Journal List > J Korean Acad Conserv Dent > v.27(3) > 1055995

Park, Choi, and Choi: Immunohistochemical study on the distribution of ion channels in rat trigeminal sensory nucleus

I. INTRODUCTION

Trigeminal nerves transmit various sensory information such as touch, pressure, pain and temperature sensations from the oromaxillofacial region to the central nervous system. The central ends terminate in the trigeminal principal nucleus and the trigeminal spinal nucleus and transmit the information through the trigeminal mesencephalic nucleus. Therefore three nucleus i.e. the trigeminal principal nucleus, the trigeminal spinal nucleus and the trigeminal mesencephalic nucleus serve as the first gates to enter sensory information, so the nucleus have fundamental roles in sensory reception in the oromaxillofacial region.
Ion channels of the sensory neurons play a pivotal role in the generation and transmission of sensory information. Generally the Na+ channels are involved in the rapid generation of action potentials. Intracellular recordings in the mammalian TRG neurons have shown that three types of action potentials exist; (1) fast spikes, most of which were completely blocked by the external application of tetrodotoxin (TTX), (2) Co2+-and TTX-resistant humped spikes (Puil et al. 1986) and (3) slowly decaying TTX-resistant and Cd2+-sensitive action potentials (Galdzicki et al. 1990). These reports suggest that voltagedependent sodium current (INa) may be involved in the electrical activities of the TRG neurons.
A proper understanding of many CNS diseases and disorders requires identification of the ion channels underlying them and knowledge of their normal physiological roles. Recently, considerable progress has been made in understanding certain forms of epilepsy (Biervert et al. 1998; De Zeeuw et al. 1995; Smart et al. 1998), and episodic ataxia (Browne et al. 1994), both of which may be caused by mutations in genes encoding voltage-gated K+ (Kv) channels. Kv channels are crucial in a wide variety of processes, including action potential preparation and lymphocyte activation, and play as essential role in neurons where they regulate the resting membrane potential, impact dendritic excitability, control the frequency and duration of action potentials, and modulate neurotransmitter release (Hille 1992).
Calcium flux through voltage-gated calcium channels has been shown to be essential for a host of important cellular functions in the nervous system. These include neurite outgrowth, gene regulation, neuronal excitability, plasticity, and neurotransmitter release. A variety of calcium channels has been described in different types of tissue (Bean 1989; Scott et al. 1991). Commonly, calcium currents (ICa) are grouped into three types on the basis of the finding of Nowycky et al. (1985) in chick dorsal root ganglia (DRG) (Fox et al. 1987a, b). They described a transient (T-type), or low voltage-activated (LVA), current; a sustained (L-type), or high voltage-activated (HVA), current; and a third current, termed an N-type, that exhibited kinetic properties between the T-type and the L-type current. Recently a fourth type, known as the P-type, has been described in Purkinje cells (Llinas et al. 1989; Regan et al. 1991).
There are several techniques to study the ion channels. The first is to isolate the ion channels from the cell membrane, and to characterize the nature and biochemical properties of them. The second is to measure the ionic currents through the ion channels by the patch clamp technique. And the third is to observe the localization of the ion channels by staining them with antibodies.
The main goals of this study were (1) to observe some kinds of ion channels (2) to observe the localized distribution of the ion channels in the trigeminal principal nucleus, the trigeminal spinal nucleus by immunohistochemical method.

II. MATERIALS & METHODS

1. Peroxidase Immunostaining

Sprague-Dawley rats of Postnatal 0 day were sacrificed by decapitation and fixed without perfusion in 4% paraformaldehyde in 0.1M Na-phosphate buffer, pH 7.4. Brains were removed and stored at 4℃ overnight in the same fixative, followed by storage for 2-3 days at 4℃ in 30% sucrose, 0.13M phosphate buffer. Frozen serial sections were cut at 30µm in the coronal orientation on a Minotome Plus microtome. Sections were pretreated with 0.3% hydrogen peroxide in 0.1M PBS containing 0.3% Triton X-100 (PBST) for 30min at room temperature to inhibit endogenous peroxidase activity. After washing with 0.1M PBST, the sections incubated in PBS containing 3% normal goat serum, 2g/l BSA and 0.1% Triton X-100 for 30-45min. Polyclonal anti-Kv 4.2, BKCa, Kir 2.1 1A, 1B, E (product no. APC-023, 021, 026 and ACC-001, 002, 006 ; Alomone Labs, Jerusalem, Israel) and anti-Na+ channel, type I (product no. #06-649 ; Argonex company) were used as primary antibodies at a dilution of 1:100 for 24 h at 4℃. After primary incubation, sections were given three 10-min washes in 0.3% Triton X-100 in PBS and then processed for peroxidase immunostaining using Vector Laboratories. Sections were incubated for 1h at room temperature in Vector goat anti-rabbit Ig-G. Three 10-min washes in 0.3% Triton X-100 in PBS. Sections were incubated under the same conditions in ABC solution. After three 10-min washes in 0.3% Triton X-100 in PBS, sections were transferred into Tris-buffered saline. Sections were incubated in 3'3-diaminobenzidine substrate for 5-10 min before the reaction was stopped in distilled water.

2. Fluorescent Immunostaining

Sprague-Dawley rats of Postnatal 0 day were sacrificed by decapitation and fixed without perfusion in 4% paraformaldehyde in 0.1M Na-phosphate buffer, pH 7.4. Brains were removed and stored at 4℃ overnight in the same fixative, followed by storage for 2-3 days at 4℃ in 30% sucrose, 0.13 M phosphate buffer. Frozen serial sections were cut at 30µm in the coronal orientation on a Minotome Plus microtome. Sections were washed in PBS containing 0.3% Triton X-100 for 30min. After washing with 0.1M PBST, the sections incubated in PBS containing 3% normal goat serum, 2g/l BSA and 0.1% Triton X-100 for 30-45 min. Polyclonal anti-Kv 4.2, BKCa, Kir 2.1 1A, 1B, E (product no. APC-023, 021, 026 and ACC-001, 002, 006 ; Alomone Labs, Jerusalem, Israel) and anti-Na+ channel, type I (product no. #06-649 ; Argonex company) were used as primary antibodies at a dilution of 1:100 for 24h at 4℃. Sections were incubated for 1h at room temperature in PBS containing anti-rabbit fluorescein-labeled IgG (dilution of 1: 100, KPL Europe). After three 10-min washes in PBS and examined with epifluorescence microscopy.

