Biochemical changes that are associated with the growth phase of stretch-induced skeletal muscle hypertrophy are better understood than events that maintain the increased muscle mass. One purpose of this study was to determine whether changes that occur during the period of rapid muscle hypertrophy persist during periods when muscle growth plateaus or the rate of enlargement slows. Serum response factor (SRF), myogenin, MyoD, and actin mRNA expression patterns were examined. SRF protein interactions with serum response element-1 (SRE1) of the chicken skeletal alpha-actin gene were also characterized. Anterior latissimus dorsi (ALD) wet weight (132% and 122%) and total RNA concentration (29% and 19%) increased after 2 and 3 weeks of stretch overload, respectively. Myogenin mRNA per microgram RNA increased after 3 (775%), 6 (1073%), 14 (227%), and 21 days (133%) of stretch overload. At 6 days, myogenin mRNA levels were increased in the distal, middle and proximal regions of the ALD. Serum response factor (SRF) mRNA per microgram total RNA was not increased after 2 or 3 weeks of stretch overload. MyoD and skeletal alpha-actin mRNAs per microgram total RNA were also unchanged after 2 and 3 weeks of stretch. Gel mobility shift assays demonstrated that SRF bound to SRE1 from 14-day-stretched ALD nuclear extracts had an increased mobility compared to control, and this difference in mobility was maintained in nuclear extracts from ALD muscle whose mass was declining. These results indicate that the expression of myogenin mRNA and total RNA remains elevated during either slow or maintenance periods of stretch-induced increases in ALD mass, when SRF mRNA has returned to control levels. Additionally, stretch-induced alterations in SRF binding to SRE1, from the skeletal alpha-actin promoter, occur regardless of the rate of stretch-induced growth.