Changes in Cellular Localization of Inter-Alpha Inhibitor Proteins after Cerebral Ischemia in the Near-Term Ovine Fetus

Inter-alpha Inhibitor Proteins (IAIPs) are key immunomodulatory molecules. Endogenous IAIPs are present in human, rodent, and sheep brains, and are variably localized to the cytoplasm and nuclei at multiple developmental stages. We have previously reported that ischemia-reperfusion (I/R) reduces IAIP concentrations in the fetal sheep brain. In this study, we examined the effect of I/R on total, cytoplasmic, and nuclear expression of IAIPs in neurons (NeuN+), microglia (Iba1+), oligodendrocytes (Olig2+) and proliferating cells (Ki67+), and their co-localization with histones and the endoplasmic reticulum in fetal brain cells. At 128 days of gestation, fetal sheep were exposed to Sham (n = 6) or I/R induced by cerebral ischemia for 30 min with reperfusion for 7 days (n = 5). Although I/R did not change the total number of IAIP+ cells in the cerebral cortex or white matter, cells with IAIP+ cytoplasm decreased, whereas cells with IAIP+ nuclei increased in the cortex. I/R reduced total neuronal number but did not change the IAIP+ neuronal number. The proportion of cytoplasmic IAIP+ neurons was reduced, but there was no change in the number of nuclear IAIP+ neurons. I/R increased the number of microglia and decreased the total numbers of IAIP+ microglia and nuclear IAIP+ microglia, but not the number of cytoplasmic IAIP+ microglia. I/R was associated with reduced numbers of oligodendrocytes and increased proliferating cells, without changes in the subcellular IAIP localization. IAIPs co-localized with the endoplasmic reticulum and histones. In conclusion, I/R alters the subcellular localization of IAIPs in cortical neurons and microglia but not in oligodendrocytes or proliferating cells. Taken together with the known neuroprotective effects of exogenous IAIPs, we speculate that endogenous IAIPs may play a role during recovery from I/R.

Immediate Insulin Treatment Prevents Diabetes-Induced Gut Region-Specific Increase in the Number of Myenteric Serotonergic Neurons

To evaluate the effects of hyperglycemia and insulin treatment on the proportion of serotonin-immunoreactive (5-HT-IR) myenteric neurons, samples were taken from the duodenum, ileum, and colon of diabetic, insulin-treated diabetic, and control rats 10 weeks after the onset of streptozotocin-induced hyperglycemia. Myenteric whole-mount preparations were immunostained with anti-5-HT and pan-neuronal anti-HuCD markers. In controls, the 5-HT-IR myenteric neurons represent a small proportion (~2.5%) of the total neuronal number in the investigated gut segments. The proportion of 5-HT-IR myenteric neurons was significantly higher in the duodenum (p < 0.01) and colon (p < 0.0001) of diabetic rats compared to the controls but exhibited a slight increase in the ileum. Immediate insulin treatment resulted in a significantly lower proportion of myenteric 5-HT-IR neurons in each segment (duodenum p < 0.0001; ileum p < 0.01; and colon p < 0.0001) compared to the untreated diabetics. Our study demonstrates that the proportion of 5-HT-IR myenteric neurons was enhanced in type 1 diabetes in a region-specific manner. Immediate insulin treatment prevents a higher hyperglycemia-induced amount of 5-HT-IR neurons and restores it to the control level in each investigated gut segment. Despite the low proportion of 5-HT-IR myenteric neurons, hyperglycemia-related changes of these neurons may play a crucial role in gastrointestinal symptoms in type 1 diabetes.

