Effect of Hypothyroidism on Different Forms of Actin in Rat Cerebral Neuronal Cultures Studied by an Improved DNase I Inhibition Assay
IR@IICB: CSIR-Indian Institute of Chemical Biology, Kolkata
View Archive Info| Field | Value | |
| Title |
Effect of Hypothyroidism on Different Forms of Actin
in Rat Cerebral Neuronal Cultures Studied by an
Improved DNase I Inhibition Assay
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| Creator |
Paul, Surojit
Das, Sumantra Sarkar, Pranab K |
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| Subject |
Cell Biology & Physiology
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| Description |
An improved DNase I inhibition assay for the
filamentous actin (F-actin) and monomeric actin (G-actin)
in brain cells has been developed. Unlike other methods, the
cell lysis conditions and postlysis treatments, established by
us, inhibited the temporal inactivation of actin in the cell
lysate and maintained a stable F-actin/G-actin ratio for at
least 4-5 h after lysis. The new procedure allowed separate
quantitation of the noncytoskeletal F-actin in the Tritonsoluble
fraction (12,000 g, 10 min supernatant) that did not
readily sediment with the Triton-insoluble cytoskeletal Factin
(12,000 g, 10 min pellet). We have applied this modified
assay system to study the effect of hypothyroidism on
different forms of actin using primary cultures of neurons
derived from cerebra of neonatal normal and hypothyroid
rats. Our results showed a 20% increase in the Triton-insoluble
cytoskeletal F-actin in cultures from hypothyroid brain relative to normal controls. In the Triton-soluble fraction,
containing the G-actin and the noncytoskeletal F-actin,
cultures from hypothyroid brain showed a 15% increase
in G-actin, whereas the F-actin remained unaltered. The
10% increase in total actin observed in this fraction from
hypothyroid brain could be totally accounted for by the
enhancement of G-actin. The mean F-actin/G-actin ratio in
this fraction was about 30% higher in the cultures from normal
brain compared to that of the hypothyroid system,
which indicates that hypothyroidism tends to decrease the
proportion of noncytoskeletal F-actin relative to G-actin.
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| Date |
1992
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| Type |
Article
PeerReviewed |
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| Format |
application/pdf
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| Identifier |
http://www.eprints.iicb.res.in/1030/1/1992%2DJN.pdf
Paul, Surojit and Das, Sumantra and Sarkar, Pranab K (1992) Effect of Hypothyroidism on Different Forms of Actin in Rat Cerebral Neuronal Cultures Studied by an Improved DNase I Inhibition Assay. Journal of Neurochemistry, 59 (2). pp. 701-707. |
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| Relation |
http://dx.doi.org/
http://www.eprints.iicb.res.in/1030/ |
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