Role of synaptic inputs in determining input resistance of developing brain stem motoneurons.

The contribution of synaptic input to input resistance was examined in 208 developing genioglossal motoneurons in 3 postnatal age groups (5-7 day, 13-16 day, and 18-24 day) using sharp electrode recording in a slice preparation of the rat brain stem. High magnesium (Mg(2+); 6 mM) media generated significant increases (21-38%) in both the input resistance (R(n)) and the first time constant (tau(0)) that were reversible. A large percent of the conductance blocked by high Mg(2+) was also sensitive to tetrodotoxin (TTX). Little increase in resistance was attained by adding blockers of specific amino acid (glutamate, glycine, and GABA) transmission over that obtained with the high Mg(2+). Comparing across age groups, there was a significantly larger percent change in R(n) with the addition of high Mg(2+) at postnatal days 13 to 15 (P13-15; 36%) than that found at P5-6 (21%). Spontaneous postsynaptic potentials were sensitive to the combined application of glycine receptor antagonist, strychnine, and the GABA(A) receptor antagonist, bicuculline. Application of either 10 microM strychnine or bicuculline separately produced a reversible increase in both R(n) and tau(0). Addition of 10 microM bicuculline to a strychnine perfusate failed to further increase either R(n) or tau(0). The strychnine/bicuculline-sensitive component of the total synaptic conductance increased with age so that this form of neurotransmission constituted the majority (>60%) of the observed percent decrease in R(n) and tau(0) in the oldest age group. The proportion of change in tau(0) relative to R(n) following strychnine or high magnesium perfusate varied widely from cell to cell and from age to age without pattern. Based on a model from the literature, this pattern indicates a nonselective distribution of the blocked synaptic conductances over the cell body and dendrites. Taken together, the fast inhibitory synapses (glycine, GABA(A)) play a greater role in determining cell excitability in developing brain stem motoneurons as postnatal development progresses. These findings suggest that synaptically mediated conductances effect the membrane behavior of developing motoneurons.

Role of potassium conductances in determining input resistance of developing brain stem motoneurons.

The role of potassium conductances in determining input resistance was studied in 166 genioglossal (GG) motoneurons using sharp electrode recording in brain stem slices of the rats aged 5-7 days, 13-15 days, and 19-24 days postnatal (P). A high magnesium (Mg(2+); 6 mM) perfusate was used to block calcium-mediated synaptic release while intracellular or extracellular cesium (Cs(+)) and/or extracellular tetraethylammonium (TEA) or barium (Ba(2+)) were used to block potassium conductances. In all cases, the addition of TEA to the high Mg(2+) perfusate generated a larger increase in both input resistance (R(n)) and the first membrane time constant (tau(0)) than did high Mg(2+) alone indicating a substantial nonsynaptic contribution to input resistance. With intracellular injection of Cs(+), GG motoneurons with lower resistance (<40 MOmega), on the average, showed a larger percent increase in R(n) than cells with higher resistance (>40 MOmega). There was also a significant increase in the effect of internal Cs(+) on R(n) and tau(0) with age. The largest percent increase (67%) in the tau(0) due to intracellular Cs(+) occurred at P13-15, a developmental stage characterized by a large reduction in specific membrane resistance. Addition of external Cs(+) blocked conductances (further increasing R(n) and tau(0)) beyond those blocked by the TEA perfusate. Substitution of external calcium with 2 mM barium chloride produced a significant increase in both R(n) and tau(0) at all ages studied. The addition of either intracellular Cs(+) or extracellular Ba(2+) created a depolarization shift of the membrane potential. The amount of injected current required to maintain the membrane potential was negatively correlated with the control R(n) of the cell at most ages. Thus low resistance cells had, on the average, more Cs(+)- and Ba(2+)-sensitive channels than their high resistance counterparts. There was also a disproportionately larger percent increase in tau(0) as compared with R(n) for both internal Cs(+) and external Ba(2+). Based on a model by Redman and colleagues, it might be suggested that the majority of these potassium conductances underlying membrane resistance are initially located in the distal dendrites but become more uniformly distributed over the motoneuron surface in the oldest animals.

Time-dependent changes in input resistance of rat hypoglossal motoneurons associated with whole-cell recording.

