Development Of Temporal Lobe Epilepsy Research Articles

Glutathione pegylated liposomal methylprednisolone administration after the early phase of status epilepticus did not modify epileptogenesis in the rat

It has been reported that glucocorticoids (GCs) can effectively control seizures in pediatric epilepsy syndromes, possibly by inhibition of inflammation. Since inflammation is supposed to be involved in epileptogenesis, we hypothesized that treatment with GCs would reduce brain inflammation and thereby modify epileptogenesis in a rat model for temporal lobe epilepsy, in which epilepsy gradually develops after electrically induced status epilepticus (SE). To prevent the severe adverse effects that are inevitable with long-term GC treatment, we used liposome nanotechnology (G-Technology(®)) to enhance the sustained delivery to the brain. Starting 4h after onset of SE, rats were treated with glutathione pegylated liposomal methylprednisolone (GSH-PEG liposomal MP) according to a treatment protocol (1× per week; 10mg/kg) that is effective in other models of neuroinflammation. Continuous electro-encephalogram (EEG) recordings revealed that SE duration and onset of spontaneous seizures were not affected by GSH-PEG liposomal MP treatment. The number and duration of spontaneous seizures were also not different between vehicle and GSH-PEG liposomal MP-treated animals. Six weeks after SE, brain inflammation, as assessed by quantification of microglia activation, was not reduced by GSH-PEG liposomal MP-treatment. Also, neuronal cell loss and mossy fiber sprouting were not affected. Our study shows that the selected GSH-PEG liposomal MP treatment regimen that was administered beyond the acute SE phase does not reduce brain inflammation and development of temporal lobe epilepsy.

The effect of paxilline on early alterations of electrophysiological properties of dentate gyrus granule cells in pilocarpine-treated rats.

The dentate gyrus of hippocampus has long been considered as a focal point for studies on mechanisms responsible for the development of temporal lobe epilepsy (TLE). Change in intrinsic properties of dentate gyrus granule cells (GCs) has been considered as an important factor responsible in temporal lobe seizures. In this study, we evaluated the intrinsic properties of GCs, during acute phase of seizure (24 h after i.p. injection of pilocarpine) compared to sham group using whole cell patch-clamp recordings. Our results showed a significant increase in the number of action potentials (APs) after applying depolarizing currents of 200 pA (p < 0.01) and 250pA (p < 0.05) compared to sham group. The evaluation of AP properties revealed a decrease in half-width of AP in GCs of seizure group (1.27 ± 0.03 ms) compared to sham group (1.60 ± 0.11). Moreover, addition of BAPTA to pipette solution prevented changes in AP half-width in seizure group (1.71 ± 0.11 ms) compared to sham group (1.91 ± 0.08 ms). In contrast, an increase in the amplitude of fast afterhyperpolarization was observed in GCs of seizure group (-11.68 ± 0.72 mV) compared to sham group (-8.28 ± 0.59 mV). Also, GCs of seizure group showed a significant increase in both firing rate and instantaneous firing frequency at depolarizing currents of 200 pA (P < 0.01) and 250 pA (P < 0.05) compared to sham group. The changes in electrophysiological properties of GCs were attenuated after bath application of paxilline suggesting possible involvement of large conductance Ca(2+)- activated K(+) channel (BK channel). Our results suggested the possible involvement of certain potassium channels in early changes of intrinsic properties of GCs which eventually facilitate TLE development.

Detection of c-Fos Expression in the Brains of Animals with a Pilocarpine Model of Temporal Lobe Epilepsy

In studies of the mechanisms of development of temporal lobe epilepsy, it is important to identify the dynamics of the involvement of various brain structures in the pathological process induced by the action of harmful factors. The present study used an experimental model of status epilepticus induced by administration of lithium chloride and pilocarpine. The indicator for brain neurons activated after the onset of status epilepticus was immunochemical detection of the immediate early gene c-fos. c-Fos expression was assessed 30, 60, and 90 min after administration of pilocarpine. Increases in immunoreactivity were seen in the neuron populations of various brain structures during the development of status epilepticus, and the activation process was found to differ in terms of extent and timing in different brain structures. The structures in which c-Fos immunoreactivity was detected earliest were the piriform cortex, the olfactory tubercle, the thalamic nuclei, the lateral nuclei of the habenula, and the caudate nucleus. Structures with later involvement were the hippocampus, septal area, amygdala, and basal ganglia. c-Fos was detected later in other structures in the hypothalamic area. These data contribute to our understanding of the mechanisms of temporal lobe epilepsy and the search for approaches to its treatment.

