Immunohistochemical phenotype of sensory neurons associated with sympathetic plexuses in the trigeminal ganglia of adult nerve growth factor transgenic mice.

Following peripheral nerve injury, postganglionic sympathetic axons sprout into the affected sensory ganglia and form perineuronal sympathetic plexuses with somata of sensory neurons. This sympathosensory coupling contributes to the onset and persistence of injury-induced chronic pain. We have documented the presence of similar sympathetic plexuses in the trigeminal ganglia of adult mice that ectopically overexpress nerve growth factor (NGF), in the absence of nerve injury. In this study, we sought to further define the phenotype(s) of these trigeminal sensory neurons having sympathetic plexuses in our transgenic mice. Using quantitative immunofluorescence staining analyses, we show that the invading sympathetic axons specifically target sensory somata immunopositive for several biomarkers: NGF high-affinity receptor tyrosine kinase A (trkA), calcitonin gene-related peptide (CGRP), neurofilament heavy chain (NFH), and P2X purinoceptor 3 (P2X3). Based on these phenotypic characteristics, the majority of the sensory somata surrounded by sympathetic plexuses are likely to be NGF-responsive nociceptors (i.e., trkA expressing) that are peptidergic (i.e., CGRP expressing), myelinated (i.e., NFH expressing), and ATP sensitive (i.e., P2X3 expressing). Our data also show that very few sympathetic plexuses surround sensory somata expressing other nociceptive (pain) biomarkers, including substance P and acid-sensing ion channel 3. No sympathetic plexuses are associated with sensory somata that display isolectin B4 binding. Though the cellular mechanisms that trigger the formation of sympathetic plexus (with and without nerve injury) remain unknown, our new observations yield an unexpected specificity with which invading sympathetic axons appear to target a precise subtype of nociceptors. This selectivity likely contributes to pain development and maintenance associated with sympathosensory coupling.

Olfactory Dysfunction and Alzheimer's Disease: A Review.

 Alzheimer's disease is the most common cause of dementia, and it is one of the leading causes of death globally. Identification and validation of biomarkers that herald the onset and progression of Alzheimer's disease is of paramount importance for early reliable diagnosis and effective pharmacological therapy commencement. A substantial body of evidence has emerged demonstrating that olfactory dysfunction is a preclinical symptom of neurodegenerative diseases including Alzheimer's disease. While a correlation between olfactory dysfunction and Alzheimer's disease onset and progression in humans exists, the mechanism underlying this relationship remains unknown. The aim of this article is to review the current state of knowledge regarding the range of potential factors that may contribute to the development of Alzheimer's disease-related olfactory dysfunction. This review predominantly focuses on genetic mutations associated with Alzheimer's disease including amyloid-ß protein precursor, presenilin 1 and 2, and apolipoprotein E mutations, that may (in varying ways) drive the cellular events that lead to and sustain olfactory dysfunction.

Evaluation of motor and sensory neuron populations in a mouse median nerve injury model.

BACKGROUND: Peripheral nerves can regenerate and restore function after injury but this process is hindered by many factors including chronic denervation, motor end-plate resorption and Schwann cell senescence. Forelimb injury models in rodents are becoming increasingly popular as they more accurately reflect the physiology and biomechanics of upper extremity nerve injuries. However several aspects of this surgical model remain poorly characterized. NEW METHOD: C57Bl/6 mice underwent enumeration of median nerve motor and sensory neuron pools using retrograde labeling with or without nerve transection. Distal histomorphometry of uninjured mouse median nerves was also examined. Baseline reference values of volitional forelimb grip strength measurements were determined and the rate of neural elongation was also estimated. RESULTS: We identified 1363 ± 165 sensory and 216 ± 16 motor neurons within the uninjured dorsal root ganglia (DRG) and ventral spinal cord, respectively. Eight days following injury, approximately 34% of motoneurons had elongated a distance of 5 mm beyond the repair site 8 days following injury. Volitional grip strength decreased 50% with unilateral median nerve transection and was negligible with contralateral flexor tendon tenotomy. COMPARISON WITH EXISTING METHOD: Our spinal cord and DRG harvesting technique presented here was technically straightforward and reliable. Estimates of motor and sensory neuron numbers for the mouse median nerve compared favourably with studies using intramuscular injection of retrograde neurotracer. Histomorphometry data was consistent with and reinforced reference values in the literature. CONCLUSIONS: This study provides data that further develops an increasingly popular surgical model for studying peripheral nerve injury and repair.

