Number Of Dorsal Root Ganglion Cells Research Articles

Genetic Tracing of Cav3.2 T-Type Calcium Channel Expression in the Peripheral Nervous System.

Characterizing the distinct functions of the T-type ion channel subunits Cav3.1, 3.2 or 3.3 has proven difficult due to their highly conserved amino-acid sequences and the lack of pharmacological blockers specific for each subunit. To precisely determine the expression pattern of the Cav3.2 channel in the nervous system we generated two knock-in mouse strains that express EGFP or Cre recombinase under the control of the Cav3.2 gene promoter. We show that in the brains of these animals, the Cav3.2 channel is predominantly expressed in the dentate gyrus of the hippocampus. In the peripheral nervous system, the activation of the promoter starts at E9.5 in neural crest cells that will give rise to dorsal root ganglia (DRG) neurons, but not sympathetic neurons. As development progresses the number of DRG cells expressing the Cav3.2 channel reaches around 7% of the DRG at E16.5, and remains constant until E18.5. Characterization of sensory neuron subpopulations at E18.5 showed that EGFP+ cells are a heterogeneous population consisting mainly of TrkB+ and TrkC+ cells, while only a small percentage of DRG cells were TrkA+. Genetic tracing of the sensory nerve end-organ innervation of the skin showed that the activity of the Cav3.2 channel promoter in sensory progenitors marks many mechanoreceptor and nociceptor endings, but spares slowly adapting mechanoreceptors with endings associated with Merkel cells. Our genetic analysis reveals for the first time that progenitors that express the Cav3.2 T-type calcium channel, defines a sensory specific lineage that populates a large proportion of the DRG. Using our Cav3.2-Cre mice together with AAV viruses containing a conditional fluorescent reporter (tdTomato) we could also show that Cre expression is largely restricted to two functionally distinct sensory neuron types in the adult ganglia. Cav3.2 positive neurons innervating the skin were found to only form lanceolate endings on hair follicles and are probably identical to D-hair receptors. A second population of nociceptive sensory neurons expressing the Cav3.2 gene was found to be positive for the calcitonin-gene related peptide but these neurons are deep tissue nociceptors that do not innervate the skin.

Thermal Stimulation of Primary Sensory Neurons in the Rat Hind Paw: Effect of Morphine on ERK1/2 Phosphorylation, TRPV1 and TRPA1 Channel Expression

Temperature-sensitive transient receptor potential (TRP) channels or ‘thermo-TRP’ were stimulated on rat sensory afferents, and the effects on the phosphorylation of ERK1/2, on the regulation of TRPV1 and TRPA1, as well as the pharmacological modulation by the opioid analgesic morphine were investigated. The thermal stimuli were applied to the rat hind paw by immersion into either hot or cold water. Phospho-ERK1/2 (p-ERK1/2) was measured by fluorescence-immunohistochemistry in the lumbar dorsal root ganglion (DRG) neurons. TRP channel mRNA expression was measured by RT-PCR in the innervating DRGs, and the protein content of TRPV1 and TRPA1 was determined by Western blot in the DRGs and in the sciatic nerve. The thermal stimuli led to a time-dependent increase in the number of DRG cells displaying cytoplasmic and nuclear staining for p-ERK1/2. Morphine partly prevented this increase in ERK1/2 phosphorylation, exerting its effect mainly on the nuclear staining. The mRNA expression for TRPV1 and TRPA1 in the DRG did not change within 24 h following the thermal stimuli. However, the protein content of both TRPV1 and TRPA1 was regulated by the thermal stimulation and by morphine. In the DRGs and in the sciatic nerve, heat or cold stimuli per se tended to decrease TRP protein levels, whereas with morphine pretreatment protein levels were raised. The present findings shed new light on the time-dependent reactions of primary sensory neurons towards irritant thermal stimuli to the skin and on their opioid modulation.