Astrocyte control of synaptic NMDA receptors contributes to the progressive development of temporal lobe epilepsy

Astrocytes modulate neuronal activity, synaptic transmission, and behavior by releasing chemical transmitters in a process termed gliotransmission. Whether this process impacts epilepsy in vivo is not known. We show that genetic impairment of transmitter release from astrocytes by the expression of a glial dominant-negative SNARE domain in mice reduced epileptiform activity in situ, delayed seizure onset after pilocarpine-induced status epilepticus, and attenuated subsequent progressive increase in seizure frequency in vivo. The reduced seizure frequency was accompanied by attenuation of hippocampal damage and behavioral deficits. As the delay in seizure onset and the reduced seizure frequency were mimicked by intracerebroventricular delivery of the NMDA receptor (NMDAR) antagonist D-(-)-2-amino-5-phosphonopentanoate in WT littermates and because dominant-negative SNARE expression leads to a hypofunction of synaptic NMDARs, we conclude that astrocytes modulate epileptogenesis, recurrent spontaneous seizures, and pathophysiological consequences of epilepsy through a pathway involving NMDARs.

Is serotonin receptor HTR1B implicated in mesial temporal lobe epilepsy development?

Is serotonin receptor HTR1B implicated in mesial temporal lobe epilepsy development?

Postnatal interleukin-1β enhances adulthood seizure susceptibility and neuronal cell death after prolonged experimental febrile seizures in infantile rats

Febrile seizures (FS) are recognized as an antecedent to the development of temporal lobe epilepsy with hippocampal sclerosis (TLE-HS), but it is unclear whether prolonged FS are a direct cause of TLE-HS. Here, we used a rat model of infantile FS to study the effects of inflammatory cytokines on seizure susceptibility and neuronal death in adults. Prolonged hyperthermia-induced seizures (pHS) were induced in male Lewis rats at post natal day (P) 10. Cytokines were administered twice intranasally, once immediately after pHS and once the following day. The effects of intranasal interleukin (IL)-1β or tumor necrosis factor (TNF) α were tested in rats undergoing a single episode of pHS (P10) and in rats undergoing repeated pHS (P10 and P12). Seizure susceptibility was tested at P70-73 by quantifying the seizure onset time (SOT) after kainic acid administration, and neuronal cell injury and gliosis in adulthood. SOT significantly reduced in rats receiving IL-1β together with repeated pHS, whereas no significant effects were seen in rats receiving IL-1β after a single pHS episode, or in rats receiving TNFα. Hippocampal neuronal cell loss was observed in the CA3 region of rats receiving IL-1β together with repeated pHS; however, there was no significant change in gliosis among each group. Our results are consistent with the hypothesis that excessive production of IL-1β after repeated prolonged FS can enhance adult seizure susceptibility and neuronal cell death, and might contribute to the development of TLE-HS.

Transient Inhibition of TrkB Kinase after Status Epilepticus Prevents Development of Temporal Lobe Epilepsy

Temporal lobe epilepsy is the most common and often devastating form of human epilepsy. The molecular mechanism underlying the development of temporal lobe epilepsy remains largely unknown. Emerging evidence suggests that activation of the BDNF receptor TrkB promotes epileptogenesis caused by status epilepticus. We investigated a mouse model in which a brief episode of status epilepticus results in chronic recurrent seizures, anxiety-like behavior, and destruction of hippocampal neurons. We used a chemical-genetic approach to selectively inhibit activation of TrkB. We demonstrate that inhibition of TrkB commencing after status epilepticus and continued for 2 weeks prevents recurrent seizures, ameliorates anxiety-like behavior, and limits loss of hippocampal neurons when tested weeks to months later. That transient inhibition commencing after status epilepticus can prevent these long-lasting devastating consequences establishes TrkB signaling as an attractive target for developing preventive treatments of epilepsy in humans.

