Shin-ichi Terashima

Active transport pumps of HVA and DOPAC in dopaminergic nerve terminals.

The effect of the membrane potential on the efflux of HVA and DOPAC from DA neurons was studied in anesthetized (1% halothane in gas mixture of 70% N2O and 30% O2) cats. Extracellular DA, HVA and DOPAC were measured continuously from the putamen, the hypothalamus, the thalamus, the raphe nuclei and the cortex using brain microdialysis technique combined with HPLC-ED monoamine measurements. HVA and DOPAC concentrations were highest in the putamen and lowest in the cerebral cortex. Extracellular HVA levels exceed those of the DOPAC. Increases in the extracellular potassium from 4 to 120 mM invariably produced decreases of the extracellular HVA and DOPAC in all the tested brain regions. These decreases were inversely proportional to the extracellular potassium concentration. Thus, it is concluded that the HVA and the DOPAC are extruded from inside the cell to the extracellular space by active mechanisms of transport similar to that reported for 5-HIAA in serotonergic neurons.

Distribution of NADPH-diaphorase in the central nervous system of an infrared-sensitive snake, Trimeresurus flavoviridis

The distribution of NADPH-diaphorase (NADPH-d) activity was studied in the central nervous system of an infrared sensitive snake. An inhibitor of nitric oxide synthase, dichloroindophenol (DPIP), was used to distinguish the characteristics of NADPH-d activity. Intensely and weakly NADPH-d-stained neurons and fibers were found in discrete regions throughout the snake brain and cervical spinal cord, such as the olfactory bulb, subcommissural organ, stratum griseum periventriculare, locus coeruleus, dorsal root, dorsal horn, and area X. It was particularly noticed that the trigeminal descending nuclei and reticular formation of the medulla oblongata contained many positive neurons and fibers, but the lateral descending nucleus and nucleus reticularis caloris (infrared sensory nuclei) certainly did not. The positive neurons and fibers were also observed in supraspinal sensory ganglia. DPIP inhibited NADPH-d activity in all regions except for the olfactory/vomeronasal nerve and glomeruli. The results prove for the first time the presence of NADPH-d activity in the ophidian brain and suggest that nitric oxide may be involved in many neural functions, but not in infrared sensory processing.

Efflux of 5-HIAA form 5-HT neurons: A membrane potential-dependent process

The effect of the membrane potential on the efflux of 5-HIAA from 5-HT neurons was studied in anesthetized (halothane: 1% in gas mixture of N2O: 70% and O2: 30%) cats. The endogenous 5-HT and its metabolite 5-HIAA were measured continuously from the cortex, the thalamus, the hypothalamus and the raphe nuclei using brain microdialysis technique combined with HPLC-ED monoamine measurements. Membrane potential variations were induced by changing the extracellular concentration of potassium through the microdialysis membrane. The levels of the extracellular 5-HIAA varied according to the different regions of the brain, being highest in the hypothalamus and lowest in the cerebral cortex. Increases in the extracellular potassium from 4 to 120 mM invariably produced a decrease of the extracellular 5-HIAA in all the tested brain regions. This decrease was inversely proportional to the logarithm of extracellular potassium concentration. Thus, it is postulated that the 5-HIAA is moved from inside the cell to extracellular space by an active mechanism of transport electrically coupled to the membrane potential.

A suspected infrared-recipient nucleus in the brainstem of the vampire bat,Desmodus rotundus

We discovered in the brainstem of infrared-sensitive vampire bats, Desmodus rotundus, a specific nucleus not known in other species of bats. Because it corresponded in location and histological features to the infrared nucleus of infrared-sensitive snakes, we suggest the probability of its being part of the infrared processing system of vampire bats.

Degeneration of infrared receptor terminals of snakes caused by capsaicin.

Capsaicin, the main pungent ingredient in hot peppers (genus Capsicum), caused degeneration of the infrared receptor terminals in infrared sensitive snakes, Trimeresurus flavoviridis, when it was applied perineurally to a branch of the trigeminal nerve. The degeneration of the terminals was found 6 h after the application. This finding suggests that capsaicin stimulates this infrared receptor terminal, a kind of warm receptor terminal.

Immunohistochemical analysis of neuronal nitric oxide synthase in the trigeminal ganglia of the crotaline snake Trimeresurus flavoviridis

The expression of neuronal nitric oxide synthase (nNOS) in the trigeminal ganglia (TG) of the infrared-sensitive crotaline snake Trimeresurus flavoviridis was studied immunohistochemically. The percentage of nNOS-positive (+) neurons in the TG was significantly higher (about 3.5-fold, P<0.001) in the mandibular division than in the infrared-sensory processing area (the maxillary division and ophthalmic ganglion). nNOS was found in varying sizes of TG neurons. However, nNOS (+) neurons were more abundant in small and large neurons than in medium-sized neurons, which include most of the infrared-sensitive neurons of the TG. These findings suggest that nNOS may be involved in normal physiological functions, such as the transmission of tactile, vibrotactile, and nociceptive sensations in the TG, rather than in infrared sensory processing in this species.

