The pectoralis major (PM) is a broad muscle commonly divided into three regions, which contribute uniquely to shoulder stability and movement. The PM muscle regions likely respond differently to stretch, but this has never been shown in vivo. We used shear wave elastography to assess the stretch response of different PM muscle regions during shoulder abduction and external rotation in 20 healthy male participants. Participants’ shoulder was passively rotated through their range of motion in 5.7° increments and shear wave velocities (SWV) were obtained for each muscle region. A piece-wise model was fitted to the SWV-joint angle data, from which slack angle, slack stiffness and elasticity coefficient were determined. For shoulder abduction, we found that the sternocostal region had a significantly smaller slack angle (p = 0.049) and greater slack stiffness (p = 0.005) than the abdominal region, but there was no difference for elasticity coefficient (p = 0.074). For external rotation, only slack stiffness was greater for the sternocostal than the abdominal region (p < 0.001) with no differences found for slack angle (p = 0.18) and elasticity coefficient (p = 0.74). However, our data indicates that neither region was slack in this condition. These findings indicate that the sternocostal and abdominal regions respond differently to passive stretch, highlighting the PM’s functional differentiation. This differentiation should be considered during treatment interventions such as PM muscle harvesting or treatments for breast cancer.