The effect of cellular dialysis associated with whole-cell recording was studied in 24 developing hypoglossal motoneurons in a rat brainstem slice preparation. In all cases, establishing whole-cell continuity with the electrode solution resulted in an increase in the input resistance measured in current clamp. The mean magnitude of this increase was 39.7% and the time course of the maximum effect was quite variable. There was no correlation found between the time to maximum effect and the magnitude of the increase in resistance. These data indicate that the passive membrane properties are not constant during whole-cell recording in mammalian motoneurons.

Physiological changes accompanying anatomical remodeling of mammalian motoneurons during postnatal development.

The development of respiratory motoneurons provides unique data that may be generalized to other mammalian motoneuron populations. Like other motoneurons, respiratory motoneurons undergo developmental changes in the shape of the action potential and their repetitive firing. The unique observations concern the postnatal change in the recruitment pattern of cat phrenic motoneurons that is correlated with a halving of mean input resistance, a stasis of growth in the cell membrane and a reduction in the complexity of the dendritic tree. A similar pattern of change was observed for hypoglossal motoneurons studied in rat brainstem slices. Without an increase in total membrane surface area, the decreased resistance must result from a reduced specific membrane resistance. Two mechanisms are proposed to explain this decrease in resistance: proliferation and redistribution of either synaptic inputs and/or potassium channels. Although there was a significant contribution of synaptic input in determining input resistance throughout postnatal development, it was the density of cesium- or barium-sensitive potassium conductances that differentiated low resistance from high resistance motoneurons. Low resistance motoneurons had more cesium- and barium-sensitive channels than their high resistance counterparts. Based on the variations in the relative changes observed in input resistance versus membrane time constant with these two potassium channel blockers (cesium and barium), it is proposed that the distribution of these potassium channels change with age. Initially, their distribution is skewed toward the dendrites but as development progresses, the distribution becomes more uniform across the motoneuron membrane. During postnatal development, the rapid decrease in input resistance results from a proliferation of potassium channels in the membrane and of synaptic inputs converging onto developing respiratory motoneurons while the membrane is being spatially redistributed but not expanded.

Relationship between membrane properties and cell size of developing rat genioglossal motoneurons studied in vitro.

Electrical properties and morphology of 21 genioglossal motoneurons were measured in a slice preparation of the rat brainstem at four different postnatal ages. The motoneurons labeled with neurobiotin were reconstructed and quantified in three-dimensional space. There was no strong correlation found between the input resistance or membrane time constant and the total membrane surface area. We conclude that there is no electrical property of these developing motoneurons that can accurately predict their anatomical size.

Dendritic morphology and its effects on the amplitude and rise-time of synaptic signals in hippocampal CA3 pyramidal cells.

Detailed anatomical analysis and compartmental modeling techniques were used to study the impact of CA3b pyramidal cell dendritic morphology and hippocampal anatomy on the amplitude and time course of dendritic synaptic signals. We have used computer-aided tracing methods to obtain accurate three-dimensional representations of 8 CA3b pyramidal cells. The average total dendritic length was 6,332 +/- 1,029 microns and 5,062 +/- 1,397 microns for the apical and basilar arbors, respectively. These cells also exhibited a rough symmetry in their maximal transverse and septotemporal extents (311 +/- 84 microns and 269 +/- 106 microns). From the calculated volume of influence (the volume of the neuropil from which the dendritic structures can receive input), it was found that these cells show a limited symmetry between their proximal apical and basilar dendrites (2.1 +/- 1.2 x 10(6) microns 3 and 3.5 +/- 1.1 x 10(6) microns 3, respectively). Based upon these data, we propose that the geometry of these cells can be approximated by a combination of two cones for the apical arbor and a single cone for the basilar arbor. The reconstructed cells were used to build compartmental models and investigate the extent to which the cellular anatomy determines the efficiency with which dendritic synaptic signals are transferred to the soma. We found that slow, long lasting signals show only approximately a 50% attenuation when they occur in the most distal apical dendrites. However, synaptic transients similar to those seen in fast glutamatergic transmission are transferred much less efficiently, showing up to a 95% attenuation. The relationship between the distance along the dendrites and the observed attenuation for a transient is described simply by single exponential functions with parameters of 195 and 147 microns for the apical and basilar arbors respectively. In contrast, there is no simple relation that describes how a transient is attenuated with respect to these cells' stratified inputs. This lack of a simple relationship arises from the radial orientation of the proximal apical and basilar dendrites. When combined, the anatomical and modeling data suggest that a CA3b cell can be approximated in three dimensions as the combination of three cones. The amplitude and time-course for a synaptic transient can then be predicted using two simple equations.

