Effect of Brn-3a deficiency on parvalbumin-, calbindin D-28k-, calretinin- and calcitonin gene-related peptide-immunoreactive primary sensory neurons in the trigeminal ganglion

doi:10.1016/S0306-4522(02)00182-3

Neuroscience

Volume 113, Issue 3, 2 September 2002, Pages 537-546

https://doi.org/10.1016/S0306-4522(02)00182-3 Get rights and content

Abstract

Immunohistochemistry for parvalbumin, calbindin D-28k, calretinin and calcitonin gene-related peptide (CGRP) was performed on the trigeminal ganglion and oro-facial tissues in Brn-3a wildtype and knockout mice at embryonic day 18.5 and postnatal day 0. In wildtype mice, the trigeminal ganglion contained abundant parvalbumin-, calbindin D-28k- and CGRP-immunoreactive neurons while the ganglion was almost devoid of calretinin-immunoreactive neurons. In Brn-3a knockout mice, a 63% decrease of parvalbumin-immunoreactive neurons was detected. In contrast, the absence of Brn-3a dramatically increased the number of calbindin D-28k-immunoreactive (3.5-fold increase) and calretinin-immunoreactive neurons (91-fold increase). The number of CGRP-immunoreactive neurons, however, was not altered by the Brn-3a deficiency. Cell size analysis indicated that loss of Brn-3a increased the proportions of small (<100 μm²) parvalbumin-, calbindin D-28k- and CGRP-immunoreactive neurons while it decreased those of large (>200 μm²) immunoreactive cells. Calretinin-immunoreactive neurons were either small or medium (100–200 μm²) in mutant mice. The oro-facial tissues contained parvalbumin-, calbindin D-28k- and CGRP-immunoreactive fibers, but not calretinin-immunoreactive ones in wildtype mice. In Brn-3a knockout mice, the number of parvalbumin-immunoreactive fibers markedly decreased in the infraorbital nerve and parvalbumin-immunoreactive endings disappeared in the vibrissa. In contrast, the number of calbindin D-28k-immunoreactive fibers increased significantly in the infraorbital and mental nerves. In addition, calbindin D-28k-immunoreactive endings appeared in the vibrissa. As well, some fibers showed calretinin-immunoreactivity in the infraorbital nerve of the mutant. However, no obvious change of CGRP-immunoreactive fibers was observed in the oro-facial region of knockout mice.

Taken together, our data suggest that Brn-3a deficiency has effects on the expression of neurochemical substances in the trigeminal ganglion.

Section snippets

Experimental procedures

Mice lacking the Brn-3a gene were derived as described by Xiang et al. (1996). Two wildtype and four knockout mice at embryonic day 18.5 (E18.5) and five wildtype and six knockout mice at postnatal day 0 (P0) were obtained from breeding of Brn-3a heterozygous mice. Animals were immersion-fixed overnight in 4% paraformaldehyde at 4°C. Subsequently, tissues were stored at 4°C in phosphate-buffered saline (PBS) containing 0.1 mM sodium azide until use. The head containing the TG, vibrissa, palate,

Trigeminal ganglion

In wildtype mice, the TG contained abundant parvalbumin-, calbindin D-28k- and CGRP-IR neurons (Fig. 1, Fig. 2) while the ganglion was almost devoid of calretinin-IR neurons (Fig. 2A). Topographically, the distribution of parvalbumin-IR neurons in the TG was not uniform, i.e., they were more concentrated in the rostral half of the ganglion (192.3±36.8 immunoreactive neurons) than in the caudal half (66±19.3 immunoreactive neurons). At P0, a 63% decrease of parvalbumin-IR neurons was detected in

Discussion

The present study demonstrated the distribution of parvalbumin-, calbindin D-28k-, calretinin- and CGRP-IR in the TG and oro-facial tissues of Brn-3a wildtype and knockout mice. The immunoreactivity was detected in cell bodies of primary sensory neurons in the TG. In the oro-facial region, the ION, MN, vibrissal pad, facial skin, lip and palate contained immunoreactive fibers. In addition, immunoreactive nerve endings were detected in the vibrissal follicle, epithelium and subepithelial