Touch and Vibrotactile Neurons in a Crotaline Snake's Trigeminal Ganglia

Thirty-five touch (M) neurons and 59 vibrotactile (V + M) neurons were recorded intrasomally in the trigeminal ganglion of a crotaline snake (the pit viper, Trimeresurus flavoviridis). The M neurons were excited by von Frey hair (5-10 mg) mechanical stimulation of the receptive field, and adapted slowly to a sustained stimulus. It was almost impossible to elicit 1:1 entrainment to sinusoidal movement. Vibration with touch was an adequate stimulus for the V + M neurons. The range of entrainment to sinusoidal movement was 5-300 Hz. Thresholds of V + M neurons to sustained mechanical stimulation could not be determined, but a response was obtained by stroking with a von Frey hair (5-10 mg). Receptive fields of both M and V + M neurons were found on the skin (scales) and the mucous membrane of the orofacial region. There was one receptive field of approximately 2 mm in diameter for each M or V + M neuron. The mean resting potentials (+/- SD) of M and V + M neurons were -57.0 +/- 5.1 mV (n = 26) and -63.7 +/- 8.2 mV (n = 49), respectively. Neurons of both modalities displayed no background discharge. The action potential of V + M neurons had a shorter mean duration than that of M neurons. The mean conduction velocities (+/- SD) of peripheral (and stem) axons of M and V + M neurons were 28.4 +/- 5.7 m/sec (n = 11) and 30.8 +/- 7.8 m/sec (n = 30), respectively. Recorded neurons were labeled with intrasomal horseradish peroxidase electrophoresis. V + M neurons had larger somata than M neurons. All axons of M and V + M neurons were myelinated and similar in diameter. M and V + M neurons had similar central projection patterns. The projection of the thick central axon divided into a thinner ascending fiber and a thick descending fiber at the entry zone of the root to the brainstem. The former ran ipsilaterally to the principal sensory nucleus of the trigeminal nerve (TPR), and the latter ran to the descending nucleus of the trigeminal nerve (TTD) and beyond, where terminal arbors and bouton swellings were observed. Smaller myelinated and unmyelinated collaterals were given off at right angles from the descending fiber of the central axon into the TTD. They projected more densely to the rostral part than to the caudal part of the TTD. All of these data were compared with data on warm-temperature neurons, previously obtained.

C mechanical nociceptive neurons in the crotaline trigeminal ganglia

Using 32 Crotaline snakes, Trimeresurus flavoviridis, intrasomal recordings were made from 44 neurons of the trigeminal ganglia in vivo. They were 10 C neurons from 9 snakes and 34 A-delta mechanical nociceptive neurons from 23 snakes. 5 of the 10 C neurons were identified as mechanical nociceptive neurons. The neurons were labeled with iontophoretically injected HRP. Each of the 5 C nociceptive neurons had one receptive field, on which 1 spike was elicited by pricking the skin or mucosa with a pin. They were sensitized after repeated stimulation. The fields were insensitive to thermal stimulation. No background discharge was observed. Average conduction velocity was 0.95 m/s (+/- 0.4 S.D., n = 5). Mean resting potential was -62.5 mV (+/- 6.0 S.D., n = 4), and mean action potential amplitude was 88.0 mV (+/- 10.9 S.D., n = 4). Two somata were successfully visualized with HRP (22 microns x 20 microns, 20 microns x 18 microns). Total lengths of labeled axons were 1260 and 1480 microns peripherally to the edge of the section, and 1810 and 770 microns centrally. Neither of the neurons had branching of the peripheral or central axons in the ganglion.

Vagal afferent C fibers projecting to the lateral descending trigeminal complex of crotaline snakes

SummaryThe primary vagal axons and terminals in the lateral descending trigeminal complex (dlv-DLV complex) in crotaline snakes were studied following HRP injections into the vagal nerve. Labeled fibers and terminals were found in the marginal neuropil, which was made up entirely of unmyelinated fibers, i.e., C fibers. The general features of vagal input to the dlv-DLV complex in snakes with infrared sensitivity (Boidae and Crotalinae) are discussed.

Immunohistochemical localization of the delta subspecies of protein kinase C in the trigeminal sensory system of Trimeresurus flavoviridis, an infrared-sensitive snake

We examined the expression of the protein kinase C (PKC) delta subspecies in the trigeminal sensory system of the infrared-sensitive snake Trimeresurus flavoviridis. In the trigeminal ganglion (TG), diffuse low-intensity PKC delta immunoreactivity was found in TG neurons and fibers, while intense reactions were observed mainly in medium-sized neurons, which include most of the infrared-sensitive neurons. In the brainstem, intense PKC delta immunoreactivity was present in the intermediate layer of the optic tectum of the midbrain and in the nucleus descendens lateralis n. trigemini of the medulla oblongata; these areas are related to the infrared sensory pathway. In the pit organ (the infrared receptor), PKC delta immunoreactivity was present in terminal nerve masses in the pit membrane. These findings suggest that the PKC delta subspecies is involved in the infrared sensory pathway in the trigeminal sensory system of the infrared-sensitive snake.