Neuron addition and neurogenesis are not interchangeable terms.
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Animals↗
Biomedical subjects
P B Farel
Publications and source records attributed to P B Farel.
Developmental regulation of sensory neuron number and limb innervation in the mouse.
Although used widely in studies of naturally occurring cell death, systematic descriptions of the time course of changes in sensory neuron number and of limb innervation in the mouse are not available. The development of sensory innervation to the mouse forelimb was traced using the lipophilic carbocyanine dye, DiI, and correlated with neuron number in dorsal root ganglia contributing to the cervical enlargement. Axon invasion of the forelimb began at E10.5. Sensory axons reached the distal margin of the forelimb by E13.5. The difficulty of identifying immature neurons precluded estimating neuron numbers during the period of limb innervation. Neuron numbers in dorsal root ganglia (DRGs) C5-C8 increased from E14 to E16 and from E18 to P4. No evidence of a decline in neuron numbers was found during the developmental periods studied. Neuron number was compared in neonates and adults to determine if sensory neurons are added as body size increases as found in the frog [J. Comp. Neurol. 314 (1991) 106] and the rat [J. Comp. Neurol. 386 (1997) 8]. In contrast to previous findings, no difference was found in sensory neuron number between neonate and adult mice in either cervical or lumbar DRGs.
Animals↗
Neuron addition and enlargement in juvenile and adult animals.
Locomotion requires bilateral symmetry of neural circuitry in the spinal cord. Although not well understood, the mechanisms responsible for establishing and maintaining this symmetry must balance the numbers, sizes, and connectivity of the neurons on both sides of the spinal cord. Those mechanisms do not cease to function after embryogenesis, since there is substantial evidence that these properties continue to change as juvenile animals grow to adult size. We review the evidence that spinal neuron number and size increase in growing juvenile frogs and mammals. We postulate that these increases are regulated by both local and systemic factors. In addition, we discuss evidence that axotomy of spinal sensory and motor neurons also enlists local and systemic regulatory factors, some of which may also be operative in normal growth and development.
Age Factors↗
University of North Carolina at Chapel Hill School of Medicine.
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Curriculum↗
Transient effects of nerve injury on estimates of sensory neuron number in juvenile bullfrog.
The effect of lumbar spinal nerve (SN) transection on estimates of neuron number was investigated in the dorsal root ganglia (DRGs) of juvenile bullfrogs (Rana catesbeiana). SN8 and SN10 were transected on one side, and SN9 was left intact. Two weeks after nerve injury, estimates of neuron number in DRG8 and DRG10 on both the operated and unoperated sides were more than twice the estimates obtained from control animals. Neuron number in the uninjured DRG9 was also elevated relative to that of control animals. Eight weeks after axotomy, differences in neuron number were less apparent. The mean cross-sectional area of DRG neurons was reduced 2 weeks after nerve injury in all DRGs. The decrease in mean area was the result of the addition of neurons to the smallest size classes. These data are discussed in the context of previous results showing that neurons are added as juvenile frogs grow to adult size (St. Wecker and Farel [1994] J. Comp. Neurol. 342:430-438). This addition results from the maturation of a population of incompletely differentiated neurons (Meeker and Farel [1997] J. Comp. Neurol. 389:569-576). The present results suggest that axotomy precipitates differentiation of these incompletely differentiated neurons, perhaps as a compensatory response to nerve injury.
Age Factors↗
Neuron addition during growth of the postmetamorphic bullfrog: sensory neuron and axon number.
Neuron addition is one means whereby the nervous system can compensate for increased body size. Neurons can be added either by mitosis of stem cells or by late differentiation of committed precursors. Previously, the doubling of hind limb dorsal root ganglion (DRG) neurons in postmetamorphic bullfrogs (Rana catesbeiana) was found to occur in the absence of neuron proliferation (St. Wecker and Farel [1994] J. Comp. Neurol. 342:430-438). In the present study, we identify a population of cells in the DRGs of juvenile frogs that lack the appearance typical of sensory neurons yet are immunoreactive to a neuron-specific probe for neurofilament protein. These less differentiated (type-L neurons) could not be labeled retrogradely with horseradish peroxidase from the periphery or dorsal root. Despite their apparent immaturity, type-L neurons appear to have extended axons both centrally and toward the periphery, because axon number in dorsal roots and peripheral nerves was similar in juvenile and adult frogs. These findings are consistent with the existence in juvenile frogs of a population of incompletely differentiated DRG neurons that lack the physiological properties and appearance typical of mature neurons.