Morphology of developing rat genioglossal motoneurons studied in vitro: relative changes in diameter and surface area of somata and dendrites.

This study describes the postnatal change in size of motoneurons in the hypoglossal nucleus that innervate the genioglossus muscle. Such anatomical information is essential for determining the cellular mechanisms responsible for the changes observed in the electrical properties of these motoneurons during postnatal development. The cells analyzed here are part of an earlier study (Núñez-Abades et al. [1994] J. Comp. Neurol. 339:401-420) where 40 genioglossal (GG) motoneurons from four age groups (1-2, 5-6, 13-15, and 19-30 postnatal days) were labeled by intracellular injection of neurobiotin in an in vitro slice preparation of the rat brainstem and their cellular morphology was reconstructed in three-dimensional space. The sequence of postnatal dendritic growth can be described in two phases. The first phase, between birth (1-2 days) and 13-15 days, was characterized by no change in either dendritic diameter (any branch order) or dendritic surface area of GG motoneurons. However, maturation of the dendritic tree produced more surface area at greater distances from the soma by redistributing existing membrane (retracting some terminal branches). During the second phase, between 13-15 days and 19-30 days, the dendritic surface area doubled as a result of an increase in the dendritic diameter across all branch orders and a generation of new terminal branches. In contrast to the growth exhibited by the dendrites, there was little discernible postnatal growth of somata. At all ages, dendrites of GG motoneurons show the largest amount of tapering in the first-and second-order dendrites. The calculated dendritic trunk parameter deviated from a value 1.0, indicating that the dendritic tree of developing GG motoneurons cannot be modeled accurately as an equivalent cylinder. However, the value of this parameter increased with age. Strong correlations were found between the diameter of the first-order dendrite and the dendritic surface area, dendritic volume, combined dendritic length, and, to a lesser extent, the number of terminal dendrites in GG motoneurons. Correlations were also found between somal and dendritic geometry but only when data were pooled across all age groups. These data support earlier studies on kitten phrenic motoneurons, which concluded that postnatal growth of motoneurons was not a continuous process. Based on the fact that there was no growth in the first 2 weeks, the changes in the membrane properties described during this phase of postnatal development (e.g., decrease in input resistance) cannot be attributed to increases in the total membrane surface area of these motoneurons.(ABSTRACT TRUNCATED AT 400 WORDS)

Morphology of developing rat genioglossal motoneurons studied in vitro: changes in length, branching pattern, and spatial distribution of dendrites.

The aim of this study is to describe the postnatal change in dendritic morphology of those motoneurons in the hypoglossal nucleus that innervate the genioglossus muscle. Forty genioglossal (GG) motoneurons from four age groups (1-2, 5-6, 13-15, and 19-30 postnatal days) were labeled by intracellular injection of neurobiotin in an in vitro slice preparation of the rat brainstem and were reconstructed in three-dimensional space. The number of primary dendrites per GG motoneuron was approximately 6 and remained unchanged with age. The development of these motoneurons from birth to 13-15 days was characterized by a simplification of the dendritic tree involving a decrease in the number of terminal endings and dendritic branches. Motoneurons lost their 6th-8th order branches, in parallel with an elongation of their terminal dendritic branches maintaining the same combined dendritic length. The elongation of terminal branches was attributed to both longitudinal growth and the apparent lengthening caused by resorption of distal branches. The elimination of dendritic branches tended to increase the symmetry of the tree, as revealed by topological analysis. Later, between 13-15 days and 19-30 days, there was a reelaboration of the dendritic arborization returning to a configuration similar to that found in the newborn. The length of terminal branches was shorter at 19-30 days, while the length of preterminal branches did not change, suggesting that the proliferation of branches at 19-30 days takes place in the intermediate parts of terminal branches. The three-dimensional distribution of dendrites was analyzed by dividing space into six equal volumes (hexants). This analysis revealed that GG motoneurons have major components of their dendritic tree oriented in the lateral, medial, and dorsal hexants. Further two-dimensional polar analysis (consisting of eight sectors) revealed a reconfiguration of the tree from birth up to 5-6 days involving resorption of dendrites in the dorsal, dorsomedial, and medial sectors and growth in the lateral sector. Later in development (between 13-15 days and 19-30 days), there was growth in all sectors, but of a greater magnitude in the dorsomedial, medial, and dorsolateral sectors.

