Spinal afferent innervation in flat-mounts of the rat stomach: anterograde tracing.

The dorsal root ganglia (DRG) project spinal afferent axons to the stomach. However, the distribution and morphology of spinal afferent axons in the stomach have not been well characterized. In this study, we used a combination of state-of-the-art techniques, including anterograde tracer injection into the left DRG T7-T11, avidin-biotin and Cuprolinic Blue labeling, Zeiss M2 Imager, and Neurolucida to characterize spinal afferent axons in flat-mounts of the whole rat stomach muscular wall. We found that spinal afferent axons innervated all regions with a variety of distinct terminal structures innervating different gastric targets: (1) The ganglionic type: some axons formed varicose contacts with individual neurons within myenteric ganglia. (2) The muscle type: most axons ran in parallel with the longitudinal and circular muscles and expressed spherical varicosities. Complex terminal structures were observed within the circular muscle layer. (3) The ganglia-muscle mixed type: some individual varicose axons innervated both myenteric neurons and the circular muscle, exhibiting polymorphic terminal structures. (4) The vascular type: individual varicose axons ran along the blood vessels and occasionally traversed the vessel wall. This work provides a foundation for future topographical anatomical and functional mapping of spinal afferent axon innervation of the stomach under normal and pathophysiological conditions.

A video protocol for rapid dissection of mouse dorsal root ganglia from defined spinal levels.

OBJECTIVE: Dorsal root ganglia (DRG) are heterogeneous assemblies of assorted sensory neuron cell bodies found in bilateral pairs at every level of the spinal column. Pseudounipolar afferent neurons convert external stimuli from the environment into electrical signals that are retrogradely transmitted to the spinal cord dorsal horn. To do this, they extend single axons from their DRG-resident somas that then bifurcate and project both centrally and distally. DRG can be dissected from mice at embryonic stages and any age post-natally, and have been extensively used to study sensory neuron development and function, response to injury, and pathological processes in acquired and genetic diseases. We have previously published a step-by-step dissection method for the rapid isolation of post-natal mouse DRG. Here, the objective is to extend the protocol by providing training videos that showcase the dissection in fine detail and permit the extraction of ganglia from defined spinal levels. RESULTS: By following this method, the reader will be able to swiftly and accurately isolate specific lumbar, thoracic, and cervical DRG from mice. Dissected ganglia can then be used for RNA/protein analyses, subjected to immunohistochemical examination, and cultured as explants or dissociated primary neurons, for in-depth investigations of sensory neuron biology.

Kambin's triangle and the position of the dorsal nerve root in the lumbar neural foramen.

INTRODUCTION: As a result of the increased utilization of neurosurgical arthroscopic techniques, investigations into population and sex-specific trends of anatomical considerations have become increasingly important. This study aimed to investigate and describe aspects of the neuroanatomical morphometry of lumbar spines in a cadaveric and magnetic resonance imaging (MRI) sample. MATERIALS AND METHODS: Twenty white adult (>18 years) cadavers (9 males; 11 females) were obtained under Ethical clearance. The lumbar regions were dissected and the position of the dorsal root ganglion (DRG) and dimensions of Kambin's triangle were determined. Twenty-six black adult (>18 years) MRI scans (17 males; 9 females) were obtained from an Academic Hospital and were used to determine the dimensions of the neural foramen and the DRGs within. RESULTS: The ganglia were mostly at the midline of the caudal pedicle. Similar to previous studies, the diagonal measurement from Kambin's triangle was the largest and the vertical measurement the shortest. Skeletal and soft-tissue measurements indicated distinct trends when moving caudolaterally in the spine. Soft-tissue parameters from the current study were within the upper limits of those from previous studies, whereas skeletal parameters were in agreement with those reported by previous authors. CONCLUSIONS: Results from this study suggest a variation of certain parameters between studies with varying population groups and therefore supports the need for and the importance of possible population-specific trends of anatomical parameters considered during surgical procedures.

Analysis of S1 DRG Programming to Determine Location of the DRG and Ideal Anatomic Positioning of the Electrode.

