A comparative study of the isozymic proteins and the effect of metachemogenesis in the tissues of Periplaneta americana (L.) Public Deposited

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  • Metachemogenesis in the tissues of Periplaneta americana (L.)
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  • An attempt was made to demonstrate isozymic forms of esterase, MDH, LDH and GPDH and the variation in enzyme pattern with increasing age in the thoracic muscle, appendage muscle and hemolymph of the American cockroach, Periplaneta americana. Sampled specimens of both sexes were aged one through seven days, two weeks and one through six months. The proteins in the muscle homogenates and hemolymph aliquots were separated with the aid of polyacrylamide gel "disc" electrophoresis and it was discovered that the protein concentration should be kept below 4 mg/ml to enable proper photopolymerization of the sample gel. Isozyme patterns were localized on the gel by utilizing histochemical staining procedures. Esterase banding was more pronounced if the pH of the incubation mixture was lowered from 7.0 to 5.6. Muscle esterase patterns exhibited six fairly consistent major bands and a number of inconsistent minor bands. One effect of aging was the increased resolution of the fastest migrating band on the gel in the muscle of specimens older than seven days. Non-specific banding in the absence of substrate accounted for the majority of the dehydrogenase activity in the three tissues examined. Muscle MDH-specific patterns were characterized by two minor bands and a major band (R[subscript f] value approximately .39) consisting of three subbands. Muscle LDH and GPDH-specific patterns each possessed one specific band with R[supscript f] values of approximately .5 and .42 respectively. The density of the main GPDH band varied somewhat in muscle patterns with female thoracic muscle displaying little or no main band activity. Three major and three to six minor esterase bands were demonstrated in hemolymph. One MDH-specific band was found in hemolymph with an R[subscript f] value approximating that of the major three band complex in muscle. Although a number of non-specific bands were displayed in hemolymph, the patterns failed to disclose any LDH and GPDH-specific bands in hemolymph. Each of the patterns were evaluated as possible biochemical systematic indicators by examining the degree of stability with increasing age. Esterase patterns, although displaying stability in main bands, showed variation in appearance of minor bands for all three tissues. MDH in all three tissues and muscle LDH displayed a great deal of stability, qualifying these tissue enzymes as candidates for further systematic comparisons. The variation in the density of the GPDH band in muscle and the lack of specific LDH and GPDH banding in hemolymph makes these tissue enzymes of questionable value in future comparisons.
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