Characterization of the dynamics and variability of neuronal subtype responses during growth, degrowth, and regeneration of Nematostella vectensis

Background The ability to regenerate body parts is a feature of metazoan organisms and the focus of intense research aiming to understand its basis. A number of mechanisms involved in regeneration, such as proliferation and tissue remodeling, affect whole tissues; however, little is known on how distinctively different constituent cell types respond to the dynamics of regenerating tissues. Preliminary studies suggest that a number of organisms alter neuronal numbers to scale with changes in body size. In some species with the ability of whole-body axis regeneration, it has additionally been observed that regenerates are smaller than their pre-amputated parent, but maintain the correct morphological proportionality, suggesting that scaling of tissue and neuronal numbers also occurs. However, the cell dynamics and responses of neuronal subtypes during nervous system regeneration, scaling, and whole-body axis regeneration are not well understood in any system. The cnidarian sea anemone Nematostella vectensis is capable of whole-body axis regeneration, with a number of observations suggesting the ability to alter its size in response to changes in feeding. We took advantage of Nematostella’s transparent and “simple” body plan and the NvLWamide-like mCherry fluorescent reporter transgenic line to probe the response of neuron populations to variations in body size in vivo in adult animals during body scaling and regeneration. Results We utilized the previously characterized NvLWamide-like::mCherry transgenic reporter line to determine the in vivo response of neuronal subtypes during growth, degrowth, and regeneration. Nematostella alters its size in response to caloric intake, and the nervous system responds by altering neuronal number to scale as the animal changes in size. Neuronal numbers in both the endodermal and ectodermal nerve nets decreased as animals shrunk, increased as they grew, and these changes were reversible. Whole-body axis regeneration resulted in regenerates that were smaller than their pre-amputated size, and the regenerated nerve nets were reduced in neuronal number. Different neuronal subtypes had distinct responses during regeneration, including consistent, not consistent, and conditional increases in number. Conditional responses were regulated, in part, by the size of the remnant fragment and the position of the amputation site. Regenerates and adults with reduced nerve nets displayed normal behaviors, indicating that the nerve net retains functionality as it scales. Conclusion These data suggest that the Nematostella nerve net is dynamic, capable of scaling with changes in body size, and that neuronal subtypes display differential regenerative responses, which we propose may be linked to the scale state of the regenerating animals.

Microglia dynamics in adolescent traumatic brain injury

Repetitive, mild traumatic brain injuries (RmTBIs) are increasingly common in adolescents and encompass one of the largest neurological health concerns in the world. Adolescence is a critical period for brain development where RmTBIs can substantially impact neurodevelopmental trajectories and life-long neurological health. Our current understanding of RmTBI pathophysiology suggests key roles for neuroinflammation in negatively regulating neural health and function. Microglia, the brain’s resident immune population, play important roles in brain development by regulating neuronal number, and synapse formation and elimination. In response to injury, microglia activate to inflammatory phenotypes that may detract from these normal homeostatic, physiological, and developmental roles. To date, however, little is known regarding the impact of RmTBIs on microglia function during adolescent brain development. This review details key concepts surrounding RmTBI pathophysiology, adolescent brain development, and microglia dynamics in the developing brain and in response to injury, in an effort to formulate a hypothesis on how the intersection of these processes may modify long-term trajectories.

Evolution of neuronal anatomy and circuitry in two highly divergent nematode species

The nematodes C. elegans and P. pacificus populate diverse habitats and display distinct patterns of behavior. To understand how their nervous systems have diverged, we undertook a detailed examination of the neuroanatomy of the chemosensory system of P. pacificus. Using independent features such as cell body position, axon projections and lipophilic dye uptake, we have assigned homologies between the amphid neurons, their first-layer interneurons, and several internal receptor neurons of P. pacificus and C. elegans. We found that neuronal number and soma position are highly conserved. However, the morphological elaborations of several amphid cilia are different between them, most notably in the absence of ‘winged’ cilia morphology in P. pacificus. We established a synaptic wiring diagram of amphid sensory neurons and amphid interneurons in P. pacificus and found striking patterns of conservation and divergence in connectivity relative to C. elegans, but very little changes in relative neighborhood of neuronal processes. These findings demonstrate the existence of several constraints in patterning the nervous system and suggest that major substrates for evolutionary novelty lie in the alterations of dendritic structures and synaptic connectivity.

