and cellular–plasmalemma barriers. Transferrin is present in blood plasma and brain extracellular fluids, and the transferrin receptor is present on brain capillary endothelial cells, choroid plexus epithelial cells, neurons, and probably also glial cells.3. The rate of Fe transport from plasma to brain is developmentally regulated, peaking in the first few weeks of postnatal life in the rat, after which it decreases rapidly to low values. Two mechanisms for Fe transport across the blood–brain barrier have been proposed. One is that the Fe–transferrin complex is transported intact across the capillary wall by receptor-mediated transcytosis. In the second, Fe transport is the result of receptor-mediated endocytosis of Fe–transferrin by capillary endothelial cells, followed by release of Fe from transferrin within the cell, recycling of transferrin to the blood, and transport of Fe into the brain. Current evidence indicates that although some transcytosis of transferrin does occur, the amount is quantitatively insufficient to account for the rate of Fe transport, and the majority of Fe transport probably occurs by the second of the above mechanisms.4. An additional route of Fe and transferrin transport from the blood to the brain is via the blood–CSF barrier and from the CSF into the brain. Iron-containing transferrin is transported through the blood–CSF barrier by a mechanism that appears to be regulated by developmental stage and iron status. The transfer of transferrin from blood to CSF is higher than that of albumin, which may be due to the presence of transferrin receptors on choroid plexus epithelial cells so that transferrin can be transported across the cells by a receptor-mediated process as well as by nonselective mechanisms.5. Transferrin receptors have been detected in neurons in vivo and in cultured glial cells. Transferrin is present in the brain interstitial fluid, and it is generally assumed that Fe which transverses the blood–brain barrier is rapidly bound by brain transferrin and can then be taken up by receptor-mediated endocytosis in brain cells. The uptake of transferrin-bound Fe by neurons and glial cells is probably regulated by the number of transferrin receptors present on cells, which changes during development and in conditions with an altered iron status.6. This review focuses on the information available on the functions of transferrin and transferrin receptor with respect to Fe transport across the blood–brain and blood–CSF barriers and the cell membranes of neurons and glial cells.     

Neural Stem/Progenitor Cells Derived from the Embryonic Dorsal Telencephalon of D6/GFP Mice Differentiate Primarily into Neurons After Transplantation into a Cortical Lesion

D6 is a promoter/enhancer of the mDach1 gene that is involved in the development of the neocortex and hippocampus. It is expressed by proliferating neural stem/progenitor cells (NSPCs) of the cortex at early stages of neurogenesis. The differentiation potential of NSPCs isolated from embryonic day 12 mouse embryos, in which the expression of green fluorescent protein (GFP) is driven by the D6 promoter/enhancer, has been studied in vitro and after transplantation into the intact adult rat brain as well as into the site of a photochemical lesion. The electrophysiological properties of D6/GFP-derived cells were studied using the whole-cell patch-clamp technique, and immunohistochemical analyses were carried out. D6/GFP-derived neurospheres expressed markers of radial glia and gave rise predominantly to immature neurons and GFAP-positive cells during in vitro differentiation. One week after transplantation into the intact brain or into the site of a photochemical lesion, transplanted cells expressed only neuronal markers. D6/GFP-derived neurons were characterised by the expression of tetrodotoxin-sensitive Na⁺-currents and K _A- and K _DR currents sensitive to 4-aminopyridine. They were able to fire repetitive action potentials and responded to the application of GABA. Our results indicate that after transplantation into the site of a photochemical lesion, D6/GFP-derived NSPCs survive and differentiate into neurons, and their membrane properties are comparable to those transplanted into the non-injured cortex. Therefore, region-specific D6/GFP-derived NSPCs represent a promising tool for studying neurogenesis and cell replacement in a damaged cellular environment.     

小鼠胚胎干细胞移植入成体大鼠脑内的区域特异性存活与分化

全能区域非特异性的胚胎干细胞是研究成体不同脑区控制干细胞分化能力的十分有力的工具。胚胎干细胞源性神经前体细胞移植入成体脑后可分化为功能性神经元,但是未分化的胚胎干细胞在成体脑内各个部位的存活、生长与分化的潜能差异尚不清楚。本文旨在探讨成体脑组织对胚胎干细胞的影响及胚胎干细胞在成体脑内的一系列行为。将少量转绿色荧光蛋白未分化的小鼠胚胎干细胞移植入成体大鼠脑内不同部位,分别于移植5、14和28d后处死大鼠,进行形态学观察及免疫组化定性,以了解未分化的小鼠胚胎干细胞在大鼠脑内不同区域的存活、生长与分化。结果发现未分化的小鼠胚胎干细胞可逐步整合入受体组织并向nestin阳性神经前体细胞分化。移植细胞及其后裔在海马生长最为旺盛,而在隔区最差（P〈0．01）;移植细胞分化为神经干细胞的效率也是在海马最高,而在隔区最低（P〈0．01）。提示只有部分脑区适合胚胎干细胞及其后裔生存,并提供促进其分化的有益环境。因此,由于位置特异的微环境因子及环境因素的存在,宿主组织特性对决定中枢神经系统疾病的细胞替代疗法策略是相当重要的。     

