Tools for efficient analysis of neurons in a 3D reference atlas of whole mouse spinal cord

Spinal neurons are highly heterogeneous in location, transcriptional identity and function. To understand their contributions to sensorimotor circuits, it is essential to map the positions of identified subsets of neurons in relation to others throughout the spinal cord (SC), but we lack tools for whole SC sample preparation, imaging and in toto analysis. To overcome this problem, we have (1) designed scaffolds (SpineRacks) that facilitate efficient and ordered cryo-sectioning of the entire SC in a single block, (2) constructed a 3D reference atlas of adult mouse SC and (3) developed software (SpinalJ) to register images of sections and for standardized analysis of cells and projections in atlas space. We have verified mapping accuracies for known neurons and demonstrated the usefulness of this platform to reveal unknown neuronal distributions. Together, these tools provide high-throughput analyses of whole mouse SC and enable direct comparison of 3D spatial information between animals and studies.

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Single-cell transcriptomic analysis of the adult mouse spinal cord reveals molecular diversity of autonomic and skeletal motor neurons

Nature Neuroscience ◽

10.1038/s41593-020-00795-0 ◽

2021 ◽

Vol 24 (4) ◽

pp. 572-583 ◽

Cited By ~ 1

Author(s):

Jacob A. Blum ◽

Sandy Klemm ◽

Jennifer L. Shadrach ◽

Kevin A. Guttenplan ◽

Lisa Nakayama ◽

...

Keyword(s):

Spinal Cord ◽

Single Cell ◽

Motor Neurons ◽

Molecular Diversity ◽

Adult Mouse ◽

Transcriptomic Analysis ◽

Mouse Spinal Cord

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Repair of a myelin lesion by Schwann cells transplanted in the adult mouse spinal cord

Journal of Neuroimmunology ◽

10.1016/0165-5728(92)90139-c ◽

1992 ◽

Vol 40 (2-3) ◽

pp. 235-242 ◽

Cited By ~ 90

Author(s):

A. Baron-Van Evercooren ◽

A. Gansmuller ◽

E. Duhamel ◽

F. Pascal ◽

M. Gumpel

Keyword(s):

Spinal Cord ◽

Schwann Cells ◽

Adult Mouse ◽

Mouse Spinal Cord ◽

Myelin Lesion

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Rapid isolation and culture of primary microglia from adult mouse spinal cord

Journal of Neuroscience Methods ◽

10.1016/j.jneumeth.2009.07.002 ◽

2009 ◽

Vol 183 (2) ◽

pp. 223-237 ◽

Cited By ~ 21

Author(s):

Ping K. Yip ◽

Timothy K.Y. Kaan ◽

Daniel Fenesan ◽

Marzia Malcangio

Keyword(s):

Spinal Cord ◽

Adult Mouse ◽

Primary Microglia ◽

Mouse Spinal Cord ◽

Rapid Isolation

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Identification of genome-wide targets of Olig2 in the adult mouse spinal cord using ChIP-Seq

PLoS ONE ◽

10.1371/journal.pone.0186091 ◽

2017 ◽

Vol 12 (10) ◽

pp. e0186091 ◽

Cited By ~ 4

Author(s):

Andrew J. Darr ◽

Matt C. Danzi ◽

Lee Brady ◽

Dorothea Emig-Agius ◽

Amber Hackett ◽

...

Keyword(s):

Spinal Cord ◽

Adult Mouse ◽

Mouse Spinal Cord ◽

Genome Wide

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Intraspinal injection of adeno-associated viruses into the adult mouse spinal cord

STAR Protocols ◽

10.1016/j.xpro.2021.100786 ◽

2021 ◽

Vol 2 (3) ◽

pp. 100786

Author(s):

Shrivas Chaterji ◽

Arnab Barik ◽

Anupama Sathyamurthy

Keyword(s):

Spinal Cord ◽

Adult Mouse ◽

Mouse Spinal Cord ◽

Intraspinal Injection

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Calpain inhibitors delay injury-induced apoptosis in adult mouse spinal cord motor neurons