III. RESULTS

1. Sodium Channels

Sodium channels exist on the membrane of the soma in the trigeminal spinal nucleus, trigeminal principal nucleus, trigeminal mesencephalic nucleus, and trigeminal motor nucleus were immunohistochemically stained with the antibody for the sodium channels. Because the primary antibodies used in this experiment are not specific for the TTX-s or TTX-r sodium channels - those are able to bind to TTX-s and TTX-r sodium channels, TTX-s and TTXr sodium channels on the membrane of TRG neurons could not be separated from each other. Sodium channels of the membrane in the nucleus were strongly stained with the antibodies. There is no difference in the degree of the staining among the tested nucleus.

2. Calcium Channels

1) N-type calcium channels

In this experiment, it is noted that there was a moderate staining of the N-type calcium channels of the boutons and somata of trigeminal spinal nucleus compared to other part of the brain. N-type calcium channels were moderately stained in the trigeminal principal nucleus.

2) P/Q-type calcium channels

In this immunohistochemical study, P/Q-type calcium channels were weakly stained in trigeminal spinal nucleus. And a very weak staining for the P/Q-type calcium channels was observed in the trigeminal principal nucleus.

3) R-type calcium channels

R-type calcium channel was not observed in trigeminal spinal nucleus and trigeminal principal nucleus.

3. Potassium Channels

1) Inwardly rectifying potassium channels

In this experiment, moderate staining of the Kir 2.1 in the trigeminal principal nucleus were observed. On the other hand, staining of the Kir 2.1 was weak trigeminal spinal nucleus.

2) Voltage-gated potassium channels

In this study, there was weak staining for the Kv 4.2 in trigeminal spinal nucleus. The Kv 4.2 channels were very weakly stained in the trigeminal principal nucleus.

3) Ca2+-activated K+ channels

In this experiment, the intensity of staining for the BKCa channels was weak in the trigeminal principal nucleus and the trigeminal spinal nucleus.