Runx1 promotes neuronal differentiation in dorsal root ganglion

Transcription factor Runx1 controls the cell type specification of peptidergic and nonpeptidergic nociceptive dorsal root ganglion (DRG) neurons by repressing TrkA and calcitonin gene-related peptide (CGRP) expression and activating Ret expression during late embryonic and early postnatal periods (Chen et al., 2006b; Kramer et al., 2006; Yoshikawa et al., 2007). Because Runx1 is expressed in DRG from early developmental stages, we examined the roles of Runx1 in the proliferation and the neuronal differentiation of DRG cells. We used transgenic Runx1-deficient (Runx1−/−::Tg) mice which are rescued from early embryonic lethality by selective expression of Runx1 in hematopoietic cells under the control of GATA-1 promoter. We found that TrkA-expressing (TrkA+) DRG neurons were decreased at embryonic day (E) 12.5 in contrast to the previous study showing that TrkA+ DRG neurons were increased at E17.5 in Runx1−/−::Tg mice (Yoshikawa et al., 2007). The number of DRG neurons which express neuronal markers Hu, NeuN and Islet1 was also reduced in Runx1−/−::Tg mice at E12.5, suggesting that the neuronal differentiation was suppressed in these mice. The cell cycle analysis using BrdU/IDU revealed that the number of DRG cells in S-phase and G2/M-phase was increased in Runx1−/−::Tg mice at E12.5, while the length of S-phase was not changed between Runx1+/+::Tg and Runx1−/−::Tg mice, suggesting that Runx1 negatively controls the proliferation of DRG progenitor cell subpopulation in early embryonic period. Hes1 is a negative regulator of neuronal differentiation (Ishibashi et al., 1995; Tomita et al., 1996), and we found that the number of Hes1+ DRG cells was increased in Runx1−/−::Tg mice at E12.5. In summary, the present study suggests a novel function that Runx1 activates the neuronal differentiation of DRG cell subpopulation through the repression of Hes1 expression in early embryonic period.

Up-regulation of low-threshold tetrodotoxin-resistant Na+ current via activation of a cyclic AMP/protein kinase A pathway in nociceptor-like rat dorsal root ganglion cells

The effects of forskolin on low-threshold tetrodotoxin-resistant (TTX-r) Na(+) currents was studied in small diameter (average ≈ 25 μm) dorsal root ganglion (DRG) cells. All DRG cells included in the study were categorized as type-2 or non-type-2 based on the expression of a low-threshold A-current. In all type-2 and some non-type-2 DRG cells held at -80 mV, the adenylyl cyclase (AC) activator forskolin (10 μM) up-regulated TTX-r Na(+) currents evoked with steps to -55 mV through -35 mV (low-threshold current). Up-regulation of low-threshold current by forskolin was mimicked by the protein kinase A (PKA) agonist Sp-cAMPs and the inflammatory mediator serotonin, and blocked by the PKA antagonist Rp-cAMPs. Forskolin-induced up-regulation of low-threshold current evoked from a holding potential of -60 mV was blocked by 40 ms steps to 0 mV, which presumably induced a long lasting inactivation of the low-threshold channels. Reducing to 3 ms the duration of steps to 0 mV, significantly increased the number of DRG cells where low-threshold current was up-regulated by forskolin, presumably by reducing the long-lasting inactivation of the low-threshold channels. In the same cells, high-threshold current, evoked by 40 ms or 3 ms steps to 0 mV, was consistently up-regulated by forskolin. The selective Na(V)1.8 channel blocker A-803467 markedly blocked high-threshold current but not low-threshold current. The different voltage protocols observed to activate and inactivate the low- and high-threshold currents, and the observation that A-803467 blocked high- but not low-threshold current suggests that the two currents were mediated by different channels, possibly Na(V)1.8 and Na(V)1.9, respectively. Inflammatory mediators may simultaneously up-regulate Na(V)1.8 and Na(V)1.9 channels in the same nociceptor via a AC/PKA signaling pathway, increasing nociceptor signaling strength, and lowering nociceptor threshold, respectively.