Accumulation of Abnormal Adult-Generated Hippocampal Granule Cells Predicts Seizure Frequency and Severity

Accumulation of abnormally integrated, adult-born, hippocampal dentate granule cells (DGCs) is hypothesized to contribute to the development of temporal lobe epilepsy (TLE). DGCs have long been implicated in TLE, because they regulate excitatory signaling through the hippocampus and exhibit neuroplastic changes during epileptogenesis. Furthermore, DGCs are unusual in that they are continually generated throughout life, with aberrant integration of new cells underlying the majority of restructuring in the dentate during epileptogenesis. Although it is known that these abnormal networks promote abnormal neuronal firing and hyperexcitability, it has yet to be established whether they directly contribute to seizure generation. If abnormal DGCs do contribute, a reasonable prediction would be that the severity of epilepsy will be correlated with the number or load of abnormal DGCs. To test this prediction, we used a conditional, inducible transgenic mouse model to fate map adult-generated DGCs. Mossy cell loss, also implicated in epileptogenesis, was assessed as well. Transgenic mice rendered epileptic using the pilocarpine-status epilepticus model of epilepsy were monitored continuously by video/EEG for 4 weeks to determine seizure frequency and severity. Positive correlations were found between seizure frequency and (1) the percentage of hilar ectopic DGCs, (2) the amount of mossy fiber sprouting, and (3) the extent of mossy cell death. In addition, mossy fiber sprouting and mossy cell death were correlated with seizure severity. These studies provide correlative evidence in support of the hypothesis that abnormal DGCs contribute to the development of TLE and also support a role for mossy cell loss.

Expression pattern of sorting nexin 25 in temporal lobe epilepsy: A study on patients and pilocarpine-induced rats

PurposeThe transforming growth factor β (TGF-β) signaling pathway is involved in the epileptogenesis. Sorting Nexin 25 (SNX25) has been recently proposed to modulate TGF-β signaling through endosomal sorting of TGF-β receptors for lysosomal degradation. The aim of the present study was to determine the expression pattern of SNX25 in brains of epilepsy patients and in animal model of epilepsy. Methods: We investigated the expression of SNX25 in the brain tissues of patients with temporal lobe epilepsy (TLE) and in the pilocarpine-induced rat model of epilepsy using western blotting, real-time quantitative RT-PCR, and double-label immunofluorescence. Results: The expression of SNX25 was significantly increased in TLE patients in comparison to controls (0.21±0.07 vs. 0.11±0.03, P<0.05). In the lithium-pilocarpine induced epileptic rats, significant elevation of SNX25 levels was detected in the chronic phase, while no SNX25 alteration occurred in the acute and latent phases. Moreover, SNX25 localized to astrocytes and neurons, in both human samples and animal models. Conclusion: Our results indicate that upregulation of SNX25 might be involved in the development of temporal lobe epilepsy.

Long‐lasting enhancement of GABAA receptor expression in newborn dentate granule cells after early‐life febrile seizures

Febrile seizures (FS) are the most common type of seizures in childhood and are suggested to play a role in the development of temporal lobe epilepsy (TLE). Animal studies demonstrated that experimental FS induce a long-lasting change in hippocampal excitability, resulting in enhanced seizure susceptibility. Hippocampal neurogenesis and altered ion channel expression have both been proposed as mechanisms underlying this decreased seizure threshold. The present study aimed to analyze whether dentate gyrus (DG) cells that were born after FS and matured for 8 weeks display an altered repertoire of ligand-gated ion channels. To this end, we applied an established model, in which FS are elicited in 10-day-old rat pups by hyperthermia (HT). Normothermia littermates served as controls. From postnatal day 11 (P11) to P16, rats were injected with bromodeoxyuridine (BrdU) to label dividing cells immediately following FS. At P66, we evaluated BrdU-labeled DG cells for coexpression with γ-aminobutyric acid-type A receptors (GABA(A)Rs) and N-methyl-D-aspartate receptors (NMDARs). In control animals, 40% of BrdU-labeled cells coexpressed GABA(A)R β2/3, whereas in rats that had experienced FS, 60% of BrdU-labeled cells also expressed GABA(A)R β2/3. The number of BrdU-NMDAR NR2A/B coexpressing cells was in both groups about 80% of BrdU-labeled cells. The results demonstrate that developmental seizures cause a long-term increase in GABA(A)R β2/3 expression in newborn DG cells. This may affect hippocampal physiology.

