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F C Zhou

Publications and source records attributed to F C Zhou.

At least 55 records · Page 3Linked to original sources

Four patterns of laminin-immunoreactive structure in developing rat brain.

Laminin has been known to promote neurite outgrowth in culture. We have recently demonstrated that laminin facilitates and guides fiber growth of grafted neurons in the brain. In this study, I report that 4 distinct patterns of laminin immunoreactivity--small and large punctiform laminin, sheath laminin, and somal laminin--were expressed in the brain, each having unique spatial and temporal distributions. They are extensively produced in the developing brain while nerve fibers and vessels are actively growing. Two patterns of laminin, small and large puncta, disappear when the brain matures; the other two, sheath laminin and somal laminin, reduce in intensity but persist through adult life. The small puncta, size less than 1 micron in diameter, frequently accumulated along acellular spaces where future fiber bundles will form, interneuronal spaces and surfaces of neurons where terminal fibers dwell, and around the microvessels where neomicrovessels form. The small puncta, which reportedly appear at the eight-cell stage, were observed in our study at stage E10. They increased in expression from E14 to 19 during the stage when many nerve fibers are navigating to their targets, and subsided in the early postnatal days (P6) when less fiber growth occurs. Large puncta, size about 2 microns in diameter, existed exclusively in the hippocampus. They appeared about E16, and ended P16. The sheath laminin is known to form wrappings on microvessels, the ependymal layer, the choroid plexus, and the surface of the brain. This pattern of laminin attaches to non-neuronal structures which associate with CNS fluid. It was detected at E10 (our youngest group), increased its distribution along with expansion of the associated structure, and persisted throughout the adult life. The somal laminin, associated mainly with the soma of neurons, was the latest appearing laminin pattern (not until E16). It also persisted through adult life in the brain. The somal laminin, which was highly associated with the appearance of neuronal groups in brain nuclei, appeared group by group along with neurons. In summary, laminin attaches with distinct groups of cells in the central nervous system with specific temporal association. This close temporal and spatial association may allow laminin to actively affect nerve growth during brain development. The detailed mapping of laminin distribution is in progress in our lab.

Animals↗

Neurotrophic factor for serotonergic neurons prevents degeneration of grafted raphe neurons in the cerebellum.

Earlier studies in this laboratory have shown that partial serotonergic (5-HT) deafferentation from the hippocampus induced a homotypic sprouting. The 5-HT-denervated hippocampus was further found to be a favorable environment for the growth of grafted 5-HT, but not for norepinephrinergic neurons. In the current study, we have extracted a trophic factor in the denervated hippocampus with hypotonic solution in high-speed supernatant fraction. The trophic extract was assayed in a unique experimental paradigm--test the growth of grafted 5-HT neurons in the cerebellum, which receives the least 5-HT innervation, and where grafted 5-HT neurons have seldom survived. The diluted trophic extract (1:200) produced a dramatic increase in (a) the survival rate, (b) the 5-HT fiber-density, and (c) the 5-HT and 5-hydroxyindoleacetic acid levels of grafted 5-HT neurons in the cerebellum. In addition, Nissl-counterstaining showed that the size of the grafted raphe also apparently increased. The concentrated trophic extract (1:1) did not increase the survival rate of the grafted raphe neurons. These results indicate that (a) the supernatant in the 5-HT-denervated hippocampus contains a potent neurotrophic factor to 5-HT neurons (b) which can either directly or indirectly also support non-5-HT raphe neurons, and (c) possibly an inhibitory factor is present which is detrimental to 5-HT neurons in high concentration.

Animals↗

Connectivities of the striatal grafts in adult rat brain: a rich afference and scant striatonigral efference.