Microglial/macrophage cells in mammalian olfactory nerve fascicles.

This is the first description of a population of Iba1- and annexin A3-immunopositive cells residing in the peripheral olfactory nerves of adult rats and adult cats. Based on their ramified appearance, positive immunostaining for the monocytic markers Iba1 and annexin A3, and reactivity to bulbectomy (in adult rats), these cells found within the olfactory nerve fascicles of both mammalian species meet several important criteria for their designation as microglia/macrophages. These Iba1-/annexin A3-immunopositive cells may be uniquely positioned to protect against the potential spread of dangerous environmental xenobiotics (such as viruses and toxins) into the brain, where such pathogens may contribute to the development of neurological diseases, such Alzheimer's and Parkinson's diseases.

Attenuation of opioid analgesic tolerance in p75 neurotrophin receptor null mutant mice.

Repeated exposure to opioid drugs can lead to the development of tolerance, which manifests as a reduction in analgesic potency, and physical dependence, a response indicated by a withdrawal syndrome. Accumulating evidence suggests that the nerve growth factor (NGF) family of neurotrophins may have an important modulatory role in the induction of opioid analgesia and opioid addiction. Because neurotrophins universally bind the p75 neurotrophin receptor (p75NTR), we investigated whether the activity of this receptor is involved in the development of opioid analgesic tolerance and physical dependence. We found that in both the wild-type and p75NTR-/- mice an acute systemic (i.p.) injection of morphine produced a maximal analgesic response as measured by the thermal tail-immersion test. Repeated injection of morphine over 5 days in wild-type mice resulted in a progressive decline of the analgesic effect and a concomitant loss of the agonist potency, reflecting development of morphine tolerance. However, the loss of morphine analgesia was not observed in p75NTR-/- mice. In the second part of this study, mice were given escalating doses of systemic (i.p.) morphine over 5 days and subsequently challenged with the opioid receptor antagonist naloxone. This challenge precipitated a robust withdrawal syndrome that was comparable in wild-type mice and p75NTR-/- mice. The findings suggest that p75NTR activity plays a critical role in the development of opioid analgesic tolerance but not in the induction or the expression of opioid physical dependence.

Student Reflections on the Queen's Accelerated Route to Medical School Programme.