Aging↗
Sensory neuron number in neonatal and adult rats estimated by means of stereologic and profile-based methods.
Postnatal neuron addition, if it occurred, would have profound implications both for the conceptualization of developmental processes and for efforts directed at replacing neurons that were lost to injury or disease. Although dorsal root ganglia (DRGs) offer the advantages of clear boundaries and functional homogeneity, studies comparing neuron number in the DRGs of animals of different ages or sizes have yielded conflicting results. In the present study, neuron number in DRGs L3-L6 was compared in neonatal (approximately 11 days old, mean weight of 24.5 g, mean volume of 25 cm3) and adult (approximately 80 days old, mean weight of 373.5 g, mean volume of 346 cm3) male Sprague-Dawley rats. Estimates of neuron number were derived by using both stereological (physical disector) and profile-counting (one or more nucleoli within a nucleus) methods. The reliability and validity of the two methods were evaluated by comparing estimates of neuron number with those derived from three-dimensional reconstruction of a subset of neurons. The recommended protocol for using the physical disector was found to give accurate estimates of neuron number, but the heterogeneous distribution of neurons in the ganglion led to sampling errors of up to 50%. Reliability was improved by increasing the number of disector pairs examined. Counts of nuclear/nucleolar profiles were more reliable, but introduced a bias that worked against the experimental hypothesis in that estimates of neuron number in neonates exceeded actual values. Nonetheless, both methods indicated that adult rats had more DRG neurons than did neonates. Profile counts were 19% higher in adults (P < .01, two-tailed t-test); and data obtained by using the physical disector showed that adult rats had 28% more neurons than did neonates (P < .05). The difference in neuron number between adults and neonates could be due either to neuron proliferation or to late differentiation of neurons that do not assume a typical appearance until adulthood.
Aging↗
Reliability and validity of the physical disector method for estimating neuron number.
The physical disector was proposed as an unbiased and efficient means to estimate neuron number; however, the validity and reliability of this method have been examined only infrequently. Estimates of neuron number in the dorsal root ganglia (DRG) of bullfrogs (Rana catesbeiana) were compared to nucleolar counts based on 3-dimensional reconstructions. Accuracy of disector estimates were not affected by size of the animal. Similarly, disector estimates were not systematically altered when area measurements were limited to cellular regions of the DRG versus inclusion of the entire cross-sectional area. However, the recommended protocol for applying the disector resulted in sampling errors that introduced considerable variability in repeated estimates of neuron number from a single ganglion. In addition to this lack of reliability, disector estimates were consistently lower than those obtained by means of a nucleolar counting method that was calibrated against 3-dimensional reconstructions of neuronal profiles. The systematic error of the disector method was greater when ganglia were cut parallel to the long axis of the DR than when they were cut perpendicular to this axis. Increasing the sample size beyond what was recommended increased the reliability of estimates obtained with the disector; however, the bias associated with the plane of section was not reduced. These results emphasize the need for empirical validation of methods used to estimate neuron number in the tissue to which they are to be applied.
Animals↗
Principal neurons of the lumbar sympathetic ganglia increase in number with body size.
Neuron number appears to be matched to body size during early development by the modulation of the processes of proliferation and naturally occurring cell death. However, body size increases rapidly as the juvenile becomes an adult, long after these processes cease to operate. The present study shows that principal neurons of lumbar sympathetic ganglia increase in number four- to fivefold during postmetamorphic life of the bullfrog. Rana catesbeiana. This increase in neuron number cannot be attributed to either counting error or selection bias and was associated with greater innervation of particular hindlimb targets, as demonstrated by retrograde labeling with horseradish peroxidase. Injection of [3H]thymidine (a marker of DNA synthesis) every third day for 20-22 weeks failed to provide evidence of neuron proliferation, although, on the basis of changes in body length during this period, substantial numbers of neurons likely were added. These results combined with previous studies of hindlimb motor and sensory neuron addition are consistent with the hypothesis that the population of sympathetic neurons is augmented by late differentiation of existing precursor cells.
