AMPK is required for uterine receptivity and normal responses to steroid hormones.

We previously demonstrated that 5'-AMP-activated protein kinase (AMPK) is essential for normal reproductive functions in female mice. Conditional ablation of Prkaa1 and Prkaa2, genes that encode the α1 and α2 catalytic domains of AMPK, resulted in early reproductive senescence, faulty artificial decidualization, uterine inflammation and fibrotic postparturient endometrial regeneration. We also noted a delay in the timing of embryo implantation in Prkaa1/2d/d female mice, suggesting a role for AMPK in establishing uterine receptivity. As outlined in new studies here, conditional uterine ablation of Prkaa1/2 led to an increase in ESR1 in the uteri of Prkaa1/2d/d mice, resulting in prolonged epithelial cell proliferation and retention of E2-induced gene expression (e.g. Msx1, Muc1, Ltf) through the implantation window. Within the stromal compartment, stromal cell proliferation was reduced by five-fold in Prkaa1/2d/d mice, and this was accompanied by a significant decrease in cell cycle regulatory genes and aberrant expression of decidualization marker genes such as Hand2, Bmp2, Fst and Inhbb. This phenotype is consistent with our prior study, demonstrating a failure of the Prkaa1/2d/d uterus to undergo decidualization. Despite these uterine defects, ovarian function seemed to be normal following ablation of Prkaa1/2 from peri-ovulatory follicles in which ovulation, luteinization and serum progesterone levels were not different on day 5 of pregnancy or pseudopregnancy between Prkaa1/2fl/fl and Prkaa1/2d/d mice. These cumulative findings demonstrate that AMPK activity plays a prominent role in mediating several steroid hormone-dependent events such as epithelial cell proliferation, uterine receptivity and decidualization as pregnancy is established.

Progesterone receptor membrane component 1 and 2 regulate granulosa cell mitosis and survival through a NFΚB-dependent mechanism†.

Progesterone receptor membrane component 1 (PGRMC1) interacts with PGRMC2, and disrupting this interaction in spontaneously immortalized granulosa cells (SIGCS) leads to an inappropriate entry into the cell cycle, mitotic arrest, and ultimately cell death. The present study revealed that PGRMC1 and PGRMC2 localize to the cytoplasm of murine granulosa cells of nonatretric follicles with their staining intensity being somewhat diminished in granulosa cells of atretic follicles. Compared to controls (Pgrmc1fl/fl), the rate at which granulosa cells entered the cell cycle increased in nonatretic and atretic follicles of mice in which Pgrmc1 was conditionally deleted (Pgrmc1d/d) from granulosa cells. This increased rate of entry into the cell cycle was associated with a ≥ 2-fold increase in follicular atresia and the nuclear localization of nuclear factor-kappa-B transcription factor P65; (NFΚB/p65, or RELA). GTPase activating protein binding protein 2 (G3BP2) binds NFΚB/p65 through an interaction with NFΚB inhibitor alpha (IκBα), thereby maintaining NFΚB/p65's cytoplasmic localization and restricting its transcriptional activity. Since PGRMC1 and PGRMC2 bind G3BP2, studies were designed to assess the functional relationship between PGRMC1, PGRMC2, and NFΚB/p65 in SIGCs. In these studies, disrupting the interaction between PGRMC1 and PGRMC2 increased the nuclear localization of NFΚB/p65, and depleting PGRMC1, PGRMC2, or G3BP2 increased NFΚB transcriptional activity and the progression into the cell cycle. Taken together, these studies suggest that PGRMC1 and 2 regulate granulosa cell cycle entry in follicles by precisely controlling the localization and thereby the transcriptional activity of NFΚB/p65.

PGRMC1/2 promotes luteal vascularization and maintains the primordial follicles of mice

To determine whether conditional depletion of progesterone receptor membrane component (PGRMC) 1 and PGRMC2 affected ovarian follicle development, follicle distribution was assessed in ovaries of young (≈3-month-old) and middle-aged (≈6-month-old) control (Pgrmc1/2fl/fl) and double conditional PGRMC1/2-knockout (Pgrmc1/2d/d) mice. This study revealed that the distribution of primary, preantral and antral follicles was not altered in Pgrmc1/2d/d mice, regardless of the age. Although the number of primordial follicles was similar at ≈3 months of age, their numbers were reduced by ≈80% in 6-month-old Pgrmc1/2d/d mice compared to age-matched Pgrmc1/2fl/fl mice. The Pgrmc1/2d/d mice were generated using Pgr-cre mice, so ablation of Pgrmc1 and Pgrmc2 in the ovary was restricted to peri-ovulatory follicles and subsequent corpora lutea (CL). In addition, the vascularization of CL was attenuated in Pgrmc1/2d/d mice, although mRNA levels of vascular endothelial growth factor A (Vegfa) were elevated. Moreover, depletion of Pgrmc1 and Pgrmc2 altered the gene expression profile in the non-luteal component of the ovary such that Vegfa expression, a stimulator of primordial follicle growth, was elevated; Kit Ligand expression, another stimulator of primordial follicle growth, was suppressed and anti-Mullerian hormone, an inhibitor of primordial follicle growth, was enhanced compared to Pgrmc1/2fl/fl mice. These data reveal that luteal cell depletion of Pgrmc1 and 2 alters the expression of growth factors within the non-luteal component of the ovary, which could account for the premature demise of the adult population of primordial follicles. In summary, the survival of adult primordial follicles is dependent in part on progesterone receptor membrane component 1 and 2.

