Molecular Genetic Characterization of Endemic Yellow Catfish, Horabagrus brachysoma (Gunther)

Horabagrus brachysoma (Gunther) an endemic, cultivable catfish belonging to Family Bagridae is found in rivers originating from southern part of the biodiversity hotspot - the Western Ghats. This species is categorized as "endangered" based on latest IUCN criteria, due to restricted distribution, loss of habitat, over exploitation, destructive fishing practices and trade (Anon., 1998). ... The species is shonlisted for taking up stock-specific, propagation-assisted rehabilitation programme in rivers where it is naturally distributed. Information on the basic genetic profile and stock structure, which is essential for the fishery management, conservation and rehahilitation of this species, is lacking. Hence, the present work was undenaken to identify molecular genetic markers like allozymes, RAPD and microsatellites in H. braclrysoma and to use them in analyzing the population genetic structure of this species, collected from three geographically isolated river systems viz., Meenachil, Chalakkudy and Nethravathi in South India. ... In allozymes, the screening was carried out to detect 25 enzymes, but only 14 showed their presence with scorable activity. Out of founeen enzymes studied, 12 enzymes were polymorphic and two enzymes were monomorphic. The founeen enzymes yielded 25 scorable loci in all populations. A total of 33 alleles were obtained in overall populations

Population Genetic Structure of Endemic and Endangered Yellow Catfish, Horabagrus brachysoma, Using Allozyme Markers

The allozyme variation and population genetic structure of Horabagrus brachysoma in three natural populations from the southern part of theWestern Ghats region, India, were investigated by polyacrylamide gel electrophoresis. Variations at 14 loci from 14 enzyme systems were analyzed. The allozyme analysis revealed a high level of genetic variation in this species, with an average of observed alleles per locus of 2.357 and observed heterozygosity of 0.178. The positive value of the fixation index (FIS = 0.507) implied a significant deficiency of heterozygosity at the population level. The highly significant probability (P<0.0001) for the overall loci suggested that the three sample sets were not part of the same gene pool

Identification of Random Amplified Polymorphic DNA (RAPD) Markers in Endemic Yellow Catfish, Horabagrus brachysoma (Gunther 1864)

Random amplified polymorphic DNA (RAPD) analysis was applied to individuals of Horabagrus brachysoma sampled from three geographic locations of Western Ghat river systems in India. Of the thirty-two 10-mer RAPD primers screened initially, ten were chosen and used in a comparative analysis of H. brachysoma collected from Chalakkudy, Meenachil and Nethravathi river systems. A total of 124 RAPD fragments were amplified, out of which 49 (39.51%) were found to be shared by individuals of all three river systems. The remaining 75 fragments were found to be polymorphic (60.48%). This confirms the suitability of RAPD markers for the study of population genetic structure in yellow catfish stocks

Two PTP receptors mediate CSPG inhibition by convergent and divergent signaling pathways in neurons

Receptor protein tyrosine phosphatase σ (PTPσ) and its subfamily member LAR act as transmembrane receptors that mediate growth inhibition of chondroitin sulfate proteoglycans (CSPGs). Inhibition of either receptor increases axon growth into and beyond scar tissues after CNS injury. However, it is unclear why neurons express two similar CSPG receptors, nor whether they use the same or different intracellular pathways. We have now studied the signaling pathways of these two receptors using N2A cells and primary neurons derived from knockout mice. We demonstrate that both receptors share certain signaling pathways (RhoA, Akt and Erk), but also use distinct signals to mediate CSPG actions. Activation of PTPσ by CSPGs selectively inactivated CRMP2, APC, S6 kinase and CREB. By contrast LAR activation inactivated PKCζ, cofilin and LKB1. For the first time, we propose a model of the signaling pathways downstream of these two CSPG receptors. We also demonstrate that deleting both receptors exhibits additive enhancement of axon growth in adult neuronal cultures in vitro. Our findings elucidate the novel downstream pathways of CSPGs and suggest potential synergy of blocking their two PTP receptors.This work was supported by research grants to SL from NIH (1R01NS079432, 1R21NS066114 and 1R01EY024575) and Shriners Research Foundation (SHC-86300-PHI and SHC-86200-PHI-16), and to MES from NIH (1R01NS092876) and Shriners Research Foundation (SHC-85400)

Identification of polymorphic allozyme markers for population structure analysis in Horabagrus brachysoma (Gunther, 1864).

Fourteen polymorphic allozyme loci were identified in yellow catfish, Horabagrus brachysoma. The genetic variation detected at each allozyme locus was assessed for samples collected from three rivers. The observed heterozygosities per locus ranged from 0.0286 to 0.4000. Significant genotype heterogeneity indicated that the samples are not drawn from same gene pool. The results suggest the potential of the identified loci to analyze stock structure of natural populations of H. brachysoma

Identification and characterization of microsatellite markers for the population genetic structure in endemic red-tailed barb, Gonoproktopterus curmuca