IV. DISCUSSION

Trigeminal sensory nerves relay mechanical, thermal, chemical and proprioceptive information from craniofacial regions. Trigeminal sensory nerves act at the focal level to maintain the integrity of craniofacial tissues through trophic influences and serve the whole animal through reflexes that protect other external sensory and internal homeostatic systems from damaging environmental changes.
There are some nuclei concerning with the trigeminal nerves: trigeminal spinal nucleus, trigeminal principal nucleus, trigeminal mesencephalic nucleus and trigeminal motor nucleus. It has been 50 years since Olszewski (1950) divided the trigeminal spinal nucleus into oralis, interpolaris and caudalis subdivisions and nearly 25 years since Gobel et al. (1977) detailed the structural homologies between caudalis and the spinal dorsal horn. While these landmark studis have shaped discussions of the special relationship between caudalis and craniofacial pain (Renehan and Jaquin 1993; Sessle 2000), this useful homology may need revision since recent evidence suggests that select portions of caudalis are organized differently form spinal systems. The subnucleus caudalis, the largest subdivision of the trigeminal spinal nucleus, consists of an elongated laminated portion that merges without clear boundaries with the cervical dorsal horn, while the rostral caudalis is displaced medially by the caudal tip of interpolaris to form a distinctive transition region. The ventral interpolaris/caudalis transition region, which is well described in rodents, consists of rostral caudalis with its fragmented laminar appearance, intersttial islands of neurons embedded in the trigeminal spinal tract, and a ventral crescent-shaped region of caudal interpolaris (Phelan and Falls 1989).
Spinal motor neurons are the final integration point for electrical signals that initiate and control skeletal muscle contraction. And trigeminal motor nucleus controls masticatory muscles. Several neuromuscular diseases result from dysfunction of the motor neurons. In at least two cases, Ca2+ channels are implicated in the disease process. Lambert-Eaton myasthenic syndrome is caused by circulating antibodies against presynaptic Ca2+ channels (Engel 1991; Sher et al. 1993). These antibodies reduce the level of presynaptic Ca2+ current and the efficiency of neurotransmitter release (Lang et al. 1983; Kim 1985). Amyotrophic lateral sclerosis (ALS) is caused by progressive death of motor neurons (Appel and Stefani 1991). One current hypothesis for the etiology of ALS implicates Ca2+ channels in motor neurons (Appel et al. 1991, 1993; Delbono 1991, 1993; Smith et al. 1992; Uchitel et al. 1992; Morton et al. 1994; Rowland 1994). These results mean that ion channels play a critical role to function in normal and disease states.
The mesencephalic trigeminal nucleus contains somata of first order neurons associate with the proprioceptors of the head regions. Proprioceptive afferents of law closing masticatory muscles, tooth mechanoceptors and extraocular muscles have been identified in the trigeminal mesencephalic nucleus (Alvarado-Mallart et al. 1975; Buisseret-Delmas et al. 1990; Cody et al. 1972; Jerge 1963; Matesz 1981). Masticatory neurons are distributed throughout the whole rostralcaudal extent of the trigeminal mesencephalic nucleus, while the periodontal and extraocular muscle sensory neurons are situated mainly in the caudal part of the nucleus (Dessem and Taylor 1989). The trigeminal mesencephalic nucleus neurons resemble dorsal root ganglion cells, but a noticeable different is the presence of axosomatic synaptic boutons (Copray et al. 1990; Liem et al. 1992)
Sodium currents through the sodium channels were first recorded by Hodgkin and Huxley, who use voltage clamp techniques to demonstrate the three key features that have come to characterize the sodium channel: (1) voltage-dependent activation, (2) rapid inactivation, and (3) selective ion conductance (Hodgkin and Huxley 1952). Detailed analysis of sodium channel function during the 1960s and 1970s using the voltage clamp method applied to invertebrate giant axons and vertebrate myelinated nerve fibers yielded mechanistic models for sodium channel function (Armstrong 1981; Hille 1984). These functional studies predicted that sodium channels would be rare membrane proteins, difficult to identify and to isolate from the many other proteins of excitable membranes. During the 1970s, a new line of research emerged, focusing on development of methods for molecular analysis of sodium channels. Biochemical methods for measurement of ion flux through sodium channels, high affinity binding of neurotoxins to sodium channels, and detergent solubilization and purification of sodium channel proteins labeled by neurotoxins were progressively developed (Ritchie and Rogart 1977; Catterall 1980). These biochemical approaches led to discovery of the sodium channel protein in 1980. Photoaffinity labeling with a photoreactive derivative of an -scorpion toxin identified the principal -subunit and the auxiliary 1 subunit of brain sodium channels (Beneski and Catterall 1980). Immediately thereafter, partial purification of tetrodotoxin binding proteins from electric eel electroplax revealed a correlation between tetrodotoxin binding activity and a protein of ~270 kDa (Agnew et al. 1980). Subsquent purification studies showed that the sodium channel from mammalian brain is a complex of (260kDa), 1 (36kDa), and 2 (33kDa) subunits (Hartshorne and Catterall 1981; Hartshorne et al. 1982), the tetrodotoxin binding component of the sodium channel purified from eel electroplax is a single protein of 270 kDa (Miller et al. 1983), and the sodium channel from skeletal muscle is a complex of and 1 subunits (Barchi 1983).
The calcium channels are known to be involved in the pacemaker depolarization, Ca2+-dependent secretion and the activation of Ca2+-dependent ionic conductance in neuronal preparations, although linking a specific type of ICa to a particular cellular process may be difficult (Tsien et al. 1988). It may be certain, however, that the subtypes of ICa contribute in different ways to the transmission of sensory signals of different modalities in primary afferent neurons. The subtypes of ICa are known to show diameter-dependent variation in acutely isolated rat DRG neurons, i. e., medium diameter neurons had a large amount of T-type ICa, whereas significantly large proportion of the whole-cell ICa was L-type in the small cells (Scroggs and Fox 1991, 1992), which shaped action potentials differently and may affect, therefore, significantly to the sensory transmission. In most cases, multiple calcium channel types coexist in the same neuron and it has been speculated that they may contribute toward different neuronal functions, possibly depending on their localization (Doughty et al. 1998; Wu et al. 1999).
