In no particular order:

Lou(ise) McCloud: The eldest of three siblings, she was left at an orphanage run by nuns when their mother died. Their father was an outlaw gunrunner.
She ran away from the orphanage at age 11 to make a living so she could raise her siblings together. A year after landing a job as a laundress at a bordello, unbeknownst to her at first, she was raped by the proprietor, Lyle Wicks. One of the prostitutes helped her escape. She cut off her hair, bought boy’s clothes and used her skill with horses to get a job riding for the Pony Express.
On her first run, she was shot by outlaws intent on stealing the horses at the Express waystation. Kid finds her and discovers her ruse. Eventually he agrees to keep her secret. The other riders found out when her father returned and took her brother and sister. In the process of rescuing them, Kid had to kill her father.

Despite a rocky courtship with Kid and occasional flirtation with Jimmy, their joint best friend, Lou eventually married Kid in the final episode.

Although a crackshot, he did not have any of the others' need to be recognized for it, just wanting to do his job well. He was the first to discover Lou’s secret and immediately fell in love with her. However, their relationship was marred by his intense sense of right and wrong and a need to protect her, complicated by her own psychological problems from a childhood rape. Eventually, they worked their problems out and married.
He was also best friends with Jimmy. Although he did not believe in slavery, Kid felt the need to return to defend his home in Virginia from invaders when the Civil War started.
It was a running joke that neither Kid, his brother or Doritha ever revealed Kid’s real name. Kid eventually whispered it in Teaspoon’s ear at his wedding, but to everyone’s, including Lou’s, disappointment Teaspoon refused to use it.

He first landed in the home of “The Judge” who encouraged him to develop his shooting skills. But, realizing the danger of the Judge’s plans for him, Jimmy left and joined the Pony Express.
He still had a deep-seated need for recognition, along with a quick temper and a hair-trigger on his pistol. The Kid’s refusal to challenge Jimmy, although he was just as good with a gun, led them to become best friends.
Being the second person to figure out Lou’s secret, and half in love with her himself, Jimmy also became her best friend. This left him caught between the Kid and Lou during their rocky courtship, as they both turned to him for advice, which often backfired.
A run-in with a dime novel writer went bad for Jimmy, when the author made up stories about him as “Wild Bill,” thrusting Jimmy into the life of a gunfighter.
A staunch abolitionist, Jimmy often butted heads with Kid and almost missed his wedding after yet another fight. However, he overcame their differences long enough to walk Lou down the aisle. Peace didn't last long, as Kid and Jimmy fought again over Noah’s body.


Landing at a Catholic Mission school, he met up with Ike and they became instant best buddies and adopted brothers. Buck taught Ike Indian Sign so he can communicate. Together they joined the Express.
Despite constantly being pulled in two directions and suffering from the prejudices of those around him, Buck eventually decided he was going to spend his life living among the white men. However, the end of the Pony Express and the beginning of the Civil War left his adopted white family scattered to the four winds.

Meeting Buck changed his life and provided him with a way to communicate using Indian Sign. The two became not only best friends but adoptive brothers. They joined the Express together.
Toward the end of the Express, Ike fell in love with Emily. He ended up being shot, trying to save her from herself. He died unexpectedly, leaving Buck feeling lost and alone and the rest of the Express family grieving.

With the coming of the Civil War, he wanted to join the Union Army, like Cody, but was rejected for being black. He and another friend followed Captain Urbach’s unit on a mission to take out Confederate bushwackers. He was killed in the ensuing ambush that nearly destroyed Urbach’s unit.
His death rang the death knell for the Express Family for the immediate future, as all split to do what their hearts told them they must.

Emma revealed to Lou that she knew the girl rider’s secret all along, but only after all the other riders had learn Lou’s secret. She promised not to tell ‘Mr. Spoon’, who remained in the dark.
Emma lost her infant son to the measles. Her husband first turned to drink, then deserted her. She began to find her footing in a new relationship with the town Marshal, Sam Cain, but her husband’s return threatened that. It turned out her husband only came back to hide from his outlaw compatriots who’d been setting up wagon trains for robbery. They eventually killed him.
This was a turning point in Emma's relationship with Sam Cain and they ended up marrying when Sam was promoted to Territorial Marshal shortly before Noah joined the family. They moved to Omaha because of Sam’s new job.