Abnormalities of granule cell dendritic structure are a prominent feature of the intrahippocampal kainic acid model of epilepsy despite reduced postinjury neurogenesis

Aberrant plastic changes among adult-generated hippocampal dentate granule cells are hypothesized to contribute to the development of temporal lobe epilepsy. Changes include formation of basal dendrites projecting into the dentate hilus. Innervation of these processes by granule cell mossy fiber axons leads to the creation of recurrent excitatory circuits within the dentate. The destabilizing effect of these recurrent circuits may contribute to hyperexcitability and seizures. Although basal dendrites have been identified in status epilepticus models of epilepsy associated with increased neurogenesis, we do not know whether similar changes are present in the intrahippocampal kainic acid model of epilepsy, which is associated with reduced neurogenesis. In the present study, we used Thy1-YFP-expressing transgenic mice to determine whether hippocampal dentate granule cells develop hilar-projecting basal dendrites in the intrahippocampal kainic acid model. Brain sections were examined 2 weeks after treatment. Tissue was also examined using ZnT-3 immunostaining for granule cell mossy fiber terminals to assess recurrent connectivity. Adult neurogenesis was assessed using the proliferative marker Ki-67 and the immature granule cell marker calretinin. Significant numbers of cells with basal dendrites were found in this model, but their structure was distinct from basal dendrites seen in other epilepsy models, often ending in complex tufts of short branches and spines. Even more unusual, a subset of cells with basal dendrites had an inverted appearance; they completely lacked apical dendrites. Spines on basal dendrites were found to be apposed to ZnT-3 immunoreactive puncta, suggestive of recurrent mossy fiber input. Finally, YFP-expressing abnormal granule cells did not colocalize Ki-67 or calretinin, indicating that these cells were more than a few weeks old, but were found almost exclusively in proximity to the neurogenic subgranular zone, where the youngest granule cells are located. Recent studies have demonstrated in other models of epilepsy that dentate pathology develops following the aberrant integration of immature, adult-generated granule cells. Given these findings, one might predict that the intrahippocampal kainic acid model of epilepsy, which is associated with a dramatic reduction in adult neurogenesis, would not exhibit these changes. Herein we demonstrate that hilar basal dendrites are a common feature of this model, with the abnormal cells likely resulting from the disruption of juvenile granule cell born in the weeks before the insult. These studies demonstrate that postinjury neurogenesis is not required for the accumulation of large numbers of abnormal granule cells.

Temporal Lobe Epilepsy after Refractory Status Epilepticus: An Illustrative Case and Review of the Literature

New onset refractory status epilepticus (NORSE) is a relatively newly defined disease entity, where otherwise healthy individuals develop unrelenting seizures that do not respond to conventional anticonvulsant therapy and may require months of therapy with anesthetic drugs. We have described a case of NORSE who subsequently developed mesial temporal lobe sclerosis (MTS) and recurrent temporal lobe seizures. We discuss the possible pathophysiological mechanisms by which refractory seizures may contribute to the development of temporal lobe epilepsy (TLE).

Natural History of Temporal Lobe Epilepsy: Antecedents and Progression

Temporal lobe epilepsy represents the largest group of patients with treatment resistant/medically intractable epilepsy undergoing epilepsy surgery. The underpinnings of common forms of TLE in many instances begin in early life with the occurrence of an initial precipitating event. The first epileptic seizure often occurs after a variable latency period following this event. The precise natural history and progression following the first seizure to the development of TLE, its subsequent resolution through spontaneous remission or the development of treatment resistant epilepsy remain poorly understood. Our present understanding of the role played by these initial events, the subsequent latency to development of temporal lobe epilepsy, and the emergence of treatment resistance remains incomplete. A critical analysis of published data suggest that TLE is a heterogeneous condition, where the age of onset, presence or absence of a lesion on neuroimaging, the initial precipitating event, association with febrile seizures, febrile status epilepticus, and neurotropic viral infections influence the natural history and outcome. The pathways and processes through which these variables coalesce into a framework will provide the basis for an understanding of the natural history of TLE. The questions raised need to be addressed in future prospective and longitudinal observational studies.