Previous work on L-type and N-type calcium channels has demonstrated different localizations in specific areas of the neuronal surface. L-type channels have been found in the neuronal somata and proximal dendrites of rat brain, spinal cord, and retina (Westenbroek et al. 1990; Ahlijanian et al. 1990), which is consistent with a role in mediating the increases in intracellular calcium occurring in the cell body in response to excitatory inputs to the dendrites. N-type channels have been detected at the release face of presynaptic nerve terminals at the frog neuromuscular junction (Robitaille et al. 1990; Cohen et al. 1991; Torri Tarelli et al. 1991), in the presynaptic calyx-type nerve terminals of chick ciliary ganglia, and in the medial nucleus of the trapezoid body in rat (Haydon et al. 1994; Wu et al. 1999). These data, together with a number of functional findings (Miller 1990; Dunlap et al. 1995; Tsien et al. 1995; Stanley 1997), clearly indicate that N-type channels play a role in supporting neurotransmitter release. In this experiment, it is noted that there was a moderate staining of the N-type calcium channels of the boutons and somata of upper and lower trigeminal spinal nucleus compared to other part of the brain.
After the initial description of P-type calcium channels in Purkinje cells (Llinas et al. 1989; Mintz et al. 1992) and the subsequent designation of the Q-type calcium channels as a separate channel category (Randall and Tsien 1995), concern has arisen regarding the distinction between them. P/Q-type calcium channels are widely expressed in the mammalian brain in a predominantly presynaptic distribution (Westenbroek et al. 1995), and have a important role in modulating neurotransmitter release (Regehr and Mintz 1994). In this immunohistochemical study, P/Q-type calcium channels were weakly stained in the upper trigeminal spinal nucleus. And a very weak staining for the P/Q-type calcium channels was observed in the trigeminal principal nucleus. R-type calcium channel was not observed in the upper trigeminal spinal nucleus.
Inwardly rectifying potassium channels (Kir) control the resting K+ conductance in many excitable and non-excitable cells (Hille 1992). They mediate a high K+ conductance at the K+ reversal potential and at voltages slightly positive to the K+ reversal potential. Thus Kir have a stabilizing effect on the resting potential. Whenever Kir are regulated by second messengers, this represents a highly effect mechanism of controlling excitability (Nilius et al. 1993). Various subtypes of the Kir family have been cloned and heterologously expressed in order to study their functional properties. One of the Kir, Kir 2.1 is widely distributed in various tissues. Kir 2.1 is found in mammalian lens epithelial cells (Cooper et al. 1991), ventricular myocytes (Josephson 1998), endothelial cells (Silver and DeCoursey 1990), and oocytes (Ruppersberg and Fakler 1996). In this experiment, moderate staining of the Kir 2.1 in the trigeminal principal nucleus were observed. On the other hand, staining of the Kir 2.1 was weak in the trigeminal spinal nucleus.
Much interest has been generated recently in the potassium channel subunit Kv 4.2 for its critical role in regulating membrane excitability. Several lines of evidence suggest that Kv 4.2 underlies the A-type current (IA) in neurons (Serodio et al. 1996; Song et al. 1998) as well as the transient outward current (Ito) in cardiac ventricular myocytes (Barry et al. 1998). In particular, Tkatch, et al. (2000) have found that Kv 4.2 mRNA abundance and A-type current amplitude are linearly related in the striatum. In the hippocampus, such A-type current has been found to dampen back-propagation of action potentials in the distal dendrites of CA1 pyramidal neurons, thereby setting up the capacity to modify the excitatory postsynaptic potential or back-propagating action potentials following synaptic activity (Hoffman et al. 1997). Immunohistochemical studies show that Kv 4.2 protein localizes abundantly to the hippocampus (Sheng et al. 1992) with particular localization to the neuronal soma and dendrites (Maletic-Savatic et al. 1995), structures in which modulating membrane excitability could have important implications for long-term potentiation and memory. Furthermore, ultrastructural studies in supraoptic neurons have shown that Kv 4.2 is localized to the postsynaptic membrane directly across from the presynaptic terminal (Alonso and Widmer 1997). In this study, there was weak staining for the Kv 4.2 in trigeminal spinal nucleus.
Ca2+-activated K+ channels (Kca) are present in a remarkable variety of animal cells, where they integrate changes in intracellular Ca2+ concentration with changes in membrane potential. They are involved in many physiologic processes including regulation of secretion, smooth muscle tone, and control of action potential shape and firing pattern in excitable cells (Marty 1989; Vergara et al. 1998). BKCa can be divided into three main subfamilies based on their electrophysiological, pharmacological, and molecular profiles: large-conductance Ca2+-activated K+ channels, intermediate-conductance Ca2+-activated K+ channels, and small-conductance Ca2+-activated K+ channels (Vergara et al. 1998). Large-conductance Ca2+-activated K+ channels (BKCa) are found in a variety of both electrically excitable and non-excitable cells (Jan and Jan 1997). Their activities are triggered by membrane depolarization and enhanced by cytosolic Ca2+, providing a link between the metabolic and electrical state of cells. The physiological roles of BKCa channels have been examined in several tissues. In smooth muscle in which they are particularly abundant, BKCa channels play a key role in setting the pace of contractile activity (Nelson et al. 1995), and in neurons, they are involved in regulation of transmitter release and repolarization of action potentials (Kaczorowski et al. 1996). In this experiment, the intensity of staining for the BKCa channels was weak in the trigeminal principal nucleus and the trigeminal spinal nucleus.
In this immunohistochemical study, thus, we could observe (1) the sodium channels in the trigeminal spinal nucleus and the trigeminal principal nucleus (2) calcium channels: N-type in the trigeminal spinal nucleus, and P/Q-type in the trigeminal spinal nucleus and the trigeminal principal nucleus (2) the potassium channels: inwardly rectifying potassium (Kir 2.1) channels in the trigeminal principal nucleus and the trigeminal spinal nucleus, voltage-gated potassium (Kv 4.2) channels in the trigeminal spinal nucleus, and very weak staining for the large-conductance calcium-activated potassium (BKCa) channels in the trigeminal spinal nucleus and the trigeminal principal nucleus.
These results means as follow.
  1. Sodium channels play important role as nociceptor in trigeminal spinal nucleus, inducer of action potential in trigeminal principal nucleus.

