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PubMed2003

Estimation of absolute microglial cell numbers in mouse fascia dentata using unbiased and efficient stereological cell counting principles.

Stereology offers a set of unbiased principles to obtain precise estimates of total cell numbers in a defined region. In terms of microglia, which in the traumatized and diseased CNS is an extremely dynamic cell population, the strength of stereology is that the resultant estimate is unaffected by shrinkage or expansion of the tissue. The optical fractionator technique is very efficient but requires relatively thick sections (e.g., > or =20 microm after coverslipping) and the unequivocal identification of labeled cells throughout the section thickness. We have adapted our protocol for Mac-1 immunohistochemical visualization of microglial cells in thick (70 microm) vibratome sections for stereological counting within the murine hippocampus, and we have compared the staining results with other selective microglial markers: the histochemical demonstration of nucleotide diphosphatase (NDPase) activity and the tomato lectin histochemistry. The protocol gives sections of high quality with a final mean section thickness of >20 microm (h=22.3 microm +/- 0.64 microm), and with excellent rendition of Mac-1+ microglia through the entire height of the section. The NDPase staining gives an excellent visualization of microglia, although with this thickness, the intensity of the staining is too high to distinguish single cells. Lectin histochemistry does not visualize microglia throughout the section and, accordingly, is not suited for the optical fractionator. The mean total number of Mac-1+ microglial cells in the unilateral dentate gyrus of the normal young adult male C57BL/6 mouse was estimated to be 12,300 (coefficient of variation (CV)=0.13) with a mean coefficient of error (CE) of 0.06. The perspective of estimating microglial cell numbers using stereology is to establish a solid basis for studying the dynamics of the microglial cell population in the developing and in the injured, diseased and normal adult CNS.

Animals↗
PubMed1992

Comparison of the effects of ethanol and chlordiazepoxide on electrophysiological activity in the fascia dentata and hippocampus regio superior.

Acute intoxicating doses of ethanol-producing blood alcohol levels of 120-200 mg% increase paired-pulse (PP) inhibition in the dentate gyrus of anesthetized rats suggesting that ethanol increases recurrent inhibitory processes (Wiesner, J.B., and S.J. Henriksen (1987) Ethanol enhances recurrent inhibition in the dentate gyrus of the hippocampus. Neurosci. Lett. 79:169-173). To further understanding of the neuronal mechanisms underlying this phenomenon, the authors studied the effects of the benzodiazepine (BZ), chlordiazepoxide, and acute intoxicating levels of ethanol on extracellular field potential recordings and single-unit activity in the dentate gyrus and area CA1 of the hippocampus. In the dentate, ethanol had no effect on population excitatory postsynaptic potential (pEPSP) amplitudes or slopes; decreased population spike (PS) amplitudes (25%); increased PP inhibition; decreased dentate granule cell (DGC) spontaneous activity (58%); had no effect on putative interneuron spontaneous activity; and markedly increased post field potential-evoked interneuron discharges (IDs, 218%). Chlordiazepoxide had no effect on pEPSP amplitudes or slopes or PS amplitudes; increased PP inhibition; decreased DGC (62%) and interneuron (72%) spontaneous activity; and markedly decreased IDs (89%). In CA1, ethanol had no effect on pEPSP amplitudes or slopes; decreased PS amplitudes (26%); had no effect on PP responses; decreased pyramidal cell (PC) spontaneous activity (39%); had no effect on interneuron spontaneous activity; and markedly increased IDs (97%). Chlordiazepoxide had no effect on pEPSP amplitudes or slopes or PS amplitudes; had no effect on PP responses; decreased PC spontaneous activity (41%); and had no effect on interneuron spontaneous activity or IDs. The results suggest that the BZs decrease principal cell excitability by postsynaptic facilitation of inhibitory processes, whereas ethanol decreases principal cell excitability indirectly by increasing the excitability of inhibitory interneurons.

Action Potentials↗
PubMed1994

Mossy cells of the rat fascia dentata are glutamate-immunoreactive.