Previous reports from this laboratory have indicated that fetal rat striatal grafts have the major types of neuronal and glial components known to be involved in Huntington's chorea. In this study a number of major afferent and efferent innervations seen in normal striatum were examined in the striatal grafts and were compared with embryonic striatal afferents. First, using immunocytochemistry and histochemistry, the host serotonergic (5-HT), dopaminergic (DA, stained with anti-tyrosine hydroxylase (TH) antiserum), and acetylcholinesterase (AChE) fibers exhibited vigorous growth into the grafts implanted in neostriatum, lateral ventricle, globus pallidus or substantia nigra within a period of 6 and 10 weeks. Individual characteristic terminal patterns formed in striatal grafts: 5-HT fibers were diffused; TH fibers became heavily packed, and AChE fibers were patchy. This peculiar patternization of 5-HT and TH growth into striatal graft appeared to be a recapitulation of the normal 5-HT and TH ingrowth into striatum in the embryonic stage. However, a significantly slow (6 week) onset of adult 5-HT and TH ingrowth into the fetal graft was noted, as compared with that of normal embryonic development (5-6 days from the appearance of 5-HT and TH neurons). With the anterograde-transport marker Phaseolus vulgaris agglutinin leuca method, host cortical neurons also projected to the graft, but in limited numbers. Finally, with the retrograde-transport marker (horseradish peroxidase method, the grafts implanted in neostriatum were found incapable of sending fibers to a major, distal target, substantia nigra. In a current evaluation of striatal transplants, it is shown that major input to the graft can be achieved over time, but output to the distal nigra seems an unrealistic expectation. These data suggest that: (1) the fetal brain tissue was found to be a strong stimulant for sprouting or regeneration of adult nerve fibers; (2) a number of functional recoveries reported on the tested behavior paradigm in this grafting model could be due to the survival of striatal graft and the establishment of input circuitries; further, (3) the data illustrate the necessity of seeking a bridge from the striatal transplant to the host nigra. If a proper functional recovery in Huntington's chorea requires complete striatonigral circuitry, then such a bridge is worthy of a major investigation.

Animals↗

Neuronal and glial elements of fetal neostriatal grafts in the adult neostriatum.

The cellular components of striatal grafts into the host striatum of rats were studied using [3H]thymidine autoradiography, histochemistry, immunocytochemistry and Golgi-staining. Autoradiography revealed that a layer of glial cells, somas smaller than 8 microns in diameter, stained positive with glial fibrillary acidic protein, and demarcating transplant from host, is derived mainly from the donor. Golgi studies revealed that many neuronal fibers fail to cross the glial layer to reach the host striatum. Migration of transplanted striatal cells into the host milieu was evident. The density of migrated cells decreased linearly as a function of distance from the transplant. Most of the far-migrated cells were glial cells. Neuronal migration was limited. In the transplant, donor cells marked by [3H]thymidine constituted at least 70% of the population. Neurons which stained positively for GABA, substance P, and acetylcholinesterase were identified in the transplant. Fibers of two of these three neuronal types, substance P and acetylcholinesterase, formed patchy patterns in the transplant. Detailed morphology on GABAergic fiber is not available to date, because of the limited antibodies or the method used. GABA is the highest population in the striatal transplant. Two types of GABA-positive cells were clearly distinguishable according to cell size. A majority resembled the medium-sized cell commonly found in striatum, while those of the other type resembled the larger GABA cells usually found in the globus pallidus.

Animals↗

Transplanted raphe and hippocampal fetal neurons do not displace afferent inputs to the dorsal hippocampus from serotonergic neurons in the median raphe nucleus of the rat.

To determine if fetal transplants can substitute for or suppress intrinsic serotonergic (5-HT) innervation, we studied the relationship between transplanted and the endogenous raphe neurons projecting to the hippocampus. Fetal raphe transplants produced a 5-HT hyperinnervation of dorsal hippocampus in adult rats. Yet, transplants of fetal raphe tissue did not affect the number of median raphe nucleus (MRN) neurons, approximately 300, which retrogradely transported HRP from the hippocampus. This provides evidence that transplanted 5-HT neurons can co-exist with intrinsic 5-HT nerve terminals in the target area for at least one month. In the second part of this study, fetal hippocampal tissue was transplanted into the host hippocampus. Intrinsic 5-HT immunoreactive fibers innervated the transplanted fetal tissue. Nevertheless, the number of MRN neurons innervating the host tissue as revealed by HRP retrograde transport remained unchanged. Changes in the innervation pattern and 5-HT level in the dorsal hippocampus occur following transplantation of fetal tissue. These changes are discussed and suggest that both the target tissue and afferent neurons readjust to accommodate extrinsic transplanted tissue.

Animals↗

Physiological and morphological characterization of striatal neurons transplanted into the striatum of adult rats.

Physiological and morphological properties of transplanted striatal neurons (TSNs) were examined in an in vitro slice preparation. Fetal striatal tissue (E13-14) was implanted as a dissociated cellular suspension into the striatum of adult rats. Intracellular records were obtained from TSNs 2-6 weeks after transplantation. TSNs exhibited biophysical, morphological, and synaptic properties characteristic of normal striatal neurons, despite the disruption involved in processing of the fetal tissue. Differences were observed, however, between the TSNs and host striatal neurons. TSNs consistently had higher input resistance values than host striatal neurons as determined by neuronal responses to intracellular current injection. Stimulation of adjacent host striatum elicited both excitatory and inhibitory postsynaptic potentials in TSNs. By contrast, the same stimuli elicited only excitatory responses in host striatal neurons. Morphologically, TSNs resembled host medium-size spiny neurons as demonstrated by intracellular injection of lucifer yellow. However, the complexity of dendritic branching and the density of spines on the dendrites were less than that observed for host striatal neurons. It was concluded that during the posttransplantation period studied, TSNs possess neuronal properties expected of developmentally immature striatal neurons.