GABBR1 gene polymorphism(G1465A)isassociated with temporal lobe epilepsy

γ-Aminobutyric acid B receptor 1(GABBR1) gene G1465A polymorphism has been considered as a potential risk factor for the development of temporal lobe epilepsy (TLE). However, the results were inconsistent. In this study, we performed a meta-analysis to assess the association between GABBR1 G1465A polymorphism and the risk of TLE. Biomedical literature databases including PubMed, ISI web of science and Embase were searched. The studies evaluating the association between GABBR1 G1465A polymorphism and TLE were included. Pooled odds ratio (OR) and 95%CI confidence interval (CI) were calculated using fixed- or random-effects model. Seven studies (1011 cases and 2184 controls) met the inclusion criteria and were included in the meta-analysis. The overall result showed that the association between GABBR1 G1465A polymorphism was statistically significant (OR=5.381, 95%CI: 1.726, 16.776, P=0.004). Subgroup analysis showed that the effect estimate was higher in the studies with high quality score (OR=14.220, 95%CI: 6.933, 29.169, P=0.000) than that in the studies with low quality score (OR=1.158, 95%CI: 0.325, 4.123, P=0.821). The present meta-analysis suggests that GABBR1 G1465A polymorphism is associated with the risk of TLE. The role of GABBR1 G1465A polymorphism in the development of TLE merits further investigation.

Irregular RNA splicing curtails postsynaptic gephyrin in the cornu ammonis of patients with epilepsy

Anomalous hippocampal inhibition is involved in temporal lobe epilepsy, and reduced gephyrin immunoreactivity in the temporal lobe epilepsy hippocampus has been reported recently. However, the mechanisms responsible for curtailing postsynaptic gephyrin scaffolds are poorly understood. Here, we have investigated gephyrin expression in the hippocampus of patients with intractable temporal lobe epilepsy. Immunohistochemical and western blot analyses revealed irregular gephyrin expression in the cornu ammonis of patients with temporal lobe epilepsy and four abnormally spliced gephyrins lacking several exons in their G-domains were isolated. Identified temporal lobe epilepsy gephyrins have oligomerization deficits and they curtail hippocampal postsynaptic gephyrin and GABA(A) receptor α2 while interacting with regularly spliced gephyrins. We found that cellular stress (alkalosis and hyperthermia) induces exon skipping in gephyrin messenger RNA, which is responsible for curtailed postsynaptic gephyrin and GABA(A) receptor α2 scaffolds. Accordingly, we did not obtain evidence for gephyrin gene mutations in patients with temporal lobe epilepsy. Cellular stress such as alkalosis, for example arising from seizure activity, could thus facilitate the development of temporal lobe epilepsy by reducing GABA(A) receptor α2-mediated hippocampal synaptic transmission selectively in the cornu ammonis.

N‐methyl‐d‐aspartate, hyperpolarization‐activated cation current (Ih) and γ‐aminobutyric acid conductances govern the risk of epileptogenesis following febrile seizures in rat hippocampus

Febrile seizures are the most common types of seizure in children, and are generally considered to be benign. However, febrile seizures in children with dysgenesis have been associated with the development of temporal lobe epilepsy. We have previously shown in a rat model of dysgenesis (cortical freeze lesion) and hyperthermia-induced seizures that 86% of these animals developed recurrent seizures in adulthood. The cellular changes underlying the increased risk of epileptogenesis in this model are not known. Using whole cell patch-clamp recordings from CA1 hippocampal pyramidal cells, we found a more pronounced increase in excitability in rats with both hyperthermic seizures and dysgenesis than in rats with hyperthermic seizures alone or dysgenesis alone. The change was found to be secondary to an increase in N-methyl-D-aspartate (NMDA) receptor-mediated excitatory postsynaptic currents (EPSCs). Inversely, hyperpolarization-activated cation current was more pronounced in naïve rats with hyperthermic seizures than in rats with dysgenesis and hyperthermic seizures or with dysgenesis alone. The increase in GABAA-mediated inhibition observed was comparable in rats with or without dysgenesis after hyperthermic seizures, whereas no changes were observed in rats with dysgenesis alone. Our work indicates that in this two-hit model, changes in NMDA receptor-mediated EPSCs may facilitate epileptogenesis following febrile seizures. Changes in the hyperpolarization-activated cation currents may represent a protective reaction and act by damping the NMDA receptor-mediated hyperexcitability, rather than converting inhibition into excitation. These findings provide a new hypothesis of cellular changes following hyperthermic seizures in predisposed individuals, and may help in the design of therapeutic strategies to prevent epileptogenesis following prolonged febrile seizures.