  2. N-type channels were stained stronger than other calcium channels. N-type channels play a role in supporting neurotransmitter release. We can estimate fibers or bodies of neuron synapse in both nucleus therefore neurotransmitter release is important in these nucleus.

  3. There were weak staining for potassium channels. In the case of BKCa, it was repored that the channel was mainly distributed in smooth muscle. Previous work it was estimated that role of Kv 4.2 channel was related to long-term memory. So Kv 4.2 protein localized abundantly to the hippocampus (Sheng et al.1992) but in our study, the protein rare localized to sensory nucleus.

V. CONCLUSION

Trigeminal sensory nerves relay mechanical, thermal, chemical and proprioceptive information from craniofacial regions. Trigeminal sensory nerves act at the focal level to maintain the integrity of craniofacial tissues through trophic influences and serve the whole animal through reflexes that protect other external sensory and internal homeostatic systems from damaging environmental changes.
Ion channels of the sensory neurons play a pivotal role in the generation and transmission of sensory information. Generally the Na+ channels are involved in the rapid generation of action potentials. Calcium influx triggers a number of cellular processes, including muscle contraction, second-messenger activation cascade, regulation of axonal guidance, control of neurotransmitter release. Generally potassium channels are involved in repolarization after depolarization, setting the pace of contractile activity, regulation of neurotransmitter release, and modulating membrane excitability.
The main goals of this study were (1) to observe some kinds of ion channels (2) to observe the localized distribution of the ion channels in the trigeminal principal nucleus, the trigeminal spinal nucleus and the trigeminal mesencephalic nucleus by immunohistochemical method.
In this immunohistochemical study we observed as follow.
  1. We observed the sodium channels in the trigeminal spinal nucleus and the trigeminal principal nucleus. We suggested that sodium channels played the role as nociceptor in trigeminal spinal nucleus, inducer of action potential in trigeminal principal nucleus.

  2. We observed the calcium channels: N-type in the trigeminal spinal nucleus, and P/Q-type in the trigeminal spinal nucleus and trigeminal principal nucleus. N-type channels were stained stronger than other calcium channels. N-type channels play a role in supporting neurotransmitter release. We can estimate fibers or bodies of neuron synapse in both nucleus therefore neurotransmitter release is important in these nucleus.

  3. We observed the potassium channels: inwardly rectifying potassium (Kir 2.1) channels in the trigeminal principal nucleus and the trigeminal spinal nucleus, voltage-gated potassium (Kv 4.2) channels in the trigeminal spinal nucleus, and very weak staining for the large-conductance calcium-activated potassium (BKCa) channels in the trigeminal spinal nucleus and trigeminal principal nucleus.