CONTEXT: Since its inception more than 150 years ago, the School of Medicine at Queen's University has aspired 'to advance the tradition of preparing excellent physicians and leaders in health care by embracing a spirit of inquiry and innovation in education and research'. As part of this continuing commitment, Queen's School of Medicine developed the Queen's University Accelerated Route to Medical School (QuARMS). As Canada's only 2-year accelerated-entry premedical programme, QuARMS was designed to reduce training time, the associated expense of medical training, and to encourage a collaborative premedical experience. Students enter QuARMS directly from high school and then spend 2 years enrolled in an undergraduate degree programme. They then are eligible to enter the first-year MD curriculum. The 2-year QuARMS academic curriculum includes traditional undergraduate coursework, small group sessions, and independent activities. The QuARMS curriculum is built on 4 pillars: communication skills, critical thinking, the role of physician (including community service learning [CSL]), and scientific foundations. Self-regulated learning (SRL) is explicitly developed throughout all aspects of the curriculum. Medical educators have defined SRL as the cyclical control of academic and clinical performance through several key processes that include goal-directed behaviour, use of specific strategies to attain goals, and the adaptation and modification to behaviours or strategies that optimize learning and performance. Based on Zimmerman's social cognitive framework, this definition includes relationships among the individual, his or her behaviour, and the environment, with the expectation that individuals will monitor and adjust their behaviours to influence future outcomes. OBJECTIVES: This study evaluated the students' learning as perceived by them at the conclusion of their first 2 academic years. METHODS: At the end of the QuARMS learning stream, the first and second cohorts of students completed a 26-item, 4-point Likert-type instrument with space for optional narrative details for each question. A focus group with each group explored emergent issues. Consent was obtained from 9 out of 10 and 7 out of 8 participants to report the 2015 survey and focus group data, respectively, and from 10 out of 10 and 9 out of 10 participants to report the 2016 survey and focus group data, respectively. Thematic analysis and a constructivist interpretive paradigm were used. A distanced facilitator, standard protocols, and a dual approach assured consistency and trustworthiness of data. RESULTS: Both analyses were congruent. Students described experiences consistent with curricular goals including critical thinking, communication, role of a physician, CSL, and SRL. Needs included additional mentorship, more structure for CSL, more feedback, explicit continuity between in-class sessions, and more clinical experience. Expectations of students towards engaging in independent learning led to some feelings of disconnectedness. CONCLUSIONS: Participants described benefit from the sessions and an experience consistent with the curricular goals, which were intentionally focused on foundational skills. In contrast to the goal of SRL, students described a need for an explicit educational structure. Thus, scaffolding of the curriculum from more structured in year 1 to less structured in year 2 using additional mentorship and feedback is planned for subsequent years. Added clinical exposure may increase relevance but poses challenges for integration with the first-year medical class.

Olfactory ensheathing cells express smooth muscle alpha-actin in vitro and in vivo.

One strategy for spinal cord repair after injury that has moved quickly from the research laboratory to the clinic is the implantation of olfactory ensheathing cells (OECs). These unique glial cells of the olfactory system have been associated with axonal remyelination and regeneration after grafting into spinalized animals. Despite these promising observations, there remains a lack of direct empirical evidence of the exact fate of OECs after intraspinal implantation, in large part because of a surprising paucity of defined biomarkers that unequivocally distinguish these cells from phenotypically similar Schwann cells. Here we provide direct neurochemical proof that OECs, both in vitro and in vivo, express smooth muscle alpha-actin. That OECs synthesize this contractile protein (and a variety of actin-binding proteins including caldesmon) provides compelling evidence that these cells are, in fact, quite different from Schwann cells. The identification of several smooth muscle-related proteins in OECs points to a new appreciation of the structural and functional features of this population of olfactory glia. These biomarkers can now be used to elucidate the fate of OECs after intraspinal implantation, in particular assessing whether smooth muscle alpha-actin-expressing OECs are capable of facilitating axon remyelination and regeneration.

Defining the role of olfactory ensheathing cells in facilitating axon remyelination following damage to the spinal cord.

Olfactory ensheathing cells (OECs) are unique cells that are responsible for the successful regeneration of olfactory axons throughout the life of adult mammals. More than a decade of research has shown that implantation of OECs may be a promising therapy for damage to the nervous system, including spinal cord injury. Based on this research, several clinical trials worldwide have been initiated that use autologous transplantation of olfactory tissue containing OECs into the damaged spinal cord of humans. However, research from several laboratories has challenged the widely held belief that OECs are directly responsible for myelinating axons and promoting axon regeneration. The purpose of this review is to provide a working hypothesis that integrates several current ideas regarding the mechanisms of the beneficial effects of OECs. Specifically, OECs promote axon regeneration and functional recovery indirectly by augmenting the endogenous capacity of host Schwann cells to invade the damaged spinal cord. Together with Schwann cells, OECs create a 3-dimensional matrix that provides a permissive microenvironment for successful axon regeneration in the adult mammalian central nervous system.

Cultures of rat olfactory ensheathing cells are contaminated with Schwann cells.