In vitro electrophysiology of developing genioglossal motoneurons in the rat.

1. Experiments were performed to determine the change in membrane properties of genioglossal (GG) motoneurons during development. Intracellular recordings were made in 127 GG motoneurons from rats postnatal ages 1-30 days. 2. The input resistance (R(in)) and the membrane time constant (t(aum)) decreased between 5-6 and 13-15 days from 84.8 +/- 25.4 (SD) to 47.0 +/- 18.9 M omega (P < 0.01) and from 10.0 +/- 4.2 to 7.3 +/- 3.3 ms (P < 0.05), respectively. During this period, the rheobase (Irh) increased (P < 0.01) from 0.13 +/- 0.07 to 0.27 +/- 0.14 nA, and the percentage of cells exhibiting inward rectification increased from 5 to 40%. Voltage threshold (Vthr) of the action potential remained unchanged postnatally. 3. There was also a postnatal change in the shape of the action potential. Specifically, between 1-2 and 5-6 days, there was a decrease (P < 0.05) in the spike half-width from 2.23 +/- 0.53 to 1.45 +/- 0.44 ms, resulting, in part, from a steepening (P < 0.05) of the slope of the falling phase of the action potential from 21.6 +/- 10.1 to 32.9 +/- 13.1 mV/ms. The slope of the rising phase also increased significantly (P < 0.01) between 1-2 and 13-15 days from 68.4 +/- 31.0 to 91.4 +/- 44.3 mV/ms. 4. The average duration of the medium afterhyperpolarization (mAHPdur) decreased (P < 0.05) between 1-2 (193 +/- 53 ms) and 5-6 days (159 +/- 43 ms). Whereas the mAHPdur was found to be independent of membrane potential, there was a linear relationship between the membrane potential and the amplitude of the medium AHP (mAHPamp). From this latter relationship, a reversal potential for the mAHPamp was extrapolated to be -87 mV. No evidence for the existence of a slow AHP was found in these developing motoneurons. 5. All cells analyzed (n = 74) displayed adaptation during the first three spikes. The subsequent firing pattern was classified into two groups, adapting and nonadapting. Cells at birth were all adapting, whereas all cells but two from animals 13 days and older were nonadapting. At the intermediate age (5-6 days), the minority (27%) was adapting and the majority (73%) was nonadapting. 6. The mean slope of primary range for the first interspike interval (1st ISI) was approximately 90 Hz/nA. This value was similar for both adapting and nonadapting cells and did not change postnatally.(ABSTRACT TRUNCATED AT 400 WORDS)

Anatomical and electrotonic coupling in developing genioglossal motoneurons of the rat.

Dye-, tracer- and electrotonic coupling were studied independently in genioglossal (GG) motoneurons using intracellular recordings in in vitro brainstem slices from rats postnatal ages 1-30 days. The subpopulation of GG motoneurons were retrogradely labeled after an injection of dextran-rhodamine into the posterior tongue. Dye-coupling was studied with Lucifer yellow injected into 55 motoneurons and tracer-coupling with neurobiotin injected into 89 presumptive GG motoneurons. Of the motoneurons injected with Lucifer yellow, only 6 of 41 cells (16.2%) exhibited dye-coupling; all occurred in animals less than 9 days old. In all but one instance, dye-coupling was restricted to only one other cell. No evidence of dye-coupling was found in the 14 cells injected in animals older than 8 days. Tracer-coupling (neurobiotin) was demonstrated in 12 of 30 cells (40%) from animals 1-2 days old and in 6 of 21 cells (28.6%) from animals 3-8 days old. Of the remaining 38 cells from animals 10 days of age and older, only one cell was found to be tracer-coupled. Cells injected with neurobiotin were coupled to an average of two other cells. Electrotonic coupling, as demonstrated with a short latency depolarization (SLD) in response to stimulation of hypoglossal axons, was found in developing GG motoneurons. These SLDs were revealed in 17 of 40 GG motoneurons (42.5%) examined in 1-8-day-old animals. There were no SLDs recorded in the 10 cells examined from animals of 10 days and older. The significance of coupling relative to patency of the newborn upper airways is discussed.