Phenotypic alterations of neurons that innervate osteoarthritic joints in rats

Pain is a prominent feature of osteoarthritis (OA). To further understand the primary mechanisms of nociception in OA, we studied the expression of the phenotype markers calcitonin gene-related peptide (CGRP), isolectin B4 (IB4), and neurofilament 200 (NF200) in sensory neurons innervating the OA knee joint in rats. OA was induced in rats by intraarticular injection of 2 mg of mono-iodoacetate (MIA) into the knee. Neurons innervating the joint were identified by retrograde labeling with fluorogold in dorsal root ganglia (DRG) and colocalized with neurochemical markers by immunofluorescence. The total number of DRG cells was determined by stereologic methods in Nissl-stained sections. A 37% decrease in the number of fluorogold-backlabeled cells was observed in rats with OA when compared with control rats, even though no decrease in the total number of cells was observed. However, an increase in the number of medium/large cell bodies and a decrease in the number of the smallest cells were observed, suggesting the occurrence of perikarya hypertrophy. The percentage of CGRP-positive cells increased significantly, predominantly in medium/large cells, suggesting the occurrence of a phenotypic switch. Colocalization of CGRP and NF200 revealed no significant changes in the percentage of double-labeled cells, but an increase in the number of medium/large double-labeled cells was observed. No differences in the expression of either IB4 or NF200 were observed in fluorogold-backlabeled cells. These results indicate that MIA-induced OA causes an up-regulation of CGRP in different subpopulations of primary afferent neurons in DRG due to a phenotypic switch and/or cell hypertrophy which may be functionally relevant in terms of the onset of pain in this pathologic condition.

Stage-Specific Control of Neural Crest Stem Cell Proliferation by the Small Rho GTPases Cdc42 and Rac1

The neural crest (NC) generates a variety of neural and non-neural tissues during vertebrate development. Both migratory NC cells and their target structures contain cells with stem cell features. Here we show that these populations of neural crest-derived stem cells (NCSCs) are differentially regulated by small Rho GTPases. Deletion of either Cdc42 or Rac1 in the NC results in size reduction of multiple NC target structures because of increased cell-cycle exit, while NC cells emigrating from the neural tube are not affected. Consistently, Cdc42 or Rac1 inactivation reduces self-renewal and proliferation of later stage, but not early migratory NCSCs. This stage-specific requirement for small Rho GTPases is due to changes in NCSCs that, during development, acquire responsiveness to mitogenic EGF acting upstream of both Cdc42 and Rac1. Thus, our data reveal distinct mechanisms for growth control of NCSCs from different developmental stages.

Up-regulation of calcitonin gene-related peptide and receptor tyrosine kinase TrkB in rat bladder afferent neurons following TNBS colitis

Colonic inflammation has profound effects on the urinary bladder physiology and produces hypersensitivity of bladder afferent neurons and neurogenic bladder overactivity. Calcitonin gene-related peptide (CGRP) expressed in dorsal root ganglia (DRG) plays an important role in mediating sensory perception following visceral inflammation. In the present study, we determined that the expression of CGRP was increased in bladder afferent neurons in lumbosacral DRG following tri-nitrobenzene sulfonic acid (TNBS)-induced colitis in rat. After colitis, the percentage of bladder afferent neurons expressing CGRP was increased in L1 (61.2 ± 2.9% in colitis vs. 37.7 ± 5.1% in controls; p < 0.05) and S1 DRG (26.3 ± 2.3% in colitis vs. 15.5 ± 1.9% in controls; p < 0.01). We also demonstrated that the expression of tyrosine kinase receptor TrkB was increased in L1 (39.7 ± 2.9% in colitis vs. 25.2 ± 4.3% in controls; p < 0.05) and S1 DRG (45.6 ± 3.8% in colitis vs. 38.3 ± 3.6% in controls; p < 0.01) following colitis. CGRP and TrkB were co-stored in a subpopulation of DRG neurons in control and colitic animals and the number of DRG cells co-expressing CGRP and TrkB was significantly increased in L1 (2.7-fold, p < 0.01) and S1 DRG (2.4-fold, p < 0.01) following colitis. In cultured DRG, exogenous BDNF application significantly increased CGRP expression, which was blocked by TrkB selective inhibitor K252a. These results suggest that up-regulation of CGRP and TrkB in bladder afferent neurons may play a role in colon-to-bladder cross-sensitization following colitis.