Analysis of glutamate receptors in primary cultured neurons from fetal rat forebrain

In order to further analyze the development of glutamatergic pathways in neuronal cells, the expression of excitatory amino acid receptors was studied in a model of neurons in primary culture by measuring the specific binding of L-[³H]glutamate under various incubation conditions in 8-day-old intact living neurons isolated from the embryonic rat forebrain, as well as in membrane preparations from these cultures and from newborn rat forebrain. In addition, the receptor responsiveness to glutamate was assessed by studying the uptake of tetraphenylphosphonium (TPP⁺) which reflects membrane polarization. In the presence of a potent inhibitor of glutamate uptake, the radioligand bound to a total number of sites of 36.7 pmol/mg protein in intact cells incubated in a Tris buffer containing Na⁺, Ca²⁺, and Cl^–, with a Kd around 2 M. In the absence of the above ions, [³H]glutamate specific binding diminished to 14.2 pmol/mg protein with a Kd-value of 550 nM. Under both of the above conditions, similar Kd were obtained in membranes isolated from cultures and from the newborn brain. However, Bmax-values were significantly lower in culture membranes than in intact cells or newborn membranes. Displacement studies showed that NMDA was the most potent compound to inhibit [³H]glutamate binding in membranes obtained from cultured neurons as well as from the newborn brain, whereas quisqualate, AMPA, kainate andtrans-ACPD were equally effective. According to these data and to the ionic dependence of glutamate binding, it was concluded that cultured neurons from the rat embryo forebrain express various glutamate receptor subtypes, mainly L-AP4 and NMDA receptors, with characteristics close to those in the newborn brain, and which display functional properties since a transient cell exposure to glutamate led to a 70% inhibition of [³H]TPP⁺ uptake.     

Neurons of the human basal ganglia (striatum and basolateral amygdala) expressing the enzyme NADPH-d

In human striatum and basolateral amygdala NADPH-d+ neurons were revealed (after Vincent et al., 1983); and in striatum strio-cortical neurons were also revealed using DiI marker (after Dahtstrom and Belichenko, 1995). The NADPH-d+ neurons were numerous in both formations. Staining of NADPH-d+ neurons with their processes, and our previous study of striatal and amygdalar human neurons by Golgi method made it possible to identify the species of neurons with their assessment as sparsely or densely branched. The main efferent neurons of striatum and basolateral amygdala (densely branched medium spiny and bushy spiny, respectively) and their densely branched interneurons were not marked. Efferent NADPH-d+ neurons included the most numerous ones in both formations. A projection of reticular striatal neurons to cortex was also shown. The NADPH-d+ interneurons belonged to sparsely branched forms. In striatum they included slender-dendritic and long-dendritic bipolars (numerous), ordinary bipolars, twisted and large poor-dendritic cells; in amygdala--the same bipolars and radial cells. Thus, the NADPH-d positive cells in the formations under study were represented by more "ancient" or less structurally complex cell forms.     

Factors Influencing the Differentiation of Dopaminergic Traits in Transplanted Neural Stem Cells

1. Our previous studies demonstrated that when neural stem cells (NSCs) of the C17.2 clonal line are transplanted into the intact or 6-hydroxydopamine (6-OHDA) lesioned rat striatum, in most, but not all grafts, cells spontaneously express the dopamine (DA) biosynthetic enzymes, tyrosine hydroxylase (TH), and aromatic L-amino acid decarboxylase (Yang, M., Stull, N. D., Snyder, E. Y., Berk, M. A., and Iacovitti, L. (2002). Exp. Neurol.).2. These results suggested that there were certain conditions which were more conducive to the development of DA traits in NSCs and possibly other neurotransmitter phenotypes.3. In the present study, we modified a number of variables in vitro (i.e. passage number, confluence) and/or in vivo (degree, type, and site of injury) before assessing the survival, migration, and differentiation of engrafted NSCs.4. We found that low confluence cultures were comprised exclusively of flattened polygonal cells, which when transplanted, migrated widely in the brain but did not express TH.5. In contrast, high confluence cultures contained both polygonal cells and an overlying bed of fusiform cells.6. When these NSCs were maintained for 12–20 passages and then transplanted, virtually all engrafted cells in 65% of the grafts expressed TH but not markers of other neurotransmitter systems.7. Importantly, all TH⁺ grafts were accompanied by significant physical damage to the brain while TH^– grafts were not, suggesting that local injury-related factors were also important.8. Of no apparent influence on TH expression, regardless of how cells were grown prior to implantation, was the site of transplantation (cortex or striatum) or the degree of chemical lesion (intact, partial or full).9. We conclude that transplanted NSCs can express traits specifically associated with DA neurons but only when cells are grown under certain conditions in vitro and then transplanted in proximity to injury-induced factors present in vivo.     

Effects of nerve growth factor on the development of the dendritic system of cholinergic neurons in dissociated culture of the rat septum]

Neurons dissociated from septal area of fetal (E18-19) rat brain were grown 14-days in culture. Cholinergic neurons were identified by cytochemical demonstration of acetyl cholinesterase. It was shown that the nerve growth factor added to the culture medium (50 u/ml) has increased the size of cell body of AchE-positive neurons, mean total length and arborization of dendrites and also the dendritic tree area.