Soumya Iyengar

Activation of apoptosis signal regulating kinase 1 (ASK1) and translocation of death-associated protein, Daxx, in substantia nigra pars compacta in a mouse model of Parkinson’s disease: protection by α-lipoic acid

Parkinsons disease (PD), a neurodegenerative disorder, causes severe motor impairment due to loss of dopaminergic neurons in substantia nigra pars compacta (SNpc). MPTP, a neurotoxin that causes dopaminergic cell loss in mice, was used in an animal model to study the pathogenic mechanisms leading to neurodegeneration. We observed the activation of apoptosis signal regulating kinase (ASK1, MAPKKK) and phosphorylation of its downstream targets MKK4 and JNK, 12 h after administration of a single dose of MPTP. Further, Daxx, the death‐associated protein, translocated to the cytosol selectively in SNpc neurons seemingly due to MPTP mediated down‐regulation of DJ‐1, the redox‐sensitive protein that binds Daxx in the nucleus. Coadministration of a‐lipoic acid (ALA), a thiol antioxidant, abolished the activation of ASK1 and phosphorylation of downstream kinases, MKK4, and JNK and prevented the down‐regulation of DJ‐1 and translocation of Daxx to the cytosol seen after MPTP. ALA also attenuated dopaminergic cell loss in SNpc seen after subchronic MPTP treatment. Our studies demonstrate for the first time that MPTP triggers death signaling pathway by activating ASK1 and translocating Daxx, in vivo, in dopaminergic neurons in SNpc of mice and thiol antioxidants, such as ALA terminate this cascade and afford neuroprotection.–Karunakaran S., Diwakar, L., Saeed, U., Agarwal, V., Ramakrishnan, S., Iyengar, S., Ravindranath V. Activation of apoptosis signal regulating kinase 1 (ASK1) and translocation of death associated protein, Daxx in substantia nigra pars compacta in a mouse model of Parkinsons disease: Protection by α‐lipoic acid. FASEB J. 21, 2226–2236 (2007)

Cortical and thalamic connections of the representations of the teeth and tongue in somatosensory cortex of new world monkeys.

Connections of representations of the teeth and tongue in primary somatosensory cortex (area 3b) and adjoining cortex were revealed in owl, squirrel, and marmoset monkeys with injections of fluorescent tracers. Injection sites were identified by microelectrode recordings from neurons responsive to touch on the teeth or tongue. Patterns of cortical label were related to myeloarchitecture in sections cut parallel to the surface of flattened cortex, and to coronal sections of the thalamus processed for cytochrome oxidase (CO). Cortical sections revealed a caudorostral series of myelin dense ovals (O1–O4) in area 3b that represent the periodontal receptors of the contralateral teeth, the contralateral tongue, the ipsilateral teeth, and the ipsilateral tongue. The ventroposterior medial subnucleus, VPM, and the ventroposterior medial parvicellular nucleus for taste, VPMpc, were identified in the thalamic sections. Injections placed in the O1 oval representing teeth labeled neurons in VPM, while injections in O2 representing the tongue labeled neurons in both VPMpc and VPM. These injections also labeled adjacent part of areas 3a and 1, and locations in the lateral sulcus and frontal lobe. Callosally, connections of the ovals were most dense with corresponding ovals. Injections in the area 1 representation of the tongue labeled neurons in VPMpc and VPM, and ipsilateral area 3b ovals, area 3a, opercular cortex, and cortex in the lateral sulcus. Contralaterally, labeled neurons were mostly in area 1. The results implicate portions of areas 3b, 3a, and 1 in the processing of tactile information from the teeth and tongue, and possibly taste information from the tongue. J. Comp. Neurol. 501:95–120, 2007.

Preterm delivery disrupts the developmental program of the cerebellum.

A rapid growth in human cerebellar development occurs in the third trimester, which is impeded by preterm delivery. The goal of this study was to characterize the impact of preterm delivery on the developmental program of the human cerebellum. Still born infants, which meant that all development up to that age had taken place in-utero, were age paired with preterm delivery infants, who had survived in an ex-utero environment, which meant that their development had also taken place outside the uterus. The two groups were assessed on quantitative measures that included molecular markers of granule neuron, purkinje neuron and bergmann glia differentiation, as well as the expression of the sonic hedgehog signaling pathway, that is important for cerebellar growth. We report that premature birth and development in an ex-utero environment leads to a significant decrease in the thickness and an increase in the packing density of the cells within the external granular layer and the inner granular layer well, as a reduction in the density of bergmann glial fibres. In addition, this also leads to a reduced expression of sonic hedgehog in the purkinje layer. We conclude that the developmental program of the cerebellum is specifically modified by events that follow preterm delivery.

