All Publications

  • Surviving Mossy Cells Enlarge and Receive More Excitatory Synaptic Input in a Mouse Model of Temporal Lobe Epilepsy HIPPOCAMPUS Zhang, W., Thamattoor, A. K., LeRoy, C., Buckmaster, P. S. 2015; 25 (5): 594-604


    Numerous hypotheses of temporal lobe epileptogenesis have been proposed, and several involve hippocampal mossy cells. Building on previous hypotheses we sought to test the possibility that after epileptogenic injuries surviving mossy cells develop into super-connected seizure-generating hub cells. If so, they might require more cellular machinery and consequently have larger somata, elongate their dendrites to receive more synaptic input, and display higher frequencies of miniature excitatory synaptic currents (mEPSCs). To test these possibilities pilocarpine-treated mice were evaluated using GluR2-immunocytochemistry, whole-cell recording, and biocytin-labeling. Epileptic pilocarpine-treated mice displayed substantial loss of GluR2-positive hilar neurons. Somata of surviving neurons were 1.4-times larger than in controls. Biocytin-labeled mossy cells also were larger in epileptic mice, but dendritic length per cell was not significantly different. The average frequency of mEPSCs of mossy cells recorded in the presence of tetrodotoxin and bicuculline was 3.2-times higher in epileptic pilocarpine-treated mice as compared to controls. Other parameters of mEPSCs were similar in both groups. Average input resistance of mossy cells in epileptic mice was reduced to 63% of controls, which is consistent with larger somata and would tend to make surviving mossy cells less excitable. Other intrinsic physiological characteristics examined were similar in both groups. Increased excitatory synaptic input is consistent with the hypothesis that surviving mossy cells develop into aberrantly super-connected seizure-generating hub cells, and soma hypertrophy is indirectly consistent with the possibility of axon sprouting. However, no obvious evidence of hyperexcitable intrinsic physiology was found. Furthermore, similar hypertrophy and hyper-connectivity has been reported for other neuron types in the dentate gyrus, suggesting mossy cells are not unique in this regard. Thus, findings of the present study reveal epilepsy-related changes in mossy cell anatomy and synaptic input but do not strongly support the hypothesis that mossy cells develop into seizure-generating hub cells. © 2014 Wiley Periodicals, Inc.

    View details for DOI 10.1002/hipo.22396

    View details for Web of Science ID 000353975200005

    View details for PubMedID 25488607

  • Increased Excitatory Synaptic Input to Granule Cells from Hilar and CA3 Regions in a Rat Model of Temporal Lobe Epilepsy JOURNAL OF NEUROSCIENCE Zhang, W., Huguenard, J. R., Buckmaster, P. S. 2012; 32 (4): 1183-1196


    One potential mechanism of temporal lobe epilepsy is recurrent excitation of dentate granule cells through aberrant sprouting of their axons (mossy fibers), which is found in many patients and animal models. However, correlations between the extent of mossy fiber sprouting and seizure frequency are weak. Additional potential sources of granule cell recurrent excitation that would not have been detected by markers of mossy fiber sprouting in previous studies include surviving mossy cells and proximal CA3 pyramidal cells. To test those possibilities in hippocampal slices from epileptic pilocarpine-treated rats, laser-scanning glutamate uncaging was used to randomly and focally activate neurons in the granule cell layer, hilus, and proximal CA3 pyramidal cell layer while measuring evoked EPSCs in normotopic granule cells. Consistent with mossy fiber sprouting, a higher proportion of glutamate-uncaging spots in the granule cell layer evoked EPSCs in epileptic rats compared with controls. In addition, stimulation spots in the hilus and proximal CA3 pyramidal cell layer were more likely to evoke EPSCs in epileptic rats, despite significant neuron loss in those regions. Furthermore, synaptic strength of recurrent excitatory inputs to granule cells from CA3 pyramidal cells and other granule cells was increased in epileptic rats. These findings reveal substantial levels of excessive, recurrent, excitatory synaptic input to granule cells from neurons in the hilus and proximal CA3 field. The aberrant development of these additional positive-feedback circuits might contribute to epileptogenesis in temporal lobe epilepsy.

    View details for DOI 10.1523/JNEUROSCI.5342-11.2012

    View details for Web of Science ID 000299801100005

    View details for PubMedID 22279204

  • Surviving Hilar Somatostatin Interneurons Enlarge, Sprout Axons, and Form New Synapses with Granule Cells in a Mouse Model of Temporal Lobe Epilepsy JOURNAL OF NEUROSCIENCE Zhang, W., Yamawaki, R., Wen, X., Uhl, J., Diaz, J., Prince, D. A., Buckmaster, P. S. 2009; 29 (45): 14247-14256


    In temporal lobe epilepsy, seizures initiate in or near the hippocampus, which frequently displays loss of neurons, including inhibitory interneurons. It is unclear whether surviving interneurons function normally, are impaired, or develop compensatory mechanisms. We evaluated GABAergic interneurons in the hilus of the dentate gyrus of epileptic pilocarpine-treated GIN mice, specifically a subpopulation of somatostatin interneurons that expresses enhanced green fluorescence protein (GFP). GFP-immunocytochemistry and stereological analyses revealed substantial loss of GFP-positive hilar neurons (GPHNs) but increased GFP-positive axon length per dentate gyrus in epileptic mice. Individual biocytin-labeled GPHNs in hippocampal slices from epileptic mice also had larger somata, more axon in the molecular layer, and longer dendrites than controls. Dual whole-cell patch recording was used to test for monosynaptic connections from hilar GPHNs to granule cells. Unitary IPSCs (uIPSCs) recorded in control and epileptic mice had similar average rise times, amplitudes, charge transfers, and decay times. However, the probability of finding monosynaptically connected pairs and evoking uIPSCs was 2.6 times higher in epileptic mice compared to controls. Together, these findings suggest that surviving hilar somatostatin interneurons enlarge, extend dendrites, sprout axon collaterals in the molecular layer, and form new synapses with granule cells. These epilepsy-related changes in cellular morphology and connectivity may be mechanisms for surviving hilar interneurons to inhibit more granule cells and compensate for the loss of vulnerable interneurons.