The mossy cells represent a prominent cell type of the hilar region. Whereas the morphology of these neurons, their synaptic connections, and physiological characteristics have been described in some detail, information about their neurotransmitter is still lacking. Using immunocytochemistry in combination with Golgi impregnation, the authors demonstrate that identified mossy cells are GABA-immunonegative but stain for glutamate. These results do not prove that these cells use glutamate as a transmitter, since glutamate is a ubiquitous metabolite. However, together with the lack of GABA staining and a recent report on asymmetric spine synapses formed by identified mossy cell axons, the present results support an excitatory nature of these neurons.

Animals↗
PubMed1994

Transient decrease in calbindin immunoreactivity of the rat fascia dentata granule cells after repeated electroconvulsive shocks.

Changes in hippocampal calbindin immunoreactivity were investigated after repeated electroconvulsive shocks. Adult rats were subjected to 10, 20, or 36 electroconvulsive shocks (50 mA, 0.5 seconds), given as on shock per day, 5 days a week. The rats were sacrificed and processed for calbindin immunohistochemistry 1, 2, and 30 days after the last electroconvulsive shock. In the rats receiving 10 or 20 electroconvulsive shocks, a selective reduction of the calbindin immunoreactivity of the dentate granule cell bodies, dendrites, and mossy fibers was noted 1 and 2 days after the last electroconvulsive shock. After 36 electroconvulsive shocks there was an almost complete loss of calbindin immunoreactivity from the granule cell bodies and dendrites, and the calbindin immunoreactivity of the mossy fibers was markedly reduced. Thirty days after the last of 36 electroconvulsive shocks, the calbindin immunoreactivity was back to normal. Besides demonstrating pronounced changes associated with repeated electroconvulsive shocks, the results confirm the transient nature of these changes.

Animals↗
PubMed2000

NMDAR1 receptor proteins and mossy fibers in the fascia dentata during rat kainate hippocampal epileptogenesis.

We examined the time course of NMDAR1 (NR1) immunoreactivity (IR) in the rat inner molecular layer of the dentate gyrus following unilateral intrahippocampal (hilar) kainic acid (KA) lesions and compared them to progressive aberrant mossy fiber (MF) sprouting into the inner molecular layer (IML). The results demonstrated that NR1 receptors in the IML of the KA side were decreased as early as 3 days after KA-induced denervation, then significantly increased at postinjection day (PID) 7. The densities of NR1 IR in the IML continued to increase up to 5 months. By comparison, MF sprouting did not occur significantly in the IML until PID 17, 10 days after NR1 IR was significantly increased. Recurrent MF-IML neoinnervation significantly increased on days 17, 60, and 150. This progressive MF innervation was significantly correlated with NR1 increases. These results suggest that NR1 receptors were decreased soon after KA-induced deafferentation of granule cell dendrites in the IML; however, they were replaced by new NR1 receptors at increased densities in the granule cell dendrites, which may have released neurotrophic factors to stimulate growth cones of MFs to reinnervate the IML. The progressive increases of NR1 and MFs in the IML suggest that such neosynaptogenesis would contribute monosynaptic recurrent excitatory mechanisms for focal hippocampal hyperexcitability and seizure onsets.

Animals↗
PubMed1979

Synaptic extensions from the mossy fibers of the fascia dentata.

In the hilar region of the rat hippocampus, filamentous extensions have been observed to originate from the en passant synaptic expansions on the so-called mossy fibers, which are the axons of the dentate granule cells. These extensions range in length from about 1 micrometer to 30 micrometer, are often branched, and appear to contact the processes of various cell types in the hilar region. In 28-day-old rats, there are between 4 and 9 such extensions from most mossy fiber expansions, and the total length of the extensions from any one expansion is on the order of 75 micrometer. Analysis of serial electron micrographs through normal and Golgi-impregnated mossy fibers has confirmed that these extensions are, indeed, presynaptic processes. Each contains one or more vesicle-rich foci along its length, and is associated with asymmetric membrane specializations. At these sites, the extensions are in synaptic contact with dendrites and dendritic spines of, as yet, unknown origin. A quantitative analysis of these extensions in Golgi material from rats at different ages indicates that they reach their greatest length around 14 days and then decline to adult values by 28 days.

Age Factors↗