PIEZO2 and perineal mechanosensation are essential for sexual function.

Despite the potential importance of genital mechanosensation for sexual reproduction, little is known about how perineal touch influences mating. We explored how mechanosensation affords exquisite awareness of the genitals and controls reproduction in mice and humans. Using genetic strategies and in vivo functional imaging, we demonstrated that the mechanosensitive ion channel PIEZO2 (piezo-type mechanosensitive ion channel component 2) is necessary for behavioral sensitivity to perineal touch.

Water exchange rates measure active transport and homeostasis in neural tissue.

For its size, the brain is the most metabolically active organ in the body. Most of its energy demand is used to maintain stable homeostatic physiological conditions. Altered homeostasis and active states are hallmarks of many diseases and disorders.

Motoneuronal Regulation of Central Pattern Generator and Network Function.

This chapter reviews recent work showing that vertebrate motoneurons can trigger spontaneous rhythmic activity in the developing spinal cord and can modulate the function of several different central pattern generators later in development. In both the embryonic chick and the fetal mouse spinal cords, antidromic activation of motoneurons can trigger bouts of rhythmic activity. In the neonatal mouse, optogenetic manipulation of motoneuron firing can modulate the frequency of fictive locomotion activated by a drug cocktail.

Optogenetic Activation of V1 Interneurons Reveals the Multimodality of Spinal Locomotor Networks in the Neonatal Mouse.

In the spinal cord, classes of interneurons have been studied to determine their role in producing or regulating locomotion. It is unclear whether all locomotor behaviors are produced by the same circuitry or engage different subsets of neurons. Here, in neonatal mice of either sex, we test this idea by comparing the actions of a class of spinal, inhibitory interneuron (V1) expressing channelrhodopsin driven by the transcription factor on the rhythms elicited by different methods.

Real-time measurement of diffusion exchange rate in biological tissue.

Diffusion exchange spectroscopy (DEXSY) provides a means to isolate the signal attenuation associated with exchange from other sources of signal loss. With the total diffusion weighting b+b=b held constant, DEXSY signals acquired with b=0 or b=0 have no exchange weighting, while a DEXSY signal acquired with b=b has maximal exchange weighting. The exchange rate can be estimated by fitting a diffusion exchange model to signals acquired with variable mixing times.

Motoneuronal Spinal Circuits in Degenerative Motoneuron Disease.

The most evident phenotype of degenerative motoneuron disease is the loss of motor function which accompanies motoneuron death. In both amyotrophic lateral sclerosis (ALS) and spinal muscular atrophy (SMA), it is now clear that dysfunction is not restricted to motoneurons but is manifest in the spinal circuits in which motoneurons are embedded. As mounting evidence shows that motoneurons possess more elaborate and extensive connections within the spinal cord than previously realized, it is necessary to consider the role of this circuitry and its dysfunction in the disease process.

Magnetic resonance measurements of cellular and sub-cellular membrane structures in live and fixed neural tissue.

We develop magnetic resonance (MR) methods for real-time measurement of tissue microstructure and membrane permeability of live and fixed excised neonatal mouse spinal cords. Diffusion and exchange MR measurements are performed using the strong static gradient produced by a single-sided permanent magnet. Using tissue delipidation methods, we show that water diffusion is restricted solely by lipid membranes.

Feedback regulation of locomotion by motoneurons in the vertebrate spinal cord.

Motoneurons are known to be an essential component of central pattern generators in invertebrates, but it is only recently that they have been shown to play a similar role in vertebrate locomotor circuits. Here, we review early experiments implicating motoneurons in the genesis of spontaneous motor activity in development and more recent experiments identifying motoneurons as important regulators of locomotor activity in the adult zebrafish and in the neonatal mouse spinal cord. We discuss the mechanisms responsible for these actions, the experimental challenges in studying the role of motoneurons in the mammalian spinal cord and the functional significance of the excitatory influence of motoneuron activity on locomotor behavior.

V1 interneurons regulate the pattern and frequency of locomotor-like activity in the neonatal mouse spinal cord.

In the mouse spinal cord, V1 interneurons are a heterogeneous population of inhibitory spinal interneurons that have been implicated in regulating the frequency of the locomotor rhythm and in organizing flexor and extensor alternation. By introducing archaerhodopsin into engrailed-1-positive neurons, we demonstrate that the function of V1 neurons in locomotor-like activity is more complex than previously thought. In the whole cord, V1 hyperpolarization increased the rhythmic synaptic drive to flexor and extensor motoneurons, increased the spiking in each cycle, and slowed the locomotor-like rhythm.