Morphometric analysis of phrenic motoneurons in the cat during postnatal development.

The dendritic geometry of 20 phrenic motoneurons from four postnatal ages (2 weeks, 1 and 2 months, and adult) was examined by using intracellular injection of horseradish peroxidase. The number of primary dendrites (approximately 11-12) remained constant throughout postnatal development. In general, postnatal growth of the dendrites resulted from an increase in the branching and in the length and diameter of segments at all orders of the dendritic tree. There was one exception. Between 2 weeks and 1 month, the maximum extent of the dendrites increased in parallel with the growth of the spinal cord; however, there was no increase in either combined dendritic length or total membrane surface area. In addition, there was a significant decrease in the number of dendritic terminals per cell (59.8 +/- 9.3 vs. 46.4 +/- 7.4 for 2 weeks and 1 month, respectively). The distance from the soma, where the peak number of dendritic terminals per cell occurred, ranged from 700-900 microns at 2 weeks and 2 months to 1,300-1,700 microns in the adult. The diameter of dendrites as a function of distance from the soma along the dendritic path increased with age. The process of maturation tended to increase the distance from the soma over which the surface area and dendritic trunk parameter (sigma d1.5/D1.5) remained constant. The three-dimensional distribution of dendrites was analyzed by dividing space into six equal volumes or hexants. This analysis revealed that the postnatal growth in surface area in the rostral and caudal hexants was proportionately larger than that in either the medial, lateral, dorsal, or ventral hexants. Strong linear correlations were found between the diameter of the primary dendrite and the combined length, surface area, volume, and number of terminals of the dendrite at all ages studied.

Electrophysiological properties of developing phrenic motoneurons in the cat.

1. Intracellular recordings were made in 427 phrenic motoneurons from kittens (in four stages of postnatal development, ranging from 2 to 14 wk) and in 72 motoneurons from adult cats. These experiments were performed to determine how the pattern of spontaneous discharge changes in phrenic motoneurons during development and how these changes might be influenced by alterations in the electrophysiological properties of these neurons. 2. The mean axonal conduction velocity increased significantly (P less than 0.0001) throughout this period of development, with the most rapid increase occurring between weeks 2 and 5 (18.5 +/- 5.4 and 32.4 +/- 5.6 m/s, respectively, mean +/- SD). 3. There was no change in the magnitude of the membrane potential, antidromic action potential, or positive overshoot; whereas there was a decrease in the half-width of the action potential from 2 (652 +/- 184 ms) to 14 (525 +/- 116 ms) wk postnatal. 4. The mean duration of the afterhyperpolarization (AHPdur) decreased from 69 +/- 20 ms at 2 wk to 60 +/- 16 ms by 9 wk, then increased to 66 +/- 18 ms by 14 wk of age and to 75 +/- 21 ms in the adult. The mean amplitude of the afterhyperpolarization (AHPamp) in the 2-wk-old group (4.9 +/- 1.8 mV) was larger than that at weeks 5 (3.9 +/- 1.7 mV) and 9 (3.9 +/- 1.6 mV), whereas the mean AHPamp of the adult (3.1 +/- 1.2 mV) was significantly smaller than the mean of any younger group. A significant negative correlation was found between AHPdur and axonal conduction velocity in all age groups studied, including the adult.(ABSTRACT TRUNCATED AT 250 WORDS)

Postnatal development of phrenic motoneurons in the cat.

The postnatal growth of phrenic motoneurons in the cat was studied using retrograde transport of horseradish peroxidase (HRP). The mean somal surface area of these developing motoneurons increased 2.5 times from day 3 to adult while the mean somal volume increased four-fold. This change in mean somal surface area during postnatal development was found to be correlated with the change in mean axonal conduction velocity measured from phrenic motoneurons.

The activity pattern of phrenic motoneurons during the aspiration reflex: an intracellular study.