A functional role for AMPK in female fertility and endometrial regeneration.

Adenosine monophosphate-activated protein kinase (AMPK) is a highly conserved heterotrimeric complex that acts as an intracellular energy sensor. Based on recent observations of AMPK expression in all structures of the female reproductive system, we hypothesized that AMPK is functionally required for maintaining fertility in the female. This hypothesis was tested by conditionally ablating the two catalytic alpha subunits of AMPK, Prkaa1 and Prkaa2, using Pgr-cre mice. After confirming the presence of PRKAA1, PRKAA2 and the active phospho-PRKAA1/2 in the gravid uterus by immunohistochemistry, control (Prkaa1/2 fl/fl ) and double conditional knockout mice (Prkaa1/2 d/d ) were placed into a six-month breeding trial. While the first litter size was comparable between Prkaa1/2 fl/fl and Prkaa1/2 d/d female mice (P = 0.8619), the size of all subsequent litters was dramatically reduced in Prkaa1/2 d/d female mice (P = 0.0015). All Prkaa1/2 d/d female mice experienced premature reproductive senescence or dystocia by the fourth parity. This phenotype manifested despite no difference in estrous cycle length, ovarian histology in young and old nulliparous or multiparous animals, mid-gestation serum progesterone levels or uterine expression of Esr1 or Pgr between Prkaa1/2 fl/fl and Prkaa1/2 d/d female mice suggesting that the hypothalamic-pituitary-ovary axis remained unaffected by PRKAA1/2 deficiency. However, an evaluation of uterine histology from multiparous animals identified extensive endometrial fibrosis and disorganized stromal-glandular architecture indicative of endometritis, a condition that causes subfertility or infertility in most mammals. Interestingly, Prkaa1/2 d/d female mice failed to undergo artificial decidualization. Collectively, these findings suggest that AMPK plays an essential role in endometrial regeneration following parturition and tissue remodeling that accompanies decidualization.

The Social Ecology of Health Beliefs and Misinformation Framework: Examining the impact of misinformation on vaccine uptake through individual and sociological factors.

BACKGROUND: Misinformation presents a critical concern for academic and public health discourse, particularly around vaccine response. Before the COVID-19 pandemic, vaccine hesitancy was responsible for decreased immunization uptake for vaccine-preventable diseases. Misinformation connected to the novel COVID-19 vaccine has further fueled vaccine hesitancy in Colorado and the United States. Our study brings together three different perspectives - physicians, public health professionals, and parents - to understand the impact of misinformation on vaccine uptake in Colorado. Our study proposes a framework for combining the Health Belief Model with the Socio-Ecological model to account for societal factors in healthcare decision making. METHODS: Semi-structured interviews and focus groups with public health professionals, physicians, and parents (n = 31) were conducted in late spring and summer 2022. Data were coded inductively using thematic analysis. Identified themes were deductively categorized according to the Socio-Ecological Model and Health Belief Model. RESULTS: Using a theoretical framework that combined the Health Belief Model and the Socio-Ecological Model, we identified seven factors that influenced vaccine hesitancy in Colorado. Intrapersonal factors included routine vaccine hesitancy connected to perceptions of severity and susceptibility, efficacy, and benefits and barriers to vaccine uptake; interpersonal factors included social networks; institutional factors included mass mediated platforms, portrayals of uncertainty, distrust in institutional sources of information, and political influences in vaccine decision making; and structural factors included economic barriers behind vaccine hesitancy. CONCLUSIONS: Our study provides a unique, triangulated, post-positivist perspective on the role of misinformation in vaccine hesitancy in Colorado. The findings provide evidence that misinformation is an important barrier to vaccination uptake and can permeate multiple socio-ecological determinants/characteristics to influence vaccination behaviors including intrapersonal, interpersonal, institutional, and structural levels. We introduce the Social Ecology of Health Beliefs and Misinformation Framework to account for how misinformation may interrupt vaccine uptake.

Undergraduate STEM Students' Science Communication Skills, Science Identity, and Science Self-Efficacy Influence Their Motivations and Behaviors in STEM Community Engagement.