INTRODUCTION: Dorsal root ganglion (DRG) stimulation has been established as a therapy in the treatment of chronic pain. Ideal electrode placement is guided by proper identification of the location of the DRG. The location of the S1 DRG is not well delineated and can be variable making ideal location of the electrode placement difficult based on fluoroscopic imaging. METHODS: This is a retrospective analysis of postoperative programming of S1 DRG leading across two centers. There were 34 lead placements in 24 patients included in this study. Programming parameters and contacts used were evaluated based on the position of the electrode in reference to the sacral border. RESULTS: The majority of the patient programming parameters were recorded at six weeks following the implant. Most commonly, the programming used a simple continuous bipole configuration. Of the 34 leads programmed, 17 (50%) had programming on the sacral border, 14 (41%) were considered posterior, and 3 (9%) were anterior to the sacral border. CONCLUSION: This analysis of S1 DRG programming demonstrates that ideal positioning of the majority of the contacts for the electrode should be posterior and along the sacral border on fluoroscopic imaging. These findings also suggest that the S1 DRG may be located most reproducibly at the border of the intraforaminal and intracanalicular region.

The Anatomical Study and Clinical Significance of the Sinuvertebral Nerves at the Lumbar Levels.

STUDY DESIGN: A dissection-based study of 10 embalmed human cadavers. OBJECTIVE: The purpose of this study was to describe the sinuvertebral nerves at the lumbar level and to discuss their possible clinical significance. SUMMARY OF BACKGROUND DATA: Discogenic low-back pain is mediated by the sinuvertebral nerves. However, the detailed descriptions of the sinuvertebral nerves at the lumbar level are lacking. METHODS: One hundred L1-L5 intervertebral foramina from 10 embalmed cadavers were studied. The presence of the sinuvertebral nerves was noted. The quantity, origin, pathway, innervation range, and spatial orientations of the sinuvertebral nerves in the L1-L5 intervertebral foramina were examined. RESULTS: A total of 450 sinuvertebral nerves were identified in the 100 lumbar intervertebral foramina; sinuvertebral nerves were observed in 100.00% of the intervertebral foramina. The sinuvertebral nerves were routinely divided into the following two types: the sinuvertebral nerve deputy branch and sinuvertebral nerve main trunk. Three hundred twelve sinuvertebral nerve deputy branches were found; on average, there were approximately 3.12 (range, 1-8) branches in each intervertebral foramen. One hundred thirty-eight sinuvertebral nerve main trunks were found, and sinuvertebral nerve main trunks were observed in 97.00% of the intervertebral foramina. The initial portion of the sinuvertebral nerve was located along the posterior-lateral edge of the disc to the spinal canal. Sixty-one (44.20%) sinuvertebral nerve main trunks originated from the starting point of the gray ramus communicans of the nerve root; 77 (55.80%) sinuvertebral nerve main trunks originated from the anterior surface of the spinal ganglia of the nerve root. CONCLUSION: This is a systematic anatomy study that describes the sinuvertebral nerve at the lumbar level and may be of clinical importance to spinal surgeons. A comprehensive understanding of the distribution of sinuvertebral nerves may lead to significant benefits for patients undergoing percutaneous endoscopic treatment for discogenic low-back pain. LEVEL OF EVIDENCE: 4.

Unique Characteristics of the Dorsal Root Ganglion as a Target for Neuromodulation.