Expression of Sonic Hedgehog During Cell Proliferation in the Human Cerebellum

The regulation of cell proliferation in the external granular layer (EGL) of the developing cerebellum is important for its normal patterning. An important signal that regulates EGL cell proliferation is Sonic hedgehog (Shh). Shh is secreted by the Purkinje cells (PC) and has a mitogenic effect on the granule cell precursors of the EGL. Deregulation of Shh signaling has been associated with abnormal development, and been implicated in medulloblastomas, which are tumors that arise from the cerebellum. Given the importance of the Shh pathway in cerebellum development and disease, there has been no systematic study of its expression pattern during human cerebellum development. In this study, we describe the expression pattern of Shh, its receptor patched, smoothened, and its effectors that belong to the Gli family of transcription factors, during normal human cerebellum development from 10 weeks of gestational age, and in medulloblastomas that represents a case of abnormal cell proliferation in the cerebellum. This expression pattern is compared to equivalent stages in the normal development of cerebellum in mouse, as well as in tumors. Important differences between human and mouse that reflect differences in the normal developmental program between the 2 species are observed. First, in humans there appears to be a stage of Shh signaling within the EGL, when the PC are not yet the source of Shh. Second, unlike in the postnatal mouse cerebellum, expression of Shh in the PC in the postnatal human cerebellum is downregulated. Finally, medulloblastomas in the human but not in patched heterozygote mouse express Shh. These results highlight cross-species differences in the regulation of the Shh signaling pathway.

Expression of δ- and μ-opioid receptors in the ventricular and subventricular zones of the developing human neocortex

Recent research has documented the involvement of the endogenous opioid system in neural development, including neurogenesis and neuronal differentiation. However, the expression of opioid receptors (ORs) in different cell types of the human ventricular and subventricular zones (VZ and SVZ) has not been studied during early gestation. In the present study, we have used immunohistochemistry and quantified the results to demonstrate that the levels of delta- and mu-OR subtypes were high in the VZ and SVZ between 11 and 16 gestation weeks (GW) and decreased by 20GW. These results have also been confirmed by studying OR mRNA expression in the VZ and SVZ. Both delta- and mu-OR subtypes were expressed by multipotential stem cells, newly differentiated neurons and developing glial cells to different extents. However, migrating neurons expressed negligible levels of both OR subtypes. Our results suggest that the opioid system may affect cellular proliferation and/or differentiation of stem cells into neurons and glia during the first and second trimesters of gestation in humans. Since layers II and III of the cerebral cortex are being formed during the second trimester, their development is most likely affected by the opioid system mediated through delta- and mu-ORs.

Opioid modulation of song in male zebra finches (Taenopygia guttata)

Endogenous opioids are known to modulate motivated behaviors. Male zebra finches produce a highly motivated behavior (directed song) to court females and also sing in isolation (undirected song). We found that adult male zebra finches sang significantly fewer directed and undirected songs after administration of low doses (2.5 mg/kg body weight) of the general opioid antagonist naloxone, even though the order of syllables in songs was not altered. Surprisingly, high doses of naloxone (10 mg/kg body weight) dramatically decreased the production of undirected songs but had no significant effects on directed songs. There were no changes in the number of calls during directed or undirected song, movement, stereotyped behaviors including pecking and preening, feeding or drinking behaviors after naloxone administration. We also found that treating zebra finches with naloxone led to a decrease in tonality (goodness of pitch), frequency and amplitude modulation and an increase in the length of intersyllable intervals. Our results suggest that the opioid system can differentially modulate directed and undirected song as well as the acoustic characteristics of birdsong, perhaps by acting on different components of the song control system.

Expression of μ- and δ-opioid receptors in song control regions of adult male zebra finches (Taenopygia guttata)

The opioid system is known to play a role in various aspects of learning and memory in diverse species of mammals and birds. Earlier studies have localized the endogenous opioids, met- and leu-enkephalin, in the song control regions of male zebra finches (Taenopygia guttata), a sexually dimorphic species of songbirds wherein only males sing. Recent research has shown that levels of enkephalin increase in some of the song control regions during singing and that blocking opioid receptors with the antagonist naloxone decreases the frequency of singing in songbirds. However, the distribution of receptors specific to the opioid system has not been studied in zebra finches. In the present study, we used reverse transcriptase PCR (RT-PCR) and quantitative real-time PCR (qRT-PCR) to demonstrate that the song control regions lateral magnocellular nucleus of the anterior nidopallium (LMAN), Area X, MSt (medial striatum), HVC and RA (robust nucleus of the arcopallium) expressed higher levels of mu-OR mRNA compared to delta-OR mRNA. In situ hybridization was used to demonstrate that neither LMAN nor Area X could be delineated from the surrounding brain regions [anterior nidopallium (ANP) and MSt, respectively], based on OR mRNA expression. However, HVC and RA neurons expressed marginally higher levels of mu-OR mRNA compared to the posterior nidopallium, which was confirmed by immunohistochemical localization. We also found that the dorsolateral subdivision of DLM (dorsolateral nucleus of the medial thalamus) demonstrated high levels of mu-OR immunoreactivity. Our results suggest that the ORs may be involved in modulating different aspects of vocalization and/or gating auditory input, as well as motor control.