Express began. He often turned to his old friend Teaspoon when he needed extra guns for a posse and Teaspoon ‘volunteered’ his riders, all handy with weapons. Sam acted as an older brother to the riders.
Sam was in love with Emma and went to great, though often faulty, lengths to win her love. After her husband returned and was killed, cementing Sam’s relationship with her, he married her and took her with him to Omaha when he was promoted to Territorial Marshal.

As the family grew closer, Teaspoon became a surrogate father to the bunch of orphaned boys. He was the last to discover Lou’s secret and was unable to figure out why he never noticed.
A former Texas Ranger and gunfighter, Teaspoon was the sole survivor of several men sent for help shortly before the fall of the Alamo.
When Sam and Emma married and moved to Omaha, Teaspoon agreed to strap on his gunfighter’s rig again and become the Marshal. Later, when the Express moved the Sweetwater station to Rock Creek, Nebraska Territory, Teaspoon finagled a transfer there as Marshal as well. He continued to serve as stationmaster at the same time.
Having been married six times, three times to white women, once to a foreigner and twice to Indian women, Teaspoon’s favorite hobby was dispensing life advice, often unintelligibly. His second wife, Polly, reappeared near the end of the Express, hoping to rekindle their relationship.
With the start of the Civil War and the end of the Express, Teaspoon announced he was thinking about returning home to Texas to fight. This greatly hurt Jimmy who had grown the closest to him due to their shared experiences as gunfighters.
Polly Hunter: She was Teaspoon’s second, and favorite, wife and the daughter of the Texas Ranger who turned Teaspoon from a petty criminal and into a lawman. She came to Rock Creek after her father’s death. She’d left Teaspoon because he was gone so much chasing criminals. After her father's death, she bought the local saloon in Rock Creek and ran it while trying to rekindle her relationship with Teaspoon.

Rachel joined the Express in response to an ad for a cook/housekeeper to replace the newlywed Emma. Her appearance at first caused a ruckus as all the boys lusted after, and Lou was jealous of, her voluptuous body. However, once she made it clear that dinner was on the table and she was not on the menu, things settled down.
Eventually, the boys began to treat her like an older sister. Toward the end of the Express, the local school master was killed in a house fire and Rachel took over teaching school.
Janusz Tartovsky: Janusz was a Polish immigrant who was falsely accused, convicted and imprisoned for his wife’s murder. He was unable to defend himself because he could not yet speak English. After serving his term, he came back to his land in Rock Creek and attempted to pick up the pieces. However, those who framed him were afraid he’d uncover their perfidy and attempted to repeat history. Instead, thanks to Janusz’ dead wife’s journal, which Janusz got Rachel to read to him, the truth is uncovered. Janusz takes over working as the town blacksmith who had been killed.

Teaspoon and the other riders took him in. First he slept in a cot in a spare cell in the Marshal’s office, before eventually moving into the bunkhouse with the other riders after Ike’s death.
Jesse was indirectly responsible for Noah’s death. He told his brother Frank about Captain Urbach’s planned mission, not realizing Frank was riding with the bushwhackers. This led to the ambush in which Noah was killed. Jesse helped capture the rest of the bushwhackers before leaving Nebraska Territory with Frank to join other Confederate sympathizers in Missouri.

However, he developed a fondness for them and when they were transferred from Sweetwater to Rock Creek it wasn't long before Thompkins' sold his store and followed them.
In the last episodes we discover that, like Emma, he always knew Lou's secret but figured it wasn't his place to say anything.

A gambler and occasional prostitute, she'd been a friend of the daughter Teaspoon didn't know he had. When his daughter, Elizabeth, died Amanda was tasked with delivering a letter from her mother to Teaspoon.
Unfortunately her protector read the letter which told Teaspoon he had inherited $10,000 from his former lover. Amanda's protector decided to trick Teaspoon out of the money. Teaspoon, along with Amanda and the rest of the Young Riders, got the upper hand and ended up sending her former protector to Hades.
With Teaspoon's support, Amanda decided to put her sordid past behind her. He gave her the inheritance money and she opened a saloon in Abilene, Kansas, with it. He remained her partner in the saloon and continued to speak of her and treat her as an adopted daughter throughout the rest of the series.
She was unsuccessfully used as leverage against the, by then, Marshal Hunter in the season 2 finale.
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