Neuroreport ◽

10.1097/01.wnr.0000220127.01597.04 ◽

2006 ◽

Vol 17 (8) ◽

pp. 761-765 ◽

Cited By ~ 8

Author(s):

Hamid R. Momeni ◽

Martin Kanje

Keyword(s):

Spinal Cord ◽

Motor Neurons ◽

Adult Mouse ◽

Mouse Spinal Cord ◽

Induced Apoptosis ◽

Calpain Inhibitors

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Rho-Associated Kinase II (ROCKII) Limits Axonal Growth after Trauma within the Adult Mouse Spinal Cord

Journal of Neuroscience ◽

10.1523/jneurosci.4650-09.2009 ◽

2009 ◽

Vol 29 (48) ◽

pp. 15266-15276 ◽

Cited By ~ 82

Author(s):

P. Duffy ◽

A. Schmandke ◽

J. Sigworth ◽

S. Narumiya ◽

...

Keyword(s):

Spinal Cord ◽

Adult Mouse ◽

Axonal Growth ◽

Mouse Spinal Cord

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Motoneuron Excitability and Muscle Spasms Are Regulated by 5-HT2B and 5-HT2C Receptor Activity

Journal of Neurophysiology ◽

10.1152/jn.00774.2010 ◽

2011 ◽

Vol 105 (2) ◽

pp. 731-748 ◽

Cited By ~ 79

Author(s):

Katherine C. Murray ◽

Marilee J. Stephens ◽

Edmund W. Ballou ◽

Charles J. Heckman ◽

David J. Bennett

Keyword(s):

Spinal Cord ◽

Calcium Currents ◽

Receptor Subtypes ◽

Constitutive Activity ◽

Spinal Neurons ◽

Cord Injury ◽

Muscle Spasms ◽

Highly Correlated ◽

And Function

Immediately after spinal cord injury (SCI), a devastating paralysis results from the loss of brain stem and cortical innervation of spinal neurons that control movement, including a loss of serotonergic (5-HT) innervation of motoneurons. Over time, motoneurons recover from denervation and function autonomously, exhibiting large persistent calcium currents (Ca PICs) that both help with functional recovery and contribute to uncontrolled muscle spasms. Here we systematically evaluated which 5-HT receptor subtypes influence PICs and spasms after injury. Spasms were quantified by recording the long-lasting reflexes (LLRs) on ventral roots in response to dorsal root stimulation, in the chronic spinal rat, in vitro. Ca PICs were quantified by intracellular recording in synaptically isolated motoneurons. Application of agonists selective to 5-HT2B and 5-HT2C receptors (including BW723C86) significantly increased the LLRs and associated Ca PICs, whereas application of agonists to 5-HT1, 5-HT2A, 5-HT3, or 5-HT4/5/6/7 receptors (e.g., 8-OH-DPAT) did not. The 5-HT2 receptor agonist–induced increases in LLRs were dose dependent, with doses for 50% effects (EC50) highly correlated with published doses for agonist receptor binding ( Ki) at 5-HT2B and 5-HT2C receptors. Application of selective antagonists to 5-HT2B (e.g., RS127445) and 5-HT2C (SB242084) receptors inhibited the agonist-induced increase in LLR. However, antagonists that are known to specifically be neutral antagonists at 5-HT2B/C receptors (e.g., RS127445) had no effect when given by themselves, indicating that these receptors were not activated by residual 5-HT in the spinal cord. In contrast, inverse agonists (such as SB206553) that block constitutive activity at 5-HT2B or 5-HT2C receptors markedly reduced the LLRs, indicating the presence of constitutive activity in these receptors. 5-HT2B or 5-HT2C receptors were confirmed to be on motoneurons by immunolabeling. In summary, 5-HT2B and 5-HT2C receptors on motoneurons become constitutively active after injury and ultimately contribute to recovery of motoneuron function and emergence of spasms.

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