Implantation of cultured olfactory ensheathing cells into the damaged spinal cord of adult rats has been reported to remyelinate central axons. This observation is curious because olfactory ensheathing cells do not myelinate axons in their native environment. We have recently determined that calponin is the first definitive phenotypic marker for olfactory ensheathing cells. Primary cultures of adult rat olfactory mucosa and olfactory bulb were immunostained for p75 neurotrophin receptor and calponin. Our results reveal that two populations of p75 neurotrophin receptor-positive cells exist in primary cultures of the olfactory mucosa and bulb: calponin-positive olfactory ensheathing cells and calponin-negative Schwann cells. As olfactory tissues likely yield a mixed glial population, the idea that olfactory ensheathing cells are capable of de novo myelin synthesis after intraspinal implantation should be re-evaluated.

Serum proteomics as a strategy to identify novel biomarkers of neurologic recovery after cardiac arrest: a feasibility study.

BACKGROUND: Serum biomarkers may play a role in prognostication after cardiac arrest. This study was designed to assess the feasibility of using two-dimensional gel electrophoresis (2D-GE) coupled with mass spectrometry (MS) as a proteomic strategy to identify novel biomarkers that may predict neurological recovery. METHODS: Adult comatose survivors of ventricular fibrillation or pulseless ventricular tachycardia were considered eligible. Blood was collected and serum separated within 6 h of hospital admission and then at 24 h afterwards. Neurological outcome was assessed at 3 months with the Cerebral Performance Category (CPC) score. Serum was assessed with 2D-GE with and without prior depletion of high abundance proteins. Protein differences between patients with good (CPC 1,2) vs. poor (CPC 3-5) neurological recovery were subsequently identified with MS. RESULTS: From August 2010 to June 2014, 11 patients meeting eligibility criteria were recruited, from which serum was available from 9 (5 with good neurological outcome). On non-depleted serum, only high abundance acute phase proteins such as haptoglobin, cell-free hemoglobin, albumin, and amyloid were detected in both patients with good and poor neurological recovery. Following depletion of high abundance proteins, proteins identified by MS in both patient populations were the acute phase reactants c-reactive protein and retinol binding protein-4. Proteins uniquely identified in the serum of patients with poor neurological recovery included 14-3-3 (epsilon and zeta isoforms) and muskelin. CONCLUSIONS: Two-D-GE coupled with MS is a feasible strategy to facilitate the identification of novel predictive biomarkers. The presence of muskelin and 14-3-3 in the serum of patients with poor neurological prognosis warrants further investigation.

Nerve growth factor-mediated collateral sprouting of central sensory axons into deafferentated regions of the dorsal horn is enhanced in the absence of the p75 neurotrophin receptor.

This study examined the growth capacity of nerve growth factor (NGF)-responsive dorsal root ganglion (DRG) central processes using mice of the following genotypes: wildtype, p75 neurotrophin receptor (p75NTR) exon III null mutant, NGF transgenic, and NGF transgenic with p75NTR exon III null mutation (NGF/p75(-/-)). In wildtype and p75NTR exon III null mutant mice calcitonin gene-related peptide (CGRP) immunoreactivity in the dorsal horn is dramatically reduced at both 3 and 28 days after rhizotomy. NGF transgenic and NGF/p75(-/-) mice also display reduced CGRP immunoreactivity 3 days after rhizotomy, but by postsurgical day 28 significant increases in the density of CGRP-positive axons are observed in the injured dorsal horns of these mice. Interestingly, NGF/p75(-/-) mice displayed significantly more new axonal growth when compared to NGF transgenic mice expressing full-length p75NTR. Immunohistochemical and ultrastructural analyses revealed that this axonal growth is not the result of regeneration but rather injury-induced sprouting by intact DRG central processes into the lesion site. This collateral growth is restricted to deafferentated areas of the dorsal horn, and we therefore propose that this is an example of compensatory sprouting by NGF-sensitive axons in the spinal cord, a response that is enhanced in the absence of NGF binding to p75NTR.

A proteomic approach to assess intraneuronal inclusions associated with neurodegenerative disorders.

