Myofilaments are the three protein filaments of myofibrils in muscle cells. The main proteins involved are myosin, actin, and titin. Myosin and actin are the contractile proteins and titin is an elastic protein. The myofilaments act together in muscle contraction, and in order of size are a thick one of mostly myosin, a thin one of mostly actin, and a very thin one of mostly titin.

Types of muscle tissue are striated skeletal muscle and cardiac muscle, obliquely striated muscle (found in some invertebrates), and non-striated smooth muscle. Various arrangements of myofilaments create different muscles. Striated muscle has transverse bands of filaments. In obliquely striated muscle, the filaments are staggered. Smooth muscle has irregular arrangements of filaments.

There are three different types of myofilaments: thick, thin, and elastic filaments.

The protein complex composed of actin and myosin, contractile proteins, is sometimes referred to as actomyosin. In striated skeletal and cardiac muscle, the actin and myosin filaments each have a specific and constant length in the order of a few micrometers, far less than the length of the elongated muscle cell (up to several centimeters in some skeletal muscle cells). The contractile nature of this protein complex is based on the structure of the thick and thin filaments. The thick filament, myosin, has a double-headed structure, with the heads positioned at opposite ends of the molecule. During muscle contraction, the heads of the myosin filaments attach to oppositely oriented thin filaments, actin, and pull them past one another. The action of myosin attachment and actin movement results in sarcomere shortening. Muscle contraction consists of the simultaneous shortening of multiple sarcomeres.

The axon terminal of a motor neuron releases the neurotransmitter, acetylcholine, which diffuses across the synaptic cleft and binds to the muscle fiber membrane. This depolarizes the muscle fiber membrane, and the impulse travels to the muscle's sarcoplasmic reticulum via the transverse tubules. Calcium ions are then released from the sarcoplasmic reticulum into the sarcoplasm and subsequently bind to troponin. Troponin and the associated tropomyosin undergo a conformational change after calcium binding and expose the myosin binding sites on actin, the thin filament. The filaments of actin and myosin then form linkages. After binding, myosin pulls actin filaments toward each other, or inward. Thus muscle contraction occurs, and the sarcomere shortens as this process takes place.

The enzyme acetylcholinesterase breaks down acetylcholine and this ceases muscle fiber stimulation. Active transport moves calcium ions back into the sarcoplasmic reticulum of the muscle fiber. ATP causes the binding between actin and myosin filaments to break. Troponin and tropomyosin revert to their original conformation and thereby block binding sites on the actin filament. The muscle fiber relaxes and the entire sarcomere lengthens. The muscle fiber is now prepared for the next contraction.

The changes that occur to the myofilament in response to exercise have long been a subject of interest to exercise physiologists and the athletes who depend on their research for the most advanced training techniques. Athletes across a spectrum of sporting events are particularly interested to know what type of training protocol will result in maximal force generation from a muscle or set of muscles, so much attention has been given to changes in the myofilament under bouts of chronic and acute forms of exercise.

While the exact mechanism of myofilament alteration in response to exercise is still being studied in mammals, some interesting clues have been revealed in Thoroughbred race horses. Researchers studied the presence of mRNA in skeletal muscle of horses at three distinct times; immediately before training, immediately after training, and four hours after training. They reported statistically significant differences in mRNA for genes specific to production of actin. This study provides evidence of the mechanisms for both immediate and delayed myofilament response to exercise at the molecular level.