Action Potentials↗

Partial recovery of gustatory function after neural tissue transplantation to the lesioned gustatory neocortex.

The ability of homotypic cortical tissue grafts to induce recovery of function after a gustatory neocortex (GN) lesion was studied using the conditioned taste aversion (CTA) paradigm. On acquisition day, 26 GN-lesioned and 8 sham-lesioned rats were presented with a saccharin solution, followed by an injection of the illness-inducing agent lithium chloride (LiCl). On the test day, 2 days later, saccharin was presented again. The GN-lesioned rats showed significantly less aversion to saccharin on the test day, indicating that the lesion impaired their ability to form taste-illness association. Nine of the lesioned rats were then bilaterally transplanted with fetal GN tissue. Nine weeks after the transplantation, the rats were presented with a LiCl solution, which served as both a tastant and an illness-inducing agent. An NaCl solution, which tasted very similar to the LiCl solution, was used to test the CTA to salt 3 days later. The nontransplanted rats consumed significantly more LiCl than the transplanted and sham-operated rats on the acquisition day, but both transplanted and nontransplanted rats consumed more NaCl than sham-operated rats on the test day. Nissl and Golgi stainings showed numerous somata and extensive arborization of neurons within the grafts. The results indicate that fetal GN grafts can restore the ability to integrate gustatory and visceral inputs but not to form long-lasting taste-illness associations.

Animals↗

Denervation of serotonergic fibers in the hippocampus induced a trophic factor which enhances the maturation of transplanted serotonergic neurons but not norepinephrine neurons.

The trophic effects of specific denervation on the growth and survival of fetal serotonergic (5-HT) or norepinephrinergic (NE) neurons grafted into the hippocampus were assessed by means of two transplantation paradigms. In the first, fetal raphe cells (containing 5-HT neurons) were transplanted into the control hippocampus. In the second, the transplantation was performed 2 weeks after the specific removal of 5-HT afferents to the hippocampus with 5,7-dihydroxytryptamine (5,7-DHT). We found that a month after transplantation, the number of 5-HT immunoreactive neurons was not significantly different between the two experimental paradigms. However, transplanted raphe neurons had 400% more 5-HT synaptosomal high-affinity uptake and 380% higher content of 5-HT in the hippocampus with prior 5,7-DHT lesion than in control hippocampus. Furthermore, immunocytochemistry showed that the transplanted 5-HT neurons had denser processes and varicosities in the hippocampus with lesion than in control hippocampus. The somatic area of the neurons with these denser processes and varicosities was 42% larger than that of control group. A greater 5-HT level could be achieved if transplanted neurons in the control hippocampus were treated with the supernatant extracted from the hippocampus with 5,7-DHT lesion. In contrast, the NE level of the implanted fetal locus ceruleus (containing NE neurons) was not significantly higher in the 5-HT denervated hippocampus than in control hippocampus a month after transplantation. These results suggest that 5-HT denervation in the hippocampus induces a trophic substance which promotes the maturation rather than survival of 5-HT neurons but not NE neurons.

5,7-Dihydroxytryptamine↗

Stimulation of serotonergic neuronal maturation after fetal mesencephalic raphe transplantation into the 5,7-DHT-lesioned hippocampus of the adult rat.

Neurotoxin lesioning of 5-HT fibers selectively induced the homotypic collateral sprouting of spared 5-HT fibers in the hippocampus. We have used this model to investigate the possibility that the neurotoxin-primed hippocampus will enhance the development of transplanted fetal serotonergic neurons in the brain. The neurotoxin 5,7-DHT, when microinjected into the FF, produced a specific and partial depletion of 5-HT in the hippocampus of adult rats. The ability of the 5,7-DHT-primed hippocampus to selectively support the neurochemical maturation of fetal serotonergic cells was tested by assaying the transplanted fetal raphe or LC 1 month after neuronal transplantation. The neurochemical maturation of fetal 5-HT and NE neurons was dramatically different when they were transplanted in the 5,7-DHT-FF-lesioned hippocampus as compared to the normal hippocampus. The transplanted 5-HT neurons had 480% more SHAU of [3H]5-HT and had a 250% greater content of 5-HT in the partially denervated hippocampus than in the normal hippocampus after 1 month. Furthermore, extracts obtained from lesioned hippocampus enhanced the 5-HT content of 5-HT neurons transplanted in the normal hippocampus, to a level similar to that seen in neurons transplanted in the lesioned hippocampus. In contrast, the implanted NE neurons of fetal LC had a lower NE level in the 5-HT partially denervated hippocampus than in normal hippocampus after 1 month in the host site. The growth of the NE transplants was not facilitated by the vacant postsynaptic space produced by the 5,7-DHT lesion. These results suggest that the 5-HT denervation triggered a trophic signal selectively enhancing the development of the 5-HT neurons but not the NE neurons. Our results are consistent with previous studies showing homotypic collateral sprouting in 5,7-DHT-primed hippocampus.

