Uncovering changes in proteomic signature of rat pelvic floor muscles in pregnancy.

BackgroundStructural and functional changes of the rat pelvic floor muscles during pregnancy, specifically, sarcomerogenesis, increase in extracellular matrix content, and higher passive tension at larger strains protect the integral muscle components against birth injury. The mechanisms underlying these antepartum alterations are unknown. Quantitative proteomics is an unbiased method of identifying protein expression changes in differentially conditioned samples. Therefore, proteomics analysis provides an opportunity to identify molecular mechanisms underlying antepartum muscle plasticity.ObjectiveTo elucidate putative mechanisms accountable for pregnancy-induced adaptations of the pelvic floor muscles, and to identify other novel antepartum alterations of the pelvic floor muscles.Materials and methodsPelvic floor muscles, comprised of coccygeus, iliocaudalis, and pubocaudalis, and nonpelvic limb muscle, tibialis anterior, were harvested from 3-month-old nonpregnant and late-pregnant Sprague-Dawley rats. After tissue homogenization, trypsin-digested peptides were analyzed by ultra-high-performance liquid chromatography coupled with tandem mass spectroscopy using nano-spray ionization. Peptide identification and label free relative quantification analysis were carried out using Peaks Studio 8.5 software (Bioinformatics Solutions Inc., Waterloo, ON, Canada). Proteomics data were visualized using the Qlucore Omics Explorer (New York, NY). Differentially expressed peptides were identified using the multi-group differential expression function, with q-value cutoff set at <0.05. Proteomic signatures of the pelvic floor muscles were compared to nonpelvic limb muscle and between nonpregnant and pregnant states.ResultsUnsupervised clustering of the data showed clear separation between samples from nonpregnant and pregnant animals along principal component 1 and between pelvic and nonpelvic muscles along principal component 2. Four major gene clusters were identified segregating proteomic signatures of muscles examined in nonpregnant vs pregnant states: (1) proteins increased in the pelvic floor muscles only; (2) proteins increased in the pelvic floor muscles and tibialis anterior; (3) proteins decreased in the pelvic floor muscles and tibialis anterior; and (4) proteins decreased in the pelvic floor muscles alone. Cluster 1 included proteins involved in cell cycle progression and differentiation. Cluster 2 contained proteins that participate in mitochondrial metabolism. Cluster 3 included proteins involved in transcription, signal transduction, and phosphorylation. Cluster 4 comprised proteins involved in calcium-mediated regulation of muscle contraction via the troponin tropomyosin complex.ConclusionPelvic floor muscles gain a distinct proteomic signature in pregnancy, which provides a mechanistic foundation for the antepartum physiological alterations acquired by these muscles. Variability in genes encoding these proteins may alter plasticity of the pelvic floor muscles and therefore the extent of the protective pregnancy-induced adaptations. Furthermore, pelvic floor muscles' proteome is divergent from that of the nonpelvic skeletal muscles

Mesh Exposure and Associated Risk Factors in Women Undergoing Transvaginal Prolapse Repair with Mesh

Objective. To determine frequency, rate, and risk factors associated with mesh exposure in women undergoing transvaginal prolapse repair with polypropylene mesh. Methods. Retrospective chart review was performed for all women who underwent Prolift Pelvic Floor Repair System (Gynecare, Somerville, NJ) between September 2005 and September 2008. Multivariable logistic regression was performed to identify risk factors for mesh exposure. Results. 201 women underwent Prolift. Mesh exposure occurred in 12% (24/201). Median time to mesh exposure was 62 days (range: 10–372). When mesh was placed in the anterior compartment, the frequency of mesh exposure was higher than that when mesh was placed in the posterior compartment (8.7% versus 2.9%, P=0.04). Independent risk factors for mesh exposure were diabetes (AOR = 7.7, 95% CI 1.6–37.6; P=0.01) and surgeon (AOR = 7.3, 95% CI 1.9–28.6; P=0.004). Conclusion. Women with diabetes have a 7-fold increased risk for mesh exposure after transvaginal prolapse repair using Prolift. The variable rate of mesh exposure amongst surgeons may be related to technique. The anterior vaginal wall may be at higher risk of mesh exposure as compared to the posterior vaginal wall

Characterizing the Maternal Adaptations of Pregnancy and Recovery Following Vaginal Delivery in the Rodent Model

