Elsevier

IBRO Reports

Volume 9, December 2020, Pages 115-131
IBRO Reports

Research Paper
Mapping of spinal interneurons involved in regulation of the lower urinary tract in juvenile male rats

https://doi.org/10.1016/j.ibror.2020.07.002Get rights and content
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Abstract

Coordination between the urinary bladder (BL) and external urethral sphincter (EUS) is necessary for storage and elimination of urine. In rats interneuronal circuits at two levels of the spinal cord (i.e., L6-S1 and L3-L4) play an important role in this coordination. In the present experiments retrograde trans-synaptic transport of pseudorabies virus (PRV) encoding fluorescent markers (GFP and RFP) was used to trace these circuits. To examine the relative localization of EUS-related and BL-related interneuronal populations we injected PRV-GFP into the EUS and PRV-RFP into the BL wall. The PRV infected populations of spinal interneurons were localized primarily in the dorsal commissure (DCM) of L6/S1 and in a hypothesized lumbar spinal coordinating center (LSCC) in L3/L4 above and lateral to central canal (CC). At both sites colocalization of markers occurred in a substantial number of labeled interneurons indicating concomitant involvement of these double-labelled neurons in the EUS- and BL-circuits and suggesting their role in EUS-BL coordination. Intense GFP or RFP fluorescent was detected in a subpopulation of cells at both sites suggesting that they were infected earlier and therefore likely to represent first order, primary interneurons that directly synapse with output neurons. Larger numbers of weakly fluorescent neurons that likely represent second order interneurons were also identified. Within the population of EUS-related first order interneurons only 3–8 % exhibited positive immunoreaction for an early transcription factor Pax2 specific to GABAergic and glycinergic inhibitory neurons suggesting that the majority of interneurons in DCM and LSCC projecting directly to the EUS motoneurons are excitatory.

Abbreviations

BCM
bulbocavernosus muscle
BL
bladder
B_G
bright green
B_R
bright red
CC -
central canal
DCM
dorsal commissure
DSD
detrusor-sphincter-dyssynergia
EMG
electromyogram
EUS
external urethral sphincter
EUS-MN
motoneuron of the external urethral sphincter
GFP
green fluorescent protein
IML
intermediolateral nucleus
IN
interneuron
Lf
lateral funiculus
LSCC
lumbar spinal coordinating center
LUT
lower urinary tract
pIN
primary interneuron
PPN
propriospinal neuron
PRV
pseudorabies virus
RFP
red fluorescent protein
SC
spinal cord
SCI
spinal cord injury
sIN
secondary interneuron
SPPN
spinal parasympathetic preganglionic neuron
VMf
ventro-medial funiculus
W_G
weak/moderate green
W_R
weak/moderate red

Keywords

Spinal cord
Transsynaptic tracing
Pseudorabies virus

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