Novel Etiological and Therapeutic Strategies for Neurodiseases: Mechanisms and Consequences of Febrile Seizures: Lessons From Animal Models

Febrile seizures (FS) are the most common type of convulsive events in infancy and childhood. Genetic and environmental elements have been suggested to contribute to FS. FS can be divided into simple and complex types, the former being benign, whereas it is controversial whether complex FS have an association with the development of temporal lobe epilepsy (TLE) in later life. In the hippocampus of TLE patients, several structural and functional alterations take place that render the region an epileptic foci. Thus, it is important to clarify the cellular and molecular changes in the hippocampus after FS and to determine whether they are epileptogenic. To achieve this goal, human studies are too limited because the sample tissues are only available from adult patients in the advanced and drug-resistant stages of the disease, masking the underlying etiology. These facts have inspired researchers to take advantage of well-established animal models of FS to answer the following questions: 1) How does hyperthermia induce FS? 2) Do FS induce neuroanatomical changes? 3) Do FS induce neurophysiological changes? 4) Do FS affect the behavior in later life? Here we introduce and discuss accumulating reports to answer these questions.

Neuronal plasticity in animal models and the epileptic human hippocampus

Prolonged status epilepticus in humans as in experimental animals can initiate the development of temporal lobe epilepsy (TLE) (Kapur, 1999). Therefore, application of potent convulsant substances such as kainic acid or pilocarpine in rats induces acute status epilepticus that, after a silent period of 1–2 weeks, is followed by spontaneous convulsions. The status epilepticus is characterized by severe limbic seizures and sequelae of neuropathologic signs including opening of the blood–brain barrier, local brain edema, bleeding into the brain, and activation of microglia and astrocytes followed by neurodegeneration in the hippocampus, amygdala, entorhinal cortex, and other brain areas (Sperk et al., 1983; Du et al., 1993; Rizzi et al., 2003). Induced by the seizure activity, neurotransmitters such as γ-aminobutyric acid (GABA), glutamate, or amine transmitters are released from their stores and mechanisms of their resynthesis are strongly activated (Sperk et al., 1983). In addition, pronounced changes in the expression of multiple functionally important proteins have been found in brains of experimental animals and humans (Herdegen et al., 1993; Sperk, 1994; McNamara, 1999; Morimoto et al., 2004).

Inflammation in rat pups subjected to short hyperthermic seizures enhances brain long-term excitability

Inflammatory processes in response to infection are involved in the pathophysiological mechanisms of febrile seizures (FS). Prolonged FS may promote the development of temporal lobe epilepsy. It has been shown in rats that prolonged hyperthermic seizures (HS) are followed by long-term modification of brain excitability. To examine whether short FS results in modification of brain excitability, we induced an inflammatory response in combination with short HS. HS were induced in rat pups at either P11 or P16 using a heating lamp with a continuous monitoring of the core temperature. Rat pups were maintained at the temperature seizure threshold during 5 min. In order to induce an inflammatory response, lipopolysaccharide (LPS, Eschericha coli 055:B5) was injected i.p. at 5 microg/kg or 50 microg/kg, 2h prior seizure induction. After 1 month, pentylenetetrazol threshold (PTZth) was used to assess the change of brain excitability. Histological studies were performed 24h after the FS (Fluorojade-B) and after the PTZth (cresyl violet). The temperature thresholds to induce the seizures were not different among the groups. The PTZth was not significantly different between sham and FS only groups, and decreased dose-dependently when LPS was combined to FS. Histological studies suggested the absence of cell injury. Lower PTZth obtained by using LPS in combination with HS in rat pups suggests a change in brain excitability. Our model with only 5 min of HS in combination with LPS suggests that an inflammatory response could, in part, explain long-term change in brain excitability following short FS.

TRIP8b Splice Variants Form a Family of Auxiliary Subunits that Regulate Gating and Trafficking of HCN Channels in the Brain

Hyperpolarization-activated cyclic nucleotide-regulated (HCN) channels, which generate the I(h) current, mediate a number of important brain functions. The HCN1 isoform regulates dendritic integration in cortical pyramidal neurons and provides an inhibitory constraint on both working memory in prefrontal cortex and spatial learning and memory in the hippocampus. Altered expression of HCN1 following seizures may contribute to the development of temporal lobe epilepsy. Yet the regulatory networks and pathways governing HCN channel expression and function in the brain are largely unknown. Here, we report the presence of nine alternative N-terminal splice forms of the brain-specific cytoplasmic protein TRIP8b and demonstrate the differential effects of six isoforms to downregulate or upregulate HCN1 surface expression. Furthermore, we find that all TRIP8b isoforms inhibit channel opening by shifting activation to more negative potentials. TRIP8b thus functions as an auxiliary subunit that provides a mechanism for the dynamic regulation of HCN1 channel expression and function.