In neuroscience, proteomic technology is being used to discover the chemical features of neurodegenerative disorders, including Parkinson's disease, Alzheimer's disease and amyotrophic lateral sclerosis. Pathologically, one hallmark feature common to these diseases is the presence of proteinaceous inclusions within affected neurons. Proteomic assessment of diseased tissues and animal models reveals that the occurrence of these protein-rich aggregations may be due to perturbed functioning of a neuron-specific ubiquitin-recycling enzyme.

The influences of p75 neurotrophin receptor and brain-derived neurotrophic factor in the sympathetic innervation of target tissues during murine postnatal development.

Post-ganglionic sympathetic neurons express the p75 neurotrophin receptor (p75NTR) and brain-derived neurotrophic factor (BDNF), which together have been implicated in controlling the degree of efferent innervation of peripheral organs [Kohn, J., Aloyz, R.S., Toma, J.G., Haak-Frendscho, M., Miller, F.D. 1999. Functionally Antagonistic Interactions between the TrkA and p75 Neurotrophin Receptors Regulate Sympathetic Neuron Growth and Target Innervation. J. Neurosci. 19, 5393-5408]. To examine this concept further, we developed null mutant mice lacking both p75NTR and BDNF, and assessed whether the loss of this receptor-ligand interaction negatively impacts the degree of sympathetic innervation to various target tissues. Between postnatal days 10 and 14, hearts, urinary bladders, kidneys, and submandibular salivary glands were isolated from p75(-/-)/BDNF-/-, p75-/-, BDNF-/-, and wild type siblings. Sympathetic axons were visualized using tyrosine hydroxylase (TH) immunohistochemistry, and TH protein levels were quantified by immunoblotting. Concerning the sympathetic innervation of the heart, urinary bladder and kidneys, no differences were seen in single and double null mutant mice, as compared with their wild type siblings. Sympathetic innervation of the submandibular salivary gland was, however, increased in both p75-/- and p75(-/-)/BDNF-/- mice over control mice. These results reveal that an absence of p75NTR and/or BDNF expression does not perturb the degree of sympathetic innervation of many peripheral tissues during postnatal development, and that a lack of p75NTR expression may actually enhance the density of these efferent fibers in other target tissues, such as the salivary glands.

Myenteric expression of nerve growth factor and the p75 neurotrophin receptor regulate axonal remodeling as a consequence of colonic inflammation in mice.

Nerve growth factor (NGF) levels increase in response to inflammation of the mammalian colon. The precise cellular sources of colonic NGF synthesis, however, remain elusive. Using lines of transgenic mice that express enhanced green fluorescent protein (EGFP) under the control of the NGF promoter, we found a subpopulation of adendritic EGFP(+) neurons in the myenteric plexus. These colonic EGFP(+) neurons display positive immunostaining for calretinin but not nitric oxide synthase 1 (NOS1) two biomarkers of mouse myenteric neurons. A loss of NGF expression in null mutant postnatal mice does not affect the survival of these EGFP(+) neurons. Induction of colonic inflammation confirms local increases in NGF mRNA/protein levels, which coincide with heightened detection of EGFP by myenteric neurons. Though NOS1(+) myenteric neurons display positive immunostaining for trkA (the receptor required for NGF binding/signaling), transgenic overexpression of NGF by smooth muscle cells in the colon does not alter the survival, somal size, or axonal density of trkA-expressing NOS1(+) myenteric neurons. Mice lacking functional p75NTR (the second receptor required for NGF binding) exhibit significantly less axonal damage among NOS1(+) myenteric neurons, in response to chemically induced colonic inflammation. Likewise, trkA-expressing sympathetic axons that innervate the myenteric ganglia display less damage in the absence of p75NTR. These data are the first to implicate calretinin(+) myenteric neurons as a source of NGF in the murine colon, and that in response to colonic inflammation, increases in NGF can exaggerate damage of intrinsic NOS1(+) axons and extrinsic sympathetic axons that co-express trkA and p75NTR.

Structural and neurochemical features of postganglionic sympathetic neurons in the superior mesenteric ganglion of spontaneously hypertensive rats.