Changes of calcitonin gene-related peptide in primary sensory neurons and their central branch after nerve root compression of the dog

Kobayashi S, Sasaki S, Shimada S, Kaneyasu M, Mizukami Y, Kitade I, Ogawa M, Kawahara H, Baba H, Yoshizawa H. Changes of calcitonin gene-related peptide in primary sensory neurons and their central branch after nerve root compression of the dog. ObjectiveTo investigate changes in axonal flow after nerve root compression by using immunohistochemical techniques to detect calcitonin gene-related peptide (CGRP), which is thought to be involved in pain sensation. DesignExperimental, controlled study. SettingUniversity medical school in Japan. AnimalsForty adult mongrel dogs (weight, 7–15kg). InterventionsIn dogs, the lumbar nerve roots were compressed using 4 types of clips with different pressures. Changes of CGRP levels in the spinal dorsal horn, dorsal root, and dorsal root ganglia (DRG) were examined immunohistochemically after compression for 24 hours or for 1 week. Main outcome measuresCGRP-positive neurons and CGRP-positive fibers. ResultsAfter compression, axonal flow in the dorsal root was impaired, accumulation of CGRP was observed distal to the site of compression, and the number of DRG cells showing positively for CGRP decreased. Compression for 1 week resulted in a decrease in the number of CGRP-positive fibers in the spinal dorsal horn. ConclusionsThese findings indicate that CGRP dynamics are modified by nerve compression.

The influence of donor age, nerve growth factor, and cografting with drosophila cells on survival of peripherally grafted embryonic or fetal rat dorsal root ganglia.

Previous studies have shown that embryonic rat and human dorsal root ganglion (DRG) cells survive grafting to the cavity of extirpated adult rat DRG. Furthermore, grafted human embryonic neurons were shown to send axons peripherally and into the spinal cord, where they establish functional synaptic connections. This study analyzed the survival of orthotopically allografted rat DRG cells from embryonic stages 15 (E15) and 20 (E20), and the influence on their survival of nerve growth factor (NGF). NGF was delivered to the DRG transplants either by pump infusion or by cotransplantation of cells from Drosophila melanogaster, transgenic for human NGF. Lumbar DRGs of adult rats were removed and a collection of E15 or E20 DRGs placed in the cavity. One month after grafting the total number of DRG cells in the grafts was counted. Differentiation of subpopulations of DRG cells was estimated by counting cells immunostained for calcitonin gene-related peptide (CGRP), Griffonia simplicifolia agglutinin isolectin B4 (GSA), or heavy neurofilament protein (antibody RT97). The results show: i) similar survival of E15 and E20 grafts, with great variability in the survival of different subpopulations in E15 transplants, but a more consistent distribution of different phenotypes in E20 transplants; ii) infusion of NGF for 2 weeks increases the survival of E15 transplants, but has a negative effect on E20 transplants; iii) Drosophila cells transfected with human NGF gene survive peripheral xenografting and have a positive effect on the survival of the GSA- and CGRP-positive populations in E15 and E20 transplants; iv) Drosophila cells without the human NGF gene increase cell survival in E20 transplants. These data suggest that i) the effect of NGF is dependent on the embryonic stage of the transplants, ii) age-dependent sensitivity to NGF influences graft survival, and iii) transgenic Drosophila cells can be cotransplanted with embryonic neural tissue to the mammalian peripheral nervous system with a positive effect on the survival of neural grafts.

Effects of the capsaicin analogue resiniferatoxin on spinal nociceptive mechanisms in the rat: behavioral, electrophysiological and in situ hybridization studies

