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

Chaterji, Shrivas; Barik, Arnab; Sathyamurthy, Anupama

doi:10.1016/j.xpro.2021.100786

Cited by 9 publications

(8 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Intraspinal injections were performed as previously described 84 , 85 . Briefly, mice were anesthetized by intraperitoneal injection of a drug cocktail containing fentanyl (0.2 mg kg), dexmedetomidine (1 mg/kg), and midazolam (5 mg/kg) dissolved in saline.…”

Section: Methodsmentioning

confidence: 99%

Single cell atlas of spinal cord injury in mice reveals a pro-regenerative signature in spinocerebellar neurons

et al. 2022

Self Cite

View full text Add to dashboard Cite

After spinal cord injury, tissue distal to the lesion contains undamaged cells that could support or augment recovery. Targeting these cells requires a clearer understanding of their injury responses and capacity for repair. Here, we use single nucleus RNA sequencing to profile how each cell type in the lumbar spinal cord changes after a thoracic injury in mice. We present an atlas of these dynamic responses across dozens of cell types in the acute, subacute, and chronically injured spinal cord. Using this resource, we find rare spinal neurons that express a signature of regeneration in response to injury, including a major population that represent spinocerebellar projection neurons. We characterize these cells anatomically and observed axonal sparing, outgrowth, and remodeling in the spinal cord and cerebellum. Together, this work provides a key resource for studying cellular responses to injury and uncovers the spontaneous plasticity of spinocerebellar neurons, uncovering a potential candidate for targeted therapy.

show abstract

Section: Methodsmentioning

confidence: 99%

Single cell atlas of spinal cord injury in mice reveals a pro-regenerative signature in spinocerebellar neurons

et al. 2022

Self Cite

View full text Add to dashboard Cite

show abstract

“…Laminectomy was performed at the thoraco-lumbar (T12–L1) part of the vertebrae 44 , where similar albeit alternative procedure has recently been suggested in mice 45 . The animals received an injection of AAV virus (1500–2000 nl) dorso-ventrally and rostro-caudally at the rate of 15 nl/min using a sharp glass capillary (inner diameter = 0.53 mm and outer diameter = 1.14 mm, Neurostar GmbH, Tubingen, Germany), which was left in place for 3–5 min after the injection to prevent the backflow 46 . Laminectomy site was covered, skin was sutured and the rats were housed in their cages for recovery.…”

Section: Methodsmentioning

confidence: 99%

Viral strategies for targeting spinal neuronal subtypes in adult wild-type rodents

Kaur

Berg

2022

Sci Rep

View full text Add to dashboard Cite

Targeting specific subtypes of interneurons in the spinal cord is primarily restricted to a small group of genetic model animals. Since the development of new transgenic model animals can be expensive and labor intensive, it is often difficult to generalize these findings and verify them in other model organisms, such as the rat, ferret or monkey, that may be more beneficial in certain experimental investigations. Nevertheless, endogenous enhancers and promoters delivered using an adeno-associated virus (AAV) have been successful in providing expression in specific subtypes of neurons in the forebrain of wildtype animals, and therefore may introduce a shortcut. GABAergic interneurons, for instance, have successfully been targeted using the mDlx promoter, which has recently been developed and is now widely used in wild type animals. Here, we test the specificity and efficiency of the mDlx enhancer for robust targeting of inhibitory interneurons in the lumbar spinal cord of wild-type rats using AAV serotype 2 (AAV2). Since this has rarely been done in the spinal cord, we also test the expression and specificity of the CamKIIa and hSynapsin promoters using serotype 9. We found that AAV2-mDlx does in fact target many neurons that contain an enzyme for catalyzing GABA, the GAD-65, with high specificity and a small fraction of neurons containing an isoform, GAD-67. Expression was also seen in some motor neurons although with low correlation. Viral injections using the CamKIIa enhancer via AAV9 infected in some glutamatergic neurons, but also GABAergic neurons, whereas hSynapsin via AAV9 targets almost all the neurons in the lumbar spinal cord.

show abstract

“…The dorsal horn of the lumbar spinal cord was injected as previously reported elsewhere 46,47 . Mice were anesthetized with 5% Isoflurane which was maintained at 1-3% through a face mask during all surgeries, analgesia (buprenorphine subcutaneously injected at 0.1-0.2 mg/kg) was provided before and after each surgery.…”

Section: Surgeriesmentioning

confidence: 99%

Descending GABAergic Neurons of the RVM That Mediate Widespread Bilateral Antinociception

Ganley

Sousa

Ranucci

et al. 2023

Preprint

View full text Add to dashboard Cite

Neurons projecting from the rostral ventromedial medulla (RVM) to the spinal dorsal horn are critical elements of endogenous pain control systems. Here, we describe a GABA/glycinergic pathway that predominantly innervates the superficial dorsal horn. Anatomical and optogenetic tracing of these neurons from a single unilateral site of the lumbar spinal cord indicated that these neurons give rise to a dense bilateral innervation of the spinal cord along its entire rostrocaudal axis. Chemogenetic activation of these neurons caused a bilateral and widespread reduction in heat, cold, and mechanical sensitivity, while their silencing with tetanus toxin induced allodynia and spontaneous pain-like aversive behaviors. Consistent with a continuous role in the prevention of spontaneous pain, many descending RVM GABAergic neurons were found to be tonically active. This pathway may therefore be relevant for widespread conditioned analgesia, while its dysfunction may underlie chronic widespread pain syndromes.

show abstract

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

Cited by 9 publications

References 27 publications

Single cell atlas of spinal cord injury in mice reveals a pro-regenerative signature in spinocerebellar neurons

Single cell atlas of spinal cord injury in mice reveals a pro-regenerative signature in spinocerebellar neurons

Viral strategies for targeting spinal neuronal subtypes in adult wild-type rodents

Descending GABAergic Neurons of the RVM That Mediate Widespread Bilateral Antinociception

Contact Info

Product

Resources

About