Animals↗
Hindlimb sensory neuron number increases with body size.
As an animal grows, its sensory systems face the task of maintaining sensitivity and discrimination in peripheral fields that are continually enlarging. Without the addition of neurons, existing cells would have to innervate a wider skin area, leading to a decrease in the precision with which stimuli are localized. Neurons were counted in the three dorsal root ganglia (DRGs) that innervate the hindlimb of the bullfrog (Rana catesbeiana). Profiles of neuronal nuclei containing the single nucleolus found in these cells were counted in every third section of serially cut ganglia. This means of assessing neuron number was validated by comparing these profile counts with three-dimensional reconstructions of sensory neurons. Large frogs (10-17 cm) had more than twice as many DRG neurons as small frogs (3.3-5 cm). The rate of increase was greatest between 3 and 8 cm, when over 1,300 hindlimb sensory neurons were added for each 1 cm increase in body length. The possibility that selective survival of frogs with many neurons biases estimates of mean neuron number was ruled out by the finding that frogs drawn from the same closed population, half of which were sacrificed immediately and half of which were sacrificed after 1 year's survival, showed expected differences in neuron number. Horseradish peroxidase applied to particular hindlimb nerves retrogradely labeled more neurons in large frogs than small frogs, supporting the hypothesis that added neurons extend their axons to the periphery.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
Coincidence of Schwann cell-derived basal lamina development and loss of regenerative specificity of spinal motoneurons.
The Schwann cell-derived basal lamina forms a tube around single peripheral axons or small groups of axons that is continuous from the spinal cord to the target. In bullfrog tadpoles (Rana catesbeiana), motor axons transected at early developmental stages regenerate to the appropriate hindlimb region. In the present paper, we found that at these stages Schwann tubes are absent by morphological criteria, and individual axons are separated only by occasional extensions of support cells. At stages when axons no longer regenerate to the correct hindlimb region, every axon is encased in a basal lamina tube. Schwann tubes persist in the distal stump after nerve transection, and regenerating axons grow within these tubes. These findings are consistent with previous results showing that the errors regenerating axons make in older animals are not random, but depend upon the course of the denervated Schwann tubes to which they have access. In order to determine whether formation of the Schwann tube itself or interaction of its molecular constituents with growing axons was associated with loss of regenerative specificity, the expression during development of two major constituents of the basal lamina, laminin and heparan sulfate proteoglycan, was investigated. Immunoreactivity to both constituents was present both before and after the transition from specific to nonspecific regeneration, indicating that their expression per se was not sufficient to limit regenerative specificity. These data support the hypothesis that the physical constraint imposed by the Schwann cell-derived basal lamina prevents regenerative specificity.
Animals↗
Size-related increase in motoneuron number: evidence for late differentiation.
The number of motoneurons in the lumbar lateral motor column (LMC) was compared in bullfrogs (Rana catesbeiana) ranging in body length from 2.5 to 19 cm. Large frogs had 36% more motoneurons than small frogs; however, within the caudal third of the LMC, large frogs had over 70% more motoneurons than small frogs. Injection of small frogs with [3H]thymidine every third day for 20-22 weeks gave no evidence of motoneuron birth. Instead, a pool of small, incompletely differentiated (type L) motoneurons appears to be converted into mature (type M) motoneurons as the animal grows. This hypothesis is supported by several lines of evidence: (1) the number of type-M motoneurons varies directly with body size while the number of type-L cells varies inversely; (2) the increase in type-M motoneurons and the decrease in type-L cells are restricted to the same regions of the LMC; and (3) type-L cells exhibited both immunoreactivity to neurofilament antibodies and histochemical evidence of acetylcholinesterase activity, a marker for spinal motoneurons.
Acetylcholinesterase↗
Developmental regulation of regenerative specificity in the bullfrog.
