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Gender roles in Mesoamerica were complementary in nature in that men and women played separate, but equally important roles. This complementarity reflects a broader Mesoamerican cosmovision that emphasizes duality. Evidence also suggests the existence of gender ambiguity and fluidity in pre-Columbian Mesoamerican societies. Gender relations and roles varied among different Mesoamerican cultures and societies, through time, and depending on social status.

With the arrival of the Spanish and their subsequent colonial rule starting in the 16th century, Mesoamerican gender relations could no longer be considered distinct cultural practices. Gender roles and gender relations instead became subject to the practices of Spanish colonial rule and the casta system which racially categorized the Mesoamericans and their indigenous and mixed descendants. However, despite suppression by Spanish colonialization, aspects of pre-Columbian Mesoamerican gender roles have survived in indigenous communities to this day.

Roles
Pre-Columbian Mesoamerican art contains evidence of a gendered division of labor, depicting women engaged in domestic labor such as weaving, childrearing, tending to animals, and giving birth. Weaving was more strongly associated with gender for the Classic Mexica than the Classic Maya, for which it indicated class. Men were depicted with weapons and in positions of religious and political authority. While evidence suggests that farming was seen as a male activity, the gendered divisions of labor may not have been so strict. Analysis of the bones of women revealed evidence of wear patterns strongly associated with the repetitive motion of grinding maize, suggesting women were primarily engaged in that labor. Anthropologists such as Miranda Stockett believe it is likely that men, women, and children all participated in farming and domestic labor to varying degrees.

The success of Mesoamerican rituals were dependent on the production of food and textiles, to which women contributed much labor. These rituals were vital to ensuring good relationships with not only the gods but also within communities. In Relación de las cosas de Yucatán, Diego de Landa observed that for nearly all rituals, Mayan women were responsible for preparing food for both offerings and consumption in addition to cloth as offerings.

Gender relations among the Mexica also suggested gender complementarity. For example, dying in battle and dying in childbirth elevated men and women respectively. In childbirth, women confront the goddess Cihuacoatl, and if they died, their bodies were considered temporarily imbued with the power of the goddess. Since parts of their bodies could be used as a protective amulet or to curse others, the husband kept vigil by her body for four nights.

Some Mesoamerican women were able to assume roles as political leaders, such as women in Maya society, others such as women in Mexica society were not. However while Mexica women couldn't serve in this capacity, they were given equal legal and economy rights and noble Mexica women could become priestesses. Additionally, two influential political figures headed the highest levels of Mexica government. One was the tlatoani, literally "the one who speaks", and another was the cihuacoatl, literally "woman snake", both representing a male/female pair. The tlatoani was responsible for military affairs and the cihuacoatl was responsible for domestic affairs like the food supply and administering justice. The position of cihuacoatl was in reality occupied by a man, but the associations with femininity were very strong, and the cihuacoatl, of the same name as the goddess, wore women's dress for ceremonies. Mesoamerican states became more centralized over time, men's power became more associated with their control over women and their capacity for productive and reproductive labor. As a result, opportunities available to women were gradually limited. In the case of the Mexica, military concerns may have eclipsed women's public significance.

Although pre-Columbian Mesoamerican art contained depictions of the body as male or female as represented by genitalia or secondary sex characteristics, it also included representations of bodies with exposed chests and waists but no visible sexual characteristics. Depictions of rituals conducted by elite Mesoamericans have included women dressed in the traditional costume of men and men dressed in the traditional costume of women. Classic Maya text also includes situations where gender is not linguistically marked or where gender is marked linguistically as male for a speaker who appears to be a woman or vice versa. This evidence suggests considerable fluidity between the genders in pre-Columbia Mesoamerican society.