Inhibition of trigeminal respiratory activity by suckling.

UNLABELLED: The trigeminal motor system is involved in many rhythmic oral-motor behaviors, such as suckling, mastication, swallowing, and breathing. Despite the obvious importance of functional coordination among these rhythmic activities, the system is not well-understood. In the present study, we examined the hypothesis that an interaction between suckling and breathing exists in the brainstem, by studying the respiratory activity in trigeminal motoneurons (TMNs) during fictive suckling using a neonatal rat in vitro brainstem preparation. The results showed that fictive suckling, which was neurochemically induced by bath application of N-methyl-D,L-aspartate and bicuculline-methiodide, or by local micro-injection of the same drugs to the trigeminal motor nucleus, inhibited the inspiratory activities in both respiration TMNs and respiratory rhythm-generating neurons. Under patch-clamp recording, fictive suckling caused membrane potential hyperpolarization of respiration TMNs. We conclude that the brainstem preparation contains an inhibitory circuit for respiratory activity in the trigeminal motor system via the rhythm-generating network for suckling. ABBREVIATIONS: BIC, bicuculline methiodide; GABA, gamma aminobutyric acid; NMA, N-methyl-D,L-aspartate; NMDA, N-methyl-D-aspartate; and TMN, trigeminal motoneuron.

Temporal patterns of trigeminal respiratory activity in rat brainstem-spinal cord in vitro.

Respiratory activity in trigeminal (V) motoneurons was studied in rhythmically active en bloc brainstem-spinal cord preparations isolated from neonatal rats (P0-P3). In the majority of preparations (83%), the temporal pattern of V activity consisted of spontaneous inspiratory phasic discharge with onset delayed or coincident with onset of phrenic motoneuron discharge. Blockade of alpha-2 noradrenergic receptor activation shifted onset of V respiratory discharges earlier than phrenic discharges, while elevation of extracellular potassium concentration or blockade of GABAergic and glycinergic inhibitory synaptic transmission had little effect on temporary pattern of V respiratory discharges. We conclude V motoneurons in the in vitro preparation generate respiratory activity during inspiratory phase, and their temporal patterns are modulated by inhibitory noradrenergic synaptic transmission.

Effect of serotonin (5-HT) on trigeminal rhythmic activities generated in in vitro brainstem block preparations.

We used rat isolated brainstem block preparations to analyze the functional roles of serotonin receptors in the generation of trigeminal rhythmic activities. We previously reported that trigeminal rhythmic activities could be induced by some pharmacological applications in an isolated brainstem preparation with a rostral boundary at the border between the inferior and superior colliculus, and a caudal border at the level of the rostral facial nucleus. However, the same stimulation did not induce trigeminal rhythmic activities in a whole brainstem block preparation with the same rostral boundary and a caudal border at the obex level. In the present study, both the 5-HT(1A) phthalimido-butyl-piperazine, and the 5-HT(2C) agonist, 1-2,5-dimethoxy-4-iodophenyl-2-aminopropane, combined with N-methyl-D,L-aspartate and bicuculline, elicited trigeminal rhythmic activities in a whole brainstem block preparation. Our results suggest that serotonin has both facilitation and inhibition effects on the generation of trigeminal rhythmic activities in an isolated brainstem block preparation in vitro.

Effect of norepinephrine receptors on trigeminal rhythm generation in newborn rats.

N-methyl-D,L-aspartate acid and bicuculline are required to enhance the trigeminal rhythmic activities in an in vitro isolated brainstem block preparation. In this study, we analyzed the effect of norepinephrine on the trigeminal neural circuit underlying rhythmic jaw movements. Rhythmic trigeminal activity is observed in brainstem preparations (inferior colliculus to obex) only following blockade of alpha(2)-adrenoceptors with idazoxan. This observation, combined with the inhibition of rhythm by alpha(2)-adrenoceptor agonists suggests endogenous alpha(2)-adrenoceptor mediated inhibition of trigeminal networks. A complex noradrenergic modulation of trigeminal systems is further supported by the prazosin-sensitive potentiation of rhythm by bath application of the alpha(1)-adrenoceptor agonist phenylephrine.

Canine parentage testing based on microsatellite polymorphisms.