    View details for DOI 10.1523/JNEUROSCI.3842-09.2009

    View details for Web of Science ID 000271664000019

    View details for PubMedID 19906972

  • Dysfunction of the Dentate Basket Cell Circuit in a Rat Model of Temporal Lobe Epilepsy JOURNAL OF NEUROSCIENCE Zhang, W., Buckmaster, P. S. 2009; 29 (24): 7846-7856


    Temporal lobe epilepsy is common and difficult to treat. Reduced inhibition of dentate granule cells may contribute. Basket cells are important inhibitors of granule cells. Excitatory synaptic input to basket cells and unitary IPSCs (uIPSCs) from basket cells to granule cells were evaluated in hippocampal slices from a rat model of temporal lobe epilepsy. Basket cells were identified by electrophysiological and morphological criteria. Excitatory synaptic drive to basket cells, measured by mean charge transfer and frequency of miniature EPSCs, was significantly reduced after pilocarpine-induced status epilepticus and remained low in epileptic rats, despite mossy fiber sprouting. Paired recordings revealed higher failure rates and a trend toward lower amplitude uIPSCs at basket cell-to-granule cell synapses in epileptic rats. Higher failure rates were not attributable to excessive presynaptic inhibition of GABA release by activation of muscarinic acetylcholine or GABA(B) receptors. High-frequency trains of action potentials in basket cells generated uIPSCs in granule cells to evaluate readily releasable pool (RRP) size and resupply rate of recycling vesicles. Recycling rate was similar in control and epileptic rats. However, quantal size at basket cell-to-granule cell synapses was larger and RRP size smaller in epileptic rats. Therefore, in epileptic animals, basket cells receive less excitatory synaptic drive, their pools of readily releasable vesicles are smaller, and transmission failure at basket cell-to-granule cell synapses is increased. These findings suggest dysfunction of the dentate basket cell circuit could contribute to hyperexcitability and seizures.

    View details for DOI 10.1523/JNEUROSCI.6199-08.2009

    View details for Web of Science ID 000267131000024

    View details for PubMedID 19535596

  • Bidirectional interactions between H-channels and Na+-K+ pumps in mesencephalic trigeminal neurons JOURNAL OF NEUROSCIENCE Kang, Y. N., Notomi, T., Saito, M., Zhang, W., Shigemoto, R. 2004; 24 (14): 3694-3702


    The Na(+)-K(+) pump current (I(p)) and the h-current (I(h)) flowing through hyperpolarization-activated channels (h-channels) participate in generating the resting potential. These two currents are thought to be produced independently. We show here bidirectional interactions between Na(+)-K(+) pumps and h-channels in mesencephalic trigeminal neurons. Activation of I(h) leads to the generation of two types of ouabain-sensitive I(p) with temporal profiles similar to those of instantaneous and slow components of I(h), presumably reflecting Na(+) transients in a restricted cellular space. Moreover, the I(p) activated by instantaneous I(h) can facilitate the subsequent activation of slow I(h). Such counteractive and cooperative interactions were also disclosed by replacing extracellular Na(+) with Li(+), which is permeant through h-channels but does not stimulate the Na(+)-K(+) pump as strongly as Na(+) ions. These observations indicate that the interactions are bidirectional and mediated by Na(+) ions. Also after substitution of extracellular Na(+) with Li(+), the tail I(h) was reduced markedly despite an enhancement of I(h) itself, attributable to a negative shift of the reversal potential for I(h) presumably caused by intracellular accumulation of Li(+) ions. This suggests the presence of a microdomain where the interactions can take place. Thus, the bidirectional interactions between Na(+)-K(+) pumps and h-channels are likely to be mediated by Na(+) microdomain. Consistent with these findings, hyperpolarization-activated and cyclic nucleotide-modulated subunits (HCN1/2) and the Na(+)-K(+) pumpalpha3 isoform were colocalized in plasma membrane of mesencephalic trigeminal neurons having numerous spines.

    View details for DOI 10.1523/JNEUROSCI.5641-03.2004

    View details for Web of Science ID 000220715400026

    View details for PubMedID 15071118

  • An involvement of trigeminal mesencephalic neurons in regulation of occlusal vertical dimension in the guinea pig JOURNAL OF DENTAL RESEARCH Zhang, W., Kobayashi, M., Moritani, M., Masuda, Y., Dong, J., Yagi, T., Maeda, T., Morimoto, T. 2003; 82 (7): 565-569


    Although the occlusal vertical dimension (OVD) is strictly controlled, the neuronal mechanism of its regulation is still unclear. We hypothesize that neurons in the trigeminal mesencephalic nucleus (MesV) play an important role in the regulation of the OVD, because the MesV receives the projection from jaw-closing muscle spindles and periodontal mechanoreceptors. We measured the temporal OVD change in the guinea pig to study the effects of MesV lesions on the OVD. OVD-raised animals without MesV lesions showed a rapid OVD decrease to the same level as that in naïve controls, followed by an OVD increase after the OVD-raising appliance was removed. In contrast, OVD-raised animals with MesV lesions showed only a slight decrease in the OVD for 15 days after removal of the appliance, and then the OVD increased. The time-course of OVD development in normal-bite animals with MesV lesions was similar to that of naïve controls. These results suggest that MesV neurons are involved in OVD regulation.

    View details for Web of Science ID 000183721400015

    View details for PubMedID 12821720