Neuronatin-mediated Aberrant Calcium Signaling and Endoplasmic Reticulum Stress Underlie Neuropathology in Lafora Disease

Background: Neuronatin was identified as a novel substrate of Lafora disease ubiquitin ligase malin; however, its role in disease biology is unknown. Results: Neuronatin causes increased intracellular Ca2+ and ER stress and accumulated as insoluble aggregates in LD brain and skin biopsy samples. Conclusion: Neuronatin-induced aberrant Ca+2 signaling might trigger LD pathogenesis. Significance: These findings provide a new insight in understanding pathogenesis of LD. Lafora disease (LD) is a teenage-onset inherited progressive myoclonus epilepsy characterized by the accumulations of intracellular inclusions called Lafora bodies and caused by mutations in protein phosphatase laforin or ubiquitin ligase malin. But how the loss of function of either laforin or malin causes disease pathogenesis is poorly understood. Recently, neuronatin was identified as a novel substrate of malin that regulates glycogen synthesis. Here we demonstrate that the level of neuronatin is significantly up-regulated in the skin biopsy sample of LD patients having mutations in both malin and laforin. Neuronatin is highly expressed in human fetal brain with gradual decrease in expression in developing and adult brain. However, in adult brain, neuronatin is predominantly expressed in parvalbumin-positive GABAergic interneurons and localized in their processes. The level of neuronatin is increased and accumulated as insoluble aggregates in the cortical area of LD brain biopsy samples, and there is also a dramatic loss of parvalbumin-positive GABAergic interneurons. Ectopic expression of neuronatin in cultured neuronal cells results in increased intracellular Ca2+, endoplasmic reticulum stress, proteasomal dysfunction, and cell death that can be partially rescued by malin. These findings suggest that the neuronatin-induced aberrant Ca2+ signaling and endoplasmic reticulum stress might underlie LD pathogenesis.

Opioid modulation of cell proliferation in the ventricular zone of adult zebra finches (Taenopygia guttata)

Besides modulating pain, stress, physiological functions, motivation, and reward, the opioid system has been implicated in developmental and adult mammalian neurogenesis and gliogenesis. In adult male songbirds including zebra finches, neurons generated from the ventricular zone (VZ) of the lateral ventricles are incorporated throughout the telencephalon, including the song control nuclei, HVC, and area X. Although the endogenous opioid met‐enkephalin is present in neurons adjacent to the VZ and is upregulated in song control regions during singing, it is not known whether the opioid system can modulate adult neurogenesis/gliogenesis in zebra finches. We used quantitative RT‐PCR and in situ hybridization to demonstrate that µ‐ and δ‐opioid receptors are expressed by the VZ of adult male zebra finches. Treating cultured VZ cells from male birds with the opioid antagonist naloxone led to an increase in cell proliferation measured by 5‐bromo‐2‐deoxyuridine incorporation, whereas administering met‐enkephalin had the opposite effect, compared with saline‐treated cultures. Systemically administering naloxone (2.5 mg/kg body wt) to adult male zebra finches for 4 d also led to a significant increase in cell proliferation in the ventral VZ of these birds, compared with saline‐treated controls. Our results show that cell proliferation is augmented by naloxone in the VZ adjacent to the anterior commissure, suggesting that the endogenous opioids modulate adult neurogenesis/gliogenesis by inhibiting cell proliferation in songbirds.—Khurshid, N., Hameed, L. S., Mohanasundaram, S., Iyengar, S. Opioid modulation of cell proliferation in the ventricular zone of adult zebra finches (Taenopygia guttata). FASEB J. 24, 3681–3695 (2010). www.fasebj.org

Avian Cognition and Consciousness—From the Perspective of Neuroscience and Behaviour

Are birds conscious? Birds and primates occupy similar ecological niches, face similar challenges in foraging for food, and live in large social groups. Despite the fact that brain evolution is divergent in birds and mammals, the evolution of brain function is convergent. A number of studies on neural networks, structure, function and behaviour have demonstrated striking similarities between the overall organisation of the brain in humans, other primates and birds. Taken together, this data suggests that both cognition and consciousness may have evolved independently and in parallel across different species of birds and mammals. The present review focuses on the remarkable cognitive abilities of different species of birds such as problem-solving, tool use, mathematical abilities and self-awareness, the neural circuits underlying these behaviours and attempts to link the avian brain and behaviour to consciousness.