Dye-coupling between neonatal spinal motoneurons and interneurons revealed by prolonged back-filling of a ventral root with a low molecular weight tracer in the mouse.

We investigated dye-coupling between motoneurons in the L6 segment of the neonatal mouse spinal cord that contains limb-innervating motoneurons and sexually dimorphic motor nuclei. Using an isolated spinal cord preparation, we back-filled the cut, L6 ventral root with the small molecule Neurobiotin, or the much larger dextran-conjugated fluorophores for 16-24 hours. Motoneurons and parasympathetic preganglionic neurons were filled with both markers, but dye-coupling was only seen with Neurobiotin fills.

Motoneurons regulate the central pattern generator during drug-induced locomotor-like activity in the neonatal mouse.

Motoneurons are traditionally viewed as the output of the spinal cord that do not influence locomotor rhythmogenesis. We assessed the role of motoneuron firing during ongoing locomotor-like activity in neonatal mice expressing archaerhopsin-3 (Arch), halorhodopsin (eNpHR), or channelrhodopsin-2 (ChR2) in Choline acetyltransferase neurons (ChAT) or Arch in LIM-homeodomain transcription factor neurons. Illumination of the lumbar cord in mice expressing eNpHR or Arch in ChAT or neurons, depressed motoneuron discharge, transiently decreased the frequency, and perturbed the phasing of the locomotor-like rhythm.

Reep1 null mice reveal a converging role for hereditary spastic paraplegia proteins in lipid droplet regulation.

Hereditary spastic paraplegias (HSPs; SPG1-76 plus others) are length-dependent disorders affecting long corticospinal axons, and the most common autosomal dominant forms are caused by mutations in genes that encode the spastin (SPG4), atlastin-1 (SPG3A) and REEP1 (SPG31) proteins. These proteins bind one another and shape the tubular endoplasmic reticulum (ER) network throughout cells. They also are involved in lipid droplet formation, enlargement, or both in cells, though mechanisms remain unclear.

Pharmacological Investigation of Fluoro-Gold Entry into Spinal Neurons.

The fluorescent tracer Fluoro-Gold has been widely used to label neurons retrogradely. Here we show that Fluoro-Gold can also enter neurons through AMPA receptor endocytosis. We found that a 30 minute application of Fluoro-Gold to the isolated spinal cord labeled neurons under control conditions and in the presence of glutamatergic agonists including NMDA and AMPA.

Metachronal propagation of motor activity.

A diverse array of biomechanical systems has evolved to satisfy locomotor requirements (reptation, swimming, walking, etc.) and in all cases, successful behabior achievement requires the integrated functioning of various segments, to ensure the appropriate positioning of the different body regions. From comparative studies on a variety of invertebrate and vertebrate organisms, it is now established that the basic motor patterns underlying limb and/or trunk movements during locomotion are driven by central networks of neurons, so-called central pattern generators (CPGs).

Sequential activation of axial muscles during different forms of rhythmic behavior in man.

In humans, studies of back muscle activity have mainly addressed the functioning of lumbar muscles during postural adjustments or rhythmic activity, including locomotor tasks. The present study investigated how back muscles are activated along the spine during rhythmical activities in order to gain insights into spinal neuronal organization. Electromyographic recordings of back muscles were performed at various trunk levels, and changes occurring in burst amplitudes and phase relationships were analyzed.

Coordinated network functioning in the spinal cord: an evolutionary perspective.

The successful achievement of harmonious locomotor movement results from the integrated operation of all body segments. Here, we will review current knowledge on the functional organization of spinal networks involved in mammalian locomotion. Attention will not simply be restricted to hindlimb muscle control, but by also considering the necessarily coordinated activation of trunk and forelimb muscles, we will try to demonstrate that while there has been a progressive increase in locomotor system complexity during evolution, many basic organizational features have been preserved across the spectrum from lower vertebrates through to humans.

Metachronal coupling between spinal neuronal networks during locomotor activity in newborn rat.

In the present study, we investigate spinal cord neuronal network interactions in the neonatal rat during locomotion. The behavioural and physiological relevance of metachronally propagated locomotor activity were inferred from kinematic, anatomical and in vitro electrophysiological data. Kinematic analysis of freely behaving animals indicated that there is a rhythmic sequential change in trunk curvature during the step cycle.