ABSTRACT Pelvic organ prolapse and urinary incontinence are common conditions in women that significantly diminish quality of life. Vaginal delivery and maternal birth injury are the number one risk factors for the development of pelvic floor disorders. The goal of this study was to characterize maternal adaptations throughout pregnancy and recovery after vaginal delivery in terms of the passive quasi-static mechanical properties of the vagina using a rodent model. Virgin (n=8), mid-pregnant (n=7, day 15-16), late-pregnant (n=7, day 20-21), immediate postpartum (n=8, <2 hours post delivery), and 4 week postpartum (n=6) Long-Evans female rats were utilized in this study. The mechanical properties (tangent modulus, tensile strength, ultimate strain, and strain energy density) were quantified by testing longitudinal sections of vaginal tissue to failure. The tangent modulus of virgin animals (25.1±5.1 MPa) was significantly higher compared to mid-pregnant (11.7±7.7 MPa, p=0.003), late-pregnant (7.9±4.0 MPa, p<0.001), and immediate postpartum (8.5±4.7 MPa, p=0.001) animals. A similar trend was also observed in the tensile strength, whereas the ultimate strain increased throughout pregnancy until the time of vaginal delivery. Recovery was observed four weeks postpartum as no significant difference was found from virgin animals for any of the parameters. This study has shown a significant decrease in the tangent modulus and tensile strength along with an increase in the ultimate strain of longitudinal sections of vaginal tissue throughout pregnancy. These maternal adaptations are likely to increase the overall distensibility of the vagina and allow for vagina delivery with minimal injury. This process appears to be effective in the rodent model as the properties recovered to virgin levels by 4 weeks. In the future, we hope to alter these adaptations or exceed them in order to study the risk and impact of birth injury in this model

The mysteries of menopause and urogynecologic health: clinical and scientific gaps.

ObjectivesA significant body of knowledge implicates menopausal estrogen levels in the pathogenesis of the common pelvic floor disorders (PFDs). These health conditions substantially decrease quality of life, increase depression, social isolation, caregiver burden, and economic costs to the individuals and society.MethodsThis review summarizes the epidemiology of the individual PFDs with particular attention to the understanding of the relationship between each PFD and menopausal estrogen levels, and the gaps in science and clinical care that affect menopausal women. In addition, we review the epidemiology of recurrent urinary tract infection (rUTI)-a condition experienced frequently and disproportionately by menopausal women and hypothesized to be potentiated by menopausal estrogen levels.ResultsThe abundance of estrogen receptors in the urogenital tract explains why the natural reduction of endogenous estrogen, the hallmark of menopause, can cause or potentiate PFDs and rUTIs. A substantial body of epidemiological literature suggests an association between menopause, and PFDs and rUTIs; however, the ability to separate this association from age and other comorbid conditions makes it difficult to draw definitive conclusions on the role of menopause alone in the development and/or progression of PFDs. Similarly, the causative link between the decline in endogenous estrogen levels and the pathogenesis of PFDs and rUTIs has not been well-established.ConclusionsInnovative human studies, focused on the independent effects of menopausal estrogen levels, uncoupled from tissue and cellular senescence, are needed

The mysteries of menopause and urogynecologic health: clinical and scientific gaps.

OBJECTIVES: A significant body of knowledge implicates menopausal estrogen levels in the pathogenesis of the common pelvic floor disorders (PFDs). These health conditions substantially decrease quality of life, increase depression, social isolation, caregiver burden, and economic costs to the individuals and society. METHODS: This review summarizes the epidemiology of the individual PFDs with particular attention to the understanding of the relationship between each PFD and menopausal estrogen levels, and the gaps in science and clinical care that affect menopausal women. In addition, we review the epidemiology of recurrent urinary tract infection (rUTI)-a condition experienced frequently and disproportionately by menopausal women and hypothesized to be potentiated by menopausal estrogen levels. RESULTS: The abundance of estrogen receptors in the urogenital tract explains why the natural reduction of endogenous estrogen, the hallmark of menopause, can cause or potentiate PFDs and rUTIs. A substantial body of epidemiological literature suggests an association between menopause, and PFDs and rUTIs; however, the ability to separate this association from age and other comorbid conditions makes it difficult to draw definitive conclusions on the role of menopause alone in the development and/or progression of PFDs. Similarly, the causative link between the decline in endogenous estrogen levels and the pathogenesis of PFDs and rUTIs has not been well-established. CONCLUSIONS: Innovative human studies, focused on the independent effects of menopausal estrogen levels, uncoupled from tissue and cellular senescence, are needed

The mysteries of menopause and urogynecologic health: clinical and scientific gaps.