OBJECTIVE: The dorsal root ganglion (DRG) is a novel target for neuromodulation, and DRG stimulation is proving to be a viable option in the treatment of chronic intractable neuropathic pain. Although the overall principle of conventional spinal cord stimulation (SCS) and DRG stimulation-in which an electric field is applied to a neural target with the intent of affecting neural pathways to decrease pain perception-is similar, there are significant differences in the anatomy and physiology of the DRG that make it an ideal target for neuromodulation and may account for the superior outcomes observed in the treatment of certain chronic neuropathic pain states. This review highlights the anatomy of the DRG, its function in maintaining homeostasis and its role in neuropathic pain, and the unique value of DRG as a target in neuromodulation for pain. METHODS: A narrative literature review was performed. RESULTS: Overall, the DRG is a critical structure in sensory transduction and modulation, including pain transmission and the maintenance of persistent neuropathic pain states. Unique characteristics including selective somatic organization, specialized membrane characteristics, and accessible and consistent location make the DRG an ideal target for neuromodulation. Because DRG stimulation directly recruits the somata of primary sensory neurons and harnesses the filtering capacity of the pseudounipolar neural architecture, it is differentiated from SCS, peripheral nerve stimulation, and other neuromodulation options. CONCLUSIONS: There are several advantages to targeting the DRG, including lower energy usage, more focused and posture-independent stimulation, reduced paresthesia, and improved clinical outcomes.

Optical recording reveals topological distribution of functionally classified colorectal afferent neurons in intact lumbosacral DRG.

Neuromodulation as a non-drug alternative for managing visceral pain in irritable bowel syndrome (IBS) may target sensitized afferents of distal colon and rectum (colorectum), especially their somata in the dorsal root ganglion (DRG). Developing selective DRG stimulation to manage visceral pain requires knowledge of the topological distribution of colorectal afferent somata which are sparsely distributed in the DRG. Here, we implemented GCaMP6f to conduct high-throughput optical recordings of colorectal afferent activities in lumbosacral DRG, that is, optical electrophysiology. Using a mouse ex vivo preparation with distal colorectum and L5-S1 DRG in continuity, we recorded 791 colorectal afferents' responses to graded colorectal distension (15, 30, 40, and 60 mmHg) and/or luminal shear flow (20-30 mL/min), then functionally classified them into four mechanosensitive classes, and determined the topological distribution of their somata in the DRG. Of the 791 colorectal afferents, 90.8% were in the L6 DRG, 8.3% in the S1 DRG, and only 0.9% in the L5 DRG. L6 afferents had all four classes: 29% mucosal, 18.4% muscular-mucosal, 34% low-threshold (LT) muscular, and 18.2% high-threshold (HT) muscular afferents. S1 afferents only had three classes: 19.7% mucosal, 34.8% LT muscular, and 45.5% HT muscular afferents. All seven L5 afferents were HT muscular. In L6 DRG, somata of HT muscular afferents were clustered in the caudal region whereas somata of the other classes did not cluster in specific regions. Outcomes of this study can directly inform the design and improvement of next-generation neuromodulation devices that target the DRG to alleviate visceral pain in IBS patients.

Pioneer axons employ Cajal's battering ram to enter the spinal cord.

Sensory axons must traverse a spinal cord glia limitans to connect the brain with the periphery. The fundamental mechanism of how these axons enter the spinal cord is still debatable; both Ramon y Cajal's battering ram hypothesis and a boundary cap model have been proposed. To distinguish between these hypotheses, we visualized the entry of pioneer axons into the dorsal root entry zone (DREZ) with time-lapse imaging in zebrafish. Here, we identify that DRG pioneer axons enter the DREZ before the arrival of neural crest cells at the DREZ. Instead, actin-rich invadopodia in the pioneer axon are necessary and sufficient for DREZ entry. Using photoactivable Rac1, we demonstrate cell-autonomous functioning of invasive structures in pioneer axon spinal entry. Together these data support the model that actin-rich invasion structures dynamically drive pioneer axon entry into the spinal cord, indicating that distinct pioneer and secondary events occur at the DREZ.

Morphological anatomy of thoracolumbar nerve roots and dorsal root ganglia.

