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Saraf, R. R.

Publications and source records attributed to Saraf, R. R..

2 recordsLinked to original sources

Spatial and temporal requirement of Mlp60A isoforms during muscle development and function in Drosophila melanogaster

Many myofibrillar proteins undergo isoform switching in a spatio-temporal manner during muscle development. The biological significance of the variants of several of these myofibrillar proteins remains elusive. One such myofibrillar protein, the Muscle LIM Protein (MLP), is a vital component of the Z-discs. In this paper, we show that one of the Drosophila MLP encoding genes, Mlp60A, gives rise to two isoforms: a short (279 bp, 10 kDa) and a long (1461 bp, 54 kDa) one. The short isoform is expressed throughout development, but the long isoform is adult-specific, being the dominant of the two isoforms in the indirect flight muscles (IFMs). A concomitant, muscle-specific knockdown of both isoforms leads to late pupal lethality, with the surviving flies being majorly flight defective. Mlp60A null flies show developmental lethality, and muscle defects in the individuals surviving till the third instar larval stage. This lethality could be rescued partially by muscle-specific overexpression of the short isoform. Almost 90% of the long isoform-specific P-element insertion mutant flies show a compromised flight ability and have reduced sarcomere length. Hence, our data shows that the two Mlp60A isoforms are functionally specialized, to ensuring normal embryonic muscle development and adult flight muscle function.

developmental biology↗

SNCAE46K transgenic Drosophila Model of Parkinsons Disease Confirmed the Causative Role of Oxidative Stress

Parkinsons disease (PD) is a class of neurodegenerative disorders in which, complex interactions of genetic and environmental agents are involved in the etiology of both sporadic and familial PD cases. -synuclein-encoding SNCA gene is known as one of the major genetic contributors of this disease. E46K mutation in SNCA gene has not been investigated as intensive as other SNCA gene mutations including A30P and A53T. In this study, to induce PD in Drosophila flies, UAS-hSNCAWT and UAS-hSNCAE46K transgenic fly lines were constructed, where SNCA gene was over-expressed in flies brains using GAL4-UAS genetic system. Western blot analysis of head samples of SNCA-expressing flies verified SNCA expression at protein level. Light and electron microscopy analysis of ommatidial structures were performed to verify neurodegeneration as a result of -synuclein gene overexpression in Drosophila transgenic flies. Confocal microscopy analysis of dopaminergic neuron clusters verified cell loss following SNCAE46K expression in the flies brain. E46K -synuclein gene over-expression resulted in an evident decline in longevity as well as climbing ability of the flies. Biochemical studies of transgenic flies showed a remarkable decline in antioxidant enzymes activity and a significant increase in oxidative markers level as well as AchE enzyme activity. Oxidative stress has been known as a causal factor in PD pathogenesis, following expression of E46K mutant version of human SNCA gene. This Drosophila model is able to facilitate comparative studies of both molecular and cellular assays implicated in the assessment of neurotoxicity of different -synuclein mutations.

neuroscience↗