ObjectivesA significant body of knowledge implicates menopausal estrogen levels in the pathogenesis of the common pelvic floor disorders (PFDs). These health conditions substantially decrease quality of life, increase depression, social isolation, caregiver burden, and economic costs to the individuals and society.MethodsThis review summarizes the epidemiology of the individual PFDs with particular attention to the understanding of the relationship between each PFD and menopausal estrogen levels, and the gaps in science and clinical care that affect menopausal women. In addition, we review the epidemiology of recurrent urinary tract infection (rUTI)-a condition experienced frequently and disproportionately by menopausal women and hypothesized to be potentiated by menopausal estrogen levels.ResultsThe abundance of estrogen receptors in the urogenital tract explains why the natural reduction of endogenous estrogen, the hallmark of menopause, can cause or potentiate PFDs and rUTIs. A substantial body of epidemiological literature suggests an association between menopause, and PFDs and rUTIs; however, the ability to separate this association from age and other comorbid conditions makes it difficult to draw definitive conclusions on the role of menopause alone in the development and/or progression of PFDs. Similarly, the causative link between the decline in endogenous estrogen levels and the pathogenesis of PFDs and rUTIs has not been well-established.ConclusionsInnovative human studies, focused on the independent effects of menopausal estrogen levels, uncoupled from tissue and cellular senescence, are needed

Isolation of muscle stem cells from rat skeletal muscles.

Muscle stem cells (MuSCs) are involved in homeostatic maintenance of skeletal muscle and play a central role in muscle regeneration in response to injury. Thus, understanding MuSC autonomous properties is of fundamental importance for studies of muscle degenerative diseases and muscle plasticity. Rat, as an animal model, has been widely used in the skeletal muscle field, however rat MuSC isolation through fluorescence-activated cell sorting has never been described. This work validates a protocol for effective MuSC isolation from rat skeletal muscles. Tibialis anterior was harvested from female rats and digested for isolation of MuSCs. Three protocols, employing different cell surface markers (CD106, CD56, and CD29), were compared for their ability to isolate a highly enriched MuSC population. Cells isolated using only CD106 as a positive marker showed high expression of Pax7, ability to progress through myogenic lineage while in culture, and complete differentiation in serum-deprived conditions. The protocol was further validated in gastrocnemius, diaphragm, and the individual components of the pelvic floor muscle complex (coccygeus, iliocaudalis, and pubocaudalis), proving to be reproducible. CD106 is an efficient marker for reliable isolation of MuSCs from a variety of rat skeletal muscles

Mechanical impact of parturition‐related strains on rat pelvic striated sphincters

AimsTo define the operational resting sarcomere length (Ls ) of the female rat external urethral sphincter (EUS) and external anal sphincter (EAS) and to determine the mechanism of parturition-related injury of EUS and EAS using a simulated birth injury (SBI) vaginal distention model.MethodsEUS and EAS of 3-month-old Sprague-Dawley control and injured rats were fixed in situ, harvested, and microdissected for Ls measurements and assessment of ultrastructure. EUS and EAS function was determined at baseline, and immediately and 4 weeks after SBI, using leak point pressure (LPP) and anorectal manometry (ARM), respectively. Operational L s was compared to species-specific optimal L s using one sample Student's t test. Data (mean ± SD) were compared between groups and time points using repeated measures one-way analysis of variance, followed by Tukey's post hoc pairwise comparisons, with significance set to 0.05.ResultsThe operational resting Ls of both sphincters (EUS: 2.09 ± 0.07 µm, EAS: 2.02 ± 0.03 µm) was significantly shorter than optimal rat Ls of 2.4 µm. Strains imposed on EUS and EAS during SBI resulted in significant sarcomere elongation and disruption, compared with the controls (EUS: 3.09 ± 0.11 µm, EAS: 3.37 ± 0.09 µm). Paralleling structural changes, LPP and ARM measures were significantly lower immediately (LPP: 21.5 ± 1.0 cmH2 O, ARM: 5.1 ± 2.31 cmH2 O) and 4 weeks (LPP: 27.7 ± 1.3cmH2 O, ARM: 2.5 ± 1.0 cmH2 O) after SBI relative to the baseline (LPP: 43.4 ± 8.5 cmH2 O, ARM: 8.2 ± 2.0 cmH2 O); P < 0.05.ConclusionsAnalogous to humans, the short resting Ls of rat EUS and EAS favors their sphincteric function. The insult experienced by these muscles during parturition leads to sarcomere hyperelongation, myofibrillar disruption, and dysfunction of the sphincters long-term