Conclusion

The present study investigated the effect of Brn-3a deficiency on parvalbumin-, calbindin D-28k-, calretinin- and CGRP-IR TG neurons. The loss of Brn-3a reduced the number of medium-sized to large parvalbumin-IR neurons which supplied the vibrissal innervation. In contrast, the numbers of small calbindin D-28k- and calretinin-IR TG neurons which innervated the vibrissa and/or facial skin increased in the knockout mouse. The lack of Brn-3a may have different effects on small and large CGRP-IR TG

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Thus, it is suggested that primary nociceptors and mechanoreceptors with medium-sized and large cell bodies in the TG require Brn-3a for their survival. On the other hand, the number of small TG neurons which contain CGRP, calbindin D-28k or calretinin is increased by the loss of Brn-3a [56]. Brn-3a may also have a function to suppress the expression of these neurochemical substances in small TG neurons.
This review presents information about the development of primary sensory neurons in the trigeminal nervous system. The deficiency of high affinity receptors for nerve growth factor (trkA) and neurotrophin-3 (trk-C) causesa reduction of primary nociceptors in the trigeminal ganglion (TG). The disruption of trkB, a receptor for brain-derived neurotrophic factor and neurotrophin-4, causes a loss of Meissner endings in the palate and Ruffini endings in the periodontal ligament. The number of Merkel cells in palatal rugae is also severely reduced by the absence of trkA, trkB or trkC. In the mesencephalic trigeminal tract nucleus (Mes5), primary proprioceptors are decreased by 50% in trkC null mutant mice. On the other hand, the deficiency of Brn-3a, a member of the POU family of transcription factors, decreases primary nociceptors and low-threshold mechanoreceptors in the TG. In the Mes5 of Brn-3a knockout mice, primary proprioceptors are completely lost. In addition, the disruption of dystonin which is a member of the plakin family of high molecular weight cytoskeletal linker proteins causes a reduction of nociceptors in the TG but not proprioceptors in the Mes5. The dependency of primary nociceptors, low-threshold mechanoreceptors and proprioceptors on neurotrophins, Brn-3a and dystonin in the trigeminal nervous system is discussed.
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ASIC3-immunoreactivity (ir) was examined in the rat vagal and glossopharyngeal sensory ganglia. In the jugular, petrosal and nodose ganglia, 24.8%, 30.8% and 20.6% of sensory neurons, respectively, were immunoreactive for ASIC3. These neurons were observed throughout the ganglia. A double immunofluorescence method demonstrated that many ASIC3-immunoreactive (ir) neurons co-expressed calcitonin gene-related peptide (CGRP)- or vanilloid receptor subtype 1 (VRL-1)-ir in the jugular (CGRP, 77.8%; VRL-1, 28.0%) and petrosal ganglia (CGRP, 61.7%; VRL-1, 21.5%). In the nodose ganglion, however, such neurons were relatively rare (CGRP, 6.3%; VRL-1, 0.4%). ASIC3-ir neurons were mostly devoid of tyrosine hydroxylase in these ganglia. However, some ASIC3-ir neurons co-expressed calbindin D-28k in the petrosal (5.5%) and nodose ganglia (3.8%). These findings may suggest that ASIC3-containing neurons have a wide variety of sensory modalities in the vagal and glossopharyngeal sensory ganglia.
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Effect of Brn-3a deficiency on parvalbumin-, calbindin D-28k-, calretinin- and calcitonin gene-related peptide-immunoreactive primary sensory neurons in the trigeminal ganglion

Abstract

Section snippets

Experimental procedures

Trigeminal ganglion

Discussion

Conclusion

Neuroscience

J. Biol. Chem.

Mech. Dev.

Dev. Biol.

Brain Res.

Brain Res.

Brain Res.

Brain Res.

Mol. Brain Res.

Exp. Neurol.

Arch. Oral Biol.

Brain Res.

Int. J. Biochem. Cell Biol.

Peptides

J. Biol. Chem.