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. J. Comp. Neurol. 386:8–15, 1997. © 1997 Wiley‐Liss, Inc.

[1]  M. Devor,et al.  Neurogenesis in adult rat dorsal root ganglia: on counting and the count. , 1991, Somatosensory & motor research.

[2]  C. Lois,et al.  Long-distance neuronal migration in the adult mammalian brain. , 1994, Science.

[3]  M. Devor,et al.  Neurogenesis in adult rat dorsal root ganglia , 1985, Neuroscience Letters.

[4]  T M Mayhew,et al.  A review of recent advances in stereology for quantifying neural structure , 1992, Journal of neurocytology.

[5]  S. Bayer,et al.  Neurons in the rat dentate gyrus granular layer substantially increase during juvenile and adult life. , 1982, Science.

[6]  S. Hatai Number and size of the spinal ganglion cells and dorsal root fibers in the white rat at different ages , 1902 .

[7]  B. Konigsmark,et al.  Methods for counting neurons , 1970 .

[8]  R. Coggeshall,et al.  Methods for determining numbers of cells and synapses: A case for more uniform standards of review , 1996, The Journal of comparative neurology.

[9]  R. Coggeshall,et al.  Do primary afferent cell numbers change in relation to increasing weight and surface area in adult rats? , 1994, Somatosensory & motor research.

[10]  C. Saper,et al.  Any way you cut it: A new journal policy for the use of unbiased counting methods , 1996, The Journal of comparative neurology.

[11]  R. Coggeshall,et al.  Absence of neurogenesis of adult rat dorsal root ganglion cells. , 1991, Somatosensory & motor research.

[12]  H. J. G. Gundersen,et al.  The new stereological tools: Disector, fractionator, nucleator and point sampled intercepts and their use in pathological research and diagnosis , 1988, APMIS : acta pathologica, microbiologica, et immunologica Scandinavica.

[13]  D. C. Sterio The unbiased estimation of number and sizes of arbitrary particles using the disector , 1984, Journal of microscopy.

[14]  Richard E. Coggeshall,et al.  A consideration of neural counting methods , 1992, Trends in Neurosciences.

[15]  Ralph Norcio,et al.  The organization of neuronal somata in the first sacral spinal ganglion of the cat , 1976, Experimental Neurology.

[16]  R. Coggeshall,et al.  Dorsal root ganglion cell death and surviving cell numbers in relation to the development of sensory innervation in the rat hindlimb. , 1994, Brain research. Developmental brain research.

[17]  I. Hardesty On the number and relations of the ganglion cells and medullated nerve fibers in the spinal nerves of frogs of different ages , 1905 .

[18]  Mark J. West,et al.  New stereological methods for counting neurons , 1993, Neurobiology of Aging.

[19]  M. Devor,et al.  Proliferation of primary sensory neurons in adult rat dorsal root ganglion and the kinetics of retrograde cell loss after sciatic nerve section. , 1985, Somatosensory research.

[20]  J. Peyronnard,et al.  Three‐dimensional computer‐aided analysis of the intraganglionic topography of primary muscle afferent neurons in the rat , 1990, The Anatomical record.

[21]  J. Ygge,et al.  Asymmetries and symmetries in the number of thoracic dorsal root ganglion cells , 1981, The Journal of comparative neurology.

[22]  S. Weiss,et al.  A multipotent EGF-responsive striatal embryonic progenitor cell produces neurons and astrocytes , 1992, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[23]  P. Graziadei,et al.  Neurogenesis and Plasticity of the Olfactory Sensory Neurons a , 1985, Annals of the New York Academy of Sciences.

[24]  P. B. Farel,et al.  Size-related increase in motoneuron number: evidence for late differentiation. , 1993, Brain research. Developmental brain research.

[25]  R. Coggeshall,et al.  The determination of an empirical correction factor to deal with the problem of nucleolar splitting in neuronal counts , 1984, Journal of Neuroscience Methods.

[26]  G J Popken,et al.  Reliability and validity of the physical disector method for estimating neuron number. , 1996, Journal of neurobiology.

[27]  G. Krauthamer,et al.  The organization of visceral sensory neurons in thoracic dorsal root gangla (DRG) of the cat studied by horseradish peroxidase (HRP) reaction using the cryostat , 1981, Brain Research.

[28]  Principal neurons of the lumbar sympathetic ganglia increase in number with body size , 1995, The Journal of comparative neurology.

[29]  P. Farel Naturally occurring cell death and differentiation of developing spinal motoneurons following axotomy , 1989, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[30]  P. B. Farel,et al.  Hindlimb sensory neuron number increases with body size , 1994, The Journal of comparative neurology.