At present, it is obviously difficult to recognize the distinct ion channel types and their function in trigeminal sensory nucleus. Based on our results we can propose possible model systems in which a detailed electrophysiological and pharmacological analysis may help clarify the functional role of ion channels with a defined subunit composition in trigeminal sensory nucleus. Additional studies will be required to explore the roles and distributions of ion channels in trigeminal sensory nucleus in relation to their subunit composition and their positions.

Figures and Tables

Fig. 1(a)
A cell stained with sodium channel antibodies in the trigeminal spinal nucleus (indicated with an arrow)
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Fig. 1(b)
A fluorescence micrograph showing stained sodium channels with sodium channel antibodies in the trigeminal spinal nucleus (indicated with an arrow)
jkacd-27-215-g002
Fig. 2(a)
A cell stained with sodium channel antibodies in the trigeminal principal nucleus (indicated with an arrow)
jkacd-27-215-g003
Fig. 2(b)
A fluorescence micrograph showing stained sodium channels with sodium channel antibodies in the trigeminal principal nucleus (indicated with an arrow)
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Fig. 3(a)
A cell stained with N-type calcium channel antibodies in the trigeminal spinal nucleus (indicated with an arrow)
jkacd-27-215-g005
Fig. 3(b)
A fluorescence micrograph showing stained N-type calcium channels with calcium channel antibodies in the trigeminal spinal nucleus (indicated with an arrow)
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Fig. 4(a)
A cell stained with N-type calcium channel antibodies in the trigeminal principal nucleus (indicated with an arrow)
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Fig. 4(b)
A fluorescence micrograph showing stained N-type calcium channels with calcium channel antibodies in the trigeminal principal nucleus (indicated with an arrow)
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Fig. 5(a)
A cell stained with P/Q-type calcium channel antibodies in the trigeminal spinal nucleus (indicated with an arrow)
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Fig. 5(b)
A fluorescence micrograph showing stained P/Q-type calcium channels with calcium channel antibodies in the trigeminal spinal nucleus (indicated with an arrow)
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Fig. 6(a)
A cell stained with P/Q-type calcium channel antibodies in the trigeminal principal nucleus (indicated with an arrow)
jkacd-27-215-g011
Fig. 6(b)
A fluorescence micrograph showing stained P/Q-type calcium channels with calcium channel antibodies in the trigeminal principal nucleus (indicated with an arrow)
jkacd-27-215-g012
Fig. 7(a)
A cell stained with R-type calcium channel antibodies in the trigeminal spinal nucleus (indicated with an arrow)
jkacd-27-215-g013
Fig. 7(b)
A fluorescence micrograph showing stained R-type calcium channels with calcium channel antibodies in the trigeminal spinal nucleus (indicated with an arrow)
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Fig. 8(a)
A cell stained with R-type calcium channel antibodies in the trigeminal principal nucleus (indicated with an arrow)
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Fig. 8(b)
A fluorescence micrograph showing stained R-type calcium channels with calcium channel antibodies in the trigeminal principal nucleus (indicated with an arrow)
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Fig. 9(a)
A cell stained with Kir 2.1 potassium channel antibodies in the trigeminal spinal nucleus (indicated with an arrow)
jkacd-27-215-g017
Fig. 9(b)
A fluorescence micrograph showing stained potassium channels with Kir 2.1 potassium channel antibodies in the trigeminal spinal nucleus (indicated with an arrow)
jkacd-27-215-g018
Fig. 10(a)
A cell stained with Kir 2.1 potassium channel antibodies in the trigeminal principal nucleus (indicated with an arrow)
jkacd-27-215-g019
Fig. 10(b)
A fluorescence micrograph showing stained potassium channels with Kir 2.1 potassium channelantibodies in the trigeminal principal nucleus (indicated with an arrow)
jkacd-27-215-g020
Fig. 11(a)
A cell stained with Kv 4.2 potassium channel antibodies in the trigeminal spinal nucleus (indicated with an arrow)
jkacd-27-215-g021
Fig. 11(b)
A fluorescence micrograph showing stained potassium channels with Kv 4.2 potassium channel antibodies in the trigeminal spinal nucleus (indicated with an arrow)
jkacd-27-215-g022
Fig. 12(a)
A cell stained with Kv 4.2 potassium channel antibodies in the trigeminal principal nucleus (indicated with an arrow)
jkacd-27-215-g023
Fig. 12(b)
A fluorescence micrograph showing stained potassium channels with Kv 4.2 potassium channel antibodies in the trigeminal principal nucleus (indicated with an arrow)
jkacd-27-215-g024
Fig. 13(a)
A cell stained with BKCa potassium channel antibodies in the trigeminal spinal nucleus (indicatedwith an arrow)
jkacd-27-215-g025
Fig. 13(b)
A fluorescence micrograph showing stained potassium channels with BKCa potassium channel antibodies in the trigeminal spinal nucleus (indicated with an arrow)
jkacd-27-215-g026
Fig. 14(a)
A cell stained with BKCa potassium channel antibodies in the trigeminal principal nucleus (indicated with an arrow)
jkacd-27-215-g027
Fig. 14(b)
A fluorescence micrograph showing stained potassium channels with BKCa potassium channel antibodies in the trigeminal principal nucleus (indicated with an arrow)
jkacd-27-215-g028
Table 1
Immunofluorescence for sodium channels and calcium channels
jkacd-27-215-i001