While numerous studies have examined how scientists perceive doing public communication and engagement, there is limited research on undergraduate science, technology, engineering, and math (STEM) student attitudes toward these meaningful activities. Undergraduate students are more diverse than STEM faculty and may serve as boundary spanners in communities, so exploring their motivations and behaviors in STEM engagement is valuable. For scientists, confidence in communication skills is one driver of public engagement behavior. In this study, we utilized a survey to examine how undergraduate STEM students' science communication skills as well as their science identity and science self-efficacy may drive motivation and behaviors in STEM community engagement. Our findings revealed that STEM students are motivated to do community engagement but lack opportunities to actually do these behaviors. Regression analyses revealed that year in academic progression did not increase STEM students' attitudes and behaviors in community engagement. However, science communication skills, science identity, and science self-efficacy were all predictors of student motivation and behaviors in STEM community engagement. These findings suggest that universities should intentionally provide training in science communication, continue providing support for students developing science identity and self-efficacy, and develop opportunities for undergraduate STEM students to do science outreach and engagement activities.

Student perceptions of inclusive pedagogy in undergraduate STEM classrooms.

In university STEM classrooms, the incorporation of inclusive practices improves student performance, decreases disparities in the academic success of underrepresented students, and increases student retention and persistence in Science, Technology, Engineering, and Mathematics (STEM) programs. Inclusive pedagogical practices include effective instructional choices like active learning, providing rubrics, and other strategies that have been shown to support students from disadvantaged backgrounds. Additionally, explicitly inclusive practices such as addressing microaggressions and sharing pronouns can promote a sense of belonging for students. While a plethora of literature has shown these impacts and faculty have access to resources and training about inclusive pedagogy, we were interested in whether students are noticing these practices and how student identities impact their observations of instructional practices. We surveyed undergraduates (n = 74) from diverse STEM disciplines at a large land-grant university regarding their observation of 11 different inclusive pedagogical practices. Overall, students observed inclusive instructional practices more often than they observed explicitly diversity, equity, and inclusion (DEI)-related practices. For explicitly DEI-related practices, white students observed more practices than Students of Color. This suggests that more work needs to be done to train faculty in explicit DEI-related practices, especially with the goal of supporting Students of Color who have been historically excluded from STEM.

A Career Preparation Course for Biomedical Science Majors Focused on Skills for Diverse Career Paths.

Training in career preparation is vital for biomedical science, microbiology, and related life science undergraduates to know the types of careers available in the field, to obtain employment after graduation, and to be successful in these careers. This is especially critical for historically marginalized students who have lower science, technology, engineering, and math (STEM) retention and lower STEM employment rates. Thus, we developed a career preparation course aimed for second- and third-year students in biomedical science, microbiology, biology, and related majors. This course introduced students to diverse careers via guest speakers and provided training and practice in key career skills, like writing CVs and cover letters. In this curriculum article, we present our course curriculum and resources, evidence of student achievement of learning objectives, and evidence that this course supported growth in constructs like science networking and confidence in future self, which are known to support student STEM retention and success.

Developing Science Literacy in Students and Society: Theory, Research, and Practice.

The subject of scientific literacy has never been more critical to the scientific community as well as society in general. As opportunities to spread misinformation increase with the rise of new technologies, it is critical for society to have at its disposal the means for ensuring that its citizens possess the basic scientific literacy necessary to make critical decisions on topics like climate change, biotechnology, and other science-based issues. As the Guest Editors of this themed issue of the Journal of Microbiology and Biology Education, we present a wide array of techniques that the scientific community is using to promote scientific literacy in both academic and nonacademic settings. The diversity of the techniques presented here give us confidence that the scientific community will rise to the challenge of ensuring that our society will be prepared to make fact-based and wise decisions that will preserve and improve our quality of life.

hMSH5 Regulates NHEJ and Averts Excessive Nucleotide Alterations at Repair Joints.

Inappropriate repair of DNA double-strand breaks (DSBs) leads to genomic instability, cell death, or malignant transformation. Cells minimize these detrimental effects by selectively activating suitable DSB repair pathways in accordance with their underlying cellular context. Here, we report that hMSH5 down-regulates NHEJ and restricts the extent of DSB end processing before rejoining, thereby reducing "excessive" deletions and insertions at repair joints. RNAi-mediated knockdown of hMSH5 led to large nucleotide deletions and longer insertions at the repair joints, while at the same time reducing the average length of microhomology (MH) at repair joints. Conversely, hMSH5 overexpression reduced end-joining activity and increased RPA foci formation (i.e., more stable ssDNA at DSB ends). Furthermore, silencing of hMSH5 delayed 53BP1 chromatin spreading, leading to increased end resection at DSB ends.

Scaffolded Curriculum for Developing Science Communication Skills in Life Science Undergraduates.

Strong communication skills are essential for future science professionals, but practical training has not been featured strongly in undergraduate curricula. To better train diverse life science majors in communication theory and skills, we created a foundational 200-level course and an advanced 400-level science communication course. Here, we outline the strategy, including lesson plans, assignments, and grading rubrics, for these courses. The science communication assignments presented are diverse in terms of audience, including communication to fellow scientists, to clinicians, and to the public, as well as in terms of format, including written, oral, and visual modes. We also provide suggestions for placing assignments designed to build upon each other into preexisting courses in a scaffolded manner to promote mastery of science communication skills.