Gonoproktopterus curmuca is an endangered red tailed barb found in Southern part of Western Ghat, India. As a part of stock-specific, propagation assisted rehabilitation and management program, polymorphic microsatellites markers were used to study the genetic diversity and population structure of this species from the three River systems of Southern Western Ghats, such as Periyar River, the Chalakkudy River, and the Chaliyar River. From selected eight polymorphic microsatellite markers, the number of alleles per locus ranged from 2 to 8, and the average number of alleles among 3 populations ranged from 5.0 to 5.75. The mean observed (Hob) and expected (Hex) heterozygosity ranged from 0.5148 to 0.5360 and from 0.5996 to 0.6067, respectively. Significant deviations from Hardy-Weinberg Equilibrium expectation were found at majority of the loci (except Gcur MFW72 and Gcur MFW19) and in all three populations in which heterozygote deficits were apparent. The analysis of molecular variance indicates that the percent of variance among populations and within populations were 6.73 and 93.27, respectively. The pairwise FST values between populations indicate that there were significant deviations in genetic differentiations for the red-tailed barb populations from these three Rivers of the Western Ghats, India. The microsatellites methods reported a low degree of gene diversity and lack of genetic heterogeneity in the population of G. curmuca, which strongly emphasize the need of fishery management, conservation and rehabilitation of G. curmuca

Aspects of astrocyte energy metabolism, amino acid neurotransmitter homoeostasis and metabolic compartmentation

Astrocytes are key players in brain function; they are intimately involved in neuronal signalling processes and their metabolism is tightly coupled to that of neurons. In the present review, we will be concerned with a discussion of aspects of astrocyte metabolism, including energy-generating pathways and amino acid homoeostasis. A discussion of the impact that uptake of neurotransmitter glutamate may have on these pathways is included along with a section on metabolic compartmentation

Methamphetamine Inhibits the Glucose Uptake by Human Neurons and Astrocytes: Stabilization by Acetyl-L-Carnitine

Methamphetamine (METH), an addictive psycho-stimulant drug exerts euphoric effects on users and abusers. It is also known to cause cognitive impairment and neurotoxicity. Here, we hypothesized that METH exposure impairs the glucose uptake and metabolism in human neurons and astrocytes. Deprivation of glucose is expected to cause neurotoxicity and neuronal degeneration due to depletion of energy. We found that METH exposure inhibited the glucose uptake by neurons and astrocytes, in which neurons were more sensitive to METH than astrocytes in primary culture. Adaptability of these cells to fatty acid oxidation as an alternative source of energy during glucose limitation appeared to regulate this differential sensitivity. Decrease in neuronal glucose uptake by METH was associated with reduction of glucose transporter protein-3 (GLUT3). Surprisingly, METH exposure showed biphasic effects on astrocytic glucose uptake, in which 20 µM increased the uptake while 200 µM inhibited glucose uptake. Dual effects of METH on glucose uptake were paralleled to changes in the expression of astrocytic glucose transporter protein-1 (GLUT1). The adaptive nature of astrocyte to mitochondrial β-oxidation of fatty acid appeared to contribute the survival of astrocytes during METH-induced glucose deprivation. This differential adaptive nature of neurons and astrocytes also governed the differential sensitivity to the toxicity of METH in these brain cells. The effect of acetyl-L-carnitine for enhanced production of ATP from fatty oxidation in glucose-free culture condition validated the adaptive nature of neurons and astrocytes. These findings suggest that deprivation of glucose-derived energy may contribute to neurotoxicity of METH abusers

Specific ion channels contribute to key elements of pathology during secondary degeneration following neurotrauma

Background: Following partial injury to the central nervous system, cells beyond the initial injury site undergo secondary degeneration, exacerbating loss of neurons, compact myelin and function. Changes in Ca 2+ flux are associated with metabolic and structural changes, but it is not yet clear how flux through specific ion channels contributes to the various pathologies. Here, partial optic nerve transection in adult female rats was used to model secondary degeneration. Treatment with combinations of three ion channel inhibitors was used as a tool to investigate which elements of oxidative and structural damage related to long term functional outcomes. The inhibitors employed were the voltage gated Ca 2+ channel inhibitor Lomerizine (Lom), the Ca 2+ permeable AMPA receptor inhibitor YM872 and the P2X 7 receptor inhibitor oxATP. Results: Following partial optic nerve transection, hyper-phosphorylation of Tau and acetylated tubulin immunoreactivity were increased, and Nogo-A immunoreactivity was decreased, indicating that axonal changes occurred acutely. All combinations of ion channel inhibitors reduced hyper-phosphorylation of Tau and increased Nogo-A immunoreactivity at day 3 after injury. However, only Lom/oxATP or all three inhibitors in combination significantly reduced acetylated tubulin immunoreactivity. Most combinations of ion channel inhibitors were effective in restoring the lengths of the paranode and the paranodal gap, indicative of the length of the node of Ranvier, following injury. However, only all three inhibitors in combination restored to normal Ankyrin G length at the node of Ranvier. Similarly, HNE immunoreactivity and loss of oligodendrocyte precursor cells were only limited by treatment with all three ion channel inhibitors in combination. Conclusions: Data indicate that inhibiting any of a range of ion channels preserves certain elements of axon and node structure and limits some oxidative damage following injury, whereas ionic flux through all three channels must be inhibited to prevent lipid peroxidation and preserve Ankyrin G distribution and OPCs