high +++, moderate ++, weak +, very weak±, negative -

Table 2
Immunofluorescence for potassium channels
jkacd-27-215-i002

high +++, moderate ++, weak +, very weak±, negative -

References

1. AGNEW WS, MOORE AC, LEVINSON SR, RAFTERY MA. Identification of a large molecular weight peptide associated with a tetrodotoxin binding proteins from the electroplax of Electrophorus electricus. Biochem Biophys Res Commun. 1980. 92:860–866.
crossref
2. Ahlijanian MK, Westenbroek RE, Catterall WA. Subunit structure and localization of dihydropyridine-sensitive calcium channels in mammalian btain, spinal cord, retina. Neuron. 1990. 4:819–832.
crossref
3. Alonso G, Widmer H. Clustering of Kv4.2 potassium channels in Phostsy. 1997.
crossref
4. ALVARADO-MALLART RM, BATINI C, BUISSERET-DELMAS C, CORVISER J. Trigeminal representations of the masticatory and extraocular proprioceptors as revealed by horseradish perioxidase retrograde transport. Exp Brain Res. 1975. 23:167–179.
crossref
5. APPEL SH, ENGELHARDT JI, BARCIA J, STEFANI E. Immunoglobulins from animal models of motor neuron disease and from human amyotrophic lateral sclerosis patients passively transfer physiological abnormalities to the neuromuscular junction. Proc Natl Acad Sci USA. 1991. 88:647–651.
crossref
6. APPEL SH, STEFANI E. Amyotrophic lateral sclerosis : etiology and pathogenesis. Curr Neurol. 1991. 11:287–310.
7. ARMSTRONG CM. Sodium channels and gating currents. Physiol Rev. 1981. 61:644–682.
crossref
8. BARCHI RL. Protein components of the purified sodium channel from rat skeletal sarcolemma. J Neurochem. 1983. 36:1377–1385.
9. Barry DM, Xu H, Schuessler RB, Nerbonne JM. Functional knockout of the transient outward current, long-QT syndrome, cardiac remodeling in mice expressing a dominant-negative Kv4 alpha subunit. Circ Res. 1998. 83:560–567.
crossref
10. Bean BP. Classes of calcium channels in vertebrate cells. Annu Rev Physiol. 1989. 51:367–384.
crossref
11. BENESKI DA, CATTERALL WA. Covalent labeling of protein components of the sodium channel with a photoactivable derivative of scorpion toxin. Proc Natl Acad Sci U S A. 1980. 77:639–643.
crossref
12. BIERVERT C, SCHROEDER BC, KUBISCH C, BERKOVIC SF, PROPPING P, JENTSCH TJ, STEINLEIN OK. A potassium channel mutation in neonatal human epilepsy. Science. 1998. 279:403–406.
crossref
13. BROWNE KL, GANCHER ST, NUTT JG, BRUNT ER, SMITH EA, KRAMER P, LITT M. Episodic ataxia/myokymia syndrome is associated with point mutations in the human potassium channel gene KCNA1. Nat Genet. 1994. 8:136–140.
crossref
14. BUISSERET-DELMAS C, BUISSERET P. Central projections of extraocular muscle afferents in cat. Neurosci Lett. 1990. 109:48–53.
crossref
15. CATTERALL WA. Neurotoxins that act on voltagesensitive sodium channels in excitable membranes. Annu Rev Pharmacol Toxicol. 1980. 20:15–43.
crossref
16. CODY FWJ, LEE RWH, TAYLOR AA. functional analysis of the components of the mesencephalic nucleus of the fifth nerve in the cat. J Physiol (London). 1972. 226:249–261.
crossref
17. Cooper K, Rae JL, Deway J. Inwardly rectifying potassium current in mammalian lens epithelial cells. Am J Physiol. 1991. 261:C115–C123.
crossref
18. COPRAY JCVM, TERHORST GJ, LIEM RSB, VAN WILLINGEN JD. Neurotransmitters and neuropeptides within the mesencepalic trigeminal nucleus of the rat : an immunohistochemical analysis. Neuroscience. 1990. 37:399–411.
crossref
19. DEZEEUW CI, BERREBI AS. Postsynaptic targets of Purkinje cell terminals in the cerebellar and vestibular nuclei of the rat. Eur J Neurosci. 1995. 7:2322–2333.
crossref
20. DELBONO O, GARCIA J, APPEL SH, STEFANI E. Ca2+ current and charge movement of mammalian muscle : action of amyotrophic lateral sclerosis immunoglobulins. J Physiol. 1991. 444:723–742.
crossref
21. DELBONO O, MAGNELLI V, SAWADA T, SMITH RG, APPEL SH, STEFANI E. Fab fragments from amyotrophic lateral sclerosis IgG affect Ca2+ channels of skeletal muscle. Am J Physiol. 1993. 264:C537–C543.
22. DESSEM D, TAYLOR A. Morphology of jawmuscle spindle afferents in the rat. J Comp Neurol. 1989. 282:389–403.
crossref
23. Doughty JM, Barnes-Davies M, Rusznak Z, Harasztosi C, Forsythe ID. Constructing Ca2+ channel subtypes at cell bodies and synaptic terminals of rat anterioventral cochlear bushy neurons. J Physiol. 1998. 512:365–376.
crossref
24. Dunlap K, Luebke JI, Turner TJ. Exocytotic Ca2+ channels in mammalian central neurons. Trends Neurosci. 1995. 18:89–98.
crossref
25. ENGEL AG. Review of evidence for loss of motor nerve terminal Ca2+ channels in Lambert-Eaton myasthenic syndrome. Ann NY Acad Sci. 1991. 635:246–258.
crossref
26. Fox AP, Nowycky MC, Tsien RW. Kinetic and pharmacological properties distinguishing three types of calcium currents in chick sensory neurones. J Physiol Lond. 1987(a). 394:149–172.
crossref
27. Fox AP, Nowycky MC, Tsien RW. Single-channel recordings of three types of calcium channels in chick sensory neurones. J Physiol Lond. 1987(b). 394:173–200.
crossref
28. Galdzicki Z, Puia G, Sciancalepore M, Moran O. Voltage-dependent calcium currents in trigeminal chick neurons. Biochem Biophys Res Comm. 1990. 167:1015–1021.
crossref
29. GOBEL S, FALLS WM, HOCKFIELD S. Anderson DJ, Matthews B, editors. The division of the dorsal and ventral horns of the mammalian caudal medulla eight layers using anatomical criteria. Pain in the trigeminal region. 1977. Amsterdam: Elsevier;443–453.
30. HARTSHORNE RP, CATTERALL WA. Purification of the saxitoxin receptor of the sodium channel from rat brain. Proc Natl Acad Sci USA. 1981. 78:4620–4624.
crossref
31. HARTSHORNE RP, MESSNER DJ, COPERSMITH JC, CATTERALL WA. The saxitation receptor of the sodium channel from rat brain. Evidence for two nonidentical beta subunits. J Biol Chem. 1982. 257:13888–13891.
crossref
32. Haydon PG, Henderson E, Stanley E. Localization of individual calcium channels at the release face of a presynaptic nerve terminal. Neuron. 1994. 13:1275–1280.
crossref
33. HILLE B. Ionic channels of excitable membranes. 1984. first edition. Sunderland, MA: Sinauer Associates, Inc.
34. Hille B. Ionic channels of Excitable Membranes. 1992. Sunderland, MA: Sinauer.
35. Hodgkin AL, Huxley AF. Currents carried by sodium and potassium ions through the membrane of the giant axon of Loligo. J Physiol Lond. 1952. 116:449–472.
crossref
36. Hoffman DA, Magee JC, Colbert CM, Johnston D. K+ channel regulation of signal propagation in dendrites of hippocampal pyramidal neurons. Nature. 1997. 387:869–875.
crossref
37. Jan LY, Jan YN. Cloned potassium channels from eukaryotes and prokaryotes. Annu Rev Neurosci. 1997. 20:91–123.
crossref
38. JERGE CR. Organization and function of the trigeminal mesencephalic nucleus. J Neurophysiol. 1963. 26:379–392.
crossref
39. Josephson IR. Properties of inwardly rectifying K+ channels in venricular myocytes. Mol Cell Biochem. 1988. 80:21–26.
crossref
40. Kaczorowski GJ, Knaus HG, Leonard RJ, Mcmanus OB, Garcia ML. High-conductance calcium-activated potassium channels : structure, pharmacology, function. J Bioenerg Biomembr. 1996. 28:255–267.
crossref
41. KIM YI. Passive transfer of the Lambert-Eaton myasthenic syndrome : neuromuscular transmission. Muscle Nerve. 1985. 8:162–172.
crossref
42. LANG B, NEWSOM-DAVIS J, PRIOR C, WRAY PW. Antibodies to motor-nerve terminals : an electrophysiological study of a human myasthenic syndrome transferred to mouse. J Physiol. 1983. 344:335–345.
crossref