The accuracy with which motor axons reinnervate the hindlimb following ventral rhizotomy was studied in the bullfrog (Rana catesbeiana) by mapping the locations within the lumbar lateral motor column of motoneurons retrogradely labeled with horseradish peroxidase. At early stages of development, axons regenerate to the correct hindlimb region. In older tadpoles and frogs, axons appear to regenerate to targets determined by basal lamina tubes, which persist in the distal nerve stump after the severed nerve fiber degenerates. Basal lamina tubes develop at the stages when regenerative specificity is no longer expressed. These results led to the hypothesis that guidance cues may be present in older animals, and axons may be capable of responding to these cues, but are prevented from doing so because they are confined in basal lamina tubes. Results obtained from crosstage hindlimb transplants are consistent with this hypothesis.
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Neuromuscular specificity following cross-stage hindlimb transplantation.
In order to determine whether spinal motoneurons can regenerate to their proper targets at stages beyond those when such specificity is typically expressed, autologous and homologous (same- and cross-stage) hindlimb transplantations were performed using bullfrog tadpoles (Rana catesbeiana). Neuromuscular specificity was assessed by applying horseradish peroxidase to the ventral thigh of the transplanted hindlimb and mapping the locations of retrogradely labeled motoneurons. Previously, we found that the hindlimb was reinnervated normally in young tadpoles whose motor nerves were transected. However, motor axons in older animals showed no evidence of target specificity when reinnervating the tadpole's own limb. In the present study, innervation was normal in young tadpoles whose hindlimb was removed and replaced in its original position. Axons of older hosts innervating hindlimbs transplanted from young tadpoles distinguished flexor from extensor limb regions, but failed to distinguish thigh from shank, demonstrating that axons in older animals can respond to at least some guidance cues. The lack of specificity along the proximal-distal axis appears to be a consequence of homologous transplantation since limbs transplanted from one young tadpole to another showed the same loss of proximal-distal specificity. One possibility is that the shank degenerates when the hindlimb is transplanted to a different host. Shank motoneurons, lacking their proper target, may then innervate the thigh. However, shank motoneurons did not innervate the thigh when motor nerve transection was combined with amputation of the hindlimb just above the presumptive knee.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
Neuron addition in the postmetamorphic frog.
Neuron number among somatic motoneurons, sensory neurons, and sympathetic postganglionic neurons that innervate the hindlimb was correlated with body length in the bullfrog, Rana catesbeiana. Two to three times more dorsal root and sympathetic ganglion neurons are found in the largest than the smallest specimens. Hindlimb motoneurons show a 20% increase in number, but this increase is restricted to the caudal third of the motor pool. Within this region, 60% more motoneurons are found among the largest frogs. Cell division does not appear to be the mechanism of neuron addition. Instead, we propose that a pool of undifferentiated neurons mature to maintain functional capabilities as the animal increases in size.
Animals↗
Apoptosis: a general comment.
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Cell Survival↗
Mauthner cells maintain their lumbar projection in adult frog.
We have used retrograde labeling with horseradish peroxidase (HRP) in the bullfrog. Rana catesbeiana, to determine whether Mauthner (M) cells maintain a projection to the lumbar spinal cord in adult bullfrogs. We found that M cells persist in the adult bullfrog and maintain a projection to the lumbar spinal cord, despite the degeneration of much of their afferent input and of their motoneuronal targets in the spinal cord.
Animals↗
Regenerative specificity of motor axons when reinnervation is partially suppressed.
We asked whether regenerating hindlimb motor axons would innervate inappropriate hindlimb regions if competition from appropriate innervation were prevented. The three ventral roots that innervate the hindlimb in the bullfrog (Rana catesbeiana) tadpole were transected, and the two more rostral roots were ligated to prevent regeneration. The most caudal root, which primarily supplies more distal limb musculature in unoperated tadpoles, was left free to regenerate. The specificity of regeneration was assessed by retrogradely labeling spinal motoneurons with HRP placed in the ventral thigh, a region that receives most of its innervation from the ligated roots. Despite the lack of competition from appropriate innervation, the regenerating root did not provide substantial innervation to proximal limb musculature. The same result was obtained in tadpoles operated upon at stages when regeneration of motor axons is specific and in tadpoles at stages when regenerating motor axons do not reinnervate their appropriate targets (Farel and Bemelmans, 1986), although the mechanisms in each case are likely different.
Animals↗