FTLD-TDP With and Without GRN Mutations Cause Different Patterns of CA1 Pathology

Heterozygous loss-of-function mutations in the GRN gene lead to progranulin (PGRN) haploinsufficiency and cause frontotemporal lobar degeneration with TDP-43 pathology type A (FTLD-TDP type A). PGRN is a highly conserved, secreted glycoprotein and functions in the central nervous system as a key modulator of microglial function. Hence, altered microglial function caused by PGRN deficiency may be tied to the pathogenesis of FTLD-TDP. Our previous studies showed that haploinsufficiency of GRN mutations extends to microglial PGRN expression in the hippocampal CA1 region. In this study, we found that the CA1 sector was associated with less neuronal loss and more frequent TDP-43 inclusions in FTLD-TDP type A cases with GRN mutations than in sporadic cases. In addition, the CA1 region in GRN mutation cases contained more rod-like microglia, which also had reduced PGRN expression. These findings suggest that the profile of TDP-43 inclusions, neuronal number, and microgliosis in the CA1 sector of FTLD-TDP type A cases may be influenced by GRN gene expression status.

Dynamics and variability of neuronal subtype responses during growth, degrowth, and regeneration

BackgroundWe are interested in nervous system dynamics in adult and regenerating animals. Preliminary studies suggest that some species alter neuronal number to scale with changes in body size. Similarly, in some species regenerates resulting from wholebody axis regeneration are smaller than their pre-amputated parent, but they maintain the correct proportionality, suggesting that tissue and neuronal scaling also occurs in regenerates. The cell dynamics and responses of neuronal subtypes during nervous system regeneration, scaling, and whole-body axis regeneration are not well understood in any system. The cnidarian sea anemone Nematostella vectensis is capable of wholebody axis regeneration, and its transparent, “simple” body plan and the availability of fluorescent reporter transgenic lines allow neuronal subtypes to be tracked in vivo in adult and regenerating animals. A number of observations suggest this anemone is able to alter its size in responses to changes in feeding. We utilized the NvLWamide-like::mCherry neuronal subtype transgenic reporter line to determine the in vivo response of neuronal subtypes during growth, degrowth, and regeneration.ResultsNematostella alters its size in response to caloric intake, and the nervous system responds by altering neuronal number to scale as the animal changes in size. Neuronal numbers in both the endodermal and ectodermal nerve nets decreased as animals shrunk, increased as they grew, and the changes were reversible. Whole-body axis regeneration resulted in regenerates that were smaller than their pre-amputated size, and the regenerated nerve nets were reduced in neuronal number. Different neuronal subtypes had several distinct responses during regeneration that included consistent, no, and conditional increases in number. Conditional responses were regulated, in part, by the size of the remnant fragment and the position of the amputation site. Regenerates and adults with reduced nerve nets displayed normal behaviors, indicating that the nerve net retains functionality as it scales.ConclusionThese data suggest that the Nematostella nerve net is dynamic, capable of scaling with changes in body size, and that neuronal subtypes display differential regenerative responses, which we propose may be linked to the scale state of the regenerating animals.

REACTION-DIFFUSION MECHANISMS UNDERLYING HIRSCHPRUNG'S DISEASE AND THEIR PRACTICAL IMPICATIONS

The erratic extent of aganglionic and hypoganglionic segments in Hirschsprung's disease (HD) makes it difficult to predict the amount of the intestine to remove in order to restore the proper functional motility. Our aim was to assess whether the embryonic rostro-caudal intestinal colonization by neuroblasts from the neural crest follows a predictable pattern in HD. In touch with Turing's reaction diffusion model (RD), which describes biological patterns (such as leopard spots and lung branching morphogenesis) in terms of interactions/competitions between activating and inhibiting factors, we hypothesized that intestinal neural density could be triggered by local gut factors that counteract the proximal-distal embryonic progression of neural progenitors. While the neuronal number is approximately the same throughout the whole intestine in healthy subjects, in HD neural density decreases rostro-caudally towards the rectal region, due to an augmented activity and concentration of distal local inhibitors. In order to prove our hypothesis of HD's nervous rostro-caudal adjustments driven by Turing-like processes, we compared the neuronal density patterns achieved through RD models' simulations with the neuronal numbers detected in different colonic regions from affected children. We showed that the virtual and the real plots display fully overlapping and matching features. The fact that neuronal decreases in impaired colons match Turing equations's previsions points towards the human intestine (both healthy and sick) as colonized through a diffusive proximal-distal neural pattern that is predictable, allowing us to straightforwardly calculate the length of the gut to resect during surgical procedures for HD.