To establish an accurate method for parentage testing in dogs, microsatellite DNA repeat length polymorphisms were examined. We selected twenty microsatellite markers reported previously and examined their application for parentage testing in Beagles and Labrador Retrievers. Heterozygosity (He), Polymorphism Information Content (PIC), the probabilities of Paternity Exclusion (PE) and the combined PE were calculated from allelic frequencies of the markers. All markers amplified by polymerase chain reactions were polymorphic and many markers showed high He and PIC in the both breeds. The final combined PEs in Beagles and Labrador Retrievers were 0.999994 and 0.999920, respectively. The results suggest that the twenty markers can be applied for routine parentage testing in dogs.

Alpha2-adrenoceptors coordinate swallowing and respiration.

Because the discoordination between swallowing and respiration may cause severe respiratory disorders such as aspiration pneumonia, understanding the neuronal mechanisms underlying such coordination is important. Recently, it was reported that medullary noradrenergic neurons are involved in evoking esophageal-gastric relaxation reflex, leading to a hypothesis that such neurons are also involved in swallowing-respiration coordination. We tested this hypothesis using an in vitro brain-stem preparation obtained from neonatal rats. A temporal inhibition of respiratory rhythm was consistently observed when swallowing activity was induced by electrical stimulations to the supralaryngeal nerve. We found that a broad adrenergic receptor agonist, norepinephrine, markedly blocked the swallowing-induced temporal inhibition of respiration. Further studies revealed that swallowing-induced respiratory inhibition is blocked by an alpha2-adrenergic receptor agonist and enhanced by an alpha2-adrenergic receptor antagonist, indicating an important role of alpha2-adrenergic receptors in regulation of the coordination between swallowing and respiration in vitro.

Regulation of trigeminal respiratory motor activity in the brainstem.

The trigeminal motor system participates in the control of respiration as well as suckling and mastication. However, the central mechanism underlying respiratory activity in trigeminal motoneurons is not well-understood. In this study, we aimed to elucidate brainstem circuitry for rhythm generation and signal transmission of trigeminal respiratory activity in in vitro neonatal rat brainstem-spinal cord preparations. We further examined the role of midline-crossing trigeminal interneurons in the bilateral synchronization of respiratory and suckling activity in trigeminal motor nerves. The results of brainstem-sectioning experiments indicated that respiratory rhythms were generated in the medulla and ipsilaterally transmitted to trigeminal motoneurons in the pons. We conclude that the trigeminal motor system, as well as the hypoglossal and phrenic motor system, is regulated by medullary respiratory networks, and that pontine interactions between bilateral trigeminal interneurons are not critical for the generation or synchronization of trigeminal respiratory activity, but are crucial for trigeminal suckling activity.

Evidence of aerosolised floating blood mist during oral surgery.

Dental surgery performed with high speed instruments, such as a dental turbine, air motor, or micro-engine handpiece, produces a large amount of splattering and particles, which can be contaminated by micro-organisms from the oral cavity. It has been speculated that such particulate mists contain blood-based elements. In the present study, we investigated whether blood-contaminated aerosol was present in a room where oral surgery was performed with high speed instruments. An extra-oral evacuator system was used for sample collection (N=132). For the experiment, a non-woven towel was set on the nozzle of the evacuator as a filter and invisible mist was collected at distances of 20, 60 and 100 cm from the surgical site. A leucomalachite green presumptive test was performed with each filter after every tooth extraction. At locations 20 and 100 cm from the surgical site, 76% and 57%, respectively, of the particulates were positive in blood presumptive tests. Based on our results, we consider that blood-contaminated materials have the potential to be suspended in air as blood-contaminated aerosol. These results indicate the risk of cross-infection at the dental practice for immunocompromised patients as well as healthy personnel.

Angiomyolipoma of the tongue.

We present a rare case of angiomyolipoma of the tongue in a 23-year-old man. The clinical appearance was of a small, solitary, well-demarcated, painless mass. The oral angiomyolipoma followed a benign course with no recurrence after excision.

Use of microsatellite markers for parentage testing of two litters of beagles born after artifcial insemination with mixed fresh semen.

This study was undertaken to determine if 20 kinds of microsatellite markers, which have already been reported, were useful for the parentage testing of 16 Beagle puppies born after artificial insemination using intentionally mixed semen from six male Beagles. Of the 20 kinds of markers used in this study, 17 kinds (the exceptions were CPH6, FH0020 and FH0419) produced 3 to 5 kinds of alleles, and using these markers, the parentage was finally established for all of the 16 puppies. These 17 kinds of markers showed sufficient amplification in PCR and had no non-specific PCR products or alleles with single-base-alterations, which makes them very useful as markers for identification of individual Beagles and useful for parentage testing.