Postganglionic sympathetic neurons, which are exquisitely sensitive to small changes in levels of target-derived nerve growth factor (NGF), express two transmembrane receptors: 1) the trkA receptor mediates neuron survival and neurite outgrowth; and 2) the p75 neurotrophin receptor (p75NTR) enhances neuronal responsiveness of trkA to NGF. Elevating levels of NGF induces several morphological and neurochemical alterations in sympathetic neurons, including axonal sprouting, increased levels of p75NTR mRNA relative to trkA mRNA, and increased accumulations of NGF in hypertrophied somata. Spontaneously hypertensive rats (SHR) display both elevated NGF levels and increased sympathetic axonal innervation of the mesenteric vasculature. In this investigation we assessed whether sympathetic neurons innervating the mesenteric vasculature of SHR display other features indicative of increased levels of target-derived NGF. In 5-week-old SHR, levels of both p75NTR and trkA mRNA in mesenteric sympathetic neurons were significantly elevated compared to levels in age-matched control rats. By 15 and 30 weeks of age, levels of p75NTR mRNA expression in mesenteric sympathetic neurons were similar between SHR and control rats. Accumulations of NGF were depleted in the sympathetic somata of 15- and 30-week-old SHR compared to age-matched control rats. Moreover, sympathetic neurons in SHR were not hypertrophied, as the sizes of somata were comparable between SHR and control rats. Our data illustrate that despite having augmented levels of NGF in the mesenteric vasculature, SHR do not display many of the morphological and neurochemical features that are associated with an enhanced responsiveness by sympathetic neurons to elevated levels of target-derived NGF.

Nerve growth factor alters p75 neurotrophin receptor-induced effects in mouse facial motoneurons following axotomy.

The p75 neurotrophin receptor (p75(NTR)) has been implicated as being detrimental for cell survival in facial motoneurons following injury. Although facial motoneurons do not respond to nerve growth factor (NGF) under normal circumstances, this study shows that NGF can interfere with p75(NTR)-mediated cell survival effects on motoneurons following injury. Twenty-five days following injury, the proportion of surviving axotomized neurons in NGF/p75(+/+) mice, which overexpress NGF, was significantly higher compared to wild-type mice, while NGF/p75(-/-) mice, which overexpress NGF but carry two mutated alleles for p75(NTR), had fewer neurons compared to wild-type and p75(-/-) mice, which carry two mutated alleles for p75(NTR), resulting in a lack of functional expression of this receptor. Sympathetic axons sprouted into the axotomized facial nucleus of both NGF/p75(+/+) and NGF/p75(-/-) following injury, due to transgene expression of NGF in reactive astrocytes. Removal of these sympathetic axons enhanced the number of surviving axotomized neurons in NGF/p75(-/-) mice but not in NGF/p75(+/+) mice. Although motoneurons do not express trkA and should therefore be unresponsive to NGF, our results reveal that NGF can influence p75-mediated motoneuron survival following axotomy.

Neuronal degeneration associated with sympathosensory plexuses in the trigeminal ganglia of aged mice that overexpress nerve growth factor.

Aberrant sympathetic sprouting is seen in the uninjured trigeminal ganglia of transgenic mice that ectopically express nerve growth factor under the control of the glial fibrillary acidic protein promoter. These sympathetic axons form perineuronal plexuses around a subset of sensory somata in 2- to 3-month-old transgenic mice. Here, we show that aged transgenic mice (i.e., 11-14 and 16-18 months old) have dystrophic sympathetic plexuses (i.e., increased densities of swollen axons), and that satellite glial cells, specifically those in contact with dystrophic plexuses in the aged mice display strong immunostaining for tumor necrosis factor alpha. The colocalization of dystrophic plexuses and reactive satellite glial cells in the aged mice coincides with degenerative features in the enveloped sensory somata. Collectively, these novel results show that, with advancing age, sympathetic plexuses undergo dystrophic changes that heighten satellite glial cell reactivity and that together these cellular events coincide with neuronal degeneration.

