Prenatal nicotine exposure alters respiratory long-term facilitation in neonatal rats.

Intermittent hypoxia can evoke persistent increases in ventilation (V (E)) in neonates (i.e. long-term facilitation, LTF) (Julien et al., 2008). Since prenatal nicotine (PN) exposure alters neonatal respiratory control (Fregosi and Pilarski, 2008), we hypothesized that PN would influence LTF of ventilation (V (E)) in neonatal rats. An osmotic minipump delivered nicotine 6 mg/kg per day or saline to pregnant dams. V (E) was assessed in unanesthetized pups via whole body plethysmography at post-natal (P) days 9-11 or 15-17 during baseline (BL, 21% O(2)), hypoxia (10 x 5 min, 5% O(2)) and 30 min post-hypoxia. PN pups had reduced BL V (E) (p<0.05) but greater increases in V (E) during hypoxia (p<0.05). Post-hypoxia V (E) (i.e. LTF) showed an agex treatment interaction (p<0.01) with similar values at P9-11 but enhanced LTF in saline (30+/-8%BL) vs. PN pups (6+/-5%BL; p=0.01) at P15-17. We conclude that the post-natal developmental time course of hypoxia-induced LTF is influenced by PN.

Graded unilateral cervical spinal cord injury and respiratory motor recovery.

We examined the potential contribution of ventromedial (VM) tissue sparing to respiratory recovery following chronic (1 mo) unilateral C2 spinal cord injury (SCI) in rats. Preserved white matter ipsilateral to the injury was quantitatively expressed relative to contralateral white matter. The ipsilateral-to-contralateral white matter ratio was 0 after complete C2 hemisection (C2HS) and 0.23+/-0.04 with minimal VM sparing. Inspiratory (breath min(-1)) and phrenic frequency (burst min(-1)), measured by plethysmography (conscious rats) and phrenic neurograms (anesthetized rats) respectively, were both lower with minimal VM sparing (p<0.05 vs. C2HS). Tidal volume also was greater in minimal VM sparing rats during a hypercapnic challenge (p<0.05 vs. C2HS). In other C2 hemilesioned rats with more extensive VM matter sparing (ipsilateral-to-contralateral white matter ratio=0.55+/-0.05), respiratory deficits were indicated at 1 mo post-injury by reduced ventilation during hypercapnic challenge (p<0.05 vs. uninjured). Anterograde (ventral respiratory column-to-spinal cord) neuroanatomical tracing studies showed that descending respiratory projections from the brainstem are present in VM tissue. We conclude that even relatively minimal sparing of VM tissue after C2 hemilesion can alter respiratory outcomes. In addition, respiratory deficits can emerge in the adult rat after high cervical SCI even when relatively extensive VM sparing occurs.

Ventilation and phrenic output following high cervical spinal hemisection in male vs. female rats.

Female sex hormones influence the neural control of breathing and may impact neurologic recovery from spinal cord injury. We hypothesized that respiratory recovery after C2 spinal hemisection (C2HS) differs between males and females and is blunted by prior ovariectomy (OVX) in females. Inspiratory tidal volume (VT), frequency (fR), and ventilation (VE) were quantified during quiet breathing (baseline) and 7% CO2 challenge before and after C2HS in unanesthetized adult rats via plethysmography. Baseline breathing was similarly altered in all rats (reduced VT, elevated fR) but during hypercapnia females had relatively higher VT (i.e. compared to pre-injury) than male or OVX rats (p<0.05). Phrenic neurograms recorded in anesthetized rats indicated that normalized burst amplitude recorded ipsilateral to C2HS (i.e. the crossed phrenic phenomenon) is greater in females during respiratory challenge (p<0.05 vs. male and OVX). We conclude that sex differences in recovery of VT and phrenic output are present at 2 weeks post-C2HS. These differences are consistent with the hypothesis that ovarian sex hormones influence respiratory recovery after cervical spinal cord injury.

Modest spontaneous recovery of ventilation following chronic high cervical hemisection in rats.

Following C2 spinal hemisection (C2HS) in adult rats, ipsilateral phrenic motoneuron (PhMN) recovery occurs through a time-dependent activation of latent, crossed-spinal collaterals (i.e., spontaneous crossed phrenic phenomenon; sCPP) from contralateral bulbospinal axons. Ventilation is maintained during quiet breathing after C2HS, but the ability to increase ventilation during a respiratory stimulation (e.g. hypercapnia) is impaired. We hypothesized that long-term expression of the sCPP would correspond to a progressive normalization in ventilatory patterns during respiratory challenge. Breathing was assessed via plethsymography in unanesthetized animals and phrenic motor output was measured in urethane-anesthetized, paralyzed and vagotomized rats. At 2-week post-C2HS, minute ventilation (VE) was maintained during baseline (room air) conditions as expected but was substantially blunted during hypercapnic challenge (68+/-3% of VE in uninjured, weight-matched rats). However, by 12 weeks the spinal-lesioned rats achieved a hypercapnic VE response that was 85+/-7% of control (p=0.017 vs. 2 wks). These rats also exhibited augmented breaths (AB's) or "sighs" more frequently (p<0.05) than controls; however, total AB volume was significantly less than control at 2- and 12-week post-injury (69+/-4% and 80+/-5%, p<0.05, respectively). We also noted that phrenic neurograms demonstrated a consistent delay in onset of the ipsilateral vs. contralateral inspiratory phrenic burst at 2-12-week post-injury. Finally, the ipsilateral phrenic response to respiratory challenge (hypoxia) was greater, though not normalized, at 4-12- vs. 2-week post-injury. We conclude that recovery of ventilation deficits occurs over 2-12-week post-C2HS; however, intrinsic neuroplasticity remains insufficient to concurrently restore a normal level of ipsilateral phrenic output.

Long-term facilitation of ipsilateral but not contralateral phrenic output after cervical spinal cord hemisection.

After chronic C2 spinal hemisection (C2HS), exposure to intermittent hypoxia (IH) evokes a persistent increase in phrenic output recorded ipsilateral to the injury (i.e., phrenic long-term facilitation, LTF; Golder and Mitchell, J. Neurosci. 25:2925-32, 2005). However, unilateral spinal cord injury induces compensatory increases in contralateral motoneuron activity that may reduce their capacity for further plasticity (i.e., a "ceiling effect"). We hypothesized that after chronic C2HS, LTF would be reduced in contralateral (vs. ipsilateral) phrenic output. Bilateral phrenic activity was recorded in three groups of anesthetized, paralyzed, vagotomized, and ventilated rats: uninjured, and 4 or 8 weeks following histologically verified C2HS. Baseline (BL) phrenic activity was established during normoxia and rats were then exposed to IH (5 x 3 min isocapnic hypoxia, 13-14% O2) followed by isocapnic normoxia; LTF was assessed 60-min post-IH. Uninjured animals showed an increase in inspiratory burst amplitude that was similar in the left (44 +/- 11%BL) and right phrenic nerves (39 +/- 13%BL). However, similar burst amplitude LTF did not occur in phrenic output recorded contralateral to C2HS at 4 (-10 +/- 7% BL) or 8 weeks post-C2HS (4 +/- 5% BL). In contrast, LTF of ipsilateral phrenic amplitude occurred at both 4 (44 +/- 11% BL) and 8 weeks post-C2HS (129 +/- 30% BL, P < 0.05). A persistent increase in phrenic burst frequency after IH (i.e., "frequency LTF") was observed in control (+9 +/- 3 burst/min, P < 0.05), but not C2HS rats. We conclude that compensatory responses to unilateral cervical spinal cord injury prevent the expression of LTF in contralateral phrenic motoneurons.