Skeletal muscle during postnatal development undergoes several structural and biochemical modifications. It is proposed that these changes are closely intertwined with the increase in load‐bearing capacity of the muscle (i.e., myofibrils) and molecular machinery to support the energy demand (i.e., mitochondria). Concomitant establishment of the sarcoplasmic reticulum (SR) and mitochondrial network seems to be a major developmental adjustment of skeletal muscle leading to adult phenotype. Here, we have studied oxidativeness, vascularization, and the changes in mitofusins (Mfn) 1–Mfn 2 expression and interaction in the due course of muscle development. Toward this, we used a series of histochemical techniques to compare neonatal and adult limb muscles (Gastrocnemius and Quadriceps) of Wistar rat (Rattus norvegicus). Additionally, we probed the proximity between Mfn 1 and Mfn 2 using a highly sensitive antibody‐based proximity ligation assay indicating the change in mitochondrial fusion pattern or mitochondria‐SR interaction. The results show that neonatal fibers bear a uniform distribution of mitochondria while a differential pattern of distribution is seen in adults. The distribution of the blood vessels is also quite distinct in adult muscles with a well‐formed capillary network but in neonates, only central blood vessels are seen. Interestingly, our Mfn 1–Mfn 2 interaction data show that this interaction is uniformly distributed throughout the neonatal fibers, while it becomes peripherally localized in fibers of adult muscles. This peripheralization of Mfn 1–Mfn 2 interaction must be an important event of muscle development and might be critical to cater to the metabolic needs of adult muscle.