The aim of this study is to ascertain the anatomic parameters of the spinal roots and dorsal root ganglia and to demonstrate their clinical significance. Samples from 24 adult autopsy subjects were obtained from roots and dorsal root ganglia at levels L1 through L5. The anatomic parameters of epidural nerve roots: the distance between the epidural nerve roots and the proximal edge of the dorsal root ganglia and the average diameter of the nerve root gradually, increased from L1 to L5. The midline nerve root angle gradually decreased from L1 to L5. The anatomic parameters of subarachnoid nerve roots: the length of subarachnoid nerve roots and both the ventral and dorsal roots' diameter, increased from L1 to L5. The number of ventral and dorsal rootlets per nerve root ranged from one to three. The anatomic parameters of dorsal root ganglia: the length and width of the thoracic and lumbar dorsal root ganglia, gradually increased from L1 to L5. The locations of the dorsal root ganglia were recorded as the intraspinal, intraforaminal and extraforaminal using some bony landmarks. Most dorsal root ganglia located intraforaminally, and the extraforaminal type is more common in the L5 root than other thoracic and lumbar roots, regardless of age. This knowledge is a must not only to avoid complications but also for the success, safety and effectiveness of microsurgical operations.

Distribution and neurochemistry of porcine urinary bladder-projecting sensory neurons in subdomains of the dorsal root ganglia: A quantitative analysis.

The aim of the present study has been to verify the inter- and intraganglionic distribution pattern of porcine urinary bladder-projecting (UBP) neurons localized in the sacral dorsal root ganglia (DRGs). The morphology and chemical phenotype of these cells have also been investigated. These neurons were visualized using the fluorescent tracer Fast Blue (FB) which was injected bilaterally into the urinary bladder wall of five juvenile female pigs. The intraganglionic distribution showed that small- and medium-sized FB+ perikarya were mainly located in the central (S3-S4) and periphero-central (S2) region of the ganglia, while large cells were heterogeneously distributed. Immunohistochemistry revealed that the most frequently observed markers in small and medium-sized UBP perikarya were: neurofilament 200, lectin from Bandeiraea simplicifolia (Griffonia simplicifolia) isolectin B4, substance P, calcitonin gene-related peptide, pituitary adenylate cyclase-activating polypeptide and transient receptor potential vanilloid 1. Moreover, UBP neurons containing these substances were also mainly observed in the central and periphero-central region of the ganglion. Differences in the percentage of traced cells and their neuropeptide content were observed between the S2, S3 and S4 DRGs. In conclusion, the present study, for the first time, describes the arrangement of UBP DRGs neurons within particular subdomains of sacral ganglia, taking into account their size and chemical phenotype.

The short and midterm outcomes of lumbar transforaminal epidural injection with preganglionic and postganglionic approach in lumbosacral radiculopathy: a systematic review and meta-analysis.

The purpose of this study was to compare clinical outcomes after preganglionic versus ganglionic epidural steroid injection (ESI) using a systematic review and network meta-analysis. A systematic review and meta-regression was performed to compare postoperative outcomes between the two difference injection techniques. Relevant randomized controlled trials were identified from Medline and Scopus up to September 24, 2016. Sixteen out of 598 studies were eligible; 3, 2, and 3 studies were included in the pooling of outcomes including effectiveness, visual analog score (VAS), and complications (nerve root, injury, dural puncture, and intraneural injection). Preganglionic ESI has a 2.38 (95% CI 1.12, 5.04) times statistically significantly higher chance of effectiveness when compared to ganglionic ESI. There were differences in pain VAS and complications in lumbar radiculopathy, but these displayed no statistical significance. This meta-analysis indicated that preganglionic ESI has a statistically significantly higher chance of effectiveness when compared to ganglionic ESI. In terms of pain score and complications, there were no statistically significant differences between the two groups. These results were generally homogeneous and with little publication bias, thus should be generalizable.

Relevant Anatomy, Morphology, and Implantation Techniques of the Dorsal Root Ganglia at the Lumbar Levels.