The effect of a single subcutaneous (s.c.) injection of the ultrapotent capsaicin analogue resiniferatoxin (RTX) on responses of adult rats to noxious thermal and mechanical stimulation was examined. The effects of RTX treatment on the nociceptive flexor reflex and activity-dependent increase in spinal excitability after conditioning C-fiber stimulation (CS) were also assessed. Finally, the expression of galanin message associated peptide (GMAP) mRNA in dorsal root ganglion (DRG) cells and the effects of the high affinity galanin receptor antagonist M35 on the flexor reflex in RTX-treated rats were evaluated. RTX, but not vehicle, produced marked thermal hypoalgesia on the hot plate test with partial recovery in about 50% of animals after about 2 weeks and no recovery in the remaining rats after 4 weeks. In all animals there was only a transient and moderate increase in paw withdrawal threshold to mechanical pressure. The flexor reflex in response to a C-fiber CS train was recorded 15–35 days after RTX or vehicle treatment. There was no difference between RTX and vehicle treated rats on baseline response, but RTX treatment lead to less wind-up during the CS and reduced hyperexcitability. This was particularly the case for rats which did not recover from RTX-induced hypoalgesia. The C-fiber mediated hyperexcitability was potentiated by the galanin receptor antagonist M35, more so in the non-recovered rats than in the partially recovered rats. The number of DRG cells expressing GMAP mRNA was significantly higher in non-recovered than in partially recovered rats. Thus, RTX produced marked and prolonged impairment of capsaicin-sensitive afferents and upregulation of the inhibitory neuropeptides GMAP and galanin in DRG neurons, which may underlie the prolonged effect of RTX.

Immunohistochemical localization of enkephalin in peripheral sensory axons in the rat

Dorsal root ganglia (DRG) contain measurable amounts of met-enkephalin (ENK), and a significant number of DRG cells contain mRNA for the manufacture of ENK. Yet almost no DRG cells are immunostained for ENK and dorsal rhizotomy does not diminish ENK staining in the dorsal horn. A hypothesis which would explain these seemingly discrepant results is the phenomenon of differential transport, where DRG cells making ENK rapidly transport the peptide only to their peripheral sensory axons. Evidence consistent with this hypothesis would be the demonstration of ENK-containing peripheral sensory axons. The present study demonstrates that approximately 17% of peripheral cutaneous axons label for ENK. The presence of a significant number of ENK-containing axons suggests an endogenous neural source of opiate ligand in the periphery and, in addition to ENK-containing inflammatory cells, this neural source may be functionally important in responses to physiologic as well as inflammatory pain.

Do Primary Afferent Cell Numbers Change in Relation to Increasing Weight and Surface Area in Adult Rats?

Adult mammalian primary afferent neurogenesis implies considerably more plasticity for the adult nervous system than is presently envisioned. One hypothesis is that such neurogenesis does not occur, because no tritiated-thymidine-labeled or mitosing neurons are observed in adults, and no increase in cells can be found in young as compared to old adults. The other hypothesis is that adult primary afferent neurogenesis does occur, but that it has not been observed because it is correlated with changes in the size of the animals. This has not been tested, and because of the implications of adult neurogenesis, it is important to see whether dorsal root ganglion (DRG) cell numbers increase with animal size. This is particularly pertinent because of recent advances in the technology for counting neurons and improved methods for resolving the cells in question. The present results indicate that DRG cell numbers are approximately the same in animals whose sizes are different by a factor of approximately 5. Thus our data are consistent with the hypothesis that numbers of adult DRG cells are stable, and we cannot demonstrate a change in relation to age or body size in our animals. Therefore, we do not find evidence of adult neurogenesis in our animals.

Expression of c-Jun as a response to dorsal root and peripheral nerve section in damaged and adjacent intact primary sensory neurons in the rat.

It was previously shown that the immediate early gene, c-jun, was highly expressed over long periods, in both peripheral sensory and motor neurons following axon damage or block of axoplasmic transport. Here we have examined the question of whether the expression of c-Jun protein is related to axon injury per se or to the process of axon growth. We have examined dorsal root ganglion (DRG) cells subjected to different manipulations which are associated with varying degrees of regrowth, as follows: (i) after peripheral nerve section, where it appears that all damaged neurons make some regenerative effort. 1-24 days after sciatic nerve section and ligation most cells in L4/L5 DRG were c-Jun-positive; (ii) after section of the central processes of the DRG cells, which then showed a slow and limited regrowth of their axons towards, but not into, the spinal cord. This resulted in a variable, but significant, expression of c-Jun in a small number of DRG cells; (iii) in intact sensory neurons that were offered the opportunity to sprout into adjacent denervated peripheral tissue. The sciatic nerve was ligated and the response of cells in the L3 ganglia (many of which project to the saphenous nerve) was measured. A small but significant number of cells were c-Jun-positive; (iv) in intact sensory neurons that were offered the opportunity to sprout centrally into partialy denervated neuropil of the spinal cord. We examined neurons in the L3 DRG after rhizotomy of the adjacent L4/L5 dorsal roots. Previous work suggests that sensory neurons show at best a very limited growth under these conditions.(ABSTRACT TRUNCATED AT 250 WORDS)