Reactive astrocytes associated with plaques in TgCRND8 mouse brain and in human Alzheimer brain express phosphoprotein enriched in astrocytes (PEA-15).

To identify potential biomarkers associated with Alzheimer's disease (AD)-like neuropathologies in the murine brain, we conducted proteomic analyses of neocortices from TgCRND8 mice. Here we found that phosphoprotein enriched in astrocytes 15 kDa (PEA-15) is expressed at higher levels in the neocortical proteomes from 6-month old TgCRND8 mice, as compared to non-transgenic mice. Immunostaining for PEA-15 revealed reactive astrocytes associated with the neocortical amyloid plaques in TgCRND8 mice and in post-mortem human AD brains. This is the first report of increased phosphoprotein enriched in astrocytes (PEA-15) expression in reactive astrocytes of an AD mouse model and human AD brains.

Olfactory ensheathing cells of hamsters, rabbits, monkeys, and mice express α-smooth muscle actin.

Olfactory ensheathing cells (OECs) are the chief glial population of the mammalian olfactory nervous system, residing in the olfactory mucosa and at the surface of the olfactory bulb. We investigated the neurochemical features of OECs in a variety of mammalian species (including adult hamsters, rabbits, monkeys, and mice, as well as fetal pigs) using three biomarkers: α-smooth muscle actin (αSMA), S100ß, and glial fibrillary acidic protein (GFAP). Mucosal and bulbar OECs from all five mammalian species express S100ß. Both mucosal and bulbar OECs of monkeys express αSMA, yet only bulbar OECs of hamsters and only mucosal OECs of rabbits express αSMA as well. Mucosal OECs, but not bulbar OECs, also express GFAP in hamsters and monkeys; mice, by comparison, have only a sparse population of OECs expressing GFAP. Though αSMA immunostaining is not detected in OECs of adult mice, GFAP-expressing mucosal OECs isolated from adult mice do coexpress αSMA in vitro. Moreover, mucosal OECs from adult mutant mice lacking αSMA expression display perturbed cellular morphology (i.e., fewer cytoplasmic processes extending among the hundreds of olfactory axons in the olfactory nerve fascicles and nuclei having degenerative features). In sum, these findings highlight the efficacy of αSMA and S100ß as biomarkers of OECs from a variety of mammalian species. These observations provide definitive evidence that mammalian OECs express the structural protein αSMA (at various levels of detection), which appears to play a pivotal role in their ensheathment of olfactory axons.

Overexpression of nerve growth factor by murine smooth muscle cells: role of the p75 neurotrophin receptor on sympathetic and sensory sprouting.

Elevating levels of nerve growth factor (NGF) can have pronounced effects on the survival and maintenance of distinct populations of neurons. We have generated a line of transgenic mice in which NGF is expressed under the control of the smooth muscle α-actin promoter. These transgenic mice have augmented levels of NGF protein in the descending colon and urinary bladder, so these tissues display increased densities of NGF-sensitive sympathetic efferents and sensory afferents. Here we provide a thorough examination of sympathetic and sensory axonal densities in the descending colon and urinary bladder of NGF transgenic mice with and without the expression of the p75 neurotrophin receptor (p75NTR). In response to elevated NGF levels, sympathetic axons (immunostained for tyrosine hydroxylase) undergo robust collateral sprouting in the descending colon and urinary bladder of adult transgenic mice (i.e., those tissues having smooth muscle cells); this sprouting is not augmented in the absence of p75NTR expression. As for sensory axons (immunostained for calcitonin gene-related peptide) in the urinary bladders of transgenic mice, fibers undergo sprouting that is further increased in the absence of p75NTR expression. Sympathetic axons are also seen invading the sensory ganglia of transgenic mice; these fibers form perineuronal plexi around a subpopulation of sensory somata. Our results reveal that elevated levels of NGF in target tissues stimulate sympathetic and sensory axonal sprouting and that an absence of p75NTR by sensory afferents (but not by sympathetic efferents) leads to a further increase of terminal arborization in certain NGF-rich peripheral tissues.