Membrane potential trajectories and discharge characteristics were measured intracellularly in 29 phrenic motoneurons of anesthesized, paralyzed and artificially ventilated cats during hypercapnic respiration and the aspiration reflex. Fifteen 'active' cells discharged spontaneously during inspiration, and the remaining 14 'quiescent' cells exhibited no discharge in spite of strong central respiratory drive. The mean membrane potential of the quiescent cells during inspiration (-62 +/- 4 mV) was significantly lower than the threshold level determined for the active cells -52 +/- 4 mV). The mean axonal conduction velocity was slower for the active (60.4 +/- 8.7 m/s) than quiescent cells (67.4 +/- 6.9 m/s). All phrenic motoneurons discharged during the aspiration reflex with maximum instantaneous frequencies ranging from 6 to 357 Hz. No differences were found for the maximum discharge frequency during the reflex between the active and quiescent cells. Although there were differences in the slopes of the depolarization during inspiration between the groups of cells, no such difference existed in the slopes during the aspiration reflex. The threshold level for the first spike during the reflex was the same as that during inspiration but the level for successive spikes became progressively less negative while spike amplitude decreased and duration increased. Stimulation of the nasopharynx to elicit the aspiration reflex was found to alter the timing of the subsequent respiratory cycles.

Morphology of developing motoneurons innervating the medial gastrocnemius of the cat.

The morphology of medial gastrocnemius (MG) motoneurons labeled by retrograde transport of horseradish peroxidase was quantified in 5 postnatal ages (3 to 79-86 days) and in adults. A bimodal distribution of somal volumes was evident at birth which permitted separating the motoneurons into alpha and gamma subpopulations for analysis. There was a significant increase in the axial dimensions, surface area and volume calculated for both alpha and gamma cell bodies between each of the age-groups studied. A greater relative growth of the major over minor axis for the gammas produced a significant decrease in the form factor (i.e. greater eccentricity) between the youngest and oldest age-groups. The number of primary dendrites observed remained constant throughout postnatal development. The surface area of alpha somata more than tripled while that of the gammas doubled from 3 days to the adult. The mean somal volume of an alpha motoneuron at birth was only 17% of its adult value while the gamma cell bodies were 33% of their adult volume. A positive correlation was found for both alpha and gamma motoneurons when their somal surface area was plotted against postnatal age and weight. The rate of growth of the MG somal surface area is compared to the changes found in axonal conduction velocity and axonal diameter for MG in the literature.

The postnatal growth of motoneurons at three levels of the cat neuraxis.

The postnatal growth of motoneuron cell bodies located in the brainstem, cervical and lumbosacral spinal cord was investigated using retrograde transport of horseradish peroxidase in kittens ages 2, 12, 30, 55, 82 and 114 postnatal days and in an adult. The motoneurons innervating an extrinsic tongue muscle, the genioglossus, reached their adult size by eight weeks after birth. In contrast, the phrenic motoneurons innervating the diaphragm achieved adult size by 12 weeks and the motoneurons innervating the medial gastrocnemius muscle continued to grow beyond the twelfth postnatal week. The sizes of these motoneurons relative to one another remained constant during periods of development.

Postnatal growth of genioglossal motoneurons.

The postnatal growth of kitten genioglossal motoneurons were examined in six different age groups (newborn, 2, 4, 8, and 12 weeks and adult) using the technique of retrograde transport of horseradish peroxidase (HRP). The cell bodies of 100-150 motoneurons in each age group were analyzed in a transverse plane of section using standard techniques. Somatic genioglossal motoneuron growth occurred primarily along the major axis, which increased from 25.2 microns to 41.3 microns between birth and 8 weeks of postnatal age, after which time there was no further increase in either major or minor dimension of the cell body. The form factor decreased from 0.94 to 0.80 from birth to adulthood indicating an increased eccentricity of the cell body. The number of primary dendrites visible with this technique remained constant throughout the postnatal period. Calculated somal surface area increased in a linear fashion from birth through 8 weeks of postnatal life. There was no further increase in surface area beyond this age. The rate of increase in somal surface area with age was significantly different from both the rate of increase of animal weight and animal surface area with age. The correlations between the demonstrated immature genioglossal morphology and its cellular electrophysiology or integrated respiratory function remain unknown. The recent demonstration of decreased activation of the genioglossus muscle following airway occlusion in premature infants with apnea suggests that the relationships between developing genioglossal motoneuron structure and function warrant further investigation.