43. LIEM RSB, COPRAY JCVM, VANWILLIGEN JD. Distribution of synaptic boutons in the mesencephalic trigeminal nucleus of the rat. A quantitative electron-microscopical study. Acta Anat (Basel). 1992. 143:74–78.
crossref
44. Llinas R, Sugimory M, Lin JW, Cherksey B. Blocking and isolation of a calcium channel from neurons in mammals and cephalopods utilizing a toxin fraction FTX from funnel-web spider poison. Proc Natl Acad Sci USA. 1989. 86:5656–5660.
45. Maletic-Savatic M, Lenn NJ, TRIMMER JS. Differential spatiotemporal expression of K+ channel polypeptides in rat hippocampal neurons developing in situ and in vitro. J Neurosci. 1995. 15:3840–3851.
crossref
46. Marty A. The physiological role of calcium-dependent channels. Trends Neurosci. 1989. 12:420–424.
crossref
47. MATESZ C. Peripheral and central distribution of fibres of the mesencephalic trigeminal root in the rat. Neurosci Lett. 1981. 27:13–17.
crossref
48. MILLER JA, AGNEW WS, LEVINSON SR. Principal glycopeptide of the tetrodotoxin/saxitoxin binding protein from Electrophorous electricus : isolation and partial chemical and physical characterization. Biochemistry. 1983. 22:462–470.
crossref
49. Miller RJ. Receptor-mediated regulation of calcium channels and neurotransmitter release. FASEB J. 1990. 4:3291–3299.
crossref
50. Mintz I, et al. P type calcium channels in rat central and peripheral neurons. Neuron. 1992. 9:85–95.
crossref
51. MORTON ME, CASSIDY TN, FROEHNER SC, GILMOUR BP, LAURENS RL. Alpha 1 and alpha 2 Ca2+ channel subunit expression in human neuronal and small cell carcinoma cells. FASEB J. 1994. 8:884–888.
crossref
52. Nelson MT, Cheng H, Rubart M, Santana LF, Bonev AD, Knot HJ, Lederer WJ. Relaxation of arterial smooth muscle by calcium sparks. Science. 1995. 270:633–637.
crossref
53. Nilius B, Schwarz G, Droogmans G. Modulation by histamine of an inwardly rectifying potassium channel in huma endotherial cells. J Physiol. 1993. 472:359–371.
crossref
54. Nowycky MC, Fox AP, Trien RW. Three types of neuronal calcium channel with different calcium agonist sensitivity. Nature Lond. 1985. 316:440–443.
crossref
55. OLSZEWSKI J. On the anatomical and functional organization of the trigeminal nucleus. J Comp Neurol. 1950. 92:401–413.
crossref
56. PHELAN KD, FALLS WM. An analysis of the cyto- and myeloarchitectonic organization of trigeminal nucleus interpolars in the rat. Somatosens Motor Res. 1989. 6:333–366.
crossref
57. Puil E, Gimbarzevsky B, Miura RM. Quantification of membrane properties of trigeminal root gangion neurons in guinea pigs. J Neurophysiol. 1986. 55:995–1016.
crossref
58. Randall A, Tsien R. Pharmacological dissection of multiple types of calcium channel currents in rat cerebellar granule neurons. J Neurosci. 1992. 15(4):2995–3012.
crossref
59. Regan LJ, Sah DWY, Bean BP. aCalcium channels in rat central and peripheral neurons : high-threshold current resistant to dihydropyridine blockers and w-conotoxin. Neuron. 1991. 6:269–280.
crossref
60. Regehr WG, Mintz IM. Participation of multiple calcium channel types in transmission at single climbing fiber to Purkinje cell synapses. Neuron. 1994. 12:605–613.
crossref
61. RENEHAN WE, JACQUIN MF. Olesen J, Tfelt-Hansen P, Welch K, editors. Anatomy of central nervous system pathways related to head pain. The headaches. 1993. New York: Raven Press;59–68.
62. RITCHIE JM, ROGART RB. The binding of saxitoxin and thetrodotoxin to excitable tissue. Rev Physiol Biochem Pharmacol. 1977. 79:1–49.
63. ROWL LP. Amyotrophic lateral sclerosis : theories and therapies. Ann Neurol. 1994. 35:129–130.
64. Scott RH, Pearson HA, Dolphin AC. Aspects of vertebrate neuronal voltage-activated calcium currents and their regulation. Prog Neurobiol. 1991. 36:485–520.
crossref
65. Scroggs RS, Fox AP. Calcium current variation between acutely isolated adult rat dorsal root ganglion neurons of different size. J Physiol Lond. 1992. 445:639–658.
crossref
66. Scroggs RS, Fox AP. Distribution of dihydropyridine and -conotoxin-sensitive calcium currents in acutely isolated rat and frog sensory neuron somata: diameter-dependent L channel expression in frog. J Neurosci. 1991. 11:1334–1346.
crossref
67. Serodio P, Vega-Saenz De Miera E, Rudy B. Cloning of a novel component of A-type K+ channels operating at subthreshold potentials with unique expression in heart and brain. J Neurophysiol. 1996. 75:2174–2179.
crossref
68. SESSLE BJ. Acute and chronic carniofacial pain : brainstem mechanisms of nociceptive transmission and neuroplasticity, their clinical correlates. Crit Rev Oral Biol Med. 2000. 11:57–91.
crossref
69. SHENG M, TSAUR MI, JAN YN, JAN LY. Subcellular segregation of two A-type K+ channel proteins in rat central neurons. Neuron. 1992. 2:271–284.
crossref
70. SHER E, CARBONE E, CLEMENTI F. Neuronal Ca2+ channels as target for Lambert-Eaton myasthenic syndrome autoantibodies. Ann NY Acad Sci. 1993. 681:373–381.
crossref
71. SILVER MR, DECOURSEY TE. Intinsic gating of inward rectifier in bovine pulmonary artery endothelial cells in the presence or absence of internal Mg2+. J Gen Physiol. 1990. 96:109–133.
crossref
72. SMART SL, LOPANTSEV V, ZHANG CL, ROBBINS CA, WANG H, CHIU SY, SCHWARTZKROIN PA, MESSING A, TEMPEL BL. Deletion of the Kv 1.1 potassium channel causes epilepsy in mice. Neuron. 1998. 20:809–819.
crossref
73. SMITH RG, HAMILTON S, HOFMANN F, SCHNEIDER T, NASTAINSZYK W, BIRNBAUMER L, STEFANI E, APPEL SH. Serum antibodies to L-type Ca2+ channels in patients with amyotrophic lateral sclerosis. N Engl J Med. 1992. 327:1721–1728.
crossref
74. SONG WJ, TKATCH T, BARANAUSKAS G, ICHINOHE N, KITAI ST, SURMEIER DJ. Somatodendritic depolarization -activated potassium currents in rat neostriatal cholinergic interneurons are predominantly of the A type and attributable to coexpression of Kv 4.2 and Kv 4.1 subunits. J Neurosci. 1998. 18:3124–3137.
crossref
75. STANLEY EF. The calcium channel and the organization of the presynaptic transmitter release face. Trends Neurosci. 1997. 20:404–409.
crossref
76. TKATCH T, BARANAUSKAS G, SURMERIER DJ. Kv4.2 mRNA abundance and A-type K(+) current amplitude are linearly related in basal ganglia and basal forebrain neurons. J Neurosci. 2000. 20:579–588.
crossref
77. TSIEN RW, LIPSCOMBE D, MANDISON DV, BLEY K, FOX A. Reflections on Ca2+ channel diversity, 1984-1994. Trends Neurosci. 1995. 18:52–54.
78. TSIEN RW, LIPSCOMBLE D, MANDISON DV, BLEY K, FOX A. Multiple types of neuronal calcium channels and their selective modulation. Trends Neurosci. 1988. 11:431–438.
crossref
79. Uchitel OD, Protti DA, Sanchez V, CHERKSEY BD, SUGIMORI M, LLINAS R. P-type voltage-dependent Ca2+ channel mediates presynaptic Ca2+ influx and transmitter release in mammalian synapses. Proc Natl Acad Sci U S A. 1992. 89:3330–3333.
crossref
80. VERGARA C, LATORRE R, MARRION NV, ADELMAN JP. Calcium-activated potassium channels. Curr Opin Neurobiol. 1998. 8:321–329.
crossref
81. WESTENBROEK RE, AHLIJANIAN MK, CATTERALL WA. Clustering of L-type Ca2+ channels at the base of major dendrites in hippocampal pyramidal neurons. Nature. 1990. 347:281–284.
crossref
82. WESTENBROEK RE, SAKURAI T, ELLIOT EM, et al. Immunochemical identification and subcellular distribution of the alpha 1A subunits of brain calcium channels. J Neurosci. 1995. 15:6403–6418.
crossref
83. WU LG, WESTENBROEK RE, BORST JGG, CATTERAL WA, SAKMANN B. Calcium channel types with distinct presynaptic localization couple differentially to transmitter release in single calyx-type synapses. J Neurosci. 1999. 19:726–736.
crossref
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