Differential HSP90α expression in fish hepatocytes from polluted estuary during summer

The molecular chaperone, heat shock protein 90α (HSP90α), plays an important role in protein folding, degradation of denatured proteins and steroid activation. It is essential for the maintenance of cellular integrity and survival when induced in response to environmental, physical and chemical stresses. In the present investigation the effect of environmental stress on HSP90α expression was examined in grey mullet Mugil cephalus living in either a contaminated (Ennore) or uncontaminated (Kovalam) estuary over two seasons: Hepatocytes were isolated from grey mullet of both estuaries. Oxidative stress was determined along with HSP90α in these fish. Additionally, immunohistochemical changes were studied to confirm the HSP90α expression. Comparison of the results revealed enhanced hepatocyte oxidative stress and HSP90α expressions in fish from Ennore to a significant extent than fish from Kovalam. Also, the results showed significant seasonal variations with maximum expression observed during summer compared to the monsoon season. Overexpression of HSP90α in hepatocytes exposed to chronic environmental stress by pollutants may confer differential effects on cell survival by protecting against oxidative stress induced changes. The results also indicate that seasonal variations have significant effect on the HSP90α expression.     

Differential regulation of pro- and antiapoptotic proteins in fish adipocytes during hypoxic conditions

Worldwide, the frequencies and magnitudes of hypoxic events in estuarine waters have increased considerably over the past two decades. Fish populations are suitable indicators for the assessment of quality of aquatic ecosystems and often comprise a variety of adaptation systems by triggering oxidants, antioxidants and hypoxia-responsive signaling proteins. Signaling pathway may lead to cell survival or cell death which is fine-tuned by both positive and negative factors, which includes hypoxia-inducible factor-1α (HIF1α), heat-shock protein-70 (HSP70), phospho-c-Jun N-terminal kinase 1/2 (p-JNK1/2) and apoptosis signal-regulating kinase-1 (ASK1). In the present study, we attempt to determine stress-mediated signaling changes and molecular mechanism behind the cell survival by comparing adipocytes of fish from field hypoxic condition and laboratory-induced hypoxic condition (in vitro hypoxia). Comparison of field and laboratory studies in fish adipocytes showed differential expression of HIF1α, HSP70, p-JNK1/2 and ASK1 with altered oxidants and antioxidants. Further, the results also suggest that in vitro hypoxic conditions mimic field hypoxic conditions. Trends of hypoxia response were same in in vitro hypoxia of control adipocytes as in Ennore estuary, and hypoxia response was more pronounced in the test adipocytes under in vitro hypoxic condition. Results of the present work suggest that hypoxia is the major crusade of water pollutants affecting fish by differential regulation of pro- and antiapoptotic proteins probably through HSP70. This may play a vital role by providing cytoprotection in pollutant-induced stressed fish adipocytes substantiated by the in vitro hypoxic studies.     

Characterization of haemocytes of the Indian edible oyster, Crassostrea madrasensis (Preston)

The haemocytes of the Indian edible oyster Crassostrea madrasensis were characterized using light and electron microscopy. The light microscopic study was conducted by staining a monolayer of the haemocytes with Geimsa. Cells without granules and with a large nucleus occupying much of the cytoplasmic area were grouped as hyalinocytes. Those with lesser amounts of basophilic cytoplasmic granules were characterized as semigranulocytes and those with large amounts of a mixture of acidophilic and basophilic granules were termed as granulocytes. Ultrastructural studies also revealed the presence of three types of haemocytes. Scanning electron microscopic studies were used to study the spreading behaviour of the haemocytes. Cytochemical studies revealed the presence of acidphosphatase, peroxidase and prophenol oxidase in the cells.     

Heat-shock protein 70 modulates apoptosis signal-regulating kinase 1 in stressed hepatocytes of Mugil cephalus

Oxidative stress causes damage at the cellular level and activates a number of signaling pathways. Heat-shock proteins (HSPs) play an important role in repair and protective mechanisms under cell response to stress conditions. HSP70 has been shown to act as an inhibitor of apoptosis. Apoptosis signal-regulating kinase-1 (ASK1) activity is regulated at multiple levels, one of which is through inhibition by cytosolic chaperons HSP70. The current study was aimed to investigate the alteration in signaling molecules that allow the fish to survive under stressed natural field conditions. The study also investigates the variation in biomolecular composition of hepatocytes by using Fourier transform infrared spectroscopy. The impact of stress on hepatocytes was assessed by measuring the level of lipid peroxides (LPO), catalase activity (CAT) and assessing the changes in hepatocytes of Mugil cephalus inhabiting Kovalam and Ennore estuaries. The expression of HSP70 and ASK1 were analyzed by immunoblot analysis and ELISA, respectively. The spectral analysis showed variations in biomolecular composition of hepatocytes at a wave number region of 4,000–400 cm^?1. There was significant decrease of CAT activity (p < 0.01) (25 %) with significant increase of LPO (p < 0.001) (35 %) and HSP70 (p < 0.001) and insignificant increase of ASK1 (p < 0.05) (16 %) in fish hepatocytes inhabiting Ennore estuary than Kovalam estuary. In conclusion, the present study suggests that the survival of fish in the Ennore estuary under stressed condition may be due to the upregulation of HSP70 that mediates the altered signal pathway which promotes cellular resistance against apoptosis.     

Revealing Genetic Relationship and Prospecting of Novel Donors Among Upland Rice Genotypes Using qDTY-Linked SSR Markers

A total of 17 simple sequence repeat (SSR) markers linked to QTLs (qDTYs) governing grain yield under reproductive stage (RS) drought stress were used to assess the genetic relationship and prospecting new donors for qDTYs among 32 popular upland rice genotypes. These SSR markers generated a total of 36 alleles with an average allele count of 2.1 per locus. Polymorphic information content value of the markers ranged from 0.376 to 0.662 with an average value of 0.484. The expected heterozyogosity ranged from 0.381 to 0.632. STRUCTURE analysis divided the 32 genotypes into three sub-populations. Subsequent phenotyping revealed that all the tolerant genotypes were grouped into one sub-population, whereas the moderately tolerant and susceptible genotypes were grouped into separate sub-populations. Phylogenetic tree constructed by the unweighted neighbour-joining method also divided the genotypes into three clusters. The grouping pattern of genotypes into the clusters was similar to that into the STRUCTURE analysis, on the basis of drought tolerance level. The average value of genetic dissimilarity coefficient among the genotypes was observed to be 0.486. Furthermore, by combining genotyping data with phenotyping data, 16 new donors for 6 qDTYs were identified.