Neural elements subserving pulmonary stretch receptor-mediated facilitation of phrenic motoneurons.

The neural elements responsible for facilitation of phrenic nerve activity by lung inflation were investigated in cats by the simultaneous recording of individual pulmonary stretch receptor afferents, respiratory neurons of the ventrolateral nucleus of the tractus solitarius and phrenic nerve activity. Monosynaptic excitation of I beta neurons by slowly adapting pulmonary stretch receptors was demonstrated by cross-correlational analysis. It was also demonstrated that the majority of these same I beta neurons projected to the contralateral C5 phrenic motoneuron pool. Thus, this study has shown that I beta neurons can act as central neural elements to mediate the facilitatory effect of lung inflation upon phrenic nerve activity.

Quantitative analysis of the dendrites of cat phrenic motoneurons stained intracellularly with horseradish peroxidase.

All the dendrites (N = 37) generated by four phrenic motoneurons were analyzed following intracellular injection of horseradish peroxidase. The dendritic arbors produced from each of these stem dendrites were studied in detail. The mean number of stem dendrites produced by a phrenic motoneuron was 9.7, their mean diameter was 6.0 micron, and their mean combined diameter was 58.3 micron. The length at which a phrenic motoneuronal dendrite terminated was 1,236 micron, with several end terminals extending more than 2 mm from the cell body. The mean value for the combined lengths of all segments originating from a single stem dendrite was 5.3 mm. A full spectrum of dendritic branching patterns was observed from simple (five unbranched) to complex, the latter producing up to ninth-order branches. Most terminal and nonterminal dendritic segments tapered, producing a mean diameter reduction of 34%, or approximately 9% per 100-micron length. All phrenic motoneurons exhibited a steady decrease in the combined dendritic parameter (sigma d3/2) with distance from the soma as a result of tapering and end-branch termination. The mean surface area and volume of a phrenic motoneuronal dendrite were 35.3 X 10(3) micron 2 and 25.9 X 10(3) micron 3, respectively. The dendrites constituted greater than 97% of the total phrenic motoneuronal surface area, with 75% of this area lying outside of a 300-micron radius from the cell body. The diameter of a stem dendrite was positively correlated with its combined dendritic length, number of terminal branches, dendritic surface area, and volume. Despite this strong correlation, the value of total dendritic surface area calculated using the power equation derived from the dendritic surface area versus stem dendritic diameter plot was not a consistent estimator of the total dendritic surface area directly measured for these four phrenic motoneurons. It is suggested that this inconsistency may be the result of a heterogeneity in the phrenic motoneuronal population and/or in the dendrites projecting to the different terminal fields.

Spatial distributions of phrenic and medial gastrocnemius motoneurons in the cat spinal cord.

The longitudinal distributions of both phrenic and medial gastrocnemius motoneurons were quantitatively studied in the cat spinal cord. Both populations of motoneurons were retrogradely labeled by applying horseradish peroxidase (HRP) to the cut central ends of the appropriate peripheral nerves. The longitudinal positions of all labeled motoneurons in each motor column were determined; these data then were used to generate longitudinal distribution histograms and spatial interval distributions (SIDs), the latter being analyzed further by means of power spectra. In three of four cats, longitudinal clustering of phrenic motoneuronal cell bodies was revealed by the presence of a narrow central peak in the SID and the presence of subsidiary peaks. In the fourth cat, only a smaller central peak was observed. Power spectral analysis of the three SIDs having subsidiary peaks revealed that the mean longitudinal distance between clusters was 0.95 mm (range 0.52 to 1.22 mm). The analyses also revealed that on average a phrenic motoneuronal cluster contained 17 motoneurons, and the mean longitudinal length of a cluster was 450 microns. Using single, small-volume injections of HRP into the diaphragm, we concluded that not all the phrenic motoneurons within a single cluster innervate muscle fibers in a discrete region of the diaphragm. Similar quantitative analysis of the distribution of medial gastrocnemius motoneurons did not reveal clustering in this motor column. We suggest that the fundamental differences in the spatial distributions of motoneurons within these two motor columns may be related to differences in the functional organizations of motoneurons innervating axial versus appendicular musculature, i.e., diaphragm versus medial gastrocnemius.