5,7-Dihydroxytryptamine↗

Induced homotypic sprouting of serotonergic fibers in hippocampus. II. An immunocytochemistry study.

The dorsal hippocampus of the rat normally receives its 5-HT innervation from two homologous groups of cells in the median raphe nucleus via the cingulum bundle-induseum griseum (CB-IG) and the fornix-fimbria (FF) (J. Comp. Neurol., 179 (1978) 641-667 and Brain Res. Bull., 10 (1983) 445-451). 5-HT immunoreactive (IR) fibers are distributed in a laminar pattern in the hippocampus. These fibers have large varicosities and are densely distributed in the infragranular layer of dentate gyrus, in the stratum lacunosum-moleculare of the cornu Ammonis and in the area fasciola cinerea (FC). The present study provides evidence that the density of the 5-HT-IR fibers in the dorsal hippocampus is greatly decreased but maintained a similar laminar pattern 3 days after lesioning by microinjection of 4 micrograms of 5,7-dihydroxytryptamine (5,7-DHT) into the CB-IG. An apparently normal density and distribution pattern of the 5-HT-IR fiber is seen by 42 days postlesion. The FC in the hippocampus is among the first regions reinnervated by 5-HT-IR fibers with very dense and large varicosities. The restitution of 5-HT-IR fibers in the dorsal hippocampus after the 5,7-DHT lesion in the CB-IG is accompanied by a marked increase in the number and intensity of 5-HT-IR fibers in the FF. No evidence of a regeneration of 5-HT-IR fibers is seen distal to the injection site in the CB-IG. These observations provide direct evidence for homotypic collateral sprouting in the CNS induced by removal of a single fiber type.

5,7-Dihydroxytryptamine↗

The effect of adrenalectomy and corticosterone on homotypic collateral sprouting of serotonergic fibers in hippocampus.

Lesioning of serotonergic (5-HT) fibers using 5,7-dihydroxytryptamine (5,7-DHT) in one of the two median raphe-hippocampal pathways induced homotypic sprouting from the other. We have utilized this model with the horseradish peroxidase (HRP) tracing technique to test the effects of an adrenal corticosteroid-corticosterone on homotypic sprouting in female adult rats. Removal of the adrenal glands does not interfere with the 5,7-DHT destruction of 5-HT fibers observed at 3 days. However, the animals without circulating adrenal steroids do not show the increase of the HRP-labeled cells normally seen 21 days after the 5,7-DHT lesions. Furthermore, s.c. implantation of corticosterone pellets was able to completely restore homotypic sprouting of 5-HT fibers in the hippocampus.

5,7-Dihydroxytryptamine↗

Induced homotypic collateral sprouting of serotonergic fibers in the hippocampus of rat.

The organization of the raphe cells efferent to the dorsal hippocampus (DHipp) has been shown to consist of two homologous groups of cells in the median raphe nucleus (MRN). Their axons project by way of the cingulum bundle (CB) and the fornix-fimbria (FF)58. A group of neurons located in the interfascicular part of the dorsal raphe nucleus (IFN) was found to use the CB route exclusively. Quantitation of the HRP-labeled cells in the MRN neurons projecting to the DHipp decreased by 55% and 92% 3 days following CB and CB-FF 5,7-DHT injections respectively. There was a significant increase in HRP-labeled neurons in the MRN at 21 and 42 days compared to 3 days after CB injection (a 2% decrease at 21 days and 38% increase at 42 days compared to sham injected). No change in the number of labeled MRN cells was seen at these long-term time points after combined CB-FF 5,7-DHT microinjections. The labeled cells in the MRN increased in size in the long survival group of CB lesioned animals. The labeled cells localized in the IFN which were lost after CB 5,7-DHT injections did not reappear after long-term survival. These results indicate that CB 5-HT fibers do not regenerate into the DHipp, but FF 5-HT fibers increase their terminal territory by collateral sprouting in response to homotypic denervation.

