There have been ongoing efforts by federal agencies and scientific communities since the early 1990's to incorporate sex and/or gender in all aspects of cardiovascular research. Scientific journals provide a critical function as change agents to influence transformation by encouraging submissions for topic areas, and by setting standards and expectations for articles submitted to the journal. As part of ongoing efforts to advance sex and gender in cardiovascular physiology research, The American Journal of Physiology: Heart and Circulatory Physiology recently launched a call for papers on Considering Sex as a Biological Variable. This call was an overwhelming success, resulting in 78 articles published in this collection. This mini-review summarizes the major themes of the collection, including sex as a biological variable within: endothelial cell and vascular physiology; cardiovascular immunity and Inflammation, metabolism and mitochondrial energy, and extracellular matrix turnover and fibrosis; neurohormonal signaling; and cardiovascular clinical and epidemiology assessments. Several manuscripts also focused on establishing rigor and reproducibility of key physiological measurements involved in cardiovascular health and disease as well as recommendations and considerations for study design. Combined, these articles summarize our current understanding of sex and gender influences on cardiovascular physiology and pathophysiology, and provide insight into future directions needed to further expand our knowledge.

Sex as a Biological Variable for Cardiovascular Physiology.

Hemispheric analysis of mitochondrial Complex I and II activity in the mouse model of ischemia-reperfusion-induced injury.

Increasing evidence indicates the role of mitochondrial and vascular dysfunction in aging and aging-associated pathologies; however, the exact mechanisms and chronological processes remain enigmatic. High energy demand organs, such as the brain, depend on the health of their mitochondria and vasculature in the maintenance of normal functions, therefore representing vulnerable targets for aging. This methodology paper describes an analysis pipeline for 3D mitochondria-associated signal geometry of two-photon image stacks of brain vasculature. The analysis methods allow the quantification of mitochondria-associated signals obtained in real time in their physiological environment. In addition, signal geometry results will allow the extrapolation of fission and fusion events under normal conditions, during aging, or in the presence of different pathological conditions; therefore, contributing to our understanding of the role mitochondria play in a variety of aging-associated diseases with vascular etiology.

3D Object Geometry of Mitochondria-Associated Signal: 3D Analysis Pipeline for Two-Photon Image Stacks of Cerebrovascular Endothelial Mitochondria.

Mitochondrial numbers and dynamics in brain blood vessels differ between young male and female rats under physiological conditions, but how these differences are affected by stroke is unclear. In males, we found that mitochondrial numbers, possibly due to mitochondrial fission, in large middle cerebral arteries (MCAs) increased following transient middle cerebral artery occlusion (tMCAO). However, mitochondrial effects of stroke on MCAs of female rats have not been studied. To address this disparity, we conducted morphological, biochemical, and functional studies using electron microscopy, western blot, mitochondrial respiration, and Ca2+ sparks activity measurements in MCAs of female, naïve or sham Sprague-Dawley rats before and 48 h after 90 min of tMCAO. Adverse changes in mitochondrial characteristics and the relationship between mitochondria and sarcoplasmic reticulum (SR) in MCAs were present on both sides. However, mitochondria and mitochondrial/SR associations were often within the range of normal appearance. Mitochondrial protein levels were similar between ipsilateral (ipsi) and contralateral (contra) sides. Non-respiratory oxygen consumption, maximal respiration, and spare respiratory capacity were similar between ipsi and contra but were reduced compared with sham. Basal respiration, proton leak, and ATP production were similar among MCAs. Ca2+ activity increased in sham and ipsi MCAs exposed to a mitochondrial ATP-sensitive potassium channel opener: diazoxide. Our results show that tMCAO has effects on mitochondria in MCAs on both the ipsi and contra sides. Mitochondrial responses of cerebral arteries to tMCAO in females are substantially different from responses seen previously in male rats suggesting the need for specific sex-based therapies.

Detrimental effects of transient cerebral ischemia on middle cerebral artery mitochondria in female rats.

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Detrimental effects of transient cerebral ischemia on middle cerebral artery mitochondria in female rats.