Respective involvement of lens capsule proteins in epithelial cell adhesion

The effects of forskolin on low-threshold tetrodotoxin-resistant (TTX-r) Na+ currents was studied in small diameter (average ≈ 25 μm) dorsal root ganglion (DRG) cells. All DRG cells included in the study were categorized as type-2 or non-type-2 based on the expression of a low-threshold A-current. In all type-2 and some non-type-2 DRG cells held at −80 mV, the adenylyl cyclase (AC) activator forskolin (10 μM) up-regulated TTX-r Na+ currents evoked with steps to −55 mV through −35 mV (low-threshold current). Up-regulation of low-threshold current by forskolin was mimicked by the protein kinase A (PKA) agonist Sp-cAMPs and the inflammatory mediator serotonin, and blocked by the PKA antagonist Rp-cAMPs. Forskolin-induced up-regulation of low-threshold current evoked from a holding potential of −60 mV was blocked by 40 ms steps to 0 mV, which presumably induced a long lasting inactivation of the low-threshold channels. Reducing to 3 ms the duration of steps to 0 mV, significantly increased the number of DRG cells where low-threshold current was up-regulated by forskolin, presumably by reducing the long-lasting inactivation of the low-threshold channels. In the same cells, high-threshold current, evoked by 40 ms or 3 ms steps to 0 mV, was consistently up-regulated by forskolin. The selective NaV1.8 channel blocker A-803467 markedly blocked high-threshold current but not low-threshold current. The different voltage protocols observed to activate and inactivate the low- and high-threshold currents, and the observation that A-803467 blocked high- but not low-threshold current suggests that the two currents were mediated by different channels, possibly NaV1.8 and NaV1.9, respectively. Inflammatory mediators may simultaneously up-regulate NaV1.8 and NaV1.9 channels in the same nociceptor via a AC/PKA signaling pathway, increasing nociceptor signaling strength, and lowering nociceptor threshold, respectively.

Absence of Neurogenesis of Adult Rat Dorsal Root Ganglion Cells

Recently, an age-related increase in the number of dorsal root ganglion (DRG) cells was reported in adult rats. This suggests neurogenesis of adult primary afferent neurons, which would be an extremely important phenomenon if it occurred. Other evidence is not compatible with this idea, however, so the issue is not settled. The primary point of contention concerns the counts of DRG cells in relation to age. In our opinion, these disagreements arise, at least in part, because different counting methods give different results for the same material. Thus, any method for determining DRG cell numbers should be calibrated. We previously calibrated some of the common methods used to count DRG cells and found that an empirical method gave accurate cell counts. In the present study, we have used this method and asked whether an age-related increase in the number of lumbar DRG cells can be demonstrated in adult rats. Our data indicate that DRG cell numbers remain essentially constant from 3 to 22 months of age. Most ancillary evidence is consistent with the hypothesis that mammalian DRG cell numbers do not change during adult life. Thus, we feel that the evidence does not support the hypothesis that there is neurogenesis of adult rat primary afferent neurons.