5,7-Dihydroxytryptamine↗

Effects of 5,7-dihydroxytryptamine on HRP retrograde transport from hippocampus to midbrain raphe nuclei in the rat.

The combination of horseradish peroxidase (HRP) retrograde tracing and specific lesioning using 5,7-dihydroxytryptamine (5,7-DHT) was applied to the midbrain raphe-hippocampal system. Serotonergic fibers from the median raphe nucleus (MRN) of the rat reach the dorsal hippocampus (HIPP) through the cingulum bundle (CB) and the fornix-fimbria (FF). Intracerebral microinjections of 5,7-DHT in these two bundles were made at various times before HRP injections into the dorsal HIPP. After both CB and FF lesion, the number of labeled cells in MRN is reduced to 49.6% at zero time (HRP injected immediately after 5,7-DHT) and to 6.5% after 2 days. There was no significant effect on the number of labeled cells in the locus ceruleus. Selective lesioning of 5-HT fibers in the CB or the FF revealed that raphe-CB-HIPP neurons and raphe-FF-HIPP neurons have a similar distribution pattern in the MRN, but that a dorsal group of neurons at the junction of MRN and dorsal raphe nucleus took the CB route exclusively to innervate the HIPP. The CB pathway was used by more neurons (55% of total number of labeled neurons) than was the FF (21%). An appreciable number of fibers (23%) appear to have branches in both pathways. Our findings are discussed with regard to the recovery of HIPP function seen after long term destruction of 5-HT fibers in the CB.

5,7-Dihydroxytryptamine↗

Alcohol deters the outgrowth of serotonergic neurons at midgestation.

We have previously demonstrated that treatment of pregnant C57BL mice from gestation days 8 to 14 with alcohol with 20% ethanol-derived calories (EDC) reduced the number of serotonin (5-HT) neurons and retarded their migration in the fetal brains. In the present study, we obtained similar results with the use of 25% EDC and extended our previous findings by demonstrating that besides the alteration of the number of 5-HT neurons, prenatal alcohol exposure also affects their projecting fibers in their early development. Pregnant C57BL mice were divided into an alcohol-exposed (ALC) group given 25% EDC (4.49%, v/v), a pair-fed group to the ethanol-fed group (PF) and a chow-fed group (Chow). The PF and Chow groups served as controls. Our results showed that in the ALC group, when compared with the control groups, prenatal alcohol exposure with 25% EDC reduced the number of 5-HT-immunoreactive neurons in both the median and dorsal raphe, and the amount of 5-HT-immunoreactive fibers in the medial forebrain bundle (MFB). The diameter of the 5-HT-immunoreactive MFB was also reduced as a result of treatment. No significant differences of the above parameters were found between the PF and Chow groups. The previous and present work confirmed that alcohol reduces the normal formation and growth of 5-HT neurons in the midbrain. Furthermore, the projection of 5-HT fibers, in density as well as in distribution, is reduced in the major trajectory bundle. This may affect the amount of 5-HT fibers available to the forebrain. In light of the importance of the 5-HT system in brain development, alcohol may affect the growth of the forebrain through its effect on 5-HT signaling.

Animals↗

Serotonergic neurons in the alcohol preferring rats.

Previously, we have shown that the serotonin (5-HT) content and fiber density in a number of terminal brain regions have been found to be decreased in the selectively bred, alcohol preferring (P) rats than in the alcohol nonpreferring (NP) rats. In this study, we further report that, compared with NP rats, there were fewer 5-HT-immunostained (5-HT-IM) neurons in the major ascending raphe nuclei of the P rats. Among the three major groups of 5-HT neurons responsible for the majority of ascending projections to forebrain, dorsal raphe (B7), median raphe (B8), and B9, there were fewer 5-HT-IM neurons in the median and dorsal raphe (not including nucleus oralis) of P rats, compared with NP rats (unpaired Student's test). No difference was observed in the B9 group. When the animals were treated with pargyline and L-tryptophan to enhance the 5-HT in the neurons, the number of 5-HT-IM neurons increased in both lines of rats. However, the difference in the number of 5-HT neurons between the rat lines remained. The intensity of 5-HT-IM was also found to be lower in the dorsal raphe neurons of the untreated P than in the untreated NP rats. The decreased 5-HT-IM was supported by high performance liquid chromatography measurement of 5-HT content, which also indicated that 5-HT content of the dorsal raphe was lower in the P than in the NP rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Alcohol Drinking↗