Diaphyseal robusticity and cross-sectional shapes of the bone tissues are influenced by mechanical loading history. It changes according to work demand on the body. It is the objective of this study to identify the shifts in the modes of subsistence, activities and mobility patterns through the comparison of the shape and strength of bone diaphyses among the Mesolithic, Chalcolithic and Harappan populations of India. For the analysis, 7 sites including 2 from Mesolithic (8000-4000 bc), 2 from Chalcolithic (2000-700 bc) and 3 from Harappan (3500-1800 bc) contexts are considered. The ratio obtained from the maximum length of the bones to their girth (at 50% or 35% from the distal end) and that from anterior-posterior and medio-lateral diameters were calculated (following Martin and Saller codes, 1957) to understand certain functional adaptations and stress markers on preserved long bones. Independent-sample T-tests and ANOVA were applied to detect mean differences of statistical significance within and between cultures. The robusticity indices obtained from the humeri increase from the hunting-gathering phase (M/F; 19.39/18.45) to the Chalcolithic phase (M/F; 21.99/19.39) showing a slight drop in the Harappan phase (M/F; 18.58/18.37). The right humeri of Mesolithic females show a directional asymmetry of length (4.08%-5.13%) while that in males reach up to 26.09% indicating right-dominant lateralization. In the Harappan phase, females show a greater value for the right ranging from 0.31% to 3.07%. The femoral robusticity of females increases from 11.42 in hunting-gathering societies to 13.28 in Harappan times while mid-shaft of the Mesolithic population clearly shows greater loading along the anterior-posterior (A-P) plane as the index in most cases exceeds 100. There are significant differences among males and females within each group in terms of the indices but significant differences could be discerned between the Mesolithic, Chalcolithic and Harappan populations in case of tibiae, radii and ulnae only. The occupation patterns certainly changed as humans became a food-producer from a hunter-gatherer. Occupations involving the extensive flexion and extension of upper limbs in the Chalcolithic period increase particularly in the Harappan phase. Sedentism gave rise to roundness of the femur and reduced the magnitude of lateralization. While equal level of mobility can be traced in case of both the sexes before the advent of agriculture, this distinction increases between the males and females in the agro-pastoral phases.