Selective neuronal destruction by Ricinus communis agglutinin I and its use for the quantitative determination of sciatic nerve dorsal root ganglion cell numbers

The selectivity of the neurotoxic lesion of Ricinus communis agglutinin I (RCAI) in rat dorsal root ganglia was examined. RCAI was injected in the sural nerve on one side. Two weeks later, the injected nerve, as well as the ipsilateral peroneal nerve, were examined in 1-μm-thick plastic embedded sections in the light microscope. The injected nerves showed a complete or almost complete Wallerian-like degeneration of myelinated fibers, but there were no signs of fiber damage in the uninjected nerves, which to a large extent originate in the same ganglia as the injected ones. We conclude that RCAI does not diffuse into and destroy ganglion cells adjacent to those that have transported the substance. We then used this selectivity in the effect of RCAI to determine indirectly the relative number of neurons in dorsal root ganglia L 4–L 6 which contribute to the sciatic nerve. Three weeks after unilateral injections of RCAI in the sciatic nerve, the L 4–L 6 dorsal root ganglion cells were counted bilaterally. On average, relative neuronal numbers between injected and uninjected sides were 0.36, 0.15 and 0.64 for L 4, L 5 and L 6 respectively. From these data we conclude that the sciatic nerve receives on average of 64%, 85% and 36%, respectively of its sensory contribution from these ganglia.

In vivo ANTI-NGF induces sprouting of sensory axons in dorsal roots.

Newborn rats were given subcutaneous injections of antibodies to mouse beta -NGF (ANTI-NGF) daily for 1 month. The number of neurons in T4-T6 dorsal root ganglia (DRG) and the numbers of myelinated and unmyelinated axons in the dorsal roots of the same segments were counted in the ANTI-NGF animals and in normal littermates. The ANTI-NGF rats had 38% fewer neurons in thoracic ganglia but 17% more myelinated and 40% more unmyelinated fibers than their untreated littermates. Dorsal root ganglion cells also have a larger average size in the ANTI-NGF animals, which we interpret as a disproportionate loss of small cells. These data are interpreted as showing that some dorsal root ganglion cells, principally small ones, die when endogenous NGF is inactivated, and that the remaining cells emit more processes than normal. Thus, removal of NGF has what appears to be a paradoxical effect, a reduction in dorsal root ganglion cell numbers but an increase in dorsal root axon numbers. The relation of myelin thickness to fiber diameter is also altered, with small fibers being more thinly myelinated in the ANTI-NGF group. Thus, Schwann cell-neuronal interactions are also affected by inactivation of NGF.

Numbers of rat dorsal root axons and ganglion cells during postnatal development

The present study demonstrates that T4 and S2 rat dorsal root axons decrease significantly from birth to adulthood with almost all of the decrease occurring in the first two weeks of life. Dorsal root ganglion cell numbers do not change during this time period. This is thus an example of postnatal axon elimination not associated with death of the cells that give rise to the axons. Presumably this regressive process is important in the formation of the normal adult nervous system. In addition, these findings raise the possibility that certain types of neonatal denervation may increase adult axon numbers by stopping a regressive process, the loss of axons, rather than initiating a progressive process, the formation of new axons.

Regulation of neuron numbers in Xenopus laevis: effects of hormonal manipulation altering size at metamorphosis.

Xenopus laevis tadpoles reared in a 0.01% solution of 6-n-propyl-2-thiouracil (PTU) are blocked in their development at larval stage 54 but continue to increase in size. When released from the effects of PTU they metamorphose into frogs of sizes significantly larger than those of their untreated siblings. Using this size difference to examine the hypothesis that neuron numbers are matched to the size of their postsynaptic targets during neuronal cell death, we measured the following on stage 66 frogs metamorphosing from PTU-treated and untreated tadpoles: lumbar lateral motor column (L-LMC) motoneuron number and mean nuclear cross-sectional area; thoracic and lumbar dorsal root ganglion (DRG) cell number and mean nuclear cross-sectional area; and muscle fiber number in two representative thigh muscles. A few measurements of neuron number and cell size were also made on untreated and PTU-treated stage 54 tadpoles. The most striking correlations observed were not between peripheral size and neuron numbers but between peripheral size and neuron size. Motoneuron numbers were not increased in the PTU-treated animals, perhaps because the increase in peripheral size involved an increase in muscle fiber diameter rather than an increase in muscle fiber number. Thoracic DRG cell number, but not the sum of thoracic and lumbar DRG cell numbers, was increased. In general, our findings do not support the hypothesis that neuron numbers are matched to peripheral size by a process regulating the amount of cell death that occurs during metamorphic stages in Xenopus laevis.