A versatile, low-cost adaptor for stereotaxic and electrophysiologic spinal preparations in juvenile and adult rodents

Cunningham, Miles G.; Donalds, Rachael A.; Scouten, Charles W.; Tresch, Matthew C.

doi:10.1016/j.brainresbull.2005.08.004

Cited by 4 publications

(6 citation statements)

References 30 publications

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“…A 1–2 cm incision was made over lumbar segments, and the connective tissue and muscle overlying the vertebrae was minimally dissected 56 to expose L1 and L2 vertebrae 57 . Vertebrae and underlying spinal segments were located by spinous process tendon attachments and spinous process shape, and confirmed by pilot injections of DiD dye.…”

Section: Methodsmentioning

confidence: 99%

Voluntary urination control by brainstem neurons that relax the urethral sphincter

et al. 2018

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Voluntary urination ensures that waste is eliminated when safe and socially appropriate, even without a pressing urge. Uncontrolled urination, or incontinence, is a common problem with few treatment options. Normal urine release requires a small region in the brainstem known as Barrington's nucleus (Bar), but specific neurons that relax the urethral sphincter and enable urine flow are unknown. Here we identify a small subset of Bar neurons that control the urethral sphincter in mice. These excitatory neurons express estrogen receptor 1 (Bar), project to sphincter-relaxing interneurons in the spinal cord and are active during natural urination. Optogenetic stimulation of Bar neurons rapidly initiates sphincter bursting and efficient voiding in anesthetized and behaving animals. Conversely, optogenetic and chemogenetic inhibition reveals their necessity in motivated urination behavior. The identification of these cells provides an expanded model for the control of urination and its dysfunction.

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Section: Methodsmentioning

confidence: 99%

Voluntary urination control by brainstem neurons that relax the urethral sphincter

et al. 2018

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“…We developed a method for simultaneously recording electrical activity across spinal cord laminae and motor behaviour in neonatal rats, based in part on previous preparations for chronic imaging of the dorsal spinal cord and spinal single-wire or patch-clamp electrophysiological recordings in anesthetized, adult rodents62636465. Specifically, prior to and during surgery, animals ( n =15) were anesthetized using isoflurane (Aerrane) and given the analgesic buprenorphine (0.03 mg Kg −1 of body weight, Buprecare).…”

Section: Methodsmentioning

confidence: 99%

Sensory feedback synchronizes motor and sensory neuronal networks in the neonatal rat spinal cord

et al. 2016

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Early stages of sensorimotor system development in mammals are characterized by the occurrence of spontaneous movements. Whether and how these movements support correlated activity in developing sensorimotor spinal cord circuits remains unknown. Here we show highly correlated activity in sensory and motor zones in the spinal cord of neonatal rats in vivo. Both during twitches and complex movements, movement-generating bursts in motor zones are followed by bursts in sensory zones. Deafferentation does not affect activity in motor zones and movements, but profoundly suppresses activity bursts in sensory laminae and results in sensorimotor uncoupling, implying a primary role of sensory feedback in sensorimotor synchronization. This is further supported by largely dissociated activity in sensory and motor zones observed in the isolated spinal cord in vitro. Thus, sensory feedback resulting from spontaneous movements is instrumental for coordination of activity in developing sensorimotor spinal cord circuits.

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“…1). The animals were suspended from a spinal‐fork stereotaxic apparatus (Harvard Apparatus) (Cunningham et al 2005), between T12 and L2 (± one vertebra). Using a surgical microscope, the muscles covering the lateral aspects of the transverse processes of T12 and L2 were identified and gently resected.…”

Section: Methodsmentioning

confidence: 99%

Long‐term in vivo imaging of normal and pathological mouse spinal cord with subcellular resolution using implanted glass windows

Fenrich

Weber

Hocine

et al. 2012

The Journal of Physiology

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Key points• Chronic in vivo imaging of cellular interactions within the adult spinal cord with subcellular resolution is important for understanding cellular physiology and disease progression.• Previous approaches for chronic in vivo spinal cord microscopy have required surgery for each imaging session.• Here we describe a novel method for implanting glass windows over the exposed spinal cords of adult mice for repeated in vivo microscopy.• We show that the windows remain clear for many months after implantation, do not damage axons or blood vessels, and are useful for studying cellular dynamics after spinal cord injury.• Our method represents an original technical breakthrough for scientists involved in spinal cord research and in vivo imaging, and is a useful tool for studying cellular physiology and disease progression.Abstract Repeated in vivo two-photon imaging of adult mammalian spinal cords, with subcellular resolution, would be crucial for understanding cellular mechanisms under normal and pathological conditions. Current methods are limited because they require surgery for each imaging session. Here we report a simple glass window methodology avoiding repeated surgical procedures and subsequent inflammation. We applied this strategy to follow axon integrity and the inflammatory response over months by multicolour imaging of adult transgenic mice. We found that glass windows have no significant effect on axon number or structure, cause a transient inflammatory response, and dramatically increase the throughput of in vivo spinal imaging. Moreover, we used this technique to track retraction/degeneration and regeneration of cut axons after a 'pin-prick' spinal cord injury with high temporal fidelity. We showed that regenerating axons can cross an injury site within 4 days and that their terminals undergo dramatic morphological changes for weeks after injury. Overall the technique can potentially be adapted to evaluate cellular functions and therapeutic strategies in the normal and diseased spinal cord.

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A versatile, low-cost adaptor for stereotaxic and electrophysiologic spinal preparations in juvenile and adult rodents

Cited by 4 publications

References 30 publications

Voluntary urination control by brainstem neurons that relax the urethral sphincter

Voluntary urination control by brainstem neurons that relax the urethral sphincter

Sensory feedback synchronizes motor and sensory neuronal networks in the neonatal rat spinal cord

Long‐term in vivo imaging of normal and pathological mouse spinal cord with subcellular resolution using implanted glass windows

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