Late-gestation heat stress alters placental structure and function in multiparous dairy cows

The placenta plays a pivotal role in fetal development and the dam's subsequent lactation performance, because it facilitates nutrient transfer, heat dissipation, and gas exchange with the growing fetus, and regulates key hormones essential for mammary gland development. Heat stress experienced during gestation and lactation can significantly reduce the placenta's capacity to perform these critical functions. To investigate the impact of heat stress, trials were conducted over the summer months of 2020, 2022, and 2023 in Florida. Multiparous pregnant Holstein cows were dried off 54 ± 5 d before their expected calving date and randomly assigned to 1 of 2 treatments for the entire dry period: active cooling (CL; access to barn shade, natural ventilation plus forced air circulation via fans, and water soakers; n = 20) or heat stress (HT; access to barn shade and natural ventilation; n = 20). Gestation length and calf birth weights were recorded. Placentas were collected from a subset of cows shortly after calving (4.00 ± 1.54 h; n = 10/treatment) and analyzed for total placental weight, as well as cotyledon weight, number, and surface area within 1 h after expulsion. A representative cotyledon sample was isolated for histological analysis. Tissues were also processed for RNA sequencing and DNA methylation analysis. DNA methylation was analyzed by double restriction enzyme reduced representation bisulfate sequencing. Differentially methylated cytosines between HT and CL were identified via logistic regression with a cut-off value of 15% methylation difference and a q-value <0.2. Morphological and histological data were analyzed using generalized linear mixed models. Results indicate that gestation length was shorter in HT cows compared with CL cows (274.2 vs. 277.2 ± 1.46 d), and heifers born to HT dams were lighter at birth (31.4 vs. 34.8 ± 1.59 kg). Placentas from HT dams tended to have lower total weight (3.54 vs. 4.54 ± 0.38 kg) and fewer cotyledons (66.2 vs. 103.3 ± 8.65). However, placental efficiency was higher in the HT versus CL group (11.5 vs. 8.52 ± 0.91%). Cotyledons from HT cows had greater vascular area (43.1% vs. 31.8% ± 10.4% of total area) and a tendency for less connective tissue (52.7% vs. 65.8% ± 5.39% of total area). A total of 289 differentially expressed genes were identified between HT and CL placentas, with 179 upregulated and 110 downregulated in the HT group. Key genes affected included NPSR1, SPATC1L, PGF, HSPB8, IL6, HBA/HBB, MMP12, PAPPA2, PAG14, and SLC7A10. Dysregulated pathways in HT placentas involved gas and oxygen transport, nutrient transport, inflammatory response, and cortisol biosynthesis. Heat stress induced hypermethylation of regulatory pathways, including collagen biosynthesis and degradation, extracellular matrix structural components, and placental tissue organization. Our findings demonstrate that late-gestation HT causes significant transcript alterations in the placenta, leading to adaptations for thermoregulation and morphological changes. These alterations negatively affect birth weight, health, and dam lactation performance, underscoring the need to address HT during late gestation to ensure optimal fetal development and postnatal outcomes. Addressing these issues can help improve dairy cow resilience to climate change, enhancing animal welfare and productivity.