OBJECTIVES: While dorsal root ganglion (DRG) stimulation has been available in Europe and Australia for the past five years and in the United States for the past year, there are no published details concerning the optimal procedures for DRG lead implantation. MATERIALS AND METHODS: We describe several techniques that can be applied to implant cylindrical leads over the DRG, highlighting some tips and tricks according to our experiences. Focus is mainly shifted toward implantations in the lumbar area. We furthermore give some insights in the results we experienced in Spain as well as some worldwide numbers. IMPLANT TECHNIQUES AND RESULTS: A 14-gauge needle is placed using a "2-Level Technique (2-LT)" or exceptionally a "1-Level Technique (1-LT)" or a "Primary- or Secondary Technique" at the level of L5. The delivery sheath, loaded with the lead, is advanced toward the targeted neural foramen. The lead is placed over the dorsal aspect of the DRG. A strain relief loop is created in the epidural space. Sheath and needle are retracted and the lead is secured using an anchor or anchorless technique. In Spain, 87.2% (N = 78) of the selected patients have been successfully implanted. Seven (8.9%) had a negative trial and three (4.2%) were explanted. Average VAS score decreased from 8.8 to 3.3 and on average 94.5% of the pain area was covered. In our center's subjects (N = 47 patients, 60.3% of all implanted patients in Spain), VAS scores decreased from an average of 8.8-1.7 and pain coverage averaged 96.4%. We used an average of 1.8 electrodes. Worldwide more than 4000 permanent cases have been successfully performed. CONCLUSIONS: We present implantation techniques whereby a percutaneous lead is placed over the DRG through the use of a special designed delivery sheath. Further investigation of the safety, efficacy, and sustainability of clinical outcomes using these devices is warranted.

Distribution pattern of dorsal root ganglion neurons synthesizing nitric oxide synthase in different animal species.

The main aim of the present review is to provide at first a short survey of the basic anatomical description of sensory ganglion neurons in relation to cell size, conduction velocity, thickness of myelin sheath, and functional classification of their processes. In addition, we have focused on discussing current knowledge about the distribution pattern of neuronal nitric oxide synthase containing sensory neurons especially in the dorsal root ganglia in different animal species; hence, there is a large controversy in relation to interpretation of the results dealing with this interesting field of research.

Variations of the accessory nerve: anatomical study including previously undocumented findings-expanding our misunderstanding of this nerve.

INTRODUCTION: The anatomy of the accessory nerve has been well described but continued new clinical and anatomical findings exemplify our lack of a full understanding of the course of this nerve. Therefore, this study aimed to expand on our knowledge of the course of the 11th cranial nerve via anatomical dissections. METHODS: Fifty-six cadavers (112 sides) underwent dissection of the accessory nerve from its cranial and spinal origins to its emergence into the posterior cervical triangle. Immunohistochemistry was performed when appropriate. RESULTS: Our findings included two cases (1.8%) where the nerve was duplicated, one intracranially and one extracranially. One accessory nerve (0.9%) was found to enter its own dural compartment within the jugular foramen. The majority of sides (80%) were found to have a cranial root of the accessory nerve. Thirty-one sides (28%) had connections to cervical dorsal roots medially and three sides (2.7%) laterally. Medial connections were most common with the C1 nerve. Medial components of these dorsal root connections were all sensory in nature. However, lateral components were motor on two sides (1.8%). Nerves traveled anterior to the internal jugular vein on 88% of sides. One (0.9%) left side nerve joined an interneural anastomosis between the dorsal rootlets. Macroganglia were found on the spinal part of the intracranial nerve on 13% of sides. The lesser occipital nerve arose directly from the accessory nerve on two sides (1.8%) and communicated with the accessory nerve on 5.4% of sides. One side (0.9%) was found to communicate with the facial nerve with both nerves innervating the sternocleidomastoid muscle. CONCLUSIONS: Additional anatomical knowledge of the variants of the accessory nerve may benefit patient care when this nerve is pathologically involved.

Fxyd2 regulates Aδ- and C-fiber mechanosensitivity and is required for the maintenance of neuropathic pain.

Identification of the molecular mechanisms governing sensory neuron subtype excitability is a key requisite for the development of treatments for somatic sensory disorders. Here, we show that the Na,K-ATPase modulator Fxyd2 is specifically required for setting the mechanosensitivity of Aδ-fiber low-threshold mechanoreceptors and sub-populations of C-fiber nociceptors, a role consistent with its restricted expression profile in the spinal somatosensory system. We also establish using the spared nerve injury model of neuropathic pain, that loss of Fxyd2 function, either constitutively in Fxyd2-/- mice or acutely in neuropathic rats, efficiently alleviates mechanical hypersensitivity induced by peripheral nerve lesions. The role of Fxyd2 in modulating Aδ- and C-fibers mechanosensitivity likely accounts for the anti-allodynic effect of Fxyd2 knockdown. Finally, we uncover the evolutionarily conserved restricted expression pattern of FXYD2 in human dorsal root ganglia, thus identifying this molecule as a potentially promising therapeutic target for peripheral neuropathic pain management.

Relationship of dorsal root ganglion to intervertebral foramen in lumbar region: an anatomical study and review of literature.

BACKGROUND: The purpose of this study was to clarify the morphologic features, location and variations of the dorsal root ganglion (DRG). METHODS: Fifteen formalin fixed cadavers for the current study were included. Total of 150 DRGs were examined from L1 to L5. The relationships of the nerve root DRGs to the intervertebral foramen were noted. Position of the DRG was classified by the location of the ganglia in relation to the pedicle. The relationship of the DRG to the intervertebral foramen was evaluated. RESULTS: The distance between the midpoint of the DRG to the cross section of the root with the medial border of the pedicle gradually increased from L1 to L5. The medial border of the foramen distances along the nerve root were L1, 1.77 mm; L2, 2.79 mm; L3, 3.23 mm; L4, 7.28 mm and L5, 8.31 mm. The mean width of the lumbar DRGs were L1, 4.36 mm; L2, 4.56 mm; L3, 4.99 mm; L4, 5.22 mm and L5, 5.82 mm. The mean length of DRGs were as follows: L1, 5.39 mm; L2, 5.83 mm; L3, 7.24 mm; L4, 7.97 mm and L5, 10.83 mm. The mean width and length of DRGs gradually increased from L1 to L5. CONCLUSIONS: The DRG in the lumbar region play a key role in the occurence of low-back pain and sciatica; therefore, it is important to understand the anatomy of DRG. The accurate anatomic information about the position of DRGs would be useful to perform a safe surgical intervention in the lumbar foraminal region.

The Relationship of the Vertebral Artery with Anatomical Landmarks in the Posterior Craniovertebral Junction of Fresh Human Cadavers in the Turkish Population.

AIM: Surgical anatomy concerning the posterior craniovertebral region in fresh human cadavers was studied to provide most accurate information for the surgical approach. MATERIAL AND METHODS: In thirty-two fresh human cadavers, the distance from the posterior tubercle to the sulcus of vertebral artery (VA), the thickness and length of the third segment of VA (V3), the distance of C1/C2 facet to V3, the length, height and shape of the C2 ganglion to the neighboring structures, the distance from medial border of C1 lateral mass to dura mater, the distance of the transverse process of atlas to mastoid tip, the thickness of C1 posterior arcus were measured. RESULTS: There were variations of sulcus of VA in 14 of 32 cadavers (43.7%), the right VA was larger in 23 cadavers (71.8%). The ganglion was found over the C1 lateral mass screw entry point in 45 of 64 ganglions (70.31%) and below the screw entry point in 19 of 64 ganglions (29.69%). The distance of the medial border of the C1 lateral mass to dural tube was 3.81±0.55 mm at the right side and 3.91±0.59 mm at the left. The thickness of C1 posterior arch was 3.73±0.75 mm at the right side and 3.75±0.77 mm at the left. The mean distance from the transverse process of C1 to the mastoid tip was 15.82±4.49 mm at the right side and 15.46±4.38 mm at the left. CONCLUSION: This is the most comprehensive and only fresh cadaver study about this region in the literature.

Mass Spectrometry Imaging and GC-MS Profiling of the Mammalian Peripheral Sensory-Motor Circuit.

Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) has evolved to become an effective discovery tool in science and clinical diagnostics. Here, chemical imaging approaches are applied to well-defined regions of the mammalian peripheral sensory-motor system, including the dorsal root ganglia (DRG) and adjacent nerves. By combining several MSI approaches, analyte coverage is increased and 195 distinct molecular features are observed. Principal component analysis suggests three chemically different regions within the sensory-motor system, with the DRG and adjacent nerve regions being the most distinct. Investigation of these regions using gas chromatography-mass spectrometry corroborate these findings and reveal important metabolic markers related to the observed differences. The heterogeneity of the structurally, physiologically, and functionally connected regions demonstrates the intricate chemical and spatial regulation of their chemical composition.

Dispersal pattern of injectate following CT-guided perineural infiltration in the canine thoracolumbar spine: a cadaver study.

An increasing proportion of canine patients are presented with chronic thoracolumbar back pain and without compressive spinal lesions. In humans, spinal perineural infiltrations have been reported to have a favorable effect on pain control. The purpose of this prospective cadaver study was to describe the dispersal pattern of injectate following CT-guided spinal perineural infiltration in the canine thoracolumbar region. Seven fresh canine cadavers were first scanned using multislice CT and then CT-guided spinal perineural infiltration was performed at 42 sites from T13/L1 to L6/L7. The injectate for each site was a mixture of new methylene blue and iodinated contrast medium. Immediately following CT-guided injection, cadavers were frozen, cut, and dissected macro- and mesoscopically (using a magnifying glass) to identify anatomic structures that were infiltrated. In the majority of sites (64.3%), complete epidural and hypaxial staining of spinal nerve components (including the spinal ganglion, trunk, and ventral branch) was successfully achieved. However, no (11.9%) or unpredictable staining (9.5%) of nervous tissue occurred in some sites despite careful CT guidance and the application of relatively large volumes of injectate. Optimal results were achieved when the needle tip was positioned periforaminally ventral to the cranial contour of the cranial articular process. Findings from this ex vivo study indicated that CT-guided spinal perineural infiltration is feasible for testing in the canine thoracolumbar region and that successful nerve tissue infiltration would likely occur in the majority of sites. Future in vivo studies are needed to determine the safety and efficacy of this technique.

Identification of different types of spinal afferent nerve endings that encode noxious and innocuous stimuli in the large intestine using a novel anterograde tracing technique.

In mammals, sensory stimuli in visceral organs, including those that underlie pain perception, are detected by spinal afferent neurons, whose cell bodies lie in dorsal root ganglia (DRG). One of the major challenges in visceral organs has been how to identify the different types of nerve endings of spinal afferents that transduce sensory stimuli into action potentials. The reason why spinal afferent nerve endings have been so challenging to identify is because no techniques have been available, until now, that can selectively label only spinal afferents, in high resolution. We have utilized an anterograde tracing technique, recently developed in our laboratory, which facilitates selective labeling of only spinal afferent axons and their nerve endings in visceral organs. Mice were anesthetized, lumbosacral DRGs surgically exposed, then injected with dextran-amine. Seven days post-surgery, the large intestine was removed. The characteristics of thirteen types of spinal afferent nerve endings were identified in detail. The greatest proportion of nerve endings was in submucosa (32%), circular muscle (25%) and myenteric ganglia (22%). Two morphologically distinct classes innervated myenteric ganglia. These were most commonly a novel class of intraganglionic varicose endings (IGVEs) and occasionally rectal intraganglionic laminar endings (rIGLEs). Three distinct classes of varicose nerve endings were found to innervate the submucosa and circular muscle, while one class innervated internodal strands, blood vessels, crypts of lieberkuhn, the mucosa and the longitudinal muscle. Distinct populations of sensory endings were CGRP-positive. We present the first complete characterization of the different types of spinal afferent nerve endings in a mammalian visceral organ. The findings reveal an unexpectedly complex array of different types of primary afferent endings that innervate specific layers of the large intestine. Some of the novel classes of nerve endings identified must underlie